Engraftable cell-based immunotherapy for long-term delivery of therapeutic proteins

Abstract

The present application relates to plasma cells and plasma cell precursors that express a macromolecule, such as a protein, protein mimetic or a peptide and compositions comprising these plasma cells or plasma cell precursors. The application further relates to methods of using and making the plasma cells and plasma cell precursors that express the macromolecule. Methods of treatment comprising administering the plasma cells or plasma cell precursors are also contemplated. ENGRAFTABLE CELL-BASED IMMUNOTHERAPY FOR LONG-TERM DELIVERY OF THERAPEUTIC PROTEINS 20 26 21 41 19 10 A ug 2 02 6 E N G R A F T A B L E C E L L - B A S E D I M M U N O T H E R A P Y F O R L O N G - T E R M D E L I V E R Y O F T H E R A P E U T I C P R O T E I N S 2 0 2 6 2 1 4 1 1 9 1 0 2 0 2 6 A u g

Description

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a divisional application of AU2024202827, the disclosure of which is expressly incorporated by reference in its entirety. REFERENCE TO SEQUENCE LISTING, TABLE, OR COMPUTER PROGRAM LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The information in the electronic format of the Sequence Listing is hereby expressly incorporated by reference in its entirety. FIELD OF THE INVENTION

[0003] A new approach for engineering human plasma cells or plasma cell precursors to express macromolecules such as proteins for therapeutic purposes, has been discovered. The alternatives described herein include methods requiring isolation and activation of B cells from subject-derived (autologous) or allogeneic peripheral blood mononuclear cells; genome engineering of these B cells or B cell precursors so that they express molecules, such as macromolecules, protein mimetics, proteins or peptides of interest in the absence of viral integration (with or without additional genetic modifications that modulate their eventual function and survival); expansion and differentiation of these cells into >1e-6 long-lived plasma cells in vitro; and administration of these autologous or allogeneic engineered protein-producing plasma cells into human recipients for therapeutic application. BACKGROUND OF THE INVENTION

[0004] To date there are limited reports of genome editing in primary B cells. These limited reports can be found, for example, in Cheong et al. 2016 and Chu et al. 2016 (“Editing of mouse and human immunoglobulin genes by CRISPR-Cas9 system.” Nat Commun. 2016 Mar 9; 7:10934. and “Efficient CRISPR-mediated mutagenesis in primary immune cells using CrispRGold and a C57BL / 6 Cas9 transgenic mouse line.” Proc Natl Acad Sci U S A. 2016 Nov 1; 113(44):12514-12519.; both incorporated by reference in their entireties herein). These reports have used either transient transfection of plasmid DNA or lentiviral vector delivery to facilitate gene disruption. However, there are no reports available in the medical literature of using nucleases to achieve homologous recombination in primary human B cells.

[0005] One of the main problems with the use of plasmids is their low efficiency. For example, plasmid-based methods of DNA delivery to primary B cells are extremely toxic, which is likely due to innate DNA sensing and they also exhibit low efficiency (<1%). Lentiviral-based gene delivery also has low efficiency in these cells, for example, in most cases less than 5% of the cells can be transduced, and gene targeting in primary B cells has historically required use of cells from animals with transgenic expression of CAS9. In contrast, the genome engineering approach used in the alternatives described herein enables one to selectively edit the genes with introduction of gene expression cassettes by homologous recombination in more than 30% of primary human B cells and to selectively expand these cells to generate an enriched cell product expressing transgenes of interest.

[0006] Selectivity is also an issue with lentiviral and plasmid DNA. For example, lentiviral vectors integrate randomly and primarily in sites of active transcription and have the potential to be oncogenic. In contrast, nuclease targeting and homology directed repair (HDR) dependent integration of payload allow for selectivity with minimal off-target effects. The need for new approaches that allow one to achieve homologous recombination in primary human B cells is manifest. SUMMARY OF THE INVENTION

[0007] Aspects of the alternatives described herein include, but are not limited to: (1) the use of blood-derived human B cells and / or B cell precursors as a starting material for a plasma cell molecule producing immunotherapy; (2) RNA- and protein-based transfection to facilitate delivery of candidate designer nucleases targeting a broad range of genetic loci in primary B cells that include, but are not limited to zinc finger nucleases, transcription activator-like effector nucleases (TALEN), homing endonucleases (HEs), combined TALEN -HE proteins (megaTALs) and CRISPR / Cas systems; (3) transfection of long single-stranded DNA oligonucleotides or transduction with recombinant adeno-associated virus to facilitate efficient delivery of donor DNA templates carrying therapeutic expression cassettes into primary human B cells and / or B cell precursors in order to facilitate efficient homologous recombination into a range of candidate genetic loci; (4) integrity measures that include, but are not limited to, methods to prevent somatic hypermutation of the B cell antibody locus during the engineering process including, but not limited, to disruption of the AID gene; (5) production enhancers that include, but are not limited to methods to introduce dimerizable drug-inducible activating proteins to enable selectable expansion of engineered human B cells and / or B cell precursors in vitro or in vivo; (6) safety measures that include, but are not limited to, introduction of sequences from cell surface proteins including, but not limited to, the CD20 protein into B cells and / or B cell precursors to enable targeted removal from recipients using rituxan or alternative therapeutic approaches; and (7) the use of a proprietary multi-step cytokine and co-culture based systems to facilitate differentiation of blood-derived B cells and / or B cell precursors into long-lived plasma cells and their survival and expansion in vitro. The human B cells as described herein, can include B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablast, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. Without being limiting, the molecule comprises macromolecules, proteins, protein mimetics and peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, of the CRISPR / Cas system described herein, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

[0008] Multiple categories of use of a plasma cell producing a molecule, such as a macromolecule, protein or a peptide in immunotherapy are also envisioned. Exemplary alternatives are listed below:

[0009] (1) Prophylactic or therapeutic protection from infection (viral, bacterial, or parasitic) in healthy subjects or following stem cell administration or solid-organ transplantation in pediatric and adult subjects including, but not limited to neutralizing antibodies that block influenza, parainfluenza, rhinovirus, Respiratory Syncitial Virus (RSV), HIV, pathogenic bacteria, and / or parasites;

[0010] (2) Protein replacement, enzyme replacement and rescue of enzyme deficiencies including, but not limited to Factor VIII (Hemophilia A), Factor IX (Hemophilia B), ADAMTS13 (Hereditary TTP), LIPA (lysosomal acid deficiency), SERPING1 (hereditary angioedema), SERPINA1 (alpha1 anti-trypsin deficiency), GLA (Fabry disease), and / or ALPL (Hypophosphatasia);

[0011] (3) Immune modulation via expressed cytokines, cytokine receptors, complement proteins or other inhibitory proteins including, but not limited to: Il1 receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes; complement inhibitory proteins (including Factor H, Factor I) for treatment or inhibition of atypical hemolytic uremic syndrome / membranoproliferative glomerulonephritis; and / or C1 inhibitor for hereditary angioedema;

[0012] (4) Anti-fibrotic molecules including, but not limited to SCGB1A1 for the treatment of pulmonary fibrosis;

[0013] (5) Therapeutic antibodies or at least one binding portion thereof for treating or ameliorating autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer including but not limited to: anti-IL1 monoclonal antibodies or a binding portion thereof for the treatment or amelioration of periodic fever / autoinflammatory syndromes; anti-TNF antibodies or a binding portion thereof for the treatment or inhibition of inflammatory arthritis / inflammatory bowel disease, anti-IL33 antibodies or a binding portion thereof for the treatment or inhibition of asthma and / or anti-C5 antibodies or a binding portion thereof for the treatment or inhibition of paroxysmal nocturnal hemoglobinuria / atypical HUS;

[0014] (6) Anti-thrombotic molecules including, but not limited to APLN to block platelet function. Antithrombotic molecules are further described by Adam et al. (“Apelin: an antithrombotic factor that inhibits platelet function.” Blood. 2016 Feb 18; 127(7):908-20.; incorporated by reference in its entirety herein);

[0015] (7) Glucose response elements upstream of insulin for the treatment or inhibition of diabetic conditions; and

[0016] (8) Therapeutic monoclonal antibodies for hyper-cholesterolemia, including anti-PCSK9 inhibitory antibodies or a binding portion thereof. Preferred alternatives include the following alternatives. In some alternatives herein, the protein is a neutralizing antibody that block influenza, parainfluenza, rhinovirus, Respiratory Syncitial Virus (RSV), HIV, pathogenic bacteria, and / or parasites. In some alternatives herein, the protein is an enzyme. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the plasma cell expresses cytokines, cytokine receptors, complement proteins or other inhibitory proteins.

[0017] In a first aspect, a method of making plasma cells or plasma cell precursors that expresses a molecule, such as a macromolecule, protein, or peptide is provided, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for expression of a molecule in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the molecule. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the protein comprises an enzyme, monoclonal antibody or a binding portion thereof, neutralizing antibodies or a binding portions thereof, therapeutic antibodies or binding portions thereof, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs), or a binding portion thereof. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed by a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a viral, fungal, parasitic or bacterial infection. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are patient derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cell. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cell comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises the steps of : (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cell short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding of the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or any other length defined by a range that is set forth by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the cells were washed with 1X PBS and seeded in medium with IL-6 (50 ng / ml), IL-15 (10 ng / ml) and human interferon-a 2B (100 U / ml, Sigma-Aldrich) for three days to stimulate plasma cell differentiation. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, antithrombotic molecule, coagulation factor, glucose response element and / or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, IL-6, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist that is used for the treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, a therapeutic antibody or binding portion thereof is used e.g., an antibody or binding portion thereof that is specific for a protein or other molecule expressed in an autoimmune disorder, autoinflammatory disorder, immune dysregulation and / or cancer. In some alternatives, the antibody or binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion thereof. In some alternatives, the protein is an antithrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic and / or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablast, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is a macromolecule, protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method further comprises purifying the plasma cells or plasma cell precursors after step e) by positive selection against CD138. In some alternatives, purifying comprises selecting the plasma cells by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection.

[0018] In a second aspect, a method of making a long lived plasma cell is provided, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for molecule expression, such as a protein, protein mimetic or a peptide in the absence of viral integration; expanding the B cells; and differentiating the B cells, thereby producing the long lived plasma cell. In some alternatives, the method further comprises selecting the long lived plasma cell by CD138 selection of the plasma cells. In some alternatives, the macromolecule is a protein, protein mimetic or peptide. In some alternatives, the protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the macromolecule comprises a carbohydrate or lipid moiety. In some alternatives, the macromolecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for the treatment or inhibition of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for the treatment or inhibition of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or a binding portion thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody or a binding portion thereof is an anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti- PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the B cells from the isolating step comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late proB cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cell. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cell comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cell or disruption of candidate loci within the B cell to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 gg / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, antithrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody or binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNABs). In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from a group selected from macromolecules, proteins, protein mimetics and peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the isolating is performed by negative selection isolation of hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the method further comprises purifying long lived plasma cells are purified by positive selection against CD138. In some alternatives, purifying comprises selecting the plasma cells by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection.

[0019] In a third aspect, a plasma cell that expresses a molecule, such as a heterologous protein, protein mimetic or a peptide is provided. In some alternatives, the cell is a B cell. In some alternatives, the heterologous protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti-thrombotic molecule, glucose response element and / or monoclonal antibody or binding portion thereof. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and cancer. In some alternatives, the antibody or binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the antithrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNABs). In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cells. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from a group of macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the plasma cell further comprises at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20.

[0020] In a fourth aspect, a composition comprising the plasma cell manufactured by any one of the alternatives provided herein or comprising the cell of any one of the alternatives herein, is provided. In some alternatives, the plasma cell is derived from a B cell. In some alternatives, the plasma cell expresses a molecule, such as a heterologous protein, protein mimetic or a peptide. In some alternatives, the cell is a B cell precursor. In some alternatives, the B cell precursor comprises hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, antithrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity.

[0021] In a fifth aspect, a method of expressing a molecule, such as a macromolecule in a subject in need is provided, wherein the method comprises: providing or administrating a plasma cell manufactured by any one of the alternative methods provided herein, the plasma cell that expresses a molecule of any one of any one of the alternative plasma cells provided herein, or the composition of any one of the alternatives described herein, into the subject in need. In some alternatives, the method further comprises purifying the plasma cells of step d), wherein the purifying is performed after the expanding step. In some alternatives, the purifying comprises positive selecting of cells against CD138. In some alternatives, the selecting the plasma cells is performed by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or a peptide. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the composition comprises the plasma cell manufactured by any one of the alternatives provided herein or comprises the cell of any one of the alternatives provide herein. In some alternatives, the cell is a B cell. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti-thrombotic molecule, glucose response element or monoclonal antibody. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, the protein is the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is an early pro-B cell, late pro-B cell, large pre-B cell, small pre-B cell, immature B cell, T1 B cell, T2 B cell, marginal zone B cell, mature B cell, naive B cell, plasmablast (short lived) cell, GC B cell, memory B cell, plasmablast cell and / or long lived plasma cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises the steps of : (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or portion thereof, wherein the antibody or portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cells. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the administrating is performed by adoptive transfer. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject has received a stem cell administration or solid organ transplantation. In some alternatives of the method of expressing protein in a subject in need, the subject suffers from influenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection. In some alternatives of the method of expressing protein in a subject in need, the subject suffers from an enzyme deficiency. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject suffers from pulmonary fibrosis. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject suffers from an autoimmune disorder, immune dysregulation or cancer. In some alternatives, the subject suffers from HIV. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject suffers from diabetes. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject suffers from hypercholesterolemia. In some alternatives of the method of expressing a molecule, such as a macromolecule in a subject in need, the subject is also receiving CAR T-cell therapy. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method comprises monitoring of the subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0022] In a sixth aspect, a composition comprising the plasma cell manufactured by any one of the alternatives herein and a second B cell, wherein the second B cell secretes a macromolecule for inducing tolerance of a peptide for immunotherapeutic application or for inducing engraftment of the plasma cell manufactured by any one of anyone of the alternatives herein is provided. In some alternatives, the plasma cell is derived from a B cell. In some alternatives, the plasma cell expresses a molecule, such as a heterologous protein, protein mimetic or a peptide. In some alternatives, the cell is a B cell precursor. In some alternatives, the B cell precursor comprises hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti thrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method further comprises purifying the plasma cells or plasma cell precursors by positive selection against CD138. In some alternatives, purifying comprises selecting the plasma cells by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection.

