Methods for detecting b cell depletion and treatment of b cell mediated diseases

Abstract

The present disclosure provides methods of treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder by administering hypoimmune allogenic CD19-directed CAR T cells. Also disclosed are pharmaceutical compositions comprising hypoimmune allogenic CD19-directed CAR T cells, for use in treating a subject having or suspected of having a B cell malignancy or a B cell mediated disorder.

Description

METHODS FOR DETECTING B CELT DEPLETION AND TRE ATMENT OF B CELL MEDIATED DISEASES BACKGROUNDI0001] B cell lymphomas encompass a group of lymphoma subtypes categorized within 2 broad categories: nonrHodgkin lymphoma (NHL) and Hodgkin lymphoma (HL), with NHLs being the majority of diagnosed cases. The therapeutic landscape varies for each distinct tumor subtype, but generally there are limited effective therapies in relapsed and / or refractory (r / r) settings across 8 cell lymphomas,|0002] Despite recent advancement, NHL remains an unmet medical need. In 2020, there were 123,000 new cases of NHL reported with 50,000 attributed deaths in Europe. NHL is a heterogeneous group of cancers originating in B lymphocytes, T lymphocytes, or natural killer (NK) cells. In the US, B cell lymphomas represent approximately 85% of all NHL cases, (0003] Treatment of autoimmune diseases often requires multiple treatments to control disease. The most common treatments have to ic; ies associated with the treatment and are often combined with immune suppression, resulting in serious infections and side effects. While many autoimmune diseases have an existing therapy available, the therapies do not result in a drug free remission of the autoimmune disease and require redosing and monitoring.

[0004] There is an urgent need to develop therapies for such patients.SUMMARY

[0005] Off-the-shelf CAR-T cells an other therapeutic cells can offer advantages over autologous cell-based strategies, including ease of manufacturing, quality control, and avoidance of malignant contamination and T cell dysfunction. However, the vigorous host-verws-graft immune response against histoineompatible T cells prevents expansion and persistence of allogeneic CAR-T cells and mitigates the efficacy of this approach,10006] There is substantial evidence in both animal models and human patients that hypoimmonogenic cell transplantation is a scientifically feasible and clinically promising approach to the treatment of numerous disorders, conditions, and diseases.[0007| There remains a need for novel approaches, compositions and methods for producing cell - based therapies that avoid detection by the recipient's immune system.:[00081 In some embodiments, described herein is a method of treating a subject having or suspected of having an IgE-mediated disease or disorder, the method including: administering to the subject a therapeutically effective amount of a composition including a population of engineered hypoimmunogenic T cells, wherein the engineered hypoimmunogenic T cells include one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibili y complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more M HC class I molecules* (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of th e one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more 'ICR molecules, (b) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not include the modification, and (c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specifici ty for an antigen expressed on the surface of an Igfi-producmg cell.

[0000] In some embodiments, described herein is a method of treatmg a subject having or suspected of having an IgE-mediated disease or disorder, the method including: administering to the subject a therapeutically effective amount of a composition including a population of engineered hypoimmunogenic T cells including: (a) reduced expression of beta-2 microglobulin (B2M), Class II Major Histocompatibility Complex Transacti ator (CIITA), and T cell receptor alpha constant (TRAC) relative to a control or wild-type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild- type T colls, and (c) expression of a CD19-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.[0010[ In some embodiments, described herein is a method of assessing B cell depletion, including obtaining a test sample from a subject and detecting a concentration of 1’gE antibodies in the test sample* wherein the subject was administered a B cell depleting composition at least 2 days priors to obtaining the test sample and a decrease In the concentration of IgE antibodies inthe test sample compared to a reference value indicates depletion of B cells by the B cell depleting composition.[(MH l| In some embodiments, described herein is a method treating a B cell mediated disease or disorder, including administering to a subject an agent or therapy that is capable of treating, preventing, delaying, or attenuating development of a B cell mediated disease or disorder, wherein the subject is determined to be at risk for maintaining or developing progressive disease by a method including, (a) obtaining a test sample from a subject and detecting a concentration of JgB antibodies in the sample, wherein the subject was administered a B cell depleting composition at least 2 days priors to obtaining the test, sample, and (b) comparing the concentration detected for the IgE antibodies io a reference value, wherein the subject is one in which the comparison indicates that the subject is at risk for maintaining or developing a progressive B cell mediated disease or disorder,[MM2] In some embodiments, described herein is a method of assessing B cell depletion, including obtaining a test sample from a subject that includes antibodies and detecting the concentration o f antibodies in the test sample that bind a major histocompatibility complex (MHC) protein and comparing the concentration of antibodies that bind a MHC protein to the concentration of antibodies that bind a MHG protein in a reference sample, wherein prior to obtaining the test sample the subject was administered a composition including a population of engineered hypoimmunogfmic T cells, wherein the hypoimjnunogenic T cells include: (a) one or more modifications that inactivate or disrupt one or more alleles of: (i) one or more major histocom atibility complex (MHC) class 1 molecules and / or one or more molecules that regulate expression of the one or more MHC class 1 molecules, (ii) one or more MHC class II molecules and / or one or more mcdecules t hat regulate expression of the one or more MHC class H molecules, (b) one or more modifications that increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not include the modification, and (c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell.In some embodiments, described herein is a method of treating a B cell disease or disorder, including, (a) administering to a subject a therapeutically effective amount of a composition including a population of engineered hypoimmunogenic T cells, wherein the hypoimmunogenic T ceils Include one or more modifications that; (1) inactivate or disrupt one or more alleles of: (I) one or more major histocompatibility complex (MHC) class 1 molecules and / or one or more molecules that regulate: expression of the one or more MHC c lass I molecules, (11) one or more MHC class II molecules and / or one «r more molecules that regulate expression of the one or more MHC class II molecules, (2) increase expressi on of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (2) is relative to a control or wild-type T cell that does not include the modification, and (3) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell; and (b) obtaining a test sample from the subject and detecting a concentration of antibodies that bind a MHC protein, wherein the test sample was obtained at least 20 days after the subject was administered the composition: including the population of engineered hypoinununogenic T cells; and: (c) comparin the concent ration of antibodies that bind a MHC protein in (b) to the concentration of antibodies that bind a MHC protein in a reference sample obtained from the subject prior to administering the composition including the population of engineered hypoimmunogenic T cells; and (d) if the comparison indicates the subject has developed anti- HC antibodies, administering to the subject a composition or therapy that is capable of depleting B cells.BRIEF DESCRIPTION OF THE DRA WINGS100141 The drawings are for illustration purposes only, not for limitation,(0015] Figure 1 shows a graph illustrating a summary of disease progression for Subjects 1-13, including the statuses of complete response (CR), partial response (PR), stable disease (SD), and progressive disease (RD).

[0016] Figure 2 shows a graph illustrating exemplary quantification of CD 19+ cells before and after treatment wi th a T cell drug product of the present d isclosure (C’D19+ eells / pL).(0017) Figure 3 shows graphs illustrating exemplary donor-specific antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure and serum IgE levels (ng / mL; Limit of Detection 0,02 ng / mL).

[0018] Figure 4 shows graphs illustrating exemplary donor-specific antibody levels against subpopulations of cel ls present in a T cell drag pro d uct of thepresent disclosure and serum IgE levels (ng / mL; Limit of Detection 0,02 ng / mL),

[0019] Figure 5 shows graphs illustrating exemplary donor-specific antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure and serum Ig'E levels (ng / mL; Limit of Detection 0,02 ng / mL).

[0920] Figure 6 shows graphs illustrating exemplary donor-specific antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure and serum IgE levels (ng / mL; Limit of Detection 0,02 ng / mL).

[0021] Figure 7 shows a graph illustrating SLEDAI-2K scores measuring disease activity for autoimmune (Al) Subject 1 after treatment with the 1’ cell drag product of the present disclosure,

[0022] Figure 8 shows a graph illustrate ng physician global assessment for AI Subject 1 after treatment with the T cell drag product of the present disclosure.

[0023] Figure 9 shows a graph illustrating fatigue subscale scores for Al Subject I after treatment with the T cell drug product of the present disclosure.

[0024] Figure 10 shows a graph illustrating SLEDA1-2K scores measuring disease activity for Al Subject 2 after treatment with the T cell drug product of the present disclosure.

[0025] Figure 11 shows a graph illustrating physician global assessment for Al Subject 2 after treatment with the T cell drag product of the present disclosure.[002<s] Figure 12 shows a graph illustrating fatigue subscale scores for Al Subject 2 after treatment with the T cell drag product of the present disclosure.

[0027] Figure 13 shows a graph illustrating exemplary quantification of CDI9 / CD20A cells before and after treatment with a T cell drag product of the present disclosure (CD19 / CD20-r ceUs / pL).(0028 j Figure 14 shows a graph illustrating percentage of GDI 9+ cell subsets from Al Subject 1 before and after treatment with a T cell drug product of the present disclosure.

[0029] Figure 15 shows graphs illustrating exemplary quantification of CD19 / CD2(H-cells (cells / gL) and BAF (pg / mL) before and after treatment with a T cell dru g product of the present disclosure.

[0030] Figure 16 shows graphs illustrating exemplary killing of subpopulations of cells present in a T cell drag product of t he presen t disclosure by a subject’s T cells.

[0031] Figure 17 shows graphs illustrating exemplary donor-specific antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure.

[0032] Figure 18 shows graphs i llustrating ki lling of subpopulations of cells present i n a T cell drag product of the present disclosure by complement-dependent cytotoxicity (CI)C ).

[0033] Figure 19 shows graphs illustrating killing of subpopulations of cells present in a T cell drag product of the present disclosure by antibody -dependent cellular cytotoxicity (ADCC).

[0034] Figure 20 sho ws graph s illustrating exemplary killing of subpopulations of cells presen t in a T cell drug product of the present disclosure by a subject’s NK. cells.

[0035] Figure 21 shows graphs illustrating exemplary killing of subpopulations of ceils present in a T cell drag product of the presen t disclosure by a subject’s cells.

[0036] Figure 22 shows graphs illustrating exemplary quantification of (A) percentage CD 19+ ceils. (B) donor-specific antibody levels against subpopulations of cells present in a T cell drug product of the present disclosure, (C) serum IgE levels (ngZmL; Limit of Detection 0.02 ng / niL), and (D) BAFF (pg / 'mL), before and after treatment with a T cell drug product of the present disclosure.

[0037] Figure 23 shows graphs illustrating exemplary killing of subpopulations of cells present in a T cell drug product of the present disclosure by a subject’s T cells.