[0023] In a seventh aspect a method of treating, ameliorating or inhibiting a disease, such as a cancer, in a subject in need, is provided, the method comprises administering a plasma cell manufactured by any one of the alternative methods herein or the plasma cell that expresses a molecule, such as a macromolecule manufactured by any one of the alternative methods herein, or the composition of any one of the alternative compositions herein into the subject in need. In some alternatives, the composition comprises the plasma cell manufactured by any one of the alternatives provided herein or comprises the cell of any one of the alternatives provided herein. In some alternatives, the cell is a B cell. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti-thrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof that is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cell to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 gg / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, antithrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives of the method of treating, inhibiting or ameliorating a disease, such as cancer, in a subject in need, the administering is performed by adoptive transfer. In some alternatives of the method of treating, inhibiting, or ameliorating a disease, such as cancer, in a subject in need, the subject has received a stem cell administration or solid organ transplantation. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from influenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from an enzyme deficiency. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from pulmonary fibrosis. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from an autoimmune disorder, immune dysregulation and / or cancer. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from diabetes. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject suffers from hypercholesterolemia. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject is receiving therapy. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject is selected to receive cancer therapy, diabetes therapy, HIV treatment, or viral therapy. In some alternatives of the method of ameliorating, treating, or inhibiting a disease in a subject in need, the subject is also receiving CAR T-cell therapy. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0024] In an eighth aspect, a method of expressing a molecule, such as a macromolecule in a subject of need is provided, wherein the method comprises: (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for molecule expression in absence of viral integration; (d) expanding the B cells; differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein; and (e) administering the B cells into the subject in need. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the method further comprises purifying the plasma cells of step d), wherein the purifying is performed after the expanding step. In some alternatives, the purifying comprises positive selecting of cells against CD138. In some alternatives, the selecting the plasma cells is performed by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises the steps of : (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a classswitch to IgG1 positive cells. In some alternatives, wherein step (a) performing a second round of genome editing on the B cells to excise a region, further comprises removing IgM positive cells. In some alternatives, the method further comprises, activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / auto-inflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the administering is performed by adoptive transfer. In some alternatives, the subject has received a stem cell by administration or solid organ transplantation. In some alternatives, the subject suffers from influenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection. In some alternatives, the subject suffers from an enzyme deficiency. In some alternatives, the subject suffers from pulmonary fibrosis. In some alternatives, the subject suffers from an autoimmune disorder, immune dysregulation and / or cancer. In some alternatives, the subject suffers from diabetes. In some alternatives, the subject suffers from hypercholesterolemia. In some alternatives, the subject is receiving a conventional therapy for the disease, malady or condition. In some alternatives, the subject is selected to receive cancer therapy, diabetes therapy, HIV treatment, or viral therapy. In some alternatives, the subject is also receiving CAR T-cell therapy. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0025] In an ninth aspect, a method of ameliorating a disease in a subject in need is provided, the method comprising: administering a plasma cell manufactured by any one of the alternatives herein or the plasma cell that expresses a molecule, such as a macromolecule manufactured by any one of the alternatives herein, or the composition of the alternatives herein into the subject in need. In some alternatives, the administering is performed by adoptive transfer. In some alternatives, the subject has received a stem cell administration or solid organ transplantation. In some alternatives, the subject suffers from influenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection. In some alternatives, the composition comprises the plasma cell manufactured by any one of the alternatives provided herein or comprises the cell of any one of the alternatives provided herein. In some alternatives, the plasma cell is derived from a B cell. In some alternatives, the plasma cell expresses a molecule, such as a heterologous protein, protein mimetic or a peptide. In some alternatives, the cell is a B cell precursor. In some alternatives, the B cell precursor comprises hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti-thrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0026] In a tenth aspect method of enhancing immune tolerance to protein therapeutics in a subject in need is provided, the method comprising: administering a plasma cell manufactured by any one of the alternatives herein or the plasma cell that a macromolecule manufactured by any one of the alternatives herein, or the composition of the alternatives herein into the subject in need. In some alternatives, the macromolecule is a protein for inducing tolerance of a protein or peptide for therapy or induces or promotes engraftment and / or survival. In some alternatives, the macromolecule is a protein. The protein may be BAFF, APRIL, IFN-alpha, IL-10 and / or IL-6. In some alternatives the protein is a viral vector capsid protein. In some alternatives, the method further comprises purifying the plasma cells of step D), wherein the purifying is performed after the differentiating step. In some alternatives, the purifying comprises positive selecting of cells against CD138. In some alternatives, the selecting the plasma cells is performed by an anti-CD138 antibody for targeted removal of the plasma cells. In some alternatives, the purifying comprises using anti-CD138 beads for plasma cell selection. In some alternatives, the viral vector capsid protein comprises one or more of the following serotypes: AAV1 (VP1, VP2 and VP3), AAV2 (VP1, VP2 and VP3), AAV4 (VP1, VP2 and VP3), AAV5 (VP1, VP2 and VP3), AAV6 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3), AAV8 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3) or AAV9 (VP1, VP2 and VP3). In some alternatives, the composition comprises the plasma cell manufactured by any one of the alternatives provided herein or comprises the cell of any one of the alternatives provided herein. In some alternatives, the plasma cell is derived from a B cell. In some alternatives, the plasma cell expresses a molecule, such as a heterologous protein, protein mimetic or a peptide. In some alternatives, the cell is a B cell precursor. In some alternatives, the B cell precursor comprises hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, anti-thrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cells. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cells. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ^g / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0027] In an eleventh aspect, a method of enhancing immune tolerance to a protein therapeutic in a subject in need is provided. The method comprises administering a B cell manufactured by any one of the alternatives provided herein or the B cell that expresses a molecule, such as a macromolecule manufactured by any one of the alternatives provided herein, or the composition of any one of the alternatives provided herein into the subject in need. In some alternatives, the subject suffers from Hemophilia A, Hemophilia B, Hereditary TPP, lysosomal acid deficiency, hereditary angioedema, alpha1 anti-trypsin deficiency, Fabry disease or hypophosphatasia. B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixture or combination thereof. In some alternatives, the composition comprises the plasma cell manufactured by any one of the alternatives provided herein or comprises the cell of any one of the alternatives provided herein. In some alternatives, the plasma cell is derived from a B cell. In some alternatives, the plasma cell expresses a molecule, such as a heterologous protein, protein mimetic or a peptide. In some alternatives, the cell is a B cell precursor. In some alternatives, the B cell precursor comprises hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the protein comprises an enzyme, neutralizing antibody, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody, antithrombotic molecule, glucose response element or a monoclonal antibody. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the plasma cell is a long lived plasma cell. In some alternatives, the plasma cell is manufactured by any one of the alternative methods provided herein. In some alternatives, the plasma cell is manufactured by a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express a protein. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the macromolecule is a protein, protein mimetic or a peptide. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activatorlike effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the plasma cell is manufactured by a method of making a long lived plasma cell, wherein the method comprises: isolating B cells; activating the B cells; a first round of genome editing of the B cells for protein expression in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 ug ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cells, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0028] In a twelfth aspect, a method of inducing tolerance of a macromolecule secreted by a B cell in a subject in need in vivo is provided, the method comprising: A) isolating B cells; B) performing a first round of genome editing of the B cells for protein expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion; C) expanding the B cells; D) differentiating the B cells, optionally, after step (c) or (d), thereby producing activated B cells or naive activated B cells that express the molecule; and E) providing or administering the B cells that expresses the macromolecule into a subject in need. In some alternatives, the macromolecule, is a protein, protein mimetic or peptide. In some alternatives, the macromolecule comprises natural amino acids and / or unnatural amino acids. In some alternatives, the protein is an enzyme. In some alternatives, the method further comprises activating the B cells prior to gene editing, where inactivating is performed using at least one stimulant. In some alternatives, the at least one stimulant is a CD40 ligand, CpG, IL2, IL10 and / or IL15. In some alternatives, cells are activated with 100 ng / ml of recombinant human MEGACD40L), 1 gg / ml of CpG oligodeoxynucleotide 2006, 50 ng / ml of IL2, 50 ng / ml of IL10 and 10 ng / ml of IL15 for two days. In some alternatives, the CD40 ligand is artificially oligermized. In some alternatives, the CD40 ligand is oligermerized with two linked CD40L trimers. In some alternatives, the cells are cultured for at least 13 days with the at least one stimulant. In some alternatives, the cells are a naive B cell phenotype, activated B cell or activated edited B cell after differentiation. In some alternatives, the cells are CD27-CD138-CD38low / -CD19highIgM+ IgD+ after differentiation. In some alternatives, the providing or administering is performed by adoptive transfer. In some alternatives, the subject has received a stem cell administration or solid organ transplantation. In some alternatives, the subject suffers from influenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or a parasite infection. In some alternatives, the subject suffers from an enzyme deficiency. In some alternatives, the subject suffers from pulmonary fibrosis. In some alternatives, the subject suffers from an autoimmune disorder, immune dysregulation or cancer. In some alternatives, the subject suffers from diabetes. In some alternatives, the subject suffers from hypercholesterolemia. In some alternatives, the subject is also receiving CAR T-cell therapy. In some alternatives, the subject suffers from HIV. In some alternatives, the subject suffers from Hemophilia A, Hemophilia B, Hereditary TPP, lysosomal acid deficiency, hereditary angioedema, alpha1 anti-trypsin deficiency, Fabry disease or hypophosphatasia. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cells and co-delivering the nuclease with a single stranded nucleic acid donor template. In some alternatives, the nuclease is a synthetic guide RNA targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixtures or combinations thereof. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0029] In a thirteenth aspect, a method of inducing tolerance of a peptide useful for immunotherapeutic application, secreted by a B cell in a subject in need in vivo and / or increasing engraftment of the B cell in a subject in need, the method comprising: A) isolating a first population of B cells; B) performing a first round of genome editing of the first population of B cells for protein or peptide expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion; C) expanding the first population of B cells; D) differentiating the first population of B cells, optionally, after step (c) or (d), thereby producing an activated first population of B cells or a naive first population of activated B cells that express the molecule; E) providing or administering the first population of B cells that expresses the peptide into a subject in need; and F) administering a second population of B cells that expresses a macromolecule, such as BAFF, APRIL, IFN-alpha, IL-10 or IL-6. In some alternatives, the immunotherapeutic application is treatment or amelioration of cancer. In some alternatives, the cancer comprises a solid tumor. In some alternatives, the peptide is an anti-PD1 antibody or a PDL-1 ligand. In some alternatives, the second population of B cells express BAFF, APRIL, IFN-alpha, IL-10 or IL-6. In some alternatives, the second population of B cells express a second peptide that is expressed and presented as a MHC class II molecule on a surface of the B cells. B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixtures or combinations thereof. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0030] In a fourteenth aspect, a method of inducing tolerance of a peptide for immunotherapeutic application, secreted by a B cell in a subject in need in vivo and / or increasing engraftment or survival of the B cells in a subject in need, the method comprising: A) isolating a first population of B cells; B) performing a first round of genome editing of the first population of B cells for protein or peptide expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion; C) expanding the first population of B cells; D) differentiating the first population of B cells, optionally, after step (c) or (d), thereby producing activated B cells or naive activated B cells that express the protein or peptide; E) providing or administering the first population of B cells that express the protein or peptide into a subject in need F) providing or administering a second population of B cells that express a macromolecule, wherein the second population of B cells allows tolerance of the protein or peptide and / or promotes engraftment. In some alternatives, the first population of B cells and the second population of B cells are co-administered. In some alternatives, the macromolecule comprises BAFF, APRIL, IL-10, IFN-alpha or IL-6. In some alternatives, the protein comprises an enzyme, neutralizing antibody or a binding portion thereof, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody or a binding portion thereof, anti-thrombotic molecule, glucose response element or monoclonal antibody or a binding portion thereof. In some alternatives the protein is a viral vector capsid protein. In some alternatives, the viral vector capsid protein comprises one or more of one of the following serotypes: AAV1 (VP1, VP2 and VP3), AAV2 (VP1, VP2 and VP3), AAV4 (VP1, VP2 and VP3), AAV5 (VP1, VP2 and VP3), AAV6 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3), AAV8 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3) or AAV9 (VP1, VP2 and VP3). In some alternatives, the protein is a receptor antagonist for the treatment or inhibition of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for the treatment or inhibition of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angioedema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the protein comprises therapeutic antibodies or a binding portion thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody or a binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof. B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixtures or combinations thereof. In some alternatives, the method comprises monitoring of subject following plasma cell transplantation. In some alternatives, the subject is determined to no longer need treatment or therapy and the method further comprises administering rituxan or an anti-CD20 antibody or binding portion thereof for targeted removal of the plasma cells.