[0030] Figure 24 shows graphs illustrating exemplary IgG donor-specific antibody levels against subpopulations of cells present in a T eel! drag product of the present disclosure.(0639| Figure 25 shows graphs illustrating exemplary IgM donor-specific antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure, [W40| Figure 26 shows graphs illustrating kilting of subpopulations of cells present in a T cell drug product of the present disclosure by complement-dependent cytotoxicity (CDC). [(>(141 | Figure 27 shows graphs illustrating kilting of subpopulations of cells present in a T cell drag product of the present disclosure by antibody-dependent cellular cytotoxicity (ADCC).(0642| Figure 28 shows graphs illustrating exemplary killing of subpopulations of cells present in a T cell drug product of the presen t disclosure by a subject’s NK. cells,(OO43| Figure 29 shows graphs illustrating exemplary killing of subpopulations of cells present In a T cell drug product of the present disclosure by a subject’s cells,[(104 1 Figure 30: show's graphs illustrating exemplary quantification of (A) percentage CD191- cells, (B) donor-specifie antibody levels against subpopulations of cells present in a T cell drag product of the present disclosure, (C) serum IgE levels (ng / mL; Limit of Detection 0,02 ng / mL), and (D) BAFF (pg / mL), before and after treatment with a T cell drug product of the present disclosure,|W45| Figures 31 A-31C show a schematic of the composition of the hypoimmune allogeneic CD19 CAR T cell product (also referred to herein as “the hypoimmune CAR T cell product”). Primary T cells were isolated from donor leukopaks, inactivation of B2M, CHTA, and TRAC using CRISPR and Cas t 2b was perforated, and the cells were transduced with a bicistronic lentivirus encoding CD19 CAR and CD47, This protocol generated a mixture of HL A replete WT CAR T cells (Figure 31 A), DKO cells (Figure 31B), and fully edited HIP CAR T cells (Figure 31C),14XM6] Figures 32 -32H show a summary of immune response data for.11 patients. Figure 32A show's IFN-garnma ELISpot data from patient peripheral blood T cells showing no activation by HIP CAR T cells or DKO cells, but strong activation by WT CAR T cells with peak spot frequency at 28 days. Figure 32B shows no T cell kill of HIP CAR T cell or DKO cell targets was observed at any time point in any patient. Figure 32C shows no K cell killing of HIP CAR T cells or WT CAR T cells to observed in any patient, but DKO cells were reliablykilled in all patients at all time points. Figure 32D shows no IgG DSAs against HIP CAR T cells or OKO cells were detected in any patient Bar graphs show mean + / ~ SO.|(KI47| Figures 33A-33H show data for patients who never achieved CD 19 aplasia.Figure 33A show the percentage of CD1 cells in peripheral blood throughout the study. Figure 33® shows circulating IgE levels over lime. Figure 33C shows IgG DS As against the 3 cell fractions of SC291 quantified by flow cytometry and expressed as MFI. figure 33D shows patient T cell activation by the 3 cell fractions of the hypoimmune CAR T cell product assessed in FXJSpot assays and is expressed as spot frequency. Figure 33F shows the cytotoxicity of patient T cells against the 3 cell fractions of SC-291, assessed in impedance cytotoxicity assays. Figure 33 F shows the cytotoxicity of pati en t NK cells against the 3 cell fractions of the hypoimmnne CAR. T cell product assessed in impedance cytotoxicity assays. Figure 33G shows for ADCC assays against the 3 cell fractions of the hypoinmwne CAR T cell product, patient NK. cells and serum were combined and killing was assessed in impedance cytotoxicity assays. In one patient (blue), the NK cells at day -5 were endogenously activated to kill DKO cells even without serum, therefore this outlier is not an indication for the presence of antibodies, Figure 33H shows CDC assays against the 3 cell tractions of the hypoimmune CAR T cell product, patient serum was used in impedance cytotoxicity assays. This.group comprised of 2 patients. Patients are color-coded and all individual patient data are shown.10848] Figures 34 -34H show data for patients with temporary CD 19 aplasia. Figure 34A shows the percentage of CD 1.9 cells in peripheral blood. Figure 34B shows circulating IgE levels over time. Figure 34C shows IgG DSAs against the 3 cell fractions of SC291 quantified by flow cytometry and expressed as MFI, Figure 34D show patient T cell activation by the 3 cell fractions of SC291 assessed, in ELISpot assays and is expressed as spot frequency. Figure 34 E shows the cytotoxicity of patient T cells against the 3 cell fractions of the hypoimmune CAR T cell product assessed in impedance cytotoxicity assays. Figure 34F shows the cytotoxicity of patient NK. cells against the 3 cell fractions of the hypoimmune CAR T cell product assessed in impedance cytotoxicity assays. Figure 34G shows ADCC assays against the 3 cell fractions of the hypoimmune CAR T cell product, patient NK cells and serum were combined and killing was assessed in impedance cytotoxicity assays. Figure 34H CDC assays against the 3 cell fractions of the hypoimmune CAR T cell product, patient serum was used inimpedance cytotoxicity assays. This group comprised of 3 patients. Patients are color-coded and all individual patient data are shown.|O049] Figures 35A-35H show data for patients with pennanent CD 19 aplasia. Figure 35 A show the percentage of CD 19 cells in peripheral blood. Figure 353 shows circulating IgE levels over time. Figure 35C shows IgG DSAs against the 3 cell fractions of the hypoimmune CAR T cell product were quantified by flow cytometry' and expressed as MFI. Figure 35D show's patient T cell activation by the 3 cell fractions of the hypoimmune CAR T cell product assessed in ELISpot assays ami is expressed as spot frequency. Figure 35E show's the cytotoxicity of patient T cells against the 3 cell fractions of the hypoimmune CAR. T cell product assessed in impedance cytotoxicity assays. One sample at 28 days (patient blue) did not contain enough T cells for the assay. Figure 35F shows the cytotoxicity of patient NK cells against the 3 cell fractions of the hypoimmune C AR T cell product assessed in impedance cytotoxicity assays. Figure 35G shows ADCC assays against the 3 cell fractions of the hypoimmune CAR T cell product, patient NK cells and serum were combined and killing was assessed in impedance cytotoxicity assays. One sample at 28 days (patient blue) did not contain enough NK cells for the assay. Figure 35H shows CDC assays against the 3 cell fractions of the hypoimmune CAR T cell product, patient serum was used in impedance cytotoxicity assays. This group comprised of 6 patients. Patients are color-coded and all individual patient data are shown.10050] Figures 3 -3611 show data for patients with baseline killing of HIP-CAR T cells. Figure 36A shows circulating IgE levels over time. Figures 3(®»36G show datafbr the first patient. Preformed DS As against all 3 cell fractions were observed on day -5 (Figure 363), Both ADCC (Figure 36C) and CDC (Figure 36D) showed cytotoxicity against all 3 cell fractions of the hypoimmune CAR. T cell product even before the treatment was started. But there was no pre-established T cell response against the 3 cell fractions of SC291 (Figure 36E, Figure 36F) and NK cells only killed DKO cells (Figure 36G). Figures 36H-36M show data for the second patient. Preformed DSAs against all 3 cell fractions were observed on day -5 (Figure 36H). Both ADCC (Figure 361) and CDC (Figure 36J) showed cytotoxicity against all 3 cell fractions of the hypoimmune CAR T cell product even before the treatment was started, But there was no pre-established T cell response against the 3 cell fractions of SC291 (Figure 36K, Figure 36L) and NK. cells only killed DKO cells (Figure 36M), Figures 36N-36U showdata for the third patient. Preformed DS As against WT CAR I cells and HIP CAR T cells were not observed using the regular secondary antibody with preferred affinity for IgG 1 (Figtire 36N but became detectable when, a secondary antibody with high affinity for lgG4 was used (Figure 360). These IgG4 antibodies mediated ADCC killing of only HIP CART cells (Figure 36P). ADCC killing, however, could be averted when recombinant CD47 was added to the assay (Figure 36Q). The IgG4 antibodies did not mediate GDC (Figure 36R). 'There was no pre-established T cell response against the 3 cell fractions of the hypoinmmne CAR T cell product (Figure 36S, Figure 36T) and NK cells only killed DKO cells (Figure 36U).|0051} Figure 37 shows deep B cell depletion data from sixteen patients treated in clinical trial NCT05878I84.DETAILED DESCRIPTIONI. INTRODUCTION(00521 B cell depletion therapy is a promising approach or the treatment of certain cancers and autoimmune diseases. The autoimmune processes are mainly mediated by IgG and IgM autoantibodies, however, autoreactive IgE i& also present in patients with autoimmune conditions (Suutmond and Diamond, 2015; Olewicz-Gawlik and Kowala-Piaskowska, Front Pharmacol. 2023. 1:1112917). IgE directed against autoantigens were observed in a number of autoimmune conditions, including systemic lupus erythematosus (SLE) (Atta et al., 2010), Bullous pemphigoid (BP) (van Seek et al., 2016), chronic spontaneous urticaria (CSU) (Panaszek et al,, 2017), autoimmune thyroiditis (AITD) (Guo et at, 1997), multiple sclerosis (Seals et al., 2022), rheumatoid arthritis (RA), and mixed connective tissue disease (Lamri et al., 2021).(0953) In addition, IgE antibodies are associated with IgE-mediated diseases or disorders. The term “IgE-mediated disease” or “IgE-mediated disorder” as used herein refers to a disease or disorder that is associated with IgE-mediated reactions of the immune system to air allergen and / or an IgE directed against an autoantigen. IgE is an immunoglobulin (1g) produced by B cells that binds with high affinity to allergens. IgE has an important role in type I h}fpersensitivity, which manifests in many allergic diseases. “Allergens” can be environmental, non-infectious proteins that invoke specific IgE immunity. Examples include peanut proteins, latex, ragweed pollen, shellfish, and bee-sting venom. In some instances, an IgE-mediateddisease or disorder can result in anaphylaxis. Allergens gain access to the body through the GI tract, the respiratory tract, and the skin. Over the past 20-plus years there has been a significant increase in children with atopic diseases and food allergies (Jimenez-Saiz, J. Allergy Clin, Immunol., 1 2(5): 1441 -1443). The IgE-expressing cells In food allergic individuals are thought to be primarily plasma cells (Science, 362:1306, 2018).|0054] The mechanisms for generation of IgE plasma cells in mouse and human appears distinct from other Ig isotypes, T helper 2 cytokines such as IL-4, IL-5 and IL- 13 skew B cell responses to IgE, Without wishing to be bound by 'theory, recent evidence suggests that memory responses for IgE are maintained primarily in IgG+ memory B cells that undergo class switching in presence of IL-4 to generate IgE plasma cells (von Borstel et al, Science Trans! Med, Feb 7 2024). It has also been reported that IgE plasma cells are CD 19+ CD38®1(van Zelm, J Allergy Clin Immimal 134:688, 2014), There is evidence for long-lived plasma cells that produce IgE and are thought to persist for up: to and longer than I - year (Xiong et al. Trant Pediatr. 2022, 10:979012,(0055] IgE has a short half-life in serum, i,e., approximately 2 days (Stone et al, J Allergy Clin laimimol, 2010, 125: S73-S80). Free IgE binds to high-affinity FceRf receptors expressed on the surface of mast cells and basophils and to low-affinity FcsRII receptors expressed on B cells and other hematopoietic cells, which significantly enhances the half-life of IgE in its bound state. It is thought that mast cells with tightly bound surface IgE have a half-life of - 3 months, |0056] Upon re-exposure to antigen, IgE-decorated mast cells undergo degranulation and massive release of many inflammatory mediators, including histamine, Tryptase, TNF-alpha, IL-3, IL-4, 1. L-5, IL-fi, IL-10, IL-13, GM-CSF, eotaxin, Heparin, prostaglandins,.and leukotrienes. Mast ceil inflammation can lead to the clinical signs and symptoms of allergy that include anaphylaxis, asthma, urticaria, atopic eczema and food allerg.16957] To date, IgE-mediated diseases have been treated with antihistamine and antiinflammatory medications, In addition, anti- IgE antibodies have been used to target and inhibit the function of IgE directly, e.g., omalizurnab. However, these therapies require regular redosing and maintenance to suppress symptoms. In addition, in some instances treatment with ornalizumab has induced adverse reactions including skin i ftflamm tion and anaphylaxis in thePage II of 277absence of anti-drug antibodies. Accordingly, there is a need for new therapies to treat l.g£ mediated diseases,Described herein are engineered or modified immune evasive cells, including but not limited to human immune evasive cells. To overcome the problem of a subject’s immune rejection of these primary and / or stem cell-derived transplants, the inventors have developed and describe herein hypoimmunogenic cells (e.g., hypoimmunogenic pluripotent cells, differentiated cells derived from such, and primary cells) that represent a viable source for any transplantable cell type. Such cells are protected from adaptive and / or i on a t c immune rejection upon administration to a recipient subject Advantageously, the cells disclosed herein are not rejected by the recipient subject’s immune system, regardless of the subject’s genetic make-up, as they are protected from adaptive and innate immune rejection upon administration to a recipient subject, In some embodiments, the engineered and / or hypoimmunogenic cells do not express major histocompatibility complex (MHC) class I and class II antigens and / or T-cell receptors. In certain embodiments, the engineered and / or hypoimmunogenic ceils do not express MHC 1 and II antigens and / or T-cell receptors and overexpress CD47 proteins. In certain embodiments, the engineered and / or hypoimmunogenic cells such as engineered and / or hypoimmunogenic T cells do not express MHC land H antigens and / or T-cell receptors, overexpress CD47 proteins and express exogenous CARs.|00S9] In some embodiments, hypoimmunogenic cells outlined herein are not subject to an innate immune cell rejection. In some instances, hypoimmunogenic cells are not susceptible to NK cell-mediated lysis. In some instances, hypoimmunogenic cells are not susceptible to macrophage engulfment In some embodiments, hypoimmunogenic cells are usefol as a source Of universally compatible cells or tissues (e.g,, universal donor cells or tissues) that are transplanted into a recipient subject with little to no immunosuppressant agent needed.The technology disclosed herein utilizes expression of tolerogenic factors and modulation (e.g,, reduction or elimination) of MHC I, MHC II, and / or TCR expression in human cells. In some embodiments, genome editing technologies utilizing rare-cutting endonucleases (e.g., CRISPR / Cas) are also used to reduce or eliminate expression of genes involved in an immune response (e.g., by deleting genomic DNA of genes involved in an immune response or by insertions of genomic DN A into such genes, such that gene expression is impacted) in thecells. In some embodiments, genome editing technologies or other gene modulation technologies are used to insert tolerance-inducing (tolerogenic) factors in human cells, rendering the cells and their progeny (include any differentiated cells prepared therefrom) able to evade immune recognition upon engrafting into a recipient subject. As such, the cells described herein exhibit modulated expression of one or more genes and fac tors that aftect MHC I, MHC II, and / or TCR expression and evade the recipient subject’s immune system.The genome editing techniques enable double-strand DN A breaks at desired locus sites. These controlled double-strand breaks promote homologous recombination at the specific locus sites. This process focuses on targeting specific sequences of nucleic acid molecules, such as chrornosornes, with endonucleases that recognize and bind to tire sequences and induce a double-stranded break in the nucleic acid molecule. The double-strand break is repaired either by an error-prone non-homologous end-joining (NHEJ) or by homologous recombination (HR), The practice of the numerous embodiments will employ, unless indicated specifically to the contrary, conventional methods of chemistry, biochemistry, organic chemistry, molecular biology, microbiology, recombinant DNA techniques, genetics, immunology, and cell biology that are within the skill of the art, many of which are described belo w for the purpose of illustration. Such techniques are explained "fully in the literature. See, e.g., Sambrook, et al., Molecular Cloning: A Laboratory Manual (3rd Edition, 2001); Sambrook, et al., Molecular Cloning: A Laboratory Manual (2nd Edition, 1989); Maniatls etal.. Molecular Cloning; A Laboratory' Manual (1982); Ausubel et at. Current Protocols in Molecular Biology (John Wiley and Sons, updated July 2008); Short Protocols in Molecular Biology; A Compendium of Methods from Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience; Glover, DMA Cloning: A. Practical Approach, vol, I & II (IR. L Press, Oxford, 1985); Anand, Techniques for the Analysis of Complex Genomes, (Academic Press, New York, 1992); Transcription and Translation (B. Hames & 8. Higgins, Eds,, 1984); Perbal, A Practical Guide to Molecular Cloning (1984); Harlow and Lane, Antibodies, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y., 1998) Current Protocols in Immunology Q. E. Coligan, A. M. Kruisbeek, D. H. Margulies, E. M. Shevach and W. Strober, eds., 1991); Annual Review of Immunology; as well as monographs in journals such as Advances in Immunology.II. DEFINITIONS

[0063] As described in the present disclosure, the following terms will be employed, and are defined as indicated below.