[0031] In a fifteenth aspect a composition is provided, wherein the composition comprises a first population of B cells manufactured by any one of the alternatives herein and a second population of B cells, wherein the second population of B cells secretes a macromolecule for inducing tolerance of a peptide for immunotherapeutic application or for inducing engraftment of the plasma cell manufactured by any one of the alternatives herein. In some alternatives, the macromolecule expressed by the second population of B cells comprises BAFF, IFN-alpha, APRIL, IL-10 or IL-6. In some alternatives the macromolecule is a viral vector capsid protein. In some alternatives, the viral vector capsid protein comprises one or more of one of the following serotypes: AAV1 (VP1, VP2 and VP3), AAV2 (VP1, VP2 and VP3), AAV4 (VP1, VP2 and VP3), AAV5 (VP1, VP2 and VP3), AAV6 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3), AAV8 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3) or AAV9 (VP1, VP2 and VP3). B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixtures or combinations thereof. In some alternatives, the method of making plasma cells or plasma cell precursors that expresses a molecule, such as a macromolecule, protein, or peptide comprises(a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the molecule. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation. In some alternatives, the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the first round of genome editing is performed by an RNA and protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cells. In some alternatives, the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease. In some alternatives, the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a CAS nuclease are delivered via translatable RNA or recombinant protein. In some alternatives, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides are unmodified. In some alternatives, the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci. In some alternatives, the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises selectively increasing a proportion of gene edited B cells. In some alternatives, the increasing the proportion of gene edited B cells comprises the steps of : (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells. In some alternatives, the first round of genome editing further comprises techniques for homology-directed repair. In some alternatives, the second round of genome editing results in the excision of the IgM constant region. In some alternatives, the second round of genome editing results in the B cells mimicking a classswitch to IgG1 positive cells. In some alternatives, step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells. In some alternatives, the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the IgM positive cells are removed by negative selection. In some alternatives, the expanding of the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1. In some alternatives, the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers. In some alternatives, the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers. In some alternatives, the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells. In some alternatives, the negative selecting of non-edited cells is performed by 6-thioguanine. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or any other length defined by a range that is set forth by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by isolation of naive or memory B cells. In some alternatives, the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element and / or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, IL-6, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist that is used for the treatment or amelioration of periodic fever / autoinflammatory syndromes or complement inhibitory proteins. In some alternatives, the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, a therapeutic antibody or binding portion thereof is used e.g., an antibody or binding portion thereof that is specific for a protein or other molecule expressed in an autoimmune disorder, autoinflammatory disorder, immune dysregulation and / or cancer. In some alternatives, the antibody or binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion thereof. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies. In some alternatives, the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic and / or bacterial infection. In some alternatives, the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria. In some alternatives, the protein comprises an antibody or binding portion thereof, wherein the antibody or binding portion thereof is specific for an antigen that is expressed in a viral, fungal, parasitic or bacterial infection. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablast, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and memory B cells. In some alternatives, the molecule is a macromolecule, protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the method of making a long lived plasma cell comprises isolating B cells; activating the B cells; a first round of genome editing of the B cells for molecule expression, such as a protein, protein mimetic or a peptide in the absence of viral integration; expanding the B cells; and differentiating the B cells. In some alternatives, the molecule is a macromolecule, such as a protein, protein mimetic or peptide. In some alternatives, the macromolecule is a prodrug. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acylated. In some alternatives, the protein is an enzyme. In some alternatives, the enzyme comprises a cofactor for enhanced enzymatic activity. In some alternatives, the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells. In some alternatives, the B cells from the isolating step comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the first round of genome editing is performed by an RNA or protein based transfection. In some alternatives, the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cells. In some alternatives, the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease. In some alternatives the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9. In some alternatives, the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci. In some alternatives, the first round of genome editing comprises transducing the B cells with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci. In some alternatives, the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells. In some alternatives, the method to prevent somatic hypermutation of an antibody locus in the B cells comprises disruption of an AID gene. In some alternatives, the method further comprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo. In some alternatives, the method further comprises introducing a nucleic acid, wherein the nucleic acid encodes at least one cell surface protein. In some alternatives, the at least one cell surface protein is CD20. In some alternatives, the first round of genome editing further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci. In some alternatives, the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values. In some alternatives, the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives, the isolating is performed by negative selection isolation of naive or memory B cells. In some alternatives, the naive or memory B cells are from healthy donor PBMCs or cells collected by apheresis. In some alternatives, the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step. In some alternatives, the activation and proliferation step is performed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15. In some alternatives, the plasmablast differentiation step is performed in the presence of any combination of IL-2, IL-6, IL-10 and / or IL-15. In some alternatives, the plasma cell differentiation step is performed in the presence of any combination of IL-6, IL-15, APRIL and / or IFNa. In some alternatives, the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein. In some alternatives, the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein. In some alternatives, the protein is IFN-alpha, Factor VIII, Factor IX, SERPING1 or SERPINA1. In some alternatives, the enzyme is ADAMTS13, LIPA, GLA, or ALPL. In some alternatives, the protein is a receptor antagonist for treatment or amelioration of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for treatment or amelioration of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema. In some alternatives, the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1. In some alternatives, the therapeutic antibodies or binding portions thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer. In some alternatives, the antibody or binding portion thereof is an anti-IL1 monoclonal antibody, anti-TNF antibody, anti-IL33 antibody or anti-C5 antibody or a binding portion of any of these antibodies. In some alternatives, the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN. In some alternatives, the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof. In some alternatives, the protein comprises broadly neutralizing HIV-1 antibodies (bNABs). In some alternatives, the protein comprises a prodrug or a proenzyme. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the molecule is selected from macromolecules, proteins, protein mimetics and / or peptides. In some alternatives, the macromolecule, protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids. In some alternatives, the molecule comprises a carbohydrate or lipid moiety. In some alternatives, the molecule comprises a cofactor. In some alternatives, the protein comprises a hydrophobic group for membrane localization. In some alternatives, the protein is acetylated. In some alternatives, the protein comprises a cofactor for enhanced enzymatic activity. In some alternatives, the isolating is performed by negative selection isolation of hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 shows a schematic of the potential clinical application of plasma cell therapy.

[0033] Figure 2 highlights the advantages of using engineered plasma cells for a protein delivery platform, such as longevity, protein production and its potential for ex vivo manipulation and generation.

[0034] Figure 3 shows candidate nuclease platforms that can be used in the alternatives described herein, such as, for example, zinc finger nucleases, CRISPR / Cas Nuclease systems, TAL effector nucleases and homing endonucleases.

[0035] Figure 4 provides a table that shows different methods of CRISPR editing of the primary B cells and the benefits of the methods.

[0036] Figure 5 shows the ribonucleoprotein strategy for the editing of B cells.

[0037] Figure 6 shows the indel in a Sanger sequencing gel. As shown, increasing the concentration of ribonuclease increased the T7 cleavage during a 5 day DNA cleavage protocol. For the experiment, B cells were isolated from peripheral blood mononuclear cells at day zero and transfected with ribonucleoproteins and a PR domain zinc finger protein at day 1. The cells were then activated with mCD40L, CpG and IL2, IL10, and IL15. At day 5, the DNA was extracted from the cell to test for DNA cleavage.

[0038] Figure 7 shows a Western blot, which illustrates results from a T7 cleavage. The antibodies were directed to PRDM1 and TUBB1.

[0039] Figure 8 shows that PRDM1 disruption limits plasmablast generation.

[0040] Figure 9 shows homologous-directed repair (HDR) using single-stranded DNA templates.

[0041] Figure 10 shows homologous-directed repair (HDR) using single-stranded DNA templates.

[0042] Figure 11 shows dose response of short oligonucleotide HDR.

[0043] Figure 12 shows low toxicity in setting of high-efficiency HDR.

[0044] Figure 13 shows long oligonucleotide templates for HDR.

[0045] Figure 14 shows that long template HDR is effective.

[0046] Figure 15 shows results from the testing of primary B cell transduction with AAV vectors. Provided in the right panel is the percent GFP expression with different AAV serotypes. From left to right are AAV serotypes by culture volume from 2%, 5% and 10%. The bottom panel shows the percent viability of the cells after 48 hours post transduction. From left to right consecutively in the graph are the AAV amount by culture volume of 2%, 5% and 10%.

[0047] Figure 16 shows results from the testing of primary B cell transduction with AAV vectors.

[0048] Figure 17 shows results from transduction of B cells using AAV6.

[0049] Figure 18 shows the strategy of homologous-directed repair (HDR) using AAV repair templates.

[0050] Figure 19 shows that CRISPR and AAV repair template co-delivery mediates HDR in primary B cells.

[0051] Figure 20 shows the strategy of Homologous-directed repair (HDR) using AAV repair templates.

[0052] Figure 21 shows CRISPR and AAV repair template co-delivery mediates HDR in primary B cells.

[0053] Figure 22 shows the culturing and genome editing strategy using negative selection isolation of B cells from healthy donor PBMCs.

[0054] Figure 23 shows B cell phenotype at end of expansion phase.

[0055] Figure 24 shows B cell expansion results. The left panel shows the cell expansion curve and the right panel shows the viability by DAPI staining.

[0056] Figure 25 shows Plasma cell differentiation using the 3-step culture system.

[0057] Figure 26 shows results of plasma cell differentiation at predifferentiation and at post-differentiation (day 12).

[0058] Figure 27 shows the demonstration of quantifiable secretion of an exogenous protein in an animal model.

[0059] Figure 28 shows the demonstration of the ability of B cells to produce functionally active Factor IX.

[0060] Figure 29A and Figure 29B shows sustained expression of human proteins in murine serum between 10 and 21 days.

[0061] Figure 30 shows the experimental design for gene disruption and protein expression.

[0062] Figure 31 shows the long lived human antibody production in mice.

[0063] Figure 32 shows expression of human IL6 in mice confers a growth advantage for transplanted ex vivo differentiated human plasma cells.

[0064] Figure 33 shows high IL6 expression correlates with high plasma cell secretion.

[0065] Figure 34 shows in vitro human APRIL treatment correlates with increased plasma cell antibody secretion in vivo.

[0066] Figure 35 shows the results of the gene disruption and protein expression.

[0067] Figure 36 shows targeting of abundant plasma cell genes using CRISPR.

[0068] Figure 37 shows multiplexed genome editing to introduce FIX and increase protein secretion in human plasma cells.

[0069] Figure 38 shows knock-out of BANK1 promotes plasma cell differentiation and antibody production.

[0070] Figure 39 shows long-lived plasma cells secrete more immunoglobulin on a per cell basis.

[0071] Figure 40A shows B cell activation cocktail leads to expansion of naive B cells in vitro. (40A-40C) Primary human CD19+ B cells were isolated from PBMCs and cultured with MEGACD40L, CpG, IL2, IL10 and IL15 for 13 days. Fresh medium with the soluble factors was replenished every one or two days to maintain a cell density of ~i*106 cells / ml. Cells were counted every day (a) and viabilities were measured using 4',6-diamidino-2-phenylindole (DAPI) staining (b) (n=2). Line graphs show means ± SEM. (c) Cell samples were stained using anti-CD27, CDi38, CDi9, CD38, IgD and IgM antibodies after 0, 2, 8, and i3 days of culture. Immunophenotypes were analyzed by flow cytometry.

[0072] Figure 41 shows primary B cells are effectively electroporated with mRNA encoding blue fluorescent protein (BFP). B cells were activated for two days in culture and transfected with BFP mRNA using the optimal electroporation settings. Percentages of BFP+ cells were measured 24 hours after transfection by flow cytometry. Left: representative flow plots showing BFP expression of mock and BFP mRNA electroporated B cells. Right: bar graph showing percentages of BFP+ cells (n=5). All values are means ± SEM.

[0073] Figure 42 shows Cas9 RNP induces site-specific indels or a precise single-nucleotide change in the presence of an ssODN in primary human B cells. (42A) CDi9+ B cells were isolated and activated in vitro for two days, mock treated or transfected with Cas9 RNPs targeting CCR5 or PRDMi (CCR5g, PRDMig-i or PRDMig-2) and cultured for five additional days. Total genomic DNA was isolated on day 5, target regions were PCR amplified and analyzed using the T7 endonuclease i (T7Ei) assay (one representative experiment shown) or sequencing for percentages of on-target indels (ii2,000 reads per experimental condition). (42B) Diagram of wild type PRDMi locus, PRDMig-2 target location and the ssODN donor template containing a single-nucleotide change. (42C, D) B cells were activated for two days and mock treated, electroporated with Cas9 RNP-PRDMig-2 alone, or with the ssODN donor template at serially increasing doses from 7.5 pmol to i20 pmol. (42C) Viabilities of B cells 2 days and 5 days after genome editing (n=3, three donors). No significant difference in viability was observed between mock and up to 30 pmol ssODN plus RNP either on day 2 or day 5. The bar graph shows means ± SEM. (42D) Percentage of HDR, indel and wildtype (WT) alleles in total genomic DNA extracted 5 days after genome editing as assessed by sequencing (greater than 1 million reads per experimental condition). N represents the number of independent experiments. One-way ANOVA with the Sidak correction for multiple comparisons is used; ***p < 0.001; ns, not significant. WT, wildtype. As shown in 42D of the bar graphs, the top black portions of the bar are wild type, light grey is HDR and the bottom grey is indel.

[0074] Figure 43A and 43B shows Cas9-mediated gene disruption is associated with protein reduction and high cell viabilities. (43A) TMD8 cells were mock treated or transfected with either of the two PRDM1-targeting RNPs and protein levels were analyzed five days later by western blot. (43B) Viabilities of primary human B cells two days or five days after genome editing.

[0075] Figure 44A-44E shows Cas9-mediated disruption of PRDM1, IRF4, PAX5, or BACH2 alters B cell phenotype. (44A) Experimental workflow of the plasma cell differentiation assay. This workflow includes a three-step differentiation culture that is divided by a B cell activation phase (phase 1), a plasmablast differentiation phase (phase 2) and a plasma cell differentiation phase (phase 3) using the different cocktails of soluble factors and cytokines specified. Cells were transfected with Cas9 RNP after two days of initial activation, indels were assessed in genomic DNA on day 5, and phenotypes and IgM / IgG secretion were quantified on day 11. (44B) Schematic diagram of the roles of transcription factors PRDM1, IRF4, PAX5, and BACH2 as established by murine studies. (44C-44E) Primary B cells were activated and either mock treated, or transfected with Cas9 RNP targeting CCR5, PRDM1, IRF4, PAX5, or BACH2 and were subsequently differentiated in vitro. (44C) Top: representative flow plots showing CD19 and CD38 expression in live singlets and bar graph showing mean percentages of CD19lowCD38high plasmablasts 11 days post transfection. Bottom: representative flow plots showing CD27 and CD138 expression and bar graph showing percentage of CD27+CD138+ plasma cells 11 days post transfection (n=5, three donors). (44D) Genomic DNA was isolated five days after RNP transfection for allelic indel analysis by sequencing (n=5). (44E) Amounts of IgM (left) and IgG (right) in each culture as measured by ELISA (n=4, three donors). All bar graphs show means ± SEM. n represents the number of independent experiments. Dotted lines mark mock levels. One -way ANOVA with the Sidak correction for multiple comparisons is used; *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant. Statistical comparisons were made compared to mock. IFN-a, interferon a; PB, plasmablast; PC, plasma cell.

[0076] Figure 45A-45C shows Cas9 RNPs targeting CCR5, PRDM1, IRF4, PAX5, and BACH2 lead to high levels of gene disruption in primary human B cells. (Figure 45A-45C) Primary B cells were activated for two days and either mock treated, or transfected with Cas9 RNP targeting CCR5, PRDM1, IRF4, PAX5, or BACH2 and were subsequently differentiated in vitro. (45A) IgG and IgM levels were quantified on day 11 by ELISA and normalized to cell numbers. IgG / IgM mass ratios were calculated and shown in bar graph (n=4). (45B) Cells were immunophenotyped on day 11 and gated on live, singlet, CD19lowCD38high plasmablasts. Left: representative CD19 histograms. Right: mean fluorescence intensities of CD19 (n=5). One-way ANOVA with the Sidak correction for multiple comparisons; *p < 0.05; ***p < 0.001; ns, not significant. Statistical comparisons were made compared to mock. As shown in Figure 45 B from top.

[0077] Figure 46A-46B shows AAV-transduced B cells are marked by rapid recovery and comparable viabilities to mock at experimental endpoint. (46A) Primary B cells were activated for two days in culture and transduced with scAAV GFP of various serotypes at 25,000 MOI. Bar graph shows viabilities two days after transduction (n=3). (46B) Primary B cells were activated for two days and gene-edited using PRDM1-targeting RNP and AAV PRDM1-GFP at different MOIs. Bar graph shows cell viabilities on day 2 and 11 in mock, BFP-no- HA control or PRDM1-GFP edited B cell cultures (n=4). All values are means ± SEM. As shown in 46B in the bar graphs are Day 2 and Day 11 consecutively.

[0078] Figure 47A-47D shows HDR-mediated GFP addition at the PRDM1 locus using Cas9 RNP and an AAV6 donor template leads to progressively increased GFP expression in a dose-dependent manner. (47A) B cells were activated for two days and were either mock treated or transduced with the GFP-expressing scAAV packaged using a comprehensive panel of AAV serotypes (1, 2, 2.5, 5, 6, 8, 9 or D-J) at a multiplicity of infection (MOI) of 25,000. Shown are the percentages of GFP+ cells (left) and mean fluorescence intensities (right) (n=3, three donors). (47B) Schematic of wild type PRDM1 locus, PRDM1g-2 target location and an AAV GFP expression cassette with 400 bp flanking PRDM1 homology arms (AAV PRDM1-GFP). The 3-base PAM sequence is deleted from the AAV template homology sequence. (47C, 47D) B cells were activated for two days and were either mock treated or transfected with Cas9 RNP-PRDM1g-2 with or without AAV transduction. Cells were subsequently cultured under the same activating condition for 11 days. (47C) Top: representative flow plots showing BFP and GFP expressions on day 2 and day 11 after genome editing. The boxed area of the bottom row second panel highlights the GFP+ population in the gene-edited B cells. Bottom: bar graph showing percentages of GFP+ cells on day 2 and day 11 after gene-editing (n=4, four donors). All bar graphs show means ± SEM. n represents the number of independent experiments. One-way ANOVA with the Sidak correction for multiple comparisons is used; *p < 0.05; ***p < 0.001; ns, not significant. MFI, mean fluorescence intensity. PAM, protospacer adjacent motif; WT, wildtype; HA, homology arm; pA, SV40 poly-adenylation signal.

[0079] Figure 48A-48B shows targeted addition of GFP at the PRDM1 locus using RNP and AAV donor templates containing either 400 bp or 1.0 kb homology arms leads to stable GFP expression. (48A) Schematic of AAV PRDM1-GFP cassettes flanked by either 400 bp (top) or 1.0 kb (bottom) homology arms. The template on the top is identical to the one shown in Figure 3. Both templates leave out the 3 bp PAM sequence. (48B) Primary B cells were activated for two days and gene-edited. Bar graph shows percentages of GFP+ cells on day 2 and day 11 after genome editing (n=3). All values are means ± SEM. Unpaired two-tailed t-test; ns, not significant. HA, homology arm; ns, not significant.

[0080] Figure 49A to 49E shows HDR-mediated integration of BAFF coding sequence at the CCR5 locus results in persistent BAFF secretion by gene-edited plasma cells and increases plasma cell differentiation and viability. (49A) Schematic of wild type CCR5 locus, CCR5g target location, an AAV construct that co-expresses GFP and BAFF via T2A linkage (AAV CCR5-GFP-BAFF) and a BAFF-expressing AAV construct (AAV CCR5-BAFF) with identical 800 bp flanking CCR5 homology arms. (49B) B cells were gene-edited after two days of in vitro activation, and were subsequently differentiated into plasma cells using the three-step culture system. Left: representative flow plots showing GFP expression on day 2 and day 11 post gene-editing in mock, CCR5-GFP-BAFF transduced cells with or without Cas9 RNP. Right: bar graph summarizing percentages of GFP+ cells on day 2 and day 11 post gene-editing (n=4, three donors). (49C) B cells were mock electroporated or transduced with AAV CCR5-GFP-BAFF or AAV CCR5-BAFF, with or without Cas9 RNP. Bar graph shows BAFF production as measured by ELISA at day 11 (n=4, three donors). (49D) Left: cells were counted at day 5 and day 11 post gene-editing. Bar graph shows fold changes in cell numbers. Right: viabilities at day 11 by flow cytometry. (n=5, four donors) (49E) Left: representative flow plots showing CD19 and CD38 expression at day 11 after genome editing using the AAV CCR5-BAFF vector. Right: bar graph summarizing percentages of CD19lowCD38high plasmablasts / plasma cells at day 11 (n=5, four donors). Both AAV CCR5-GFP-BAFF and CCR5-BAFF were added at 20,000 MOI. All bar graphs show means ± SEM. N represents the number of independent experiments. One-way ANOVA was used with the Sidak correction for multiple comparisons; **p < 0.01; ***p < 0.001. PAM, protospacer adjacent motif; IL2ss, IL2 signal sequence; WT, wildtype; HA, homology arm; pA, SV40 poly-adenylation signal.

[0081] Figure 50 shows that using AAV without CCR5 homology leads to minimal fluorochrome expression. B cells were either mock electroporated or co-treated with CCR5-targeting RNP and an AAV BFP-noHA control after two days of in vitro activation, and were subsequently differentiated into plasma cells using the three-step culture system. Representative flow plots on day 2 and day 11 (left) and bar graph showing mean percentages of BFP+ cells ± SEM (n=4).

[0082] Figure 51A-51D shows HDR-mediated integration of FIX coding sequence at the CCR5 locus leads to high levels of FIX secretion by gene-edited plasma cells ex vivo. (51A) Schematic of wild type CCR5 locus, CCR5g target location, and a FIXexpressing AAV construct with 800 bp flanking CCR5 homology arms (AAV CCR5-FIX). (51B-51D) Primary B cells were gene-edited using the AAV CCR5-FIX donor template and CCR5-targeting RNP (CCR5g RNP) with or without PAX5-targeting RNP (PAX5g RNP). B cells were subsequently differentiated in vitro. (51B) Frequency of on-target FIX integration in total alleles on day 11 after genome editing as assessed by digital droplet PCR. (51C) CD19 and CD38 expression on day 11. Left: representative flow plots; right: mean percentages of CD19lowCD38high plasmablasts (n=2). (51D) FIX production at day 11 after genome editing as measured by ELISA (n=3, two donors). AAV was added at 20,000 MOI. Bar graphs show means ± SEM. n represents the number of independent experiments. Oneway ANOVA with the Sidak correction for multiple comparisons were used; **p < 0.01; ***p < 0.001. PAM, protospacer adjacent motif; WPRE3, a shortened woodchuck hepatitis virus posttranscriptional regulatory element 55; WT, wildtype; HA, homology arm; pA, SV40 poly-adenylation signal.

[0083] Figure 52A to 52C shows Outline of flow cytometry gating strategy. (52A-52B) Activated B cells were analyzed by flow cytometry. (52A) Preliminary gates for live cells (left) and singlets (middle) were used to define a viable cell population in unstained B cells before measuring fluorescence (GFP shown on right). (52B) Cells were stained with DAPI to mark dead cells. Viability levels were confirmed by quantifying percentages of DAPI- cells. (52C) Demonstration of general flow cytometry gating strategy for analyzing B cell immunophenotypes. B cells were stained with fluorochrome-conjugated anti-CD27, CD138, CD19, CD38 and live / dead stain-Alexa Fluor 350. Live singlets were defined using FSC / SSC, FSC-W / FSC-H and AF350- gates, and the resulting cell population was analyzed for expression of CD markers. Positive and negative populations were separated based on unstained controls. DAPI, 4,6-diamidino-2-phenylindole.

[0084] Figure 53A-53D shows a description of library preparation and molecular analysis of gene disruption and ssODN editing studies. (53A) Genomic DNA flanking the cut site was initially amplified using sequence specific primers to introduce molecular adapters (orange bars). After bead-based purification to remove primers and enzyme, these sequences were amplified with primers containing molecular indices unique to each experiment (red bar) and cluster-generating sequences (purple bars). The samples were purified, pooled and analyzed using a NextSeq 500 Mid-kit. Finally, the data were de-multiplexed, indel and HDR frequencies were quantified using the CRISPResso software package. (53B) An example from one replicate showing indel frequency and read-depth for the IRF4 guide. (53C) A histogram showing the percent insertion, deletion and substitutions plotted relative to the guide cut site (dotted line) in the 150 bp read sequence. (53D) The number of sequences corresponding to each indel size quantified in this experiment (negative and positive numbers represent deletions and insertions, respectively).

[0085] Figure 54 shows Gene-edited, BAFF-expressing plasma cells stably secrete BAFF and human immunoglobulins in NSG mice. (54A) Experimental layout of NSG mouse transplant. Gene-edited B cells were generated as before after two days of in vitro B cell activation and were subsequently differentiated into plasma cells using the three-step culture system. 11 days post genome editing, cells were delivered intravenously into NSG mice (conditioned with Busulfan a day before transplant) at 10 million cells / animal. Blood samples were collected at day 10 and, finally, at day 21 when mice were sacrificed. (54B-54C) Blood serum proteins were quantified by ELISA at day 10 and day 21 (n=3). Shown are serum BAFF levels (54B) and serum human IgM and IgG levels (54C). All graphs represent means ± SEM. n represents the number of mice per group. One-way ANOVA with the Sidak correction for multiple comparisons between groups was used, and paired two-tailed t-test for comparisons between two time points (day 10 vs 21); *p<0.05; **p < 0.01; ***p < 0.001. NSG, NOD / SCID / gamma-c null; PCs, plasma cells; IV, intravenous.

[0086] Figure 55 shows data that demonstrates that in vivo delivery of human APRIL or IL6 enables increased long-term secretion by edited long-lived plasma cells. Primary human B cells were expanded in culture and edited as described in the alternatives herein. Following differentiation into plasma cells, 10 million were injected into NSG mice. Human BAFF was quantified using ELISA. Each dot represents data from an individual recipient mouse at the indicated time point. Definitions

[0087] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[0088] As used herein, “a” or “an” may mean one or more than one.

[0089] About” as used herein when referring to a measurable value is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±l%, and still more preferably ±0.1 % from the specified value.

[0090] Nucleic acid” or “nucleic acid molecule” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, polynucleotides, such as deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), oligonucleotides, fragments generated by the polymerase chain reaction (PCR), and fragments generated by any of ligation, scission, endonuclease action, and exonuclease action. Nucleic acid molecules can be composed of monomers that are naturally-occurring nucleotides (such as DNA and RNA), or analogs of naturally-occurring nucleotides (e.g., enantiomeric forms of naturally-occurring nucleotides), or a combination of both. Modified nucleotides can have alterations in sugar moieties and / or in pyrimidine or purine base moieties. Sugar modifications include, for example, replacement of one or more hydroxyl groups with halogens, alkyl groups, amines, and azido groups, or sugars can be functionalized as ethers or esters. Moreover, the entire sugar moiety can be replaced with sterically and electronically similar structures, such as aza-sugars and carbocyclic sugar analogs. Examples of modifications in a base moiety include alkylated purines and pyrimidines, acylated purines or pyrimidines, or other well-known heterocyclic substitutes. Nucleic acid monomers can be linked by phosphodiester bonds or analogs of such linkages. Analogs of phosphodiester linkages include phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoranilidate, phosphoramidate, and the like. The term “nucleic acid molecule” also includes so-called “peptide nucleic acids,” which comprise naturally-occurring or modified nucleic acid bases attached to a polyamide backbone. Nucleic acids can be either single stranded or double stranded. In some alternatives, a nucleic acid sequence encoding a protein is provided. In some alternatives, the nucleic acid is RNA or DNA.

[0091] Macromolecule” has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a large molecule, such as a protein. The most common macromolecules in biochemistry can include nucleic acids, proteins, peptides, carbohydrates and macrocycles, for example. In the alternatives herein, plasma cells or plasma cell precursors are provided, wherein the plasma cells or plasma cell precursors express a macromolecule, such as a protein, protein mimetic, or a peptide. In some alternatives, the macromolecule is a prodrug.