[0064] The term ’’autoimmune disease” refers to any disease or disorder in which the subject mounts an immune response against its own tissues and / or cells. Autoimmune disorders can affect almost every organ system in the subject (e.g., human), including, but not limited to, diseases of the respiratory, nervous, gastrointestinal, and endocrine systems, as well as skin and other connective tissues, eyes, blood and blood: vessels. Examples of autoimmune diseases include, but are not limited to Hashimoto's thyroiditis, Systemic lupus erythematosus, Sjogren’s syndrome, Graves' disease, Scleroderma. Rheumatoid arthritis, Multiple sclerosis, Myasthenia gravis and Diabetes.

[0065] The term “best response rate’' or “BOR” is defined as an incidence of subjects with a complete response, partial response, stable disease, and / or progressi ve disease.

[0066] The term “biomarker” refers to a distinctive biological or biologically deri ved indicator of a process, event or conditi ons, hi certain embodiments, the biornarker is a gene or gene product (e.g.. a polypeptide),

[0067] The term "cancer” as used herein is defined as a hyperproliferation of cells whose unique trait (e.g., loss of normal controls) results in unregulated growth, lack of differentiation, iocal tissue invasion, and metastasis. With respect to the inventive methods, the cancer can be any cancer, including any of acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bladder cancer, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the eye, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbl adder, or pleura, cancer of the nose, nasal cavity, or middle ear, cancer of the oral cavity, cancer of the vulva, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, esophageal cancer, cervical cancer, fibrosarcoma, gastrointestinal carcinoid tumor, Hodgkin lymphoma, hypopharynx cancer, kidney cancer, larynx cancer, leukemia, liquid tumors, liver cancer, lung cancer, lymphoma, malignant mesothelioma, mastocytoma, melanoma, multiple myeloma, nasopharynx cancer, non-Hodgkin lymphoma, ovarian cancer, pancreatic cancer, peritoneum, omentum, and mesentery cancer, pharynx cancer, prostate cancer, rectal cancer, renal cancer, skin cancer, small intestine cancer, soft, tissue cancer, solid turners, stomachcancer, testicular cancer, thyroid: cancer, ureter cancer, and / or urinary bladder cancer. As u ed herein, the term "tumor’' refers to an abnormal growth of cells or tissues of the malignant type, unless otherwise specifically indicated and does not include a benign type tissue.

[0068] The term "chronic infectious disease” refers to a disease caused by an infectious agent wherein the infection has persisted. Such a disease may include hepatitis (A, B, er C), herpes virus (e.g., VZV, HSV-1, HSV-6, HSV CMV, and EBV), and HIV / A1DS. Nomviral examples may include chronic fungal diseases such Aspergillosis, Candidiasis, Coccidioidomycosis, and diseases associated with Cryptococcus and Histoplasmosis. None limiting examples of chronic bacterial infectious agents may be Chlamydia pneumoniae, Listeria monocytogenes, and Mycobacterium tuberculosis. In some embodiments, the disorder is human immunodefieiency virus (HIV) infection, hi some embodiments, the disorder is acquired immunodeficiency syndrome (AIDS).

[0069] As used herein, “clinically effective amount” refers to an amount sufficient to provide a clinical benefit in the treatment and / or management of a disease, disorder, or condition. In some embodiments, a clinically effective amount is an amount that has been shown to produce at least one improved clinical endpoint to the standard of care for the disease, disorder, or condition. In some embodiments, a clinically effective amount is an amount that has been demonstrated, for example in a clinical trial, to be sufficient to provide statistically significant and meaningful effectiveness for treating the disease, disorder, or condition. In some embodiments, the clinically effective amount is also a therapeutically effective amount. In other embodiments, the clinically effective amount is not a therapeutically effective amount.

[0070] In some embodiments, an alteration or modification (including, for example, genetic alterations or modifications) describedherein results in reduced expression of a target or selected polynucleotide sequence. In some embodiments, an alteration or modification described herein results in reduced expression of a target or selected polypeptide sequence. In some embodiments, an alteration or modification described herein results in increased expression of a target or selected polynucleotide sequence. In some embodiments, an alteration or modificatiou described herein results in increased expression of a target or selected polypeptide sequence.

[0071] The terms "decrease," "reduced,” ’’reduction,’' and "decrease” are all used herein generally to mean a decrease by a statistically significant amount. However, for avoidance of Page IS of 277doubt, decrease," "reduced,” "reduction,” "decrease” means a decrease by at least 10% as compared to a reference level, for example a decrease by at least about 20%, or at least about 30%, or at least about 40%, ar at least about 50%, or at least about 60%, or at least about 70%, or at feast about 80%, or at least about 90% or up to and including a 100% decrease (i.e. absent level as compared to a reference sample), or any decrease between 10-100% as compared to a reference level. In some embodiments, the cells are engineered to have reduced expression of one or more targets relative to an unaltered or unmodified wild-type cell.

[6072] The term “deep B cell depletion” refers to a reduction in the number of or an absence of detectable B cells in a subj ect detected in circulation and detected in a tissue sample. In some embodiments, the reduction in number of B cells is based on a comparison to a reference value, in some instances, the reference value is based on an established normal range of B cells in a healthy subject. In some instances, the reference value is the number of B cells in the subject detected in circulation and detected in a comparable tissue sample before a B cell depleting agent or composition is administered to the subject. In some embodiments, the tissue sample is a tissue biopsy. Examples of tissues include a lymph node, germinal center, gastrointestinal tract, liver, spleen, gall bladder, lung, kidney, and / or bone marrow. Methods of detecting B cells in circulation are known. For example, B cells may be detected a blood sample using flow cytometry and staining B cell markers. Method of detecting B ceils in a tissue sample (e.g., biopsy ) are known. For example, a tissue sample may be sectioned and B cells identified using immunohistochemistry' to stain B cell markers. Non-Hr ting examples of B cell markers include CD19 and CD20. In some embodiments, the number of B cells detected in a biopsy is from 0-20 c ls / mnr, from 0-15 eells / mm2, from 0-10 cells / mm2, from 0-5 cells / mm2, or 0 cells / mm2. Deep B cell depletion is discussed in Robinson, et at Front, Immunol, 2024, 15: 1454747 and Tur C, et al. Ann R ieun? Dfr. 2024. 0:1-8, both of which are incorporated by reference in their entirety herein,100731 The term “donor subject” refers to an animal, for example, a human from whom cells can be obtained, The “aan-human animals” and “noa-humanmammals” as used interchangeably herein, includes mammals such as rats, mice, rabbits, sheep, cats, dogs, cows, pigs, and nomhuman primates. The term “donor subject” also encompasses any vertebrate including but not limited to mammals, reptiles, amphibians and fish. However, advantageously,the denar subject is a mammal such as a human, or other mammals such as a domesticated mammal, e.g., dog, eat, horse, and the like, or production mammal, e.g.,, cow, sheep, pig, and the like. A “donor subject” can also refer to more than one donor, for example one or more humans or non-human animals or non-human mammals,[007 1 The term “duration of response” or “DOR” i s defined for subjects who experience a best objective response (complete response or partial response) and is the time from the first objective response to disease progression or death due to any cause, whichever occurs first. In some embodiments, if the progression or death is not observed, the DOR. will be censored at the last evaluable assessment date on or prior to the censoring events defined as (i) ongoing without event, (ii) lost to follow-up, (iii ) withdrew consent, (iv) new anticancer therapy (including stern cell transplant), and / or (v) adequate assessments no longer available.[0075| The term “endogenous" refers to a referenced molecule or polypeptide that is naturally present in the cell. Similarly, the term when used in reference to expression of an encoding nucleic acid refers to expression of an encoding nucleic acid naturally contained within the cell and not exogenously introduced. Similarly, the term when used in reference to a promoter sequence refers to a promoter sequence naturally contained within the cell and not exogenously introduced.

[0076] The term "engineered cell" as used herein refers to a cell that has been altered in at least some way by human intervention, incto ing, for example, by genetic alterations or modiflcafions such that the engineered cell differs from a wild-type cell.[0®77| The term “event-free survival” or “EFS” is the time from infusion of hypoimmune allogenic CD19-directcd CAR T cells of the present disclosure to the earliest of date of death from any cause, disease progression or relapse, or new anticancer therapy (including stem cell transplant). In some embodiments, when a subject does not have any of the above events prior to data cutoff, EFS is censored at the last adequate response assessment date on or prior to the earliest censoring event (except for stem cell transplant). In some embodiments, the censoring reason could be (i) ongoing without event, (ii) lost to follow-up, (iii) withdrew consent, (iv) stem cell transplant (subjects who proceed to stem cell transplant after infusion of hypoimmune allogenic CD19-dlrecled CAR T cells of the present disclosure will be censored at the time of stern cell transplant), and / or (v) event after at least 2 missing scheduled disease assessments,

[0978] As used herein, the term "exogenous" in the context of a polynucleotide or polypeptide being expressed is intended to mean that the referenced molecule or the referenced polypeptide is introduced into the cell of interest. The polypeptide can be introduced, for example, by introduction of an encoding nucleic acid into the genetic material of the cells such as by integration into a chromosome or as non-chromosomal genetic material such as a plasmid or expression vector. Therefore, the term as it is used i n reference to expression of an encoding nucleic acid refers to introduction of the encoding nucleic acid in an expressible form into the cell. A n exogenous polynucleotide can be inserted into at least one allele of the cell using viral transduction, for example, with a vector. In some embodiments, the vector is a pseudotyped, selfinactivating feutrviral vector that canies exogenous polynucleotide. In some embodiments, the vector is a self-inactivating lentivira.l vector pseudotyped with a vesicular stomatitis VSV-G envelope, and which carries the exogenous polynucleotide. In some embodiments, the exogenous polynucleotide is inserted into at least one allele of the cell using viral transduction, In some embodiments, exogenous polynucleoti e is inserted into at least one allele of the cell using a lent i virus based viral vector. In some embodiments, the exogenous polynucleotide is inserted into a safe harbor or target locus of at least one allele of the cell.|fiG79] At "exogenous" molecule is a molecule, construct, factor and the like that is not normally present in a cell, but can be introduced into a cell by one or more genetic, biochemical or other methods. ’’Normal presence in the cell" is determined with respect to the particular developmental stage and environmental conditions of the cell. Thus, for example, a molecule that is present only during embryonic development of neurons is an exogenous molecule with respect to an adult neuron cell An exogenous molecule can comprise, for example, a functioning version of a malfunctioning endogenous molecule or a malfunctioning version of a normally-fimctiou g endogenous molecule.