[0092] “Prodrug” has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, an inactive molecule, or macromolecule that is metabolized into a pharmacologically active drug. After administration, the prodrug is metabolized into a pharmacologically active drug. Prodrugs can be classified into two types, Type I prodrugs may be bioactivated inside the cells (intracellularly). Examples of these are anti-viral nucleoside analogs that must be phosphorylated and the lipid-lowering statins. Type II prodrugs are bioactivated outside cells (extracellularly), especially in digestive fluids or in the body's circulatory system, particularly in the blood. Examples of Type II prodrugs are salicin (described above) and certain antibody-, gene- or virus-directed enzyme prodrugs used in chemotherapy or immunotherapy.

[0093] “Proenzyme” has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a precursor of an enzyme. In some alternatives, the macromolecule is a protein, such as a proenzyme. In some alternatives, the proenzyme is Factor XI. In some alternatives, the proenzyme is from the family of the coagulation system. In some alternatives the proenzyme comprises fibrinogen.

[0094] Protein” has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, large macromolecules that comprise long chains of amino acid residues. Protein, polypeptide, and peptide can be ambiguous terms and can overlap in meaning. The term “protein” can refer to the complete biological molecule in a stable conformation, whereas “peptide” can be reserved for short amino acid oligomers that can lack a stable three-dimensional structure. The boundary between the two terms is not well defined and the number of amino acids can be close to about 20-30 residues for a peptide. The term “polypeptide” refers to a single linear chain of amino acids, usually regardless of length. Without being limiting proteins can be a serum protein, glycoprotein, lipoprotein, enzyme, nucleoprotein, structural protein, antibody, or prodrug, for example.

[0095] B-cell activating factor” (BAFF), can also be referred to as “tumor necrosis factor ligand superfamily member 13B.” BAFF has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a protein that is encoded by the TNFSF 13B gene. BAFF is a cytokine that belongs to the tumor necrosis factor (TNF) ligand family. In some alternatives herein, a B cell or plasma cell is provided, wherein the B cell or plasma cell expresses BAFF. BAFF has been shown to interact with and activate noncanonical NF-kB signaling pathways. This interaction triggers signals essential for the formation and maintenance of B cell, thus it is important for a B-cell survival. In some alternatives, the cells are manufactured to secrete BAFF to improve or enhance B-cell survival.

[0096] MHC class II molecules, have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a class of major histocompatibility complex (MHC) molecules normally found only on antigenpresenting cells such as dendritic cells, mononuclear phagocytes, some endothelial cells, thymic epithelial cells, and B cells. These cells are important in initiating immune responses.

[0097] A protein, polypeptide or peptide can also be found with a functional group or an additional moiety for functional purposes such as an enzyme co-factor, for example. Without being limiting a protein, polypeptide or peptide can comprise a hydrophobic group for membrane localization. Hydrophobic groups can be added by myristoylation, palmityolation, isoprenylation, prenylation, farnesyltion, gerangylgeranylation or glypiation, for example.

[0098] Cofactors are another functional group, which are used for enhanced enzymatic activity. Without being limiting, cofactors can include lipolyation, Flavin moiety, heme C attachment, phophopantetheinylation, retinylidene formations.

[0099] Proteins and peptides can also comprise modified amino acids or nonnatural amino acids. In some alternatives, the macromolecule comprises a protein mimetic. In some alternatives, the macromolecule is a prodrug.

[0100] In some alternatives herein, a method for of making plasma cells or plasma cell precursors that expresses a molecule, such as a protein, peptide or macromolecule is provided, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the molecule. In some alternatives, the molecule is a prophylactic or therapeutic protection from infection (viral, bacterial, or parasitic) following stem cell administration or solid-organ transplantation in pediatric and adult subjects including, but not limited to neutralizing antibodies that block influenza, parainfluenza, rhinovirus, Respiratory Syncitial Virus (RSV), HIV, pathogenic bacteria, and / or parasites. In some alternatives, the molecule is a macromolecule, such as a protein or an enzyme. In some alternatives, the protein or enzyme is for protein replacement, enzyme replacement and rescue of enzyme or protein deficiencies. In some alternatives, the macromolecule comprises Factor VIII, Factor IX, ADAMTS13, LIPA, SERPING1, SERPINA1, GLA, and / or ALPL. In some alternatives, the macromolecule is a prodrug.

[0101] “Cytokines” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a group of proteins that function in cell signaling. For example cytokines can be involved in autocrine signaling, paracrine signaling and endocrine signaling as immunomodulating agents. Cytokines are important in health and disease, for example, playing arole in host responses to infection, immune responses, inflammation, trauma, sepsis, cancer, and reproduction. In some alternatives herein, a method for of making plasma cells or plasma cell precursors that expresses a molecule, such as a protein, peptide or macromolecule is provided, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the molecule. In some alternatives, the molecule is a macromolecule, such as a cytokine, cytokine receptor, complement protein or other inhibitory protein including, but not limited to: Il1 receptor antagonist for treatment or amelioration of periodic fever / autoinflammatory syndromes; complement inhibitory proteins (including Factor H, Factor I) for treatment or inhibition of atypical hemolytic uremic syndrome / membranoproliferative glomerulonephritis; and / or C1 inhibitor for hereditary angioedema.

[0102] Anti-fibrotic molecule” has its plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a molecule that is used in the treatment of fibrosis, or excess fibrous connective tissue in an organ or tissue in a reparative or reactive process. Fibrosis can include pulmonary fibrosis, cystic fibrosis, Idiopathic pulmonary fibrosis, liver cirrhosis, atrial fibrosis, endomyocardial fibrosis, myocardial infarction, glial scars of the brain, arterial stiffness, Arthrofibrosis (knee, shoulder, other joints), Crohn's Disease (intestine), Dupuytren's contracture (hands,fingers), Keloid (skin), Mediastinal fibrosis (soft tissue of the mediastinum), Myelofibrosis (bone marrow), Peyronie's disease, Nephrogenic systemic fibrosis (skin), Progressive massive fibrosis (lungs), Retroperitoneal fibrosis (soft tissue of the retroperitoneum), Scleroderma / systemic sclerosis (skin, lungs) and / or some forms of adhesive capsulitis (shoulder). Without being limiting, anti-fibrotic molecules can include but is not limited to SCGB1A1 for the treatment of pulmonary fibrosis.

[0103] Antibodies” also known as “immunoglobulins” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, large Y-shaped proteins that are secreted by plasma cells to neutralize pathogens. In some alternatives herein, a method for of making plasma cells or plasma cell precursors that expresses a molecule, such as a protein, peptide or macromolecule is provided, wherein the method comprises (a) isolating B cells; (b) developing the B cells; (c) performing a first round of genome editing of the B cells for protein expression in absence of viral integration; (d) expanding the B cells; and (e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the molecule. In some alternatives, the molecule is an antibody, or a portion thereof. In some alternatives herein, the plasma cell or plasma cell precursor expresses an antibody.

[0104] “Coding for" or “encoding” are used herein, has itsplain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, the property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other macromolecules such as a defined sequence of amino acids. Thus, a gene codes for a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system.

[0105] Vector,” “Expression vector” or “construct” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a nucleic acid used to introduce heterologous nucleic acids into a cell that has regulatory elements to provide expression of the heterologous nucleic acids in the cell.

[0106] B cells” or “B lymphocytes” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a white blood cell type of the lymphocyte subtype. B cells are unlike lymphocytes such as T cells and natural killer cells, as B cells express B cell receptors on their cell membrane. The B cell receptors allow the B cell to bind a specific antigen, which will initiate an antibody response. B cells develop from hematopoietic stem cells. As described herein, B cells can include B cell precursors, stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, activated B cells derived from any starting B cell population, plasmablasts (short-lived) cells, GC B cells, memory B cells, and / or long- or short-lived plasma cells and / or any mixtures or combinations thereof depending on the context.

[0107] B cell precursors include the cells from which the B cells are derived. Like T cells, B cells are lymphatic cells that are originated form the bone marrow, where they can reside until they are mature. The B cells, as described in the alternatives herein, include stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablasts (short-lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells. In some alternatives of the plasma cell for expressing a molecule such as a macromolecule, protein or a peptide, the plasma cell is derived from a B cell. In some alternatives, the B cell is a memory B cell. In some alternatives, the B cell is a stem cell, early pro-B cell, late pro-B cell, large pre-B cell, small pre-B cell, immature B cell, T1 B cell, T2 B cell, marginal zone B cell, mature B cell, naive B cell, plasmablasts (short-lived) cell, GC B cell, memory B cell, plasmablast cell or a long lived plasma cell. In some alternatives, the B cells comprise B cell precursors such as hematopoietic stem cells (HSCs), multipotent progenitor (MPP) cells, lymphoid progenitor (CLP) cells, naive B cells, GC B cell, plasmablasts, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells and / or memory B cells. In some alternatives, the macromolecule is a prodrug.

[0108] “Memory B cells” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, the B cell subtypes that are formed within germinal centers following primary infection and are important in generating an accelerated and more robust antibody-mediated immune response in the case of re-infection. The B lymphocytes form the memory cells that can remember the same pathogen for future antibody production during future infections. In some alternatives of the plasma cell for expressing a molecule such as a macromolecule, protein or a peptide, the plasma cell is derived from a B cell. In some alternatives, the B cell is a memory B cell. In some alternatives, the macromolecule is a prodrug.

[0109] “Naive B cell” has itsplain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a B cell that has not yet been exposed to an antigen. Once exposed to an antigen, the naive B cell becomes a memory B cell. In some alternatives of the plasma cell for expressing a molecule such as a macromolecule, protein or a peptide, the plasma cell is derived from a B cell. In some alternatives, the B cell is a memory B cell. In some alternatives, the macromolecule is a prodrug. “Peripheral blood mononuclear cells” (PBNC) as described herein are peripheral blood cells having a round nucleus. These cells consist of lymphocytes (T cells, B cells, NK cells) and monocytes, whereas erythrocytes and platelets have no nuclei, and neutrophils, basophils, and eosinophils have multi-lobed nuclei. In the alternatives described herein, the B cells are subject derived or are allogeneic peripheral blood mononuclear cells. In some alternatives, the B cells are blood-derived human B cells.

[0110] Plasma cells” as described herein, are also called plasma B cells, plasmocytes, plasmacytes, or effector B cells. Plasma cells have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, white blood cells that secrete antibodies and are transported by the blood plasma and the lymphatic system.

[0111] Plasma cell precursor” can begin as an immature plasma cell. The most immature blood cell of the plasma cell lineage is called the plasmablast which can differentiate into a mature fully differentiated plasma cells. Plasmablasts can secrete more antibodies than a B cell, but less than a plasma cell. In some alternatives, a method of making a plasma cell that expresses a molecule is provided. In some alternatives, the plasma cell is a plasma cell precursor. In some alternatives, the plasma cell precursor is a plasmablast.

[0112] Cell isolation,” also referred to as “isolating of cells,” have their plain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a cell separation technique. Such techniques are important in immunology and the techniques can be appreciated by those of skill in the art. Isolation or the separation of the cells can be performed, for example, by the use of antibodies against differentially expressed cell-surface molecules of targets. Without being limiting, B cells can be isolated by using a commercially available kit. Without being limiting, there are commercially available kits for B cell isolation by Miltenyi Biotec, EasySepTM Human B Cell Isolation Kit by StemCellTM Technologies, and many others. In some alternatives herein, a method of making plasma cells or plasma cell precursors that express a molecule, such as macromolecule is provided, wherein the method comprises a step for isolating B cells. In some alternatives, the macromolecule is a protein, a protein mimetic or a peptide. In some alternatives, a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule is provided, the method comprising isolating B cells, developing the B cells, performing a first round of genome editing of the B cells for protein expression in absence of viral integration, expanding the B cells; and differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells that express the molecule.

[0113] “Development” of a cell describes a cells differentiation to a cell type. For example, development into a B cell can occur in several stages, wherein each stage is marked by various gene expression patterns and immunoglobulin H chain and L chain gene loci arrangement. For example the B cells undergo VJ recombination as they develop. B cells develop from hematopoietic stem cells that originate from bone marrow. In some alternatives herein, a method of making plasma cells or plasma cell precursors that express a molecule such as a macromolecule is provided, wherein the method comprises a step for developing the B cells. In some alternatives, the cells are memory B cells or naive B cells. In some alternatives, the macromolecule is a prodrug.

[0114] B cells undergo two types of selection while developing in the bone marrow to ensure proper development. For example, positive selection occurs through antigen independent signaling. Negative selection occurs through the binding of self-antigen with the B cell receptor.

[0115] Genome editing” has itsplain and ordinary meaning when read in light of the specification, and may include but is not limited to, for example, a process that include methods for genetic engineering in which DNA is inserted, deleted or replaced in the genome of a living organism. Editing a gene is also known as gene editing. In some alternatives described herein, a method of making plasma cells or plasma cell precursors that express a molecule, such as a macromolecule is provided, in which B cells or B cell precursors are subjected to at least one round of genome editing. Methods of genome editing can include, but is not limited to nucleic acid being inserted, deleted or replaced in the genome of a cell. In some alternatives, a nuclease is used to achieve this process. In some alternatives, the nuclease is engineered. In some alternatives, the methods include inducing double strand breaks that are repaired through nonhomologous end-joining (NHEJ) or homologous recombination (HR). In some alternatives, the step of genome editing is performed by introduction of a single stranded nucleic acid. In some alternatives, the at least one round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci. In some alternatives, the genome editing of the B cells for protein expression is performed in the absence of viral integration. In some alternatives, a second round of genome editing is performed to excise a region. In some alternatives, a third round of genome editing is performed to result in expression of a drug activatable growth enhancer. In some alternatives herein, the genome editing is performed by nonpathogenic AAV mediated editing by direct homolougous recombination.

[0116] Genome editing can also employ the use of RNA and protein based transfection. For example the CRISPR / Cas system can be modified to edit genomes. This technique requires the delivery of the Cas nuclease complexed with a synthetic guide RNA (gRNA) into a cell, thus the cell’s genome can be cut at a specific location and allow existing genes to be removed and / or add new ones. Thus, CRISPR / Cas and related programmable endonuclease systems are rapidly becoming significant genome editing tools of the biomedical research laboratory, with their application for gene disruption and / or gene targeting as demonstrated in a variety of cultured cell and model organism systems. In some alternatives, of the CRISPR / Cas system described herein, the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

[0117] The basic components of CRISPR / Cas system comprise a target gene, a protospacer adjacent motif (PAM), a guide RNA, Cas endonuclease. An important aspect of applying CRISPR / Cas for genome editing is the need for a system to deliver the guide RNAs efficiently to a wide variety of cell types. This could, for example, involve delivery of an in vitro generated guide RNA as a nucleic acid (the guide RNA generated by in vitro transcription or chemical synthesis). In some alternatives, the nucleic acid could be rendered nuclease resistant by incorporation of modified bases.

[0118] The CRISPR-Cas system falls into two classes. The Class 1 system has a complex of multiple Cas proteins for the degradation of foreign nucleic acids. The Class 2 system has a single large Cas protein for a same purpose for the degradation of foreign nucleic acids. There are a 93 cas genes that are grouped into 35 families. 11 of the 35 families from a cas core which includes the protein families CAS1 to CAS9. As described herein, Cas comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

[0119] Gene editing may also be performed by a novel non-nuclease based gene editing platform. A novel family of AAVs were previously isolated from human hematopoietic stem cells. These nonpathogenic AAVs are naturally present in healthy individuals and may possess unique gene editing and gene transfer properties. This technique is also described as AAV mediated editing by direct homolougous recombination (AmENDRTM). This process is homolougous recombination by a natural biological mechanism that is used by cells to ensure highly precise DNA repair.

[0120] AAV mediated editing by direct homolougous recombination is initiated by design of homology sequence “arms” that are specific to a region of the genome and results in a permanent correction in the DNA when administered to cells. In some alternatives herein, the gene editing is performed by nonpathogenic AAV mediated editing by direct homolougous recombination. The identification of novel AAV genomes are described in Smith et al. (Mol Ther. 2014 Sep; 22(9): 1625-1634; incorporated by reference in its entirety herein). The novel AAVs described by Smith et al., represents a new class of genetic vector for the manipulation of HSC g...

Claims

1. A method of making plasma cells or plasma cell precursors that express a macromolecule, the method comprising:(a) isolating B cells;(b) developing the B cells;(c) performing a first round of genome editing of the B cells for molecule expression in absence of viral integration;(d) expanding the B cells; and(e) differentiating the B cells, optionally, after step (c) or (d), thereby producing plasma cells or plasma cell precursors that express the macromolecule.

2. The method of Claim 1, wherein the macromolecule is a protein, protein mimetic or peptide.

3. The method of Claim 2, wherein the protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids.

4. The method of any one of Claims 1-3, wherein the macromolecule comprises a carbohydrate or lipid moiety.

5. The method of any one of Claims 1-4, wherein the macromolecule comprises a cofactor.

6. The method of any one of Claims 2-5, wherein the protein comprises a hydrophobic group for membrane localization.

7. The method of any one of Claims 2-6, wherein the protein is acylated.

8. The method of any one of Claims 2-7, wherein the protein comprises an enzyme, monoclonal antibody or a binding portion thereof, neutralizing antibodies or a binding portions thereof, therapeutic antibodies or binding portions thereof, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, antithrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein.

9. The method of any one of Claims 2-8, wherein the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1.

10. The method of any one of Claims 2-9, wherein the enzyme is ADAMTS13, LIPA, GLA, or ALPL.

11. The method of any one of Claims 2-10, wherein the protein is a receptor antagonist for treatment of periodic fever / autoinflammatory syndromes or complement inhibitory proteins.

12. The method of Claim 11, wherein the compliment inhibitory protein is Factor H, Factor I or a C1 inhibitor.

13. The method of any one of Claims 2-8, wherein the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1.

14. The method of Claim 8, wherein the therapeutic antibody or a binding portion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer.

15. The method of Claim 8 or 14, wherein the antibody is an anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies.

16. The method of any one of Claims 2-8, wherein the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN.

17. The method of any one of Claims 2-8, wherein the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof.

18. The method of any one of Claims 2-8, wherein the protein comprises broadly neutralizing HIV-1 antibodies (bNAbs), or a binding portion thereof.

19. The method of any one of Claims 2-8 or 18, wherein the protein is an engineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection.

20. The method of Claim 19, wherein the engineered protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed by a virus, fungus, parasite or bacteria.

21. The method of any one of Claims 2-8, wherein the protein comprises an antibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed on a viral, fungal, parasitic or bacterial infection.

22. The method of any one of Claims 2 or 8-21, wherein the protein is an enzyme.

23. The method of any one of Claims 2 or 8-22, wherein the enzyme comprises a cofactor for enhanced enzymatic activity.

24. The method of any one of Claims 1-23, wherein the B cells in step (a) comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells.

25. The method of any one of Claims 1-24, wherein the B cells in step (a) comprise memory B cells and / or naive B cells.

26. The method of any one of Claims 1-25, wherein the developing of the B cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation.

27. The method of Claim 26, wherein the B cell is arrested as an early pro-B cell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell.

28. The method of any one of Claims 1-27, wherein performing the first round of genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid.

29. The method of any one of Claims 1-28, wherein the B cells are subject derived or are allogeneic peripheral blood mononuclear cells.

30. The method of any one of Claims 1-29, wherein the B cells are blood-derived human B cells.

31. The method of any one of Claims 1-30, wherein the first round of genome editing is performed by an RNA and protein based transfection.

32. The method of any one of Claim 31, wherein the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell.

33. The method of Claim 32, wherein the at least one genetic loci comprises JCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1.

34. The method of Claim 32 or 33, wherein the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease.

35. The method of Claim 34, wherein the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

36. The method of any one of Claims 32-35, wherein the nuclease or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to the CAS nuclease are delivered via translatable RNA or recombinant protein.

37. The method of any one of Claims 1-36, wherein the first round of genome editing comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci.

38. The method of Claim 37, wherein the single stranded DNA oligonucleotides are unmodified.

39. The method of Claim 37, wherein the single stranded DNA oligonucleotides are modified with 5’ or 3’ phosphorothioate linkages as repair templates.

40. The method of any one of Claims 1-39, wherein the first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci.

41. The method of Claim 40, wherein the recombinant adeno-associated virus vector is single-stranded, double stranded or self-complementary.

42. The method of any one of Claims 1-41, wherein the method further comprises applying methods to prevent somatic hypermutation of an antibody locus in the B cell.

43. The method of Claim 42, wherein the method to prevent somatic hypermutation of an antibody locus in the B cell comprises disruption of an AID gene.

44. The method of any one of Claims 1-43, wherein the method further comprises selectively increasing a proportion of gene edited B cells.

45. The method of Claim 44, wherein the increasing the proportion of gene edited B cells comprises: (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genome editing on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells.

46. The method of Claim 45, wherein the first round of genome editing further comprises techniques for homology-directed repair.

47. The method of Claims 45 or 46, wherein the second round of genome editing results in the excision of the IgM constant region.

48. The method of any one of Claims 45-47, wherein the second round of genome editing results in the B cells mimicking a class-switch to IgG1 positive cells49. The method of any one of Claims 45-48, wherein step (a) of the step increasing the proportion of gene edited B cells further comprises removing IgM positive cells.

50. The method of any one of Claims 48 or 49, wherein the method further comprises activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1.

51. The method of Claim 49 or 50, wherein the IgM positive cells are removed by negative selection.

52. The method of Claim 50 or 51, wherein the expanding the IgG1 positive cells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1.

53. The method of any one of Claims 45-52, wherein the drug activatable growth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers.

54. The method of any one of Claims 45-53, wherein the activatable growth enhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers.

55. The method of any one of Claims 45-54, wherein the inserting genetic modifications that artificially induce triggerable, non-transformative expansion of edited cells results in disruption of a HPRT locus and negative selection of non-edited cells.

56. The method of Claim 55, wherein the negative selecting of non-edited cells is performed by 6-thioguanine.

57. The method of any one of Claims 1-56, further comprising introducing nucleic acid, wherein the nucleic acid encodes at least one cell surface protein.

58. The method of Claim 57, wherein the at least one cell surface protein is CD20.

59. The method of any one of Claims 37-58, wherein the first round of genome editing further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci.

60. The method of any one of Claims 37-59, wherein the single stranded DNA oligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned lengths.

61. The method of any one of Claims 1-60, wherein the isolating is performed by isolation of early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells.

62. The method of any one of Claims 1-61, wherein the isolating is performed by isolation of naive or memory B cells.

63. The method of Claim 62, wherein the isolating is performed from isolation from healthy donor PBMCs or cell apheresis collections.

64. The method of any one of Claims 1-63, wherein the differentiating step is performed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step.

65. The method of Claim 64, wherein the activation and proliferation step is performed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15.

66. The method of Claims 64 or 65, wherein the plasmablast differentiation step is performed in the presence any combination of IL-2, IL-6, IL-10 and / or IL-15.

67. The method of any one of Claims 64-66, wherein the plasma cell differentiation step is performed in the presence any combination of IL-6, IL-15, APRIL and / or IFNa.

68. The method of any one of Claims 37-67, wherein the single stranded DNA oligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein.

69. The method of any one of Claims 1-68, wherein the method further comprises purifying the plasma cells or plasma cell precursors after step e) by positive selection against CD138.

70. The method of Claim 69, wherein purifying comprises selecting the plasma cells by an anti-CD138 antibody for targeted removal of the plasma cells.

71. The method of Claim 69, wherein the purifying comprises using anti-CD138 beads for plasma cell selection.

72. A method of making a long lived plasma cell, the method comprising:isolating B cells;activating the B cells;a first round of genome editing of the B cells for macromolecule expression in the absence of viral integration;expanding the B cells; anddifferentiating the B cells, thereby producing the long lived plasma cell.

73. The method of Claim 72, further comprising selecting the long lived plasma cell by CD138 selection of the plasma cells.

74. The method of Claim 72 or 73, wherein the macromolecule is a protein, protein mimetic or peptide.

75. The method of Claim 74, wherein the protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids.

76. The method of any one of Claims 72-75, wherein the macromolecule comprises a carbohydrate or lipid moiety.

77. The method of any one of Claims 72-76, wherein the macromolecule comprises a cofactor.

78. The method of any one of Claims 74-77, wherein the protein comprises a hydrophobic group for membrane localization.

79. The method of any one of Claims 74-78, wherein the protein is acylated.

80. The method of Claim 74-79, wherein the protein comprises an enzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein.

81. The method of Claim 74-80, wherein the protein is Factor VIII, Factor IX, SERPING1 or SERPINA1.

82. The method of Claim 80, wherein the enzyme is ADAMTS13, LIPA, GLA, or ALPL.

83. The method of Claim 74-80, wherein the protein is a receptor antagonist for the treatment or inhibition of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for the treatment or inhibition of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema.

84. The method of Claim 74-80, wherein the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1.

85. The method of Claim 80, wherein the therapeutic antibodies or a binding portion thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer.

86. The method of Claim 80, wherein the antibody or a binding portion thereof is an antiIL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies.

87. The method of any one of Claims 74-80, wherein the protein is an anti-thrombotic molecule, wherein the anti-thrombotic molecule is APLN.

88. The method of any one of Claims 74-80, wherein the protein comprises an anti-PCSK9 inhibitory antibody or a binding portion thereof.

89. The method of any one of Claims 72-88, wherein the B cells in the isolating step comprise B cell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells.

90. The method of any one of Claims 72-89, wherein the B cells in the isolating step comprises naive B cells and / or memory B cells.

91. The method of any one of Claims 72-90, wherein the B cells are subject derived or are allogeneic peripheral blood mononuclear cells.

92. The method of any one of Claims 72-91, wherein the B cells are blood-derived human B cells.

93. The method of any one of Claims 72-92, wherein the first round of genome editing is performed by an RNA or protein based transfection.

94. The method of any one of Claims 72-93, wherein the first round of genome editing comprises delivering a nuclease, wherein the nuclease targets at least one genetic locus in the B cell.

95. The method of Claim 94, wherein the nuclease is a zinc finger nuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease.

96. The method of Claim 95, wherein the Cas nuclease comprises Cas1, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

97. The method of any one of Claims 72-96, wherein the first round of genome editing comprises transfecting single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci.

98. The method of any one of Claims 72-97, wherein first round of genome editing comprises transducing the B cell with a recombinant adeno-associated virus vector for homologous recombination into a candidate genetic loci.

99. The method of any one of Claims 72-98, wherein the method further comprises applying methods to prevent somatic hypermutation of an antibody locus and other target loci in the B cells.

100. The method of Claim 99, wherein the method to prevent somatichypermutation of an antibody locus in the B cells comprises disruption of an AID gene.

101. The method of any one of Claims 72-100, wherein the method furthercomprises introducing production enhancers into the B cells or disruption of candidate loci within the B cells to enable selectable expansion of the B cells in vitro or in vivo.

102. The method of any one of Claims 72-101, further comprising introducingnucleic acid, wherein the nucleic acid encodes at least one cell surface protein.

103. The method of Claim 102, wherein the at least one cell surface protein isCD20.

104. The method of any one of Claims 97-103, wherein the first round of genomeediting further comprises cycling the cells for homologous recombination of the single stranded DNA oligonucleotides or the recombinant adeno-associated virus vector into the candidate genetic loci.

105. The method of any one of Claims 97-104, wherein the single stranded DNAoligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values.

106. The method of any one of Claims 72-105, wherein the isolating is performedby negative selection isolation of hematopoietic stem cells, early pro-B cells, late proB cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells.

107. The method of any one of Claims 72-106, wherein the isolating is performedby negative selection isolation of naive or memory B cells.

108. The method of Claim 107, wherein the naive or memory B cells are fromhealthy donor PBMCs or cells collected by apheresis.

109. The method of any one of Claims 72-108, wherein the differentiating step isperformed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step and a plasma cell differentiation step.