[9880] An exogenous molecule or factor can be, among other things, a small molecule, such as is generated by a combinatorial chemistry process, or a macromolecule such as a protein, nucleic acid, carbohydrate, lipid, glycoprotein, lipoprotein, polysaccharide, any modified derivative of the above molecules, or any complex comprising one or more of the above molecules. Nucleic acids include DN A and RNA, can be single- or double-stranded; can be linear, branched or circular; and can be of any length. Nucleic acids include those capable offorming duplexes, as well as triplex-forming nucleic acids. See, for example, U. S. Pat. Nos, 5,176,996 and 5,422,251. Proteins include, but are not limited to, DNA-binding proteins, transcription factors, chromatin remodeling factors, methylated DNA binding proteins, polymerases, methylases, demethylases, acetylases, deacetylases, kinases, phosphatases, integrases, recombinases, ligases, topoisomerases, gyrases and helicases,10081] An exogenous molecule or construct can be the same type of molecule as an endogenous molecule, e.g,, an exogenous protein or nucleic acid. In such instances, the exogenous molecule is introduced into the cell at greater concentrations than that of the endogenous molecule in the cell, In some instances, an exogenous nucleic acid can comprise an infecting viral genome, a plasmid or epi some introduced into a cell, or a chromosome that is not normally present in the cell. Methods for the introduction of exogenous molecules into cells are known to those of skill in foe art and include, but are not limited to, lipid-mediated transfer (he., liposomes, including neutral and cationic lipids), electroporation, direct injection, cell fusion, particle bombardment, calcium phosphate co-precipitation, DEAE -dextran-mediated transfer and viral vector-mediated transfer.W82 A "gene,” for the purposes of the present disclosure, includes a DNA region encoding a gene product, as well as all DN A regions which regulate the production of the gene product, whether or not such regulatory sequences are adjacent to coding and / or transcribed sequences. Accordingly, a gene includes, but is not necessarily limited to, promoter sequences, terminators, translational regulatory sequences such as ribosome binding sites and internal ribosome entry sites, enhancers, silencers, insulators, boundary elements, replication origins, matrix attachment sites and / or locus control regions,(00S3] ’’Gene expression” refers to the conversion of the information, contained in a gone, into a gene produet, A gene product can be the direct transcriptional product of a gene (e.g., mRNA, tRNA, rRN A, antisense RNA, ribozyme, structural RNA or any other type of RNA) or a protein produced by translation of an mRN A. Gene products also include R As which are modified, by processes such as capping, polyadenylatiom methy lation, and editing, and proteins modified by, for example, methylation, acetylation, phosphorylation, ubiquitination, ADP-ribosylation, myristoylation, and / or glycosylation.(00S4| The term “genetic modification” and: its grammatical equivalents as used herein can refer to one or more alterations of a nucleic acid, c.g., the nucleic acid within an organism's genome. For example, genetic modification can refer to alterations, additions, and / or deletion of genes or portions of genes or other nucleic aci d sequences, A genetically modified cell can also refer to a cell with an added, deleted and / or altered gene or portion of a gene. A genetically modified cell can also refer to a cell with an added nucleic acid sequence that is not a gene or gene portion. Genetic modifications include, for example, both transient knock-in or knock-down mechanisms, and mechanisms that result in permanent knock-in, knock-down, or knock-out of target genes or portions of genes or nucleic acid sequences Genetic modifications include, for example, both transient knock-in and mechanisms that resul t in permanent knock-in of nucleic acids sequences Genetie modifications also include, for example, reduced or increased transcription, reduced or increased mRNA stability, reduced or increased translation, and reduced or increased protein stability <;00H5| As used herein, the terms "grafting’’, "administering," "introducing1’, "implanting” and "transplanting" as well as grammatical variations thereof are u sed interchangeably in the context of the placement of cells (e.g,, cells described herein) into a subject, by a method or route which results in localizati on or at least partial localization of the introduced cells at a desired site or systemic introduction (e.,, into circulation). The cells can be implanted directly to the desired site, or alternatively be administered by any appropriate route which results in delivery to a desired location in the subject where at least a portion of the implanted cells or components of the cells remain viable. The period of viability of the cells after administration to a subject can be as short as a few hours, e, g. twenty-four hoars, to a few days, to as long as several years. In some embodiments, the cells can also be administered (e.g., injected) a location other than the desired site, such as in the brain or subcutaneously, for example, in a capsule to maintain the implanted cells at the implant location and avoid migration of the implanted cells,By "HLA” or "human leukocyte antigen" complex is a gene complex encoding the MHC proteins in humans. These cell-surface proteins that make up the HLA complex are responsible for the regulation of the immune response to antigens. In humans, there are two MHCs, class I and class 11, "HLA-I" and "HLA-II". HLA- 1 includes three proteins, HLA-A, HLA-B and HLA-C, which present peptides from the inside of the cell, and antigens presentedby the HLA-1 complex attract killer T-cells (also known as CD8+ T-cells or cytotoxic I cells). The HLA-I proteins are associated with 0-2 microglobulin (B2M). HLA-I1 includes five proteins, HLA-DP, HLA-DM, MLA-DOB, HLA-DQ and HLA-DR, which present antigens from outside the cell to T lymphocytes. This stimulates CD4+ cells (also known as T-helper cells). It should be understood that the use of either ”MHC '* or "HLA” is not meant to be limiting, as it depends on whether the genes axe from humans (HLA) or murine (MHC). Thus, as it relates to mammalian cells, these terms may be used interchangeably herein.(0087] As used herein to characterize a cell, the term ’’hypoimmunogenic'' generally means that such cell is less prone to innate or adaptive immune rejection by a subject into which such cells are transplanted, e.g., the cell is less prone to allorejection by a subject into which such cells are transplanted. For example, relative to a cell of the same cell type that does not comprise the modifications, such a hypoimmunogemc cell may be about 2.5%, 5%, 10%. 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97.5%, 99% or more less prone to innate or adaptive immune rejection by a subject into which such cells are transplanted. In some embodiments, genome editing technologies arc used to modulate the expression of MHC 1 and MHC II genes, and thus, contribute to generation of a hypoi nnnunogenic cell. In some embodiments, a hypoimmunogenic cell evades immune rejection in an MHC-mismatched allogeneic recipient. In some instance, differentiated cells produced from the hypoimmunoge c stem cells outlined herein evade immune rejection when administered (e.g., transplanted or grafted) to an MHC-mismatched allogeneic recipient. In some embodiments, a hypoimrnunogenic cell is protected from T cell-mediated adaptive immune rejection and / or innate immune cell rejection. Detailed descriptions of hypoimmunogenic cells, methods of producing thereof, and methods of using thereof are found in WO2016183041 filed May 9, 201.5; WQ2018132783 filed January 14, 2018; WO2018176390 filed March 20, 2018; W02020018615 filed July 17, 2019; W020200I8620 filed July 17, 2019; PCT7US202O / 44635 filed July 31, 2020; WO2021022223 filed July 31, 2020; WO2021041316 filed August 24, 2020;WO20 1222285 filed April 27, 2021, 2020; and WO2021222285 fifed April 27, 2021, the disclosures including the examples, sequence listings and figures are incorporated herein by reference in their entirety.[00881 Hypoimmunogenicity of a cell can be determined by evaluating the immunogenicity' of the cell such as the cell’s ability to elicit adaptive and innate immune responses or to avoid eliciting such adaptive and innate immune responses. Such immune response can be measured using assays recognized by those skilled in the art. In some embodiments, an immune response assay measures the effect of a hypoimmunogenic cell on T cell proliferation, T cell activation, T cell killing, donor specific antibody generation, NK cel! proliferation, NK cell activation, and macrophage activity. In some cases, hypoimmurmgenie cells and derivatives thereof undergo decreased killing by T cells and / or NK cells upon administration to a subject. In some instances, the cells and derivatives thereof show decreased macrophage engulfment compared to an unmodified or wild-type cell. In some embodiments, a hypoimmunogenic cell elicits a reduced or diminished immune response in a recipient subject compared to a corresponding unmodified wild-type cell. In some embodiments, a hypoimmunogenic cell is nonimmunogenic ar fails to elicit an immune response in a recipient subject.The term percent "identity, ” in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLAST? and BLASTN or other algorithms available to persons of skill) or by visual inspection. Depending oa the application, the percent "identity" can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences io be compared. For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algori thm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence: identity for the test sequenee(s) relative to the reference sequence, based on the designated program parameters.0890 Optimal alignment of sequences tor comparison can be conducted* e,g.}by the local homoiogy algorithm of Smith & Waterman, Adv. AppL Math. 2:482 (1981), by thehomology alignment algorithm of Needlemau & Wunsch, J. MoL Biol 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat’l. Acad. Sei. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr,, Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).10091] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algori hm, which is described in Altschul et at, J. Mol, Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology' Information.(0092] ’’Immune signaling factor'' as used herein refers to, in some eases, a molecule, protein, peptide and the like that activates immune signaling pathways.|0093| "Immunosuppressive factor” or "immune regulatory' factor" or ’’tolerogenic factor" as used herein include hypoimmunity factors, complement inhibitors, and other factors that modulate or affect the ability of a cell to be recognized by the immune system of a host or recipient subject upon admimstratim, trarssplantation, or engraftment. These may be in combination with additional genetic modifications.(0094] The terms ’’increased", ’’increase*' or "enhance” or "activate” are all used herein to generally mean an increase by a statically significant amount; for the avoidance of any doubt, the terms "increased ”, "increase” or "enhance" or "activate" means an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a -fold, or at least about a 4-fold, c>r at least about a 5 -fold or at least about a 10-fold increase, or any increase between 2-fold and 10- fold or greater as compared to a reference level. In some embodiments, the reference level, also referred to as the basal level, is 0.|0095| In some embodiments, the alteration is an indel. As used herein, "indel" refers to a mutation resulting from an insertion, deletion, or a combination thereof. As will be appreciated by those skilled in the art, an indel in a coding region of a genomic sequence will result in a ftameshift mutation, unless the length of the indel is a. multiple of three. In some embodiments,the alteration is a point mutation. As used herein, ’’point mutation” refers to a substitution that replaces one of the nucleotides, A gene editing (e.g.. CRISPR / Cas) system of the present disclosure can be used to induce an indel of any length or a point mutation in a target polynucleotide sequence.As used herein, “knock down” refers to a red action in expression of the target mRNA or the corresponding target protein. Knock down is commonly reported relative to levels present following administration or expression of a noncontrol molecule that does notmediate reduction in expression levels of RNA (e.g,, a non-targeting control shRNA, siRNA, or miRNA). In some embodiments, knock down of a target, gene is achieved by genetic modification, including use of gene editing systems (e.g., CRISPR / Cas).|0097| Knock down is commonly assessed by measuring the mRNA levels using quantitative polymerase chain reaction (qPCR) amplification or by measuring protein levels by western blot or enzyme-linked immunosorbent assay (ELISA). Analyzing the protein level provides an assessment of both mRN cleavage as well as translation inhibition. Further techniques for measuring knock down include RNA solution hybridization, nuclease protection, northern hybridization, gene expression monitoring with a microarray, antibody binding, radioimmunoassay, and fluorescence activated cell analysis. Those skilled in the art will readily appreciate how to use the gene editing systems (e.g,, CRISPR / Cas) of the present disclosure to knockout a target polynucleotide sequence or a portion thereof based upon the details described herein.[0098} By "knock in" or “knock-in” herein is meant a genetic modification resulting from the insertion of a DNA sequence into a chromosomal locos in a host celt This causes initiation of ar increased levels of expression of the knocked in gene, portion of gene, or nucleic acid sequence inserted product, e.g., an increase in R A transcript levels and / or encoded protein levels. As will be appreciated by those in the art, this can be accomplished in several ways, including inserting or adding one or more additional copies of the gene or portion thereof to the host cell or altering a regulatory component of the endogenous gene increasing expression of the protein is made or inserting a specific nucleic acid sequence whose expression is desired. This may be accomplished by modifying a promoter, adding a different promoter, adding an enhancer, adding other regulatory elements, or modifying other gene expression sequences.[W99] As used herein, "knock out" or “knock-out” includes deleting all or a portion of a target polynucleotide sequence in a way that interferes with the translation or function of the target polynucleotide sequence. For example, a knock out can be achieved by altering a target polynucleotide sequence by inducing an insertion or a deletion f ‘Indel”) in the target polynucleotide sequence, including in a functional domain of the target polynucleotide sequence (e.g., a DNA binding domain). Those skilled in the art will readily appreciate how to use the gene editing systems (e.g., CRISPR / Cas) of the present disclosure to knock out a target polynucleotide sequence or a portion thereof based upon the details described herein.

[0100] In some embodiments, a genetic modification or alteration results in a knock out or knock down of the target polynucleotide sequence or a portion thereof. Knocking out a target polynucleotide sequence or a portion thereof using a: gene editing system (e.g., CRISPR / Cas) of the present disclosure can be useful for a variety of applications. For example, knocking out a target polynucleotide sequence in a cell can be performed in vitro for research purposes. For ex vivo purposes, knocking out a target polynucleotide sequence in a cell can be usefill for treating or preventing a disorder associated with expression of the target polynucleotide sequence (e.g., by knocking out a mutant allele in a cell ex vivo and introducing those cells comprising the knocked out mutant allele into a subject) or for changing the genotype or phenotype of a cell.

[0101] "Modulation*' of gene expression refers to a change in the expression level of a gene. Modulation of expression can include, but is not limited to, gene acti vation and gene repression. Modulation may also be complete, i.e., wherein gene expression is totally inactivated or is activated to wild-type levels or beyond; or it may be partial, wherein gene expression is partially reduced, or partially activated to some fraction of wild-type levels,

[0102] The methods provided herein can be used to alter a target polynucleotide sequence in a cell The present disclosure contemplates altering target polynucleotide sequences t» a cell for any purpose. In some embodiments, foe target polynucleotide sequence in a cell is altered to produce a mutant cell. As used herein, a "mutant cell” refers to a cell with a resulting genotype that differs from its original genotype. In some instances, a "mutant cell” exhibits a mutant phenotype, for example when a normally functioning: gene is altered using the gene editing systems (e.g., CRISPR as) systems of foe present disclosure. In other instances, a "mutant cell” exhibits a wild-type phenotype, for example when a gene editing system (e.g,,CRISPR / Cas) system of the present disclosure is used to correct a mutant genotype. In some embodiments, the target polynucleotide sequence in a cell is altered to correct or repair a genetic mutation (e.g., to restore a normal phenotype to the cell). In some embodiments, the target polynucleotide sequence in a cell is altered to induce a genetic mutation (e.g., to disrupt the function of a gene or genomic element).10103] The term “native cell” as used herein refers to a cell that is net otherwise modified (e.g., engineered). In some embodiments, a native cell is a naturally occurring wild-type or a control celt(0104] The term “objective response rate” or “ORR” is defined as a proportion of subjects with a: best overall response of complete response or partial response as determined by an investigator and / or central assessment. Subjects who do not meet the criteria for an objective response by an analysis cutoff date will be considered non-responders. In some embodiments, disease assessments obtained after infusion and up through an observation of progression or start of new anti-cancer therapy will be used. In some embodiments, response assessment in nonHodgkin lymphoma subjects will be based on the Lugano classification criteria and in chronic lymphocytic leukemia subjects based on the International Workshop on Chronic Lymphocytic Leukemia criteria.(0105] The term "operatively linked” or "operably linked'' are used interchangeably with reference to a j uxtaposition of two or more componen ts (such as sequence elements), in which the components are arranged such that both components function normally and allow the possibility that at least one of the components can mediate a function that is exerted upon at least one of the other components. By way of illustration, a transcriptional regulatory sequence, such as a promoter, is operatively linked to a coding sequence if the transcriptional regulatory sequence controls the level of transcription of the coding sequence in response to the presence or absence of one or more transcriptional regulatory factors. A transcriptional regulatory sequence is generally operatively linked in cis with a coding sequence, but need not be directly adjacent to it. For example, an enhancer is a transcriptional regulatory sequence that is operatively linked to a coding sequence, even though they w not contiguous.|0106| The term “overall survi val” or “OS” is defined as the time from infusion of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure to death from anycause. In some embodiments, subjects who have not died by the analysis data cutoff date will have survival time censored at their last date known to be alive. In some embodiments, for subjects alive or dead after the data cutoff date, survival time will be censored at the data cutoff date.|(| W7| The term “progression free survival” or “PFS” is defined as the time of infusion of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure to disease progression or death from any cause, whichever occurs first. In some embodiments, if the progression or death is not observed, the PFS will be censored at the last evaluable assessment date on or prior to the censoring events defined as (1) ongoing without event, (ii) lost to follow up, (11 i) withdrew consent, (iv) new anticancer therapy (including stem cell transplant), and / or (v) adequate assessments no longer available,|O108| As used herein, "promoter," "promoter sequence,’* or "promoter region" refers to a DMA regulatory region / sequence: capable of binding RNA polymerase and involved in initiating transcription of a downstream coding or non-coding sequence. In some examples, the promoter sequence includes the transcription initiation site and extends upstream to include the minimum number of bases or elements necessary to initiate transcription at levels detectable above background. In some embodiments, the promoter sequence includes a transcription initiation site, as well as protein binding domains responsible for the binding of RN polymerase. Eukaryotic promoters will often, but not always, contain "TATA ” boxes and "CAT” boxes.In some embodiments, the engineered and hypoimmanogenic cells described are propagated from a primary T cell or a progeny thereof As used herein, the term “propagated from a primary' T cell or a progeny thereof” encompasses the initial primary T cell that is isolated from the donor subject and any subsequent progeny thereof. As used herein, the term “progeny” encompasses, e.g,, a first-generation progeny, re,, the progeny is directly derived from, obtained from, obtainable from or derivable from the initial primary T cell by, e.g., traditional propagation methods. The term “progeny” also encompasses further generations such as second, third, fourth, fifth, sixth, seventh, or more generations, i.e., generations of cells which are derived from, obtained from, obtainable from or derivable from the former generation by, e.g., traditional propagation methods. The term “progeny” also encompasses modified cells that result from the modification or alteration of the initial primary T cell or a progeny thereof.

[9119] Th© term “recipient patient” refers to an animal, for example, a human to whom treatment, including prophylactic treatment, with the cells as described herein, is provided. For treatment of those infections, conditions or disease states, which are specific for a specific animal such as a human patient, the term patient refers to that specific animal. Hie term ‘’recipient patient” also encompasses any vertebrate including but not limited to mammals, reptiles, amphibians and fish. However, advantageously, the recipient patient is a mammal such as a human, or other mammals such as a domesticated mammal, e.g., dog, cat, horse, and the like, or production mammal, e.g., cow, sheep, pig, and the like.[6111| As used herein, the “reference value” refers to a value used in comparison to fest value or lest result to interpret the result. In some instances* a reference value is based on the results that are seen in 95% of a population. In some instances, refence is a range of values, e.g., a reference interval or a reference range. In some instances, a reference value is based on the results of an assay, test, or assessment performed on a subject or sample prior to administration of a composition, therapy, intervention, or the like. For example, a reference value cart be the number of B cells in a sample obtained from a subject before the subject is administered a B cell depleting therapy or composition. As another example, a reference value can be the concentration of antibodies (e.g., IgG, IgM, IgE) in a sample obtained from a subject before the subject is administered a B cell depleting therapy or composition. In some instances, a reference value is an established range typically seen in a population having a disease or disorder, e.g., allergy, autoimmune disease, and / or cancer. In some instances, a reference value is an established range seen in approximately 95% of a healthy population. A population can be based on. one or more characteristics of individual subjects, e.g., male, female, age, genetic profile, ethnicity, disease, and or disorder.

[0112] As used herein, the terms ’’regulatory sequences,” '’regulatory elements," and "control elements’' arc interchangeable and refer to polynucleotide sequences that are upstream (5* non-coding sequences), within, or downstream (3’ non-translated sequences) of a polynucleotide target to be expressed. Regulatory sequences influence, for example but are not limited to, the timing of transcription, amount or level of transcription, RNA processing or stability, and / or translation o f the re lated structural nucleotide sequence. Regulatory sequences may include activator binding sequences, enhancers, introns, poly adenylation recognitionsequences, promoters, repressor binding sequences, stem-loop structures, translational initiation sequences, translation leader sequences, transcription termination sequences, translation termination, sequences, primer binding sites, and the like. It is recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, nucleotide sequences of different lengths may have identical regulatory or promoter activity.10113] As used herein, a “target” can refer to a gene, a portion of a gene, a portion of the genome, or a protein that is subject to regulatable reduced expression by the methods described herein,|0114] As used herein, ‘therapeutically effective amount” refers to an amount sufficient to provide a therapeutic benefit in the treatment and / or management of a disease, disorder, or condition. In some embodiments, a therapeutically effective amount is an amount sufficient to ameliorate, palliate, stabilize, reverse, slow, attenuate or delay the progression of a disease, disorder, or condition, or of a sy mptom or side effect of the disease, disorder, or condition. In some embodiments, the therapeutical ly effective amount is also a clinically effective amount, In other embodiments, the therapeutically effective amount is not a clinically effective amount. |G115] The term “time to next treatment” or ‘T NT” is defined as the time from infusion of hypoimmune allogenic CD19-directed C AR T cells of tile present disclosure to the next anticancer treatment (including stem cell transplant) or death from any cause, whichever occurs first. In some embodiments, when subsequent anticancer treatment is not received (e.g., continuing on current treatment) and subjects are still alive by the analysis data cutoff date, subjects will be censored at their last available follow-up date before the cutoff date.10116] As used herein, the term ’’treating*' and ’'treatment** includes administering to a subject a therapeutically or clinically effective amount of cells described herein so that the subject has a reduction in at least one symptom of the disease or an improvement in the disease, for example, beneficial or desired therapeutic or clinical results. For purposes of this technology, beneficial or desired therapeutic or clinical results include, but are not limited to, alleviation of one or more symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. Treating can refer to prolonging survival as compared to expected survival if not receiving treatment. Thus, one ofskill in the art realizes that a treatment may improve the disease condition, but may not be a complete cure for the disease. In some embodiments, one or more symptoms of a condition, disease or disorder are alleviated by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% upon treatment of the condition, disease or disorder.

[0117] For purposes of this technology, beneficial or desired therapeutic or clinical results of disease treatment include, but are not limited to, alleviation of one or more symptoms, dimimshment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable,

[0118] A ’’vector" or "construct” is capable of transferring gene sequences to target cells. Typically, ’'vector construct," "expression vector," and "gene transfer vector," mean any nucleic acid construct capable of directing: the expression of a gene of interest and which can transfer gene sequences to target cells. Thus, the term includes cloning, and expression vehicles, as well as integrating vectors. Methods for the introduction of vectors or constructs into cells are known to those of skill in the art and include, but are not limited to, lipid-tnediate transfer (i.e,, liposomes, including neutral and cationic lipids), electroporation, direct injection, cell fusion, particle bombardment, calcium phosphate eo-precipitation, DE E-dextran-mediated transfer and / or viral vector-mediated transfer.

[0119] In some embodiments, the cells are engineered to have reduced or increased expression of on© or more targets relati ve to an unaltered or unmodified wi i- type cell In some embodiments, the cells are engineered to have constitutive reduced or increased expression of one or more targets relative to an unaltered or unmodified wild-type cell. In some embodiments, the cells are engineered to have regulatable reduced or increased expression of one or more targets relative to an unaltered or unmodified wild -type cell. In some embodiments, the cells comprise increased expression of CD47 relative to a wild-type cell or a control cell of the same cell type. By '‘wil -t pe" or “wt” or “control” in the context of a cell means any cell found in nature. Examples of wild type or control cells include primary cells and T cells found in nature. However, by way of example, in the context of an engineered cell, as used herein, “wild-type” or “control” can also mean an engineered cell that may contait nucleic acid changes resulting in reduced expression of M’HC I and / or II and / or T-cell receptors, but did not undergo the geneediting procedures to result in overexpression of CD47 proteins. For example, as used herein, “wild-type” or ‘’control” means an engineered cell that comprises reduced or knocked out expression of B2M, CUT A, and / or TRAC. Also as used herein, “wild-type” or “control” means an engineered cell that comprises reduced or knocked out expression of B2M CUTA, 'FRAC, and / or TRBC. As used herein, “wild-type” or “control” also means an engineered cell that may contain nucleic acid changes: resulting in overexpression of CD47 proteins, but did not undergo: the gene editing procedures to result in reduced expression of MHC I and / or II and / or T-cell receptors. In the contex t of an iPSC or a progeny thereof / “wild-type” or “control” also means an iPSC or progeny thereof that may contain nucleic acid changes resulting in pluripotency but did notundergo the gene editing procedures of the present disclosure to achieve reduced expression of MHC I and / or II and or T-cell receptors, and / or overexpression of CD47 proteins. Tn the context of a primary T cell or a progeny thereof, “wild-type” or “control” also means a primary T cell or progeny thereof that may contain nucleic acid changes resulting in reduced expression of MHC I and / or II and / or T-cell receptors, but did not undergo the gene editing procedures to result In overexpression of CD47 proteins. For example, as used herein, “wild-type’’ or “control” means a primary T cell or progeny thereof that comprises reduced or knocked out expression of B2M, CUT A, and / or TRAC. Also as used herein, “wild-type’* or “control” means a primary T cell or progeny thereof that comprises reduced or knocked out expression of B2M, CUT A, TRAC, and / or TRBC. Also in the context of a primary T cell or a progeny thereof, “wild-type” or “control” also means a primary T cell or progeny thereof that may contain nucleic acid changes resulting in overexpression of CD47 proteins, but did not undergo the gene editing procedures to result in reduced expression of MHC I and / or II and / or T-cell receptors. In some embodiments, the cells are engineered to have regulatable reduced or increased expression of one or more targets relative to a cell of the same cell type that does not comprise the modifications. In some embodiments, the wild-type cell or the control cell is a starting material. In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. For example, unmodified T ceils obtained from a donor is a starting material that are considered wild-type or control cells as contemplated herein. In another example, an iPSC cell line starting material is a starting material that is considered a wild-type or control cell ascontemplated herein. In some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered celt | 120| It is noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusi ve terminology as “solely,” “only,” and the like in connection with the recitation of claim elements, or use of a “negative” limitation. As will be apparent to those of skil l in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure, Any recited method may be carried out in the order of e vents recited or in any other order that is logically possible. Although any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present disclosure, representotivc illustrative methods and materials are now described.[0121) 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 this technology belongs. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that, stated range, is encompassed within the present disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the present disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the present disclosure. Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating unrecited number may be a number, which, in the context presented, provides the substantial equivalent of the specifically recited number. The term about is used herein to mean plus or minus ten percent (Whs) of a value. For example, “about 100” refers to any number between 90 and 110.1012 ] All publications, patents, and patent applications cited in this specification are incorporated herein by reference to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference. Furthermore, each cited publication, patent, or patent application is incorporated herein by reference to disclose and describe the subject matter in connection with which tire publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the technology descri bed herein is not enti tled to antedate such publication by virtue of prior technology. Further, the dates of publication provided might be different from the actual publication dates, which may need to be independently confirmed. |01231 Before the technology is further described, it is to be understood that this technology is not limited to particular embodiments described, as such may, of course, vary'- It « also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be li ited only by the appended claims. It should also be understood that the headers used herein are not limiting and are merely intended to orient the reader, but the subject matter generally applies to the technology disclosed herein.Ill, DETAIEEB DESCRIPTION OF THE EMBODIMENTSA. Methods of Treatment with Hypoimmmiogeniic T Cells(01241 As is described in further detail herein, provided herein are methods for treating a patient with a disease, disorder, or condition through administration of hypoimmunogenic cells, particularly hypo immunogenic T cells. As will be appreciated, far all the multiple embodiments described herein related to the timing and / or combinations of therapies, administration of the cells is accomplished by a method or route which results in at least partial l ocalization of the introduced cells at a desired site. Cells can be infused, implanted, or transplanted directly to the desired site, or alternatively be administered by any appropriate route which results in delivery to a desired location in the subject where at least a portion of the implanted cells or components of the cells remain viable.16125] In some embodiments, described herein is a method of treating a subject having or suspected of having an IgE-mediated disease or disorder, the method including: administering to the subject a therapeutically effective amount of a composition including a population of engineered hypoi munogenic T cells, wherein the engineered hypoinununogenic T cells include one or more modifications that: (a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHO) class I molecules and / or one or more molecules that regulate expression of the one or more M I IC class I molecules, (ii) one or more M. HC class II molecules and / or one or more molecules that regulate expression of the one or more MI-IC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, (h) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not include the modification, and (c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface o f an IgE -producing cell. In some embodiments, the antigen expressed on the surface of an IgE-prodricing cell includes CD 19, CD20, CD22, BCMA, GPRC5D, CD38, or any combination thereof. In some embodiments, the engineered T cells include one or more of a CD19-specific CAR, a CDSO-specific CAR, a CD22-specific CAR., a BCMA-specific CAR, a GPRC5D-speeific CAR, a CD38-specifie CAR, or any combination thereof.