110. The method of Claim 109, wherein the activation and proliferation step isperformed in the presence any combination of MCD40L, CpG, IL-2, IL-10 and / or IL-15.

111. The method of Claims 109 and 110, wherein the plasmablast differentiationstep is performed in the presence any combination of IL-2, IL-6, IL-10 and / or IL-15.

112. The method of any one of Claims 109-111, wherein the plasma celldifferentiation step is performed in the presence any combination of IL-6, IL-15, APRIL, and / or IFNa.

113. The method of any one of Claims 74-112, wherein the single stranded DNAoligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein.

114. The method of any one of Claims 73-113 wherein the method furthercomprises purifying long lived plasma cells are purified by positive selection against CD138.

115. The method of Claim 114, wherein purifying comprises selecting the plasmacells by an anti-CD138 antibody for targeted removal of the plasma cells.116.The method of Claim 114, wherein the purifying comprises using anti-CD138beads for plasma cell selection.

117. A plasma cell that expresses a macromolecule, such as a heterologous protein,protein mimetic or a peptide.

118. The plasma cell of Claim 117, wherein the cell is a B cell or a B cellprecursor.

119. The plasma cell of Claim 117 or 118, wherein the protein comprises anenzyme, neutralizing antibody or a binding portion thereof, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody or a binding portion thereof, anti-thrombotic molecule, glucose response element or monoclonal antibody or a binding portion thereof.

120. The plasma cell of Claim 119, wherein the protein is Factor VIII, Factor IX,SERPING1 or SERPINA1.

121. The plasma cell of Claim 119, wherein the enzyme is ADAMTS13, LIPA,GLA, or ALPL.

122. The plasma cell of Claim 119, wherein the protein is a receptor antagonist forthe treatment or inhibition of periodic fever or autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for the treatment or inhibition of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angiodema.

123. The plasma cell of Claim 119, wherein the protein is an anti-fibrotic molecule,wherein the anti-fibrotic molecule is SCGB1A1.

124. The plasma cell of Claim 119, wherein the therapeutic antibodies or a bindingportion thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer.

125. The plasma cell of Claim 119, wherein the antibody or a binding portionthereof is an anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies.

126. The plasma cell of Claim 119, wherein the protein is an anti-thromboticmolecule, wherein the anti-thrombotic molecule is APLN.

127. The plasma cell of Claim 119, wherein the protein comprises an anti-PCSK9inhibitory antibody or a binding portion thereof.

128. The plasma cell of Claim 119, wherein the protein comprises broadlyneutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof.

129. The plasma cell of any one of Claims 117-128, further comprising at least onecell surface protein.

130. The plasma cell of Claim 129, wherein the at least one cell surface protein isCD20.

131. A composition comprising the plasma cell manufactured by any one of Claims1-116 and a second B cell, wherein the second B cell secretes a macromolecule for inducing tolerance of a peptide for immunotherapeutic application or for inducing engraftment of the plasma cell manufactured by any one of Claims 1-116.

132. The composition of Claim 131, wherein the macromolecule expressed by thesecond B cell comprises IFN-alpha, BAFF, APRIL, IL-10 or IL-6.

133. A composition comprising the plasma cell manufactured by any one of Claims1-116 or comprising the cell of any one of Claims 117-130.

134. A method of expressing a molecule, such as a macromolecule in a subject inneed, the method comprising:transplanting a plasma cell manufactured by any one of Claims 1-116, the plasma cell that expresses protein of any one of Claims 117-130, or the composition of any one of Claims 119 or 120 131-133 into the subject.

135. The method of Claim 134, wherein the administering is performed by adoptivetransfer.

136. The method of any one of Claims 134-135, wherein the subject has received astem cell administration or solid organ transplantation or is identified or selected as one to receive such therapies.

137. The method of any one of Claims 134-136, wherein the subject suffers frominfluenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection.

138. The method of any one of Claims 134-137, wherein the subject suffers froman enzyme deficiency.

139. The method of any one of Claims 134-138, wherein the subject suffers frompulmonary fibrosis.

140. The method of any one of Claims 134-139, wherein the subject suffers froman autoimmune disorder, immune dysregulation or cancer.

141. The method of any one of Claims 134-140, wherein the subject suffers fromdiabetes.

142. The method of any one of Claims 134-141, wherein the subject suffers fromhypercholesterolemia.

143. The method of any one of Claims 134-142, wherein the subject is alsoreceiving CAR T-cell therapy or is identified or selected to receive such a therapy.

144. The method of any one of Claims 134-143, wherein the subject suffers fromHIV.

145. The method of any one of Claims 134-144, further comprising monitoring ofsubject following plasma cell transplantation.

146. The method of Claim 145, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

147. A method of ameliorating a disease in a subject in need, the methodcomprising:administering a plasma cell manufactured by any one of Claims 1-116 or the plasma cell that expresses a molecule, such as a macromolecule manufactured by any one of Claims 117-130, or the composition of any one of Claims 131-133 into the subject in need.

148. The method of Claim 147, wherein the administering is performed by adoptivetransfer.administration or solid organ transplantation.

150. The method of any one of Claims 147-149, wherein the subject suffers frominfluenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection.

151. The method of any one of Claims 147-150, wherein the subject suffers froman enzyme deficiency.

152. The method of any one of Claims 147-151, wherein the subject suffers frompulmonary fibrosis.

153. The method of any one of Claims 147-152, wherein the subject suffers froman autoimmune disorder, immune dysregulation or cancer.

154. The method of any one of Claims 147-153, wherein the subject suffers fromdiabetes.

155. The method of any one of Claims 147-154, wherein the subject suffers fromhypercholesterolemia.

156. The method of any one of Claims 147-155, wherein the subject is alreadyreceiving a therapy for the disease or infection.

157. The method of any one of Claims 147-156, wherein the subject is selected toreceive cancer therapy, diabetes therapy, HIV treatment, or viral therapy.

158. The method of any one of Claims 147-157, wherein the subject is alsoreceiving CAR T-cell therapy or is selected to receive such a therapy.

159. The method of any one of Claims 147-158, further comprising monitoring ofthe subject following plasma cell administration.

160. The method of Claim 159, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

161. A method of expressing a molecule, such as a macromolecule in a subject ofneed, the method comprising:(a) isolating B cells;(b) developing the B cells;(c) performing a first round of genome editing of the B cells for molecule expression in absence of viral integration;(d) expanding the B cells; differentiating the B cells, optionally, after step (c) or (d), thereby producing differentiated B cells or plasma cells that express the molecule; and(e) administering the differentiated B cells or plasma cells produced in step d) into the subject in need.

162. The method of Claim 161,163.      The method of any one of Claims 162-164, wherein the purifying comprisesusing anti-CD138 beads for plasma cell selection.

164. The method of any one of Claims 161-165, wherein the molecule is amacromolecule, such as a protein, protein mimetic or a peptide.

165. The method of any one of Claims 161-166, wherein the B cells comprise Bcell precursors such as hematopoietic stem cells, early pro-B cells, late pro-B cells, large pre-B cells, small pre-B cells, immature B cells, T1 B cells, T2 B cells, marginal zone B cells, mature B cells, naive B cells, plasmablast (short lived) cells, GC B cells, memory B cells, plasmablast cells and / or long lived plasma cells.

166. The method of any one of Claims 161-167, wherein the macromolecule,protein, protein mimetic, or peptide comprises natural amino acids and / or unnatural amino acids.

167. The method of any one of Claims 161-168, wherein the molecule comprises acarbohydrate or lipid moiety.

168. The method of any one of Claims 161-169, wherein the molecule comprises acofactor.

169. The method of any one of Claims 161-170, wherein the protein comprises ahydrophobic group for membrane localization.

170. The method of any one of Claims 166-171, wherein the protein is acylated.

171. The method of any one of Claims 166-172, wherein the protein is an enzyme.

172. The method of Claim 173, wherein the enzyme comprises a cofactor forenhanced enzymatic activity.cells is performed after the B cells are arrested at a specific phase of development and / or activating the B cells such that the B cells are permissive for recombination without further B cell differentiation.

174. The method of Claim 175, wherein the B cells are arrested as an early pro-Bcell, a late pro-B cell, a large pre-B cell, a small pre-B cell, an immature B cell, a T1 B cell, a T2 B cell, a marginal-zone B cell, a mature B cell or a memory B cell.

175. The method of any one of Claims 161-176, wherein performing the first roundof genome editing of the B cells for protein expression is performed in the absence of viral integration.

176. The method of any one of Claims 161-177, wherein performing the first roundof genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid.

177. The method of any one of Claims 161-178, wherein the B cells are subjectderived (autologous) or are allogeneic peripheral blood mononuclear cells.

178. The method of any one of Claims 161-179, wherein the B cells are blood-derived human B cells.

179. The method of any one of Claims 161-180, wherein the first round of genomeediting is performed by an RNA and protein based transfection.

180. The method of Claim 181, wherein the first round of genome editingcomprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell.

181. The method of Claim 182, wherein the at least one genetic loci comprisesJCHAIN, IGKC, IGMC, PON3, PRG2, FKBP11, SDC1, SLPI, DERL3, EDEM1, LY6C2, CRELD2, REXO2, PDIA4, PRDM1, CARD11, CCR5 or SDF2L1.

182. The method of Claim 182 or 183, wherein the nuclease is a zinc fingernuclease, transcription activator-like effector nuclease (TALEN), homing endonuclease (HEs), combined TALEN-HE protein (megaTALs) or synthetic guide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease.Cas3, Cas4, Cas5, Cas6, Cas7, Cas8 or Cas9.

184. The method of any one of Claims 182-185, wherein the nuclease or syntheticguide RNAs targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to the CAS nuclease are delivered via translatable RNA or recombinant protein.

185. The method of any one of Claims 181-186, wherein the first round of genomeediting comprises transfecting with single stranded DNA oligonucleotides for homologous recombination into a candidate genetic loci.

186. The method of Claim 187, wherein the single stranded DNA oligonucleotidesare unmodified.

187. The method of Claim 188, wherein the single stranded DNA oligonucleotidesare modified with 5’ or 3’ phosphorothioate linkages as repair templates.

188. The method of any one of Claims 181-189, wherein the first round of genomeediting comprises transducing the B cells with a recombinant adeno-associated virus vector to serve as a donor template for homologous recombination into a candidate genetic loci.

189. The method of Claim 190, wherein the recombinant adeno-associated virusvector is single-stranded, double stranded or self-complementary.

190. The method of any one of Claims 161-191, wherein the method furthercomprises applying methods to prevent somatic hypermutation of an antibody locus in the B cells.

191. The method of Claim 192, wherein the method to prevent somatichypermutation of an antibody locus in the B cells comprises disruption of an AID gene.

192. The method of any one of Claims 161-193, wherein the method furthercomprises selectively increasing a proportion of gene edited B cells.

193. The method of Claim 194, wherein the increasing the proportion of geneedited B cells comprises the steps of : (a) performing a second round of genome editing on the B cells to excise a region; (b) performing a third round of genomeediting on the B cells, wherein the third round of genome editing results in expression of drug activatable growth enhancers; (c) RNA transfecting into the B cells short lived drug activatable growth enhancers; and (d) inserting genetic modifications that artificially induce non-transformative expansion of gene edited B cells.

194. The method of Claim 195, wherein the first round of genome editing furthercomprises techniques for homology-directed repair.

195. The method of Claims 195 or 196, wherein the second round of genomeediting results in the excision of the IgM constant region.

196. The method of any one of Claims 195-197, wherein the second round ofgenome editing results in the B cells mimicking a class-switch to IgG1 positive cells197.      The method of any one of Claims 195-188, wherein step (a) further comprisesremoving IgM positive cells.

198. The method of any one of Claims 198 or 199, wherein the method furthercomprises, activating the IgG1 positive cells and expanding the IgG1 positive cells, wherein the expanding is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1.

199. The method of Claim 199 or 200, wherein the IgM positive cells are removedby negative selection.

200. The method of Claim 200 or 201, wherein the expanding the IgG1 positivecells is performed by exogenous antigens that specifically bind the inserted surface-expressed IgG1.

201. The method of any one of Claims 195-202, wherein the drug activatablegrowth enhancers expressed after the third round of genome editing are rapamycin-inducible FKBP11 dimers.

202. The method of any one of Claims 195-203, wherein the activatable growthenhancers that are RNA transfected into the B cells of step (c) are rapamycin-inducible FKBP11 dimers.

203. The method of any one of Claims 195-204, wherein the inserting geneticmodifications that artificially induce triggerable, non-transformative expansion ofedited cells results in disruption of a HPRT locus and negative selection of non-edited cells.

204. The method of Claim 206, wherein the negative selecting of non-edited cellsis performed by 6-thioguanine.

205. The method of any one of Claims 161-206, further comprising introducingnucleic acid, wherein the nucleic acid encodes at least one cell surface protein.

206. The method of Claim 207, wherein the at least one cell surface protein isCD20.

207. The method of any one of Claims 187-208, wherein the first round of genomeediting further comprises cycling the B-cells for homologous recombination of the single stranded DNA oligonucleotides or recombinant adeno-associated virus into the candidate genetic loci.

208. The method of any one of Claims 206-209, wherein the single stranded DNAoligonucleotides or recombinant adeno-associated virus comprises a length of 0.2kb, 0.5 kb, 1 kb, 2 kb, 3 kb, 4 kb or 5kb or a length within a range defined by any two of the aforementioned values.

209. The method of any one of Claims 161-210, wherein the isolating is performedby isolation of naive or memory B cells.

210. The method of Claim 211, wherein the isolating is performed from isolationfrom healthy donor PBMCs or cell apheresis collections.

211. The method of any one of Claims 161-212, wherein the differentiating step isperformed in a three-step culture system comprising an activation and proliferation step, a plasmablast differentiation step, and a plasma cell differentiation step.

212. The method of Claim 213, wherein the activation and proliferation step isperformed in the presence of any combination of MCD40L (CD40 trimer), CpG, IL-2, IL-10 and / or IL-15.