[0126] In some embodiments, described herein is a method of treating a subject having or suspected of having: an IgE-mediated disease or disorder, the method including: administering to the subject a therapeutically effective amount of a composition including a population of engineered hypoimmunogenic T cells including: (a) reduced expression of beta-2 microglobulin (B2M), Class II Major Histocompatibility Complex Transactivator (CUT A), and T cell receptor alpha constant (TRAC) relative to a control or wild -type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CDI9-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

[0127] In some embodiments, the IgE-mediated disease or disorder is asthma, Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chronic spontaneous (idiopathic) urticaria, chronic rhinos usitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angioederna, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or any combination thereof.

[0120] In some embodiments, the IgE-mediated disease or disorder is asthma. In some embodiments, the asthma is allergic asthma, steroid refractory asthma, severe asthma, and / or difficult to treat asthma, or any combination thereof. In some embodiments, the asthma is severe,refractory T Helper 2 (TH2)~high IgE-mediated allergic asthma. Examples of categories, diagnoses, and characteristics of asthma can be found in US 2.022 / 0169739, the disclosure of which is incorporated herein by reference in its entirety. Further guidelines for definition and evaluation of severe asthma are: providing in Chung et al,, Ruropecm Respiratory Jaurrai. 2013.43(2):343-373, the disclosure of which is incorporated herein by reference in its entirety’.

[0129] In some embodiments, described herein is a method, wherein the food allergy is a severe, refractory food allergy. In some embodiments, the food allergy is an allergy to peanut, shellfish, egg, soy, wheat, fish, and / or tree nut, or a polypeptide allergen thereof.

[0130] In some embodiments, the methods of treatment described herein result in deep B cell depletion. In some embodiments, the method results B cell depletion in circulation and a tissue. In some embodiments, the method results in B cell depletion in a test tissue sample, and the test tissue sample is or includes tissue from a lymph node, genninal center, gastrointestinal tract, liver, spleen, gall bladder, lung, kidney, and / or bone marrow. In some embodiments, the test tissue sample is a tissue biopsy. In some embodiments, the B ceils express CDl 9 and / or CD20 and the CD19-expressing and / or CD20rexpressing B cells are detected in the tissue biopsy by specifically staining the CDl 9-expressing and / or CD20~expre$smg cells. Methods detecting or staining B cells and / or makers (e.g., CDl 9, CD20) are known. Examples of methods of detecting B cells include immunohistochemical staining. Au example of detecting deep B cell depiction is provided in Tur C, et al. Am Rheum Dis 2024: 1-8, which is incorporated herein by reference in its entirety. In some embodiments, the number of B cells detected is from 0-20 cells / mrir, from 0-15 cells / rmu2, from ( O ceils / mm3, from 0-5 cells / mnr, or 0 cells / mm2. Insome embodiments, the tissue biopsy is from a lymph node, fa some embodiments, the test tissue sample is collected at least 2 weeks after administration of the composition including a population of engineered hypoimnmnogenic T cells. In some embodiments, the test tissue sample is collected no more than 3 months after administration: of the composition including a population of engineered hypoimmunogenic T cells. In some embodiments, the test tissue sample is collected between 2 weeks and 3 months after the administration of the composition including a population of engineered hypoimmunogenic: T cells. In some embodiments, the test tissue sample is collected about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, and / or about 12 weeks after the administration of the composition including a population of engineered hypoimmunogerdc T cells. In some embodiments, the test tissue sample is collected about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 7 weeks to about 8 weeks, about 8 weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, and / or about 11 weeks ft) about 12 weeks after the administration of the composition including a population of engineered hypoimmunogenic T cells.[0131) In some embodiments, the methods o f trea tm ent described herein result reduced or depleted IgE antibodies fa a test sample from the sub ect In some embodiments, the test sample is a blood sample or serum sample. In some embodiments, the test sample includes a concentration of IgE antibodies that is decreased compared to a reference value. In some embodiments, the reference value is the concentration of IgE antibodies detected in a reference sample obtained from the subject prior to administering: the composition including a population of engineered hypolmmunogenic T cells. In some embodiments, the reference value is a concentration of IgE associated with and / or diagnostic of an IgE-mediated disease or disorder. In some embodiments, the reference value is a concentration of IgE antibodies from 200 ng / mL to 1000 ng / mL, from 200 ng / mL to 900 ng / mL, from 200 ng / mL to 800 ng / mL, from 200 ng / mL to 700 ng / mL, from 200 ng / mL to 600 ng / mL, from 200 ng / mL to 500 ng / mL, from or 200 n / mL to 400 ng / mL. Without wishing to be bound by theory, it is thought that a scrum concentration of IgE antibodies above 200 ng / mL is elevated above IgE concentrations observed in healthy subjects and / or subjects that have and / or are diagnosed with an IgE-mediated disease or disorder. In some embodiments, the concentration of IgE antibodies in the test sample is within a rangefrom 0 ng / mL to 200 ng / mL, In some embodiments, the concentration of IgE antibodies in the test sample is up to 200 ng / mL, ISO ng / mL, 160 ngmt, 140 ng / mL, 120 cgniL. 100 ug'mL, 80 ng / mL, 60 ng / mL, 40 ug-'mL, 20 ng / mL, I ngmL, 0, 1 ng / mL, 0.09 ng L, 0,08 ng / mL, 0.07 ng / mL, 0.06 ng / mL, 0,05 ng / mL, 0.04 ng / mL, 0.03 ng / mL, 0.02 ng / mL, or 0.01 ng / mL. In some embodiments, the test sample includes no IgE antibodies or IgE antibodies are undetectable. Without wishing to be bound by theory, It is thought that a serum coneentration of igh antibodies below.200 ng / mL (e.g., 0-200 ng / L) is within a normal range found in healthy subjects and / or not found in subjects that have and / or are diagnosed with an IgE-mediated disease or disorder. In some embodiments, the test sample is collected between 2 days and 3 months after administration of the composition including a population of engineered hypoimmunogenic T cells. In some embodiments, the test sample is collected from the subject about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, and / or about 1 weeks after administration of the composition including a population, of engineered hypoimmtmogemc T cells. In some embodiments, the test sample is collected from the subject about 1 week to about 2 weeks, about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 7 weeks to about 8 weeks, about 8 ’weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, and / or about 11 weeks to about 12 weeks after administration of the composition including a population of engineered hypoimmunogenfc T cells.(0132] In some embodiments, B cell counts will substantially recover to normal levels after about 3, 4, 5, 6, 7, S, 9, 10, T I, or 12 months. In some embodiments, recovered B cells will show a sustained naive phenotype profile, indicating the potential for an immune reset and a long-term functional cure of the subject. Without wishing to be bound by theory, it is contemplated that the B cell depletion caused by the CAR T cells may lead to an immune reset, e.g., as evidenced by the durable absence of symptoms of the disease even in the presence of reconstituted B cell numbers.(0133] In some embodiments, described herein is a method, wherein the engineered hypoimmunogenic T cells are B2M knock out, CIITA knock out, and / or TRAC knock out cells, optionally wherein the engineered hypoimmuuogenic T cells are B2M knock out, CIITA knockout, and TRAC knock out ceils. In some embodiments, the engineered hypoimmunogeuic T cells am BSM”^® ®5, CHTA”llfe^fldiii5and / or TRAC®^^ cells. In some embodiments, the engineered hypoimmunogenic T cells are the engineered hypoimmunogemc T cells areCinA*!':i:;y and T'RAC!::'te?:;iffc;ceils.

[0134] In some embodiments, the one or more tolerogenic factors include or are selected from the group consisting of CD47, SIRPalpha engagers, CD24, CD27, CD35, CD46, CD55, CD59, CD200, HLA-C, IILA-E, HLA-E heavy chain, HLA-G, PD-LI, [DOI, CTLA4-Ig, Cl-Inhibitor, IL-10, IL-35, FasL, CCL2L CCL22, MfgeS, A20 / TNFAIP3. CD39, CR. L HLA-F, IL15-RF, MA. NF, and / or Serpmb9» optionally wherein the one or more tolerogenic factors include CD47 or an engineered CD47. Examples of engineered CD47 variants can be found in WO2023158836 and PCT / US2024 / 043615, herein incorporated by reference. In some embodiments, the engineered T cells evade NK cell-mediated cytotoxicity upon administration to the subject, hi some embodiments, the engineered T cells are protected from cell lysis by mature NK cells upon administration to the subject. In some embodiments, the engineered T cells evade macrophage-mediated cytotoxicity, optionally wherein the macrophage-mediated cytotoxicity involves phagocytosis and / or reactive oxygen species.

[0135] In some embodiments, file subject was treated with an immunodepleting therapy prior to administering the engineered T cells.

[0136] In some embodiments, the composition comprising a population of engineered hypoi mmunogenic T cells is administered to the subject at a dosage of about 30 x. 10* CART cells to about 1000x106CAR+ cells. In some embodiments, the composition comprising a population of engineered, hypoimmunogenic T cells is administered, to the subject at a dosage of at least about 30x10* CAR* cells, at. least about 60x 10* CART cells, at least about 90x10* CAR+ cells, at least about 120x10* C R* cells, at least about 200 10* CART cells, at least about 400 xlO* CART cells, at least about 600 x 10* CART cells, at least about 800 x 10* CART cells, or at least about 1000 xlO* CAR* cells.

[0137] In some embodiments, the control or wild-type cell is a starting material,

[0138] In some embodiments, described herein is a method of assessing B cell depletion, including obtaining a test sample from a subject and detecting a concentration of IgE antibodies in the test sample, wherein the subject was administered a B cell depleting composition at least 2days priors to obtaining the test sample and a decrease in the concentration oflgE antibodies in the test sample compared to a reference value indicates depletion of B cells by the B cell depleting composition.139] In some embodiments, described herein Is a method treating a B cell mediated disease or disorder, including administering to a subject an agent or therapy that is capable of treating, preventing, delaying, or atenuating development of a B cell mediated disease or disorder, wherein the subject is determined to be at risk for maintaining or developing progressive disease by a method including, (a) obtaining a test sample from a subject and detecting a concentration oflgE antibodies in thesample, wherein the subject was administered a B cell depleting composition at least 2 days priors to obtaining the test sample, and (b) comparing the concentration detected for the IgE antibodies to a reference value, wherein the subject is one in which the comparison indicates that the subject is at risk for maintaining or developing a progressive B cell mediated disease or disorder.|0140| In some embodiments, the reference value is the concentration of IgE antibodies detected in a reference sample obtained prior to administering the B ceil depleting composition. In some embodiments, the reference value is a concentration of IgE antibodies from 10 ng / mL to 200 ng / mL, from 50 ng / mL to 200 ng / mL, front 75 ng / mL to 200 ng / mL, from 100 ng / mL to 200 ng / mL, from 50 ng / ml. to 600 ng / mL, from 75 ng / mL to 600 ng / mL, from 100 ng / mL to 600 ng / mL, front 100 ng / mL to 500 ng / mL, from 100 ng mL to 400 ng / mL, from 200 ng / mL to 600 ng / mL, from 200 ng / L to 500 ng / mL, and / or from 200 ng / mL to 400 ng / mL. In some embodiments, the test sample and / or the reference sample is blood or serum.0141] In some embodiments, the B cell depleting composition includes a therapeutically effective amount of acomposition including a population of engineered hypoimmunogenic T cells, wherein the hypoimmuno genic T cells include one or more modifications that: (a) inactivate or disrupt one ar more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more mo lecules that regulate expression of the one or more MHC class 1 molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class 11 molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, (b) increase expression of a tolerogenic factor encoded by afirst exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not inclode the modification, and (c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of an IgE-producmg cell. In some embodiments, the antigen expressed on the surface of an IgE-producing cell includes CD19, CD20, CD22, BCMA, GPRC5D, CD38, or any combination thereof. In some embodiments, the engineered T cells include one or more of a GDI 9~speclfrc CAR, a CD20-specifc CAR, a CD22-specific CAR, a BCMA-spe fic CAR, a GPRC5D-specific CAR, a CD38~specifi.c C R, or any combination thereof.|0142] In some embodiments, foe B cell depleting composition includes a therapeutically effective amount of a composition including a population of engineered hypoimmunogenic T cells including; (a) reduced expression of beta-2 microglobulin (B2M), Class II Major Histocompatibility Complex Transacfi vator (CHI' A), and T cell receptor alpha constant (TRAC) relative to a coiitrol or wild-type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD19-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.|0143| In some embodiments, described herein is a method, wherein B cell mediated disease or disorder is an IgE-mediated disease or disorder. In some embodiments, the IgE-mediated disease or disorder is asthma, Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chronic spontaneous (idiopathic) urticaria., chronic rh osimjsitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angioedema, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or any combination thereof. |6144| In some embodiments, B cell mediated disease or disorder is a B cell malignancy. In some embodiments, the B cell malignancy is a lymphoma or a leukemia. In some embodiments, the B cell malignancy includes or is selected from the group consisting of: Non-Hodgkin's Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (EBCL), transplant ineligible large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), highgrade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle celllymphoma (MC'L), follicular lymphoma (FL), marginal zone lymphoma (MZL), and / or small lymphocytic lymphoma (SLL).|0145 In some embodiments, described herein is a method, wherein B cell mediated disease or disorder is an autoimmune condition. In some embodiments, the autoimmune condition includes or is selected from the group consisting of: systemic lupus erythematosus (SLE), extrarenal systemic lupus erythematosus (ERL), lupus nephritis (LN), CNS lupus, vasculitis, granulomatosis vasculitis, poly angiitis & microscopic polyangiitis vasculitis, anti-neutrophilic cytoplasmic autoantibody (ANCA) associated vasculitis (AAV), granulomatous polyangiitis, microscopic polyangiitis, multiple sclerosis (relapsing an / or progressive), pemphigus vulgaris, autoimmune blistering skin diseases, membranous nephropathy (MN), anti-NMDA receptor neuropathy', neuromyelitis optica, idiopathic thrombocytopenic purpura, autoimmune hepatitis, type 1 diabetes mellitus, rheumatoid arthritis, juvenile rheumatoid arthritis, chronic inflammatory demyelinating polyneuropathy, poly yositis / dermatomyositis, stiff ersons disease, anti-'NMDA receptor encephalitis, anti -synthetase autoimmune syndromes, anti-phospholipid antibody syndrome, Sjogren’s syndrome, cryoglotml mentis. focal segmental glomenikrsclerosis, rapidly progressive glomerulopathy, autoimmune hemolytic anemia, amyloidosis, scleroderma, idiopathic mflammatory myositis, and / or immune-mediated necrotizing myopath.

[0146] In some embodiments, described herein is a method of assessing B cell depletion, iflcludiftg obtaining a test sample from a subject that includes antibodies and detecting the concentration of antibodies in the fest sample that bind a major histocompatibility complex (MHC) protein and comparing the concentration of antibodies that bind a MHC protein to the concentration of anti bodies that bind a MHC protein in a reference sample, wherein prior to obtaining the test sample the subject was administered a composition including a population of engineered hypoimmunogemc T cells, wherein the hypoimmunogeuic T cells include; (a) one or more modifications that inactivate or disrupt one or more alleles of: (i) one or more major histocompatibiliiyfcomplex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class 11 molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, (b) one or more modifications that Increase expression of a tolerogenic factorencoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not include the modification, and (e) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell. In some embodiments, the reference sample was obtained from the subj ect prior to administering the composition including the population of engineered hypoimmunogenic T cells. In some embodiments, the test sample is obtained at least 20 days after the subject was administered the composition including a population of engineered hypoimtnunogenic T cells. In some embodiments., the test sample and / or the reference sample is blood or serum. In some embodiments, absence of antibodies that bind a MHC protein indicates deep B cell depletion. In some embodiments, the.antibodies that bind a MHC protein are IgG and / or IgM isotypes.

[0147] In some embodiments, described herein is a method of treating a B celt disease or disorder, including,(a) administering to a subject a therapeutically effective amount of a composition including a population of engineered hypoimtnunogenic T cells, wherein the hypolmmunogenic T cell s Include one or more modifications that: (1) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class 11 molecules, (2) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (2) is relative to a control or wild-type T cell that does not include the modification, and (3) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR includes an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell; and (b) obtaining a test sample from the subject and detecting a concentration of antibodies that bind a MHC protein, wherein the test sample was obtained at least 20 days after the subject was administered the composition including the population of engineered hypoimtnimogenic T cells; and(c) comparing the concentration of antibodies that bind a MHO protein in (b) to the concentration of antibodi es that bind a MHC pro tein in a reference sample obtained fem the sub ject: prior to administering the composition including the population of engineered hypoimmunogenic T cells; and(d) if the comparison indicates the subject has developed anti-MHC antibodies, administering to the subject a composition or therapy that is capable of depleting B cells.| 148| In some embodiments, the population of engineered hypoimmunogenic T cells further include one or more modifications that inactivate or disrupt one or more alleles of (hi) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules. In some embodiments, the antigen expressed on the surface of a B cell includes CD 19, CD20, CD22, BCMA, GPRC5D, CD38, or any combination thereof. In some embodiments, the engineered T cells include one or more of a CD19-specific CAR, a CD20~specific CAR, a CD22~specific CAR, a BCMA-specific CAR, a GPRC5D-specifie CAR, a CD38“Specific CAR, or any combination thereof.10149] In some embodiments, the B cell depleting composition includes a therapeutically effecti ve amount of a composition including a populati on of engineered hypoimmunogenic T cells including: (a) reduced expression of bet -2 microglobulin (B2M), Class 11 Major Histocompatibility Complex Transacti vator (CRT A), and T cell receptor alpha constant (TRAC) relati ve tn a control or wild-type T cells, (b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD19-speeific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide,f0150] In some embodiments, B cell mediated disease or disorder is an IgE-mediated disease or disorder. In some embodiments, the IgE-mediated disease or disorder is asthma.Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chronic spontaneous (idiopathic) urticaria, chronic rhinosinusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary' aspergillosis, recurrent idiopathic angioedema, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or aty combination thereof In some embodiments, the B cell mediated disease or disorder is a B cell malignancy. In someembodiments, the B cell malignancy is a lymphoma or a leukemia. In some embodiments, the B cell malignancy includes or is selected from the group consisting of: Non-Hodgkii s Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), transplant Ineligible large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), and / or small lymphocytic lymphoma (SLL). In some embodiments, B cell mediated disease or disorder is an autoimmune condition. In some embodiments, the autoimmune condition includes or is selected from the group consisting of: systemic lupus erythematosus (SLE), extrarenal systemic lupus erythematosus (ERL), lupus nephritis (LN). CNS lupus, vasculitis, granulomatosi vasculitis, polyangiitis & microscopic polyangiitis vasculitis, anti-neutr philic cytoplasmic autoantibody (AN'CA) associated vasculitis (AAV), granulomatous polyangiitis, microscopic polyangiitis, multiple sclerosis (relapsing and / or progressive), pemphigus vulgaris, autoimmune blistering skin diseases, membranous nephropathy (MN), anti-NMDA receptor neuropathy, neuromyelitis optica, idiopathic thrombocytopenic purpura, autoimmune hepatitis, type 1 diabetes mellitus, rheumatoid arthritis, juvenile rheumatoid arthritis, chronic inflammatory demyelinating polyneuropathy, polymyositis / dennatomyositis,: stiff persons disease, anti-NMDA receptor encephalitis, anti-synthetase autoimmune syndromes, anti -phospholipid antibody syndrome, Sjogren’s syndrome, cryoglobulinemia, focal segmental glomerulosclerosis, rapidly progressive glomerulopathy, autoimmune hemolytic anemia, amyloidosis, scleroderma, idiopathic inflammatory myositis, and / or immune-mediated necrotizing myopathy.(0151 ] Provided herein are methods for treating a patient with a disease, disorder, or condition of the present disclosure, which includes administration of a population of hypoimmunogenic cells (e.g., primary T cells) to a subject, e.g., a human patient. For instance, a population of hypoimmunogenic primary T cells such as, but limited to, CD3+ T cells, CD4+ T cells, CD8 T cells, naive T cells, regulatory' T (Treg) cells, non-regulatory T cells, Thl ceils, Th2 cells, Th9 cells. Thl? cells, T-follicular helper (Tfh) cells, cytotoxic T lymphocytes (CTL), effector T (Teff) cells, central memory T (Tcm) cells, effector memory T (Tem) cells, effector memory T cells that express CD45RA (TEMRA cells), tissue-resident memory (1 rm) cells, virtual memory T cells, innate memory T cells, memory stem cell (Tse), y T cells, and anyother subtype of T cell is administered to a patient to treat a disease, disorder, or condition of the present disclosure.0152 In some embodiments, pharmaceutical compositions as described herein are co administered with a therapeutic agent that that binds to and / or interacts with one or more receptors selected from the group consisting of CD94, KIR2DL4, PD-1, an inhibitory NK cell receptor, and an activating NK receptor. In some instances, a therapeutic agent binds to a receptor on the surface of an NK cell, including one or more subpopulations of NK cells. In some embodiments, a therapeutic agent is selected from the group consisting of an antibody and fragments and variants thereof, an antibody mimetic, a small molecule, a blocking peptide, and a receptor antagonist. In some embodiments, pharmaceutical compositions as described herein are administered as pan of a combination therapy comprising a Bruton tyrosine kinase (BTK) inhibitor. In some embodiments, a BTK inhibitor is selected from a group consisting of: ibratimb, aealabratinib, zanubrutmih. and phtorutinib.1. Dosing and Fermulations