213. The method of Claims 213 or 214, wherein the plasmablast differentiationstep is performed in the presence any combination of IL-2, IL-6, IL-10 and / or IL-15.differentiation step is performed in the presence any combination of IL-6, IL-15, APRIL and / or IFNa.

215. The method of any one of Claims 166-216, wherein the single stranded DNAoligonucleotides or the recombinant adenovirus comprises a sequence encoding the protein.

216. The method of any one of Claims 166-217, wherein the protein comprises anenzyme, monoclonal antibody, neutralizing antibodies, therapeutic antibodies, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, anti-thrombotic molecule, coagulation factor, glucose response element or a synthetic engineered protein.

217. The method of any one of Claims 166-218, wherein the protein is IFN-alpha,Factor VIII, Factor IX, SERPING1 or SERPINA1.

218. The method of Claim 218, wherein the enzyme is ADAMTS13, LIPA, GLA,or ALPL.

219. The method of anyone of Claims 166-220, wherein the protein is a receptorantagonist for the treatment or inhibition of periodic fever or auto-inflammatory syndromes or complement inhibitory proteins.

220. The method of Claim 221, wherein the compliment inhibitory protein is FactorH, Factor I or a C1 inhibitor.

221. The method of anyone of Claims 166-218, wherein the protein is an anti-fibrotic molecule, wherein the anti-fibrotic molecule is SCGB1A1.

222. The method of Claim 2218, wherein the therapeutic antibody or a bindingportion thereof is specific for a protein or other molecule expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer.

223. The method of Claim 218, wherein the antibody or a binding portion thereof isan anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies.

224. The method of anyone of Claims 166-218, wherein the protein is an antithrombotic molecule, wherein the anti-thrombotic molecule is APLN.anti-PCSK9 inhibitory antibody or a binding portion thereof.

226. The method of anyone of Claims 166-218, wherein the protein comprisesbroadly neutralizing HIV-1 antibodies (bNAbs) or a binding portion thereof.

227. The method of anyone of Claims 166-218, wherein the protein is anengineered protein, wherein the engineered protein is protective in viral, fungal, parasitic or bacterial infection.

228. The method of Claim 229, wherein the engineered protein comprises anantibody or portion thereof, wherein the antibody or portion thereof is specific for an antigen that is expressed on a virus, fungus, parasite or bacteria.

229. The method of anyone of Claims 166-218, wherein the protein comprises anantibody or a binding portion thereof, wherein the antibody or a binding portion thereof is specific for an antigen that is expressed by a viral, fungal, parasitic or bacterial infection.

230. The method of any one of Claims 161-231, wherein the administering isperformed by adoptive transfer.

231. The method of any one of Claims 161-232, wherein the subject has received astem cell administration or solid organ transplantation.

232. The method of any one of Claims 161-233, wherein the subject suffers frominfluenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection.

233. The method of any one of Claims 161-234, wherein the subject suffers froman enzyme deficiency.

234. The method of any one of Claims 161-235, wherein the subject suffers frompulmonary fibrosis.

235. The method of any one of Claims 161-236, wherein the subject suffers froman autoimmune disorder, immune dysregulation or cancer.

236. The method of any one of Claims 161-237, wherein the subject suffers fromdiabetes.hypercholesterolemia.

238. The method of any one of Claims 161-239, wherein the subject is receivingtherapy.

239. The method of any one of Claims 161-240, wherein the subject is selected toreceive cancer therapy, diabetes therapy, HIV treatment, or viral therapy.

240. The method of any one of Claims 161-241, wherein the subject is alsoreceiving CAR T-cell therapy or is selected to receive such a therapy.

241. The method of any one of Claims 161-242, further comprising monitoring ofthe subject following plasma cell transplantation.

242. The method of Claim 243, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

243. A method of ameliorating a disease in a subject in need, the methodcomprising:administering a plasma cell manufactured by any one of Claims 1-116 or the plasma cell that expresses a molecule, such as a macromolecule manufactured by any one of Claims 117-130, or the composition of any one of Claims 131-133 into the subject in need.

244. The method of Claim 245, wherein the administering is performed by adoptivetransfer.

245. The method of Claim 245 or 246, wherein the subject has received a stem celladministration or solid organ transplantation.

246. The method of any one of Claims 245-247, wherein the subject suffers frominfluenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite.

247. The method of any one of Claims 245-248, further comprising monitoring ofthe subject following plasma cell transplantation.

248. The method of Claim 249, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

249. A method of enhancing immune tolerance to protein therapeutics in a subjectin need, the method comprising:administering a plasma cell manufactured by any one of Claims 1-116 or the plasma cell that expresses a molecule, such as a macromolecule manufactured by any one of Claims 117-130, or the composition of any one of Claims 131-133 into the subject in need.

250. The method of Claim 251, wherein the subject suffers from Hemophilia A,Hemophilia B, Hereditary TPP, lysosomal acid deficiency, hereditary angioedema, alpha1 anti-trypsin deficiency, Fabry disease or hypophosphatasia.

251. The method of any one of Claims 251-252, further comprising monitoring ofthe subject following plasma cell transplantation.

252. The method of Claim 253, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

253. A method of inducing tolerance of a macromolecule secreted by a B cell in asubject in need in vivo, the method comprising:A) isolating B cells;B) performing a first round of genome editing of the B cells for protein expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion;C) expanding the B cells;D) differentiating the B cells, optionally, after step (c) or (d), thereby producing an activated B cell or naive activated B cell that express the molecule; andE) providing or administering the B cell that expresses the macromolecule into a subject in need.plasma cells of step D), wherein the purifying is performed after the differentiating step.

255. The method of Claim 256, wherein the purifying comprises positive selectingof cells against CD138.

256. The method of Claim 256 or 257, wherein the selecting the plasma cells isperformed by an anti-CD138 antibody for targeted removal of the plasma cells.

257. The method of any one of Claims 256-258, wherein the purifying comprisesusing anti-CD138 beads for plasma cell selection.

258. The method of anyone of Claims 255-259, wherein the macromolecule, is aprotein, protein mimetic or peptide.

259. The method of Claim 260, wherein the macromolecule comprises naturalamino acids and / or unnatural amino acids.

260. The method of any one of Claims 260-261, wherein the protein is an enzyme.

261. The method of any one of Claims 255-262, wherein the method furthercomprises activating the B cells prior to gene editing, wherein activating is performed using at least one stimulant.

262. The method of Claim 263, wherein the at least one stimulant is CD40 ligand,CpG, IL-2, IL-10 and / or IL-15.

263. The method of Claim 264, wherein the CD40 ligand is artificially oligermized.

264. The method of Claim 265, wherein the CD40 ligand is oligermerized with twolinked CD40L trimers.

265. The method of any one of Claims 263-266, wherein the cells are cultured forat least 13 days with the at least one stimulant.

266. The method of any one of Claims 255-267, wherein the cells are a naive B cellphenotype, activated B cell or activated edited B cell after differentiation.

267. The method of any one of Claims 239-248, the cells are CD27-CD138-CD38low / -CD19highIgM+ IgD+ after differentiation.

268. The method of any one of Claims 255-269, wherein the providing oradministering is performed by adoptive transfer.stem cell administration or solid organ transplantation.

270. The method of any one of Claims 255-271, wherein the subject suffers frominfluenza, parainfluenza, rhinovirus, respiratory syncytial virus, HIV, or other pathogenic bacteria, virus, fungus or parasite infection.

271. The method of any one of Claims 255-272, wherein the subject suffers froman enzyme deficiency.

272. The method of any one of Claims 255-273, wherein the subject suffers frompulmonary fibrosis.

273. The method of any one of Claims 255-274, wherein the subject suffers froman autoimmune disorder, immune dysregulation or cancer.

274. The method of any one of Claims 255-275, wherein the subject suffers fromdiabetes.

275. The method of any one of Claims 255-276, wherein the subject suffers fromhypercholesterolemia.

276. The method of any one of Claims 255-277, wherein the subject is alsoreceiving CAR T-cell therapy or is selected to receive such a therapy.

277. The method of any one of Claims 255-278, wherein the subject suffers fromHIV.

278. The method of any one of Claims 255-279, wherein the subject suffers fromHemophilia A, Hemophilia B, Hereditary TPP, lysosomal acid deficiency, hereditary angioedema, alpha1 anti-trypsin deficiency, Fabry disease or hypophosphatasia.

279. The method of any one of Claims 255-280, wherein performing the first roundof genome editing of the B cells for protein expression is performed in the absence of viral integration.

280. The method of any one of Claims 255-281, wherein performing the first roundof genome editing of the B cells for protein expression is performed by introduction of a single stranded nucleic acid.

281. The method of any one of Claims 255-282, wherein the first round of genomeediting is performed by an RNA and protein based transfection.comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell.

283. The method of any one of Claims 255-284, wherein the first round of genomeediting comprises delivering a nuclease, wherein the nuclease targets at least one genetic loci in the B cell and co-delivering the nuclease with a single stranded nucleic acid donor template.

284. The method of any one of Claims 255-285, wherein the nuclease is a syntheticguide RNA targeting clustered regularly interspersed short palindromic repeat DNA (CRISPR) coupled to a Cas nuclease.

285. The method of any one of Claims 255-286, further comprising monitoring ofthe subject following plasma cell transplantation.

286. The method of Claim 287, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the plasma cells.

287. A method of inducing tolerance of a peptide useful for immunotherapeuticapplication, secreted by a B cell in a subject in need in vivo and / or increasing engraftment of the B cell in a subject in need, the method comprising:A) isolating a first population of B cells;B) performing a first round of genome editing of the first population of B cells for protein or peptide expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion;C) expanding the first population of B cells;D) differentiating the first population of B cells, optionally, after step (c) or (d), thereby producing an activated first population of B cells or naive activated first population of B cells that express the peptide;E) providing or administering the first population of B cells that expresses the peptide into a subject in need; andF) administering a second population of B cells that expresses a macromolecule for tolerance, survival of a B cell or engraftment.treatment or amelioration of a cancer.

289. The method of Claim 290, wherein the cancer comprises a solid tumor.

290. The method of any one of Claims 289-291, wherein the peptide is an anti-PD1antibody or a PDL-1 ligand.

291. The method of Claim 292, wherein the second B cell expresses BAFF,APRIL, IFN-alpha, IL-10 or IL-6.

292. The method of Claim 292, wherein the second B cell expresses a secondpeptide that is expressed and presented as a MHC class II molecule on a surface of the B cell.

293. The method of Claim 292, wherein the second B cell expresses a viral vectorcapsid protein.

294. The method of any one of Claims 289-295, further comprising monitoring ofthe subject following B cell transplantation.

295. The method of Claim 296, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the B cells.

296. The method of Claim 295, wherein the viral vector capsid protein comprisesone or more of the following serotypes: AAV1 (VP1, VP2 and VP3), AAV2 (VP1, VP2 and VP3), AAV4 (VP1, VP2 and VP3), AAV5 (VP1, VP2 and VP3), AAV6 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3), AAV8 (VP1, VP2 and VP3), AAV1 (VP1, VP2 and VP3) and / or AAV9 (VP1, VP2 and VP3).

297. A method of inducing tolerance of a peptide for immunotherapeuticapplication, secreted by B cells in a subject in vivo and / or increasing engraftment or survival of the B cells in the subject, the method comprising:A) isolating a first population of B cells;B) performing a first round of genome editing of the first population of B cells for protein or peptide expression in absence of viral integration, wherein the performing is under conditions of rapid B cell expansion;C) expanding the first population of B cells;D) differentiating the first population of B cells, optionally, after step (c) or (d), thereby producing an activated first population of B cells or naive activated first population of B cells that express the protein or peptide;E) providing or administering the first population of B cells that expresses the protein or peptide into a subject in need;F) providing or administering a second population of B cells that expresses a macromolecule, wherein the second population of B cells allows tolerance of the protein or peptide and / or promotes engraftment and / or survival.

298. The method of Claim 297, wherein the first population of B cells and thesecond population of B cells are co-administered.

299. The method of Claim 297 or 298, wherein the macromolecule comprisesBAFF, APRIL, IFN-alpha, IL-10 or IL-6.

300. The method of any one of Claims 297-299, wherein the protein comprises anenzyme, neutralizing antibody or a binding portion thereof, cytokine, cytokine receptor, complement protein, inhibitory protein, anti-fibrotic molecule, therapeutic antibody or a binding portion thereof, anti-thrombotic molecule, glucose response element or monoclonal antibody or a binding portion thereof.

301. The method of Claim 300, wherein the protein is SERPING1 or SERPINA1.

302. The method of Claim 300, wherein the protein is a receptor antagonist for thetreatment or inhibition of periodic fever / autoinflammatory syndromes, complement inhibitory proteins (including Factor H, Factor I) for the treatment or inhibition of atypical hemalytic uremic syndrome / membranoproliferative glomerulonephritis or a C1 inhibitor for hereditary angioedema.

303. The method of Claim 302, wherein the protein is an anti-fibrotic molecule,wherein the anti-fibrotic molecule is SCGB1A1.

304. The method of Claim 302, wherein the protein comprises therapeuticantibodies or a binding portion thereof are specific for a protein expressed in autoimmune disorders, autoinflammatory disorders, immune dysregulation and / or cancer.an anti-IL-1 monoclonal antibody, anti-TNF antibody, anti-IL-33 antibody or anti-C5 antibody or a binding portion of any one or more of these antibodies.

306. The method of Claim 302, wherein the protein is an anti-thrombotic molecule,wherein the anti-thrombotic molecule is APLN.

307. The method of Claim 302, wherein the protein comprises an anti-PCSK9inhibitory antibody or a binding portion thereof.

308. The method of Claim 302, wherein the protein comprises broadly neutralizingHIV-1 antibodies (bNAbs) or a binding portion thereof.

309. The method of any one of Claims 299-308, further comprising monitoring ofthe subject following B cell administration.

310. The method of Claim 310, wherein the subject is determined to no longer needtreatment and the method further comprises administering rituxan or an anti-CD20 antibody for targeted removal of the B cells.

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