[0153] Any tlrerapeu tically effective amount of cells described herein can be included in a pharmaceutical composition of the present disclosure, depending on the indication being treated. Non-limiting examples of tire cells include primary T cells. In some embodiments, a pharmaceutical composition includes at least about lx.103, 5x 102, IxlO3, 5x10s, IxlO4, 5xI04, 1x10s, 5x10s, 1x10s, 5x10s, IxlO7,5x107, 1x10s, 5x10*, IxlO9, 5x1 *, lx10i: G, or 5x10s0cells. In some embodiments, the pharmaceutical composition includes up to about 1x102, 5xl02, 1x10s, 5xl()\ 1 xlO4, 5x 10'\ 1x10s, 5 x10s, 1x10s, 5x10s, ixiOy 5xl(f, 1x10s, 5x10, 1x1 O', 5xl09, lx.10, or 5xlO10cells. In some embodiments, the pharmaceutical composition includes up to about 6,0 x 10scells. In some embodiments, the pharmaceutical composition includes up to about S>0 x 10scells. In some embodiments, the pharmaceutical composition includes at least 30x10scells. In some embodiments, the pharmaceutical competition includes approximately 30xl06cells. In some embodiments, the pharmaceutical composition Includes at least 60x1 (P cells. In some embodiments, the pharmaceutical composition includes approximately 60x10scells. In some embodiments, the pharmaceutical composition includes at least 90x10scells. In some embodiments, the pharmaceutical composition includes approximately 90 106cells. In some embodiments, the pharmaceutical composition includes at least 120x10scells. In someembodiments, the pharmaceutical composition includes approximately 120 10° cells. In some embodiments, the pharmaceutical composition includes at least 200x10° cells. In some embodiments, the pharmaceutical composition includes approximately 200x10° cells. In some embodiments, the pharmaceutical competition includes at least 400 10° cells. In some embodiments, the pharmaceutical composition includes approximately 400x10° cells. In some embodiments, the pharmaceutical composition includes at least 1000x10° cells. In some embodiments, the pharmaceutical composition includes approximately 1000x10° cells. In some embodiments, the pharmaceutical composition includes at least about I IO2to SxlO2, 5xI02to 1x10s, 1x103to 5x1 (, 5x10® to IxIO4, IxIO4to 5xl04, 5xlO to 1x10s, 1x10sto 5x10% 5x10sto 1x10°, 1x10° to 5x10°, 5x10° to 1x10% 1x10sto 5x10s, 5xlO7to 1x10s, 1x10sto 5x1025x10® to l io 1x109to 5x10°, 5x10° to IxIO50or I xl. Oi0to 5x! O50cells. In some embodiments, the pharmaceutical composition includes from about 1.0x10° to about 2.5x10® cells. In some embodiments, the pharmaceutical composition includes from about 2.0x10° to about 2.0x10® cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises from about 30x10° cells to about 200 10° cells. In some embodiments, a pharmaceutical composition of tbe present disclosure comprises from about 20x10° cells to about 400x10° cells. |01S4] In some embodiments, a pharmaceutical composition includes at least about lx It)2, 5x10% 1 x 10\ 5x10% 1x1 (A 5x10 1x10% 5 l(P. I lO6, 5> J0(\ Ix107, 5x102 1x10®, 5x10s, IxIO®, 5x109, 1x1 O10, or 5x10i0CAR* cells. In some embodiments, the pharmaceutical composition includes up to about 1x10 5x10 IxIO3, 5x10s, IxIO4, 5xlC) IxIO5, 5x10% 1x10°, 5x10°, IxIO7, 5x IO7, I IO8, 5x10®, IxIO9, 5xl09, I IO10, or 5xlOsoCAR + cells. In some embodiments, the pharmaceutical composition includes up to about 6.0 10sCAR* cells. Jn some embodiments, the pharmaceutical composition includes up to about 8.0x10® CARA cells. In some embodiments, the pharmaceutical composition includes at least 30x10° CAR* cells. In some embodiments, the pharmaceutical composition includes approximately 30 10° CARA cells. In some embodiments, the pharmaceutical composition includes at least 60x10° CARA cells. In some embodiments, the pharmaceutical composition includes approximately 60x10° CAR* cells. In some embodiments, the pharmaceutical composition includes at least 120x10° CAR i cells. In some embodiments, the pharmaceutical composition includes approximately 120 10° CARA cells. In some embodiments, the pharmaceutical composition includes at least 200x10° CAR* cells. In some embodiments, the pharmaceutical composition includes approximately 200x10°CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 400 10® C ARA cells. In some embodiments, the pharmaceutical composition includes approximately 400x10® CAR+ cells. In some embodiments, the pharmaceutical composition includes at least 1000x106CAR cells. In some embodiments, the pharmaceutical composition includes approximately 1000x10® CARA cells. In some embodiments, the phawnaceutical composition includes at least: about I x ID2to 5 x 102, 5 x 10" to 1 x 10s, 1 x 10’ to 5 x 10 \ 5 x 10}to 1 x W4, I x I4to 5 x 104, 5 x IO4to 1 x 10s, I x 10sto 5 x 10s, 5 x 10sto 1 x 10®, 1 x 10sto 5 x 10®, 5 x 10® to 1 x TO7, 1 x IO7to 5 x 10 5 x 107to 1 x 10s, 1 x 10® to 5 x 10s, 5 x 10* to 1 x IO9, 1 x W9to 5 x 109, 5 x 109to 1 x 10w, or 1 x 10’° to 5 x 10-° CARA cells. In some embodiments, the pharmaceutical composition includes from about 1.0 x IO6to about 2,5 x 10® CAR + cells. In seme embodiments, the pharmaceutical composition includes from about 2.0 x til® to about 2,0 x 10sCARA cells, hi some embodiments, a pharmaceutical composition of the present disclosure comprises from about 30x10® CAR- cells to about 200x10® CARA cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5x10* CAR+ cells to at least about 400x10® CARA cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 20x10® CAR- cells to about 400x 106C R A cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises at least about 30x10® CARr cells, at least about 60x10® CAR- cells, at least about 120x 10" CARt- cells, or at least about 200x10® CARA cells, least about 400x10® CA cells, least about 1000x10® CARA cells. In some embodiments, a pharmaceutical camposition of the present disclosure comprises no more than about 200x10® CARA cells. In some embodiments, a pharmaceutical composition10®, 76 x: 1.0®, 77 x 10®, 78 x 10®, 79 x 10®, 80 x 10®, 81 x 10®, 82 x 10®, 83 x 10®, 84 x 10®, 85 x 10®, 86 x 10®, 87 x 10®, 88 x 10®, 89 x 10®, 90 x 10®, 91 x 10®, 92 x 10®, 93 x 10®, 94 x 10®, 95 x 10®, 96 x 10®, 97 x 10®, 98 x 10®, 99 x 10®, 100 x TO®, 101 x 10®, 102 x 10®, 103 x 10®, 104 x 10®, 105 x.10®, 106 x 10®, 107 x 10®, 108 x 10®, 109 x 10®, 1 10 x 10®, 111 x 10®, 112 x 10®, 113 x 10®, 114 x 10®, 115 x 10®, 116 x 10®, 117 x 10®, 118 x 10®, 1 19 x 10®, 120 x 10®, 121 x 10®, 122 x 10®, 123 x 10®, 124 x 10®, 125 x 10®, 126 x 10®, 127 x 10®, 128 x 10®, 129 x 10®, 130 x 10®, 131 x 10®, 132 x 10®, 133 x 10®, 134 x 10®, 135 x 10®, 136 x 10®, 137 x 10®, 138 x 10®, 139 x 10®, 140 x 10®,141 x ICC 142 x 10% 143 x 10s, 144 x 10% 145 x 10% 146 x 10% 147 x 10% 14§ x 10% 149 x 10% 150 x IO, 151 x 10s. 152 x 10% 153 x 10% 154 x 10% 155 x 10s, 156 x 10% 157 x 10% 158 x 10% 159 x 10s, 160 x 10s, 161 x 10s, 162 x 10% 163 x 10s, 164 x 10% 165 x 10s, 166 x 10% 167 x 10s, 168 x 10% 169 x 10% 170 x 10% 171 x 10s, 172 x 10% 173 x 10s, 174 x 10% 175 x 10% 176 x 10% 177 x 10% 178 x 10% 179 x 10% 180 x 10% 181 x 10% 182 x 10% 183 x 10s, 184 x 10% 185 x 10% 186 x 10% 187 x 10s, 188 x 10% 189 x 1 % 190 x 10% 191 x 10s, 192 x 10% 193 x 10% 194 x 10% 195 x 10 196 x 10% 197 10% 198 x 10% 199 x 10% or 200 x 10sCAR? cells.[0155 fa some embodiments, a pharmaceutical composition of the present disclosure comprises fully edited hypoimmunogenic CAR- cells as described herein. In seme embodiments, folly edited hypoiinmutogetic (’ARA cells comprise one or more modifications that: (a) inactivate or disrupt one or more alleles of: ( i ) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and (ill) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules, (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or iki-type T cell that does not comprise the modification^, and (c) express a CDl9~specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide. In some embodiments, a pharmaceutical coinposition of the present disclosure compri ses partially edited CAR A cells as described herein. In some embodiments, partially edited CAR t cells are missing one or more mo ifications that: (a) inactivate or disrupt one or more alleles of: (1) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expressi n of the one or more TCR molecules, and / or (b) increase expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type T cell that does not comprise the modification, relative to fully edited hypoimmunogenic CAR+ ceils as described herein. In some embodiments, partially edited CAR-r cells (i) are missing one or moremodifications thatinacti vate or disrupt one or more alleles of one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of th e one or more MHC class 1 molecules, and (ii) comprise one or more modifications that inactivate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR* cells as described herein. In some embodiments, partially edited CAR* cells (I) comprise one or more modifications that inactivate or disrupt one or more alleles of one or more majorhislocon tpafibi 1 ity complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, and (ii) are missing one or more modifications that inacti vate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR* cells as 'described herein. In some embodiments, partially edited CAR* cells (i) are missing one or more modifications that inactivate or disrupt one or more alleles of one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class 1 molecules, and (ii) are missing one or more modifications that inactivate one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, relative to fully edited hypoimmunogenic CAR* cells as described herein. In some embodiments, a pharmaceutical composition of the present disclosure comprises felly edited hypoimmunogenic CAR* cells and partially edited CAR* cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises at least 20%, at leas t 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92*14, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% fully edited hypoimmunogen ie CAR* cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises 100% felly edited hypoimmunogenic CAR* cells. In some embodiments, a. pharmaceutical composition of the present disclosure comprises about 20% to about 40% felly edited hypoimmunogenic CAR* cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 30% to about 50% folly edited hypoimmunogenic CAR* cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 40% to about 60% folly edited hypoimmunogenic CAR*cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 50% to about 70% fully edited hypoimmtmogemc CAR-+- cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 603 to about 80% fully edited hypoimmunogenic CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 70% to about 90% fully edited hypoirnmunogenic CAR -cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 80% to about 99% fully edited h ypoiirnnunogenic C AI% cells, hi some embodiments, a pharmaceutical composition of the present disclosure comprises less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40% partially edited CAR.+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 1 % to about 10% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 10% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 15% partially edited CAR+ cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 5% to about 20% partially edited CAR* cells. In some embodiments, a pharmaceutical composition of the present disclosure comprises about 10% to about 30% partially edited CAH+ cells. In some embodiments, a pharmaceutical composition of the present discl osure comprises about 20% to about 40% partially edited CAR* cells.|B156] In some embodiments, a pharmaceutical composition has a volume of at least 1.0, 1.25, 1,5, 1.7.5, 2.0. 2.5, 3M 3.5, 4.0, 4.5, 5, 10. 15, 20, 5, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, or 500 mL, In exemplary embodiments, a phanuaceutical composition has a volume of up to about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70. 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, 300, 350, 400, or 500 mb. In exemplary embodiments, a pharmaceutical composition has a volume of about 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 1'70, 180, 190, 200, 250, 300, 350, 400, cr 500 mL. In some embodiments, a pharmaceutical composition has a volume of from about I -50 mL, 50-100 mL. 100-150 mL, 150-200 mL, 200-250 mL, 250-300 mL, 300-350 mL, 350-400 mL, 400-450 L, or 450-500 L. In some embodiments, a pharmaceutical composition has a volume of from about 1-50 mL, 50-100 mL, 100-150 ml... 150-200 mL, 200-250 mL, 250-.’00 mL, 300-350 mi,, 350-400 mL, 400-450 m. L. or 450-500 mL. In some embodiments, a pharmaceutical composition has a volume of from about 1-10 mL, 10-20 mL, 20-30 L, 30-40 ml... 40-50 ml, 50-00 L, 60-70 mL, 70-80 mL, 70-80 mL, 80-90 mL, or 90- 100 mL, In some embodiments, a pharmaceutical composition has a volume that ranges from about 1.25mL to about lOOmL. In some embodiments, a pharmacal composition has a volume that ranges from about 5 mL to about 80 mL. In some embodiments,: a pharmaceutical composition has a volume that ranges from about 10 L to about 70 mL. In some embodiments, a pharmaceutical composition has a volume that ranges from about 10 mL to about 50 mL. In some embodiments, a pharmacal composition has a volume of at least about 7.5 mL, at least about 15 ml.., at least about.30 ml... or at least about 50 mL.10157] In some embodiments, a specific amount / dosage regimen will vary depending on the weight, gender, age and health of a subject; the formulation, the biochemical nature, bioacti vity, bioavailability and the side effects of the cells and the number and identity of the cells in the complete therapeutic regimen.|0158] In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition includes about l. Oxl (P to about 2.5x1 (Iscells at a volume of about 10 mL to 50 mL and the pharmaceutical composition is administered as a single therapeutic ally effective dose or clinically effect ve dose. In some eases., the therapeutically effective dose or clinically effective dose includes about 1,0x10sto about 2.5x10sprimary T cells described herein at a volume of about 10 nil to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose includes about 1.0 10sto about 2.5x10sprimary T cells as disclosed herein at a volume of about 10 i»L to 50 mL.(0159 j In some embodiments, a therapeutically effecti ve dose or a clinically effective dose of the phanBaceutwal composition comprises a concentration of engineered hypoimmunogenic T cells of at least 1.25 106cells / mL. In some embodiments, a therapeutically effective dose or a clinically effective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x 10scells / mL. In some embodiments, a therapeutically effecti ve dose or a clinically effective dose of the pharmaceutical composition compri ses a concentration of engineered hypoimmunogenic T cells of at least 1.2xl()6CAR+ eells / mL. In some embodiments, a therapeutically effective dose or a clinicallyeffective dose of the pharmaceutical composition comprises a concentration of engineered hypoimmunogenic T cells of at least 4.0x10* CAR;cells / mL.160 In some embodiments, the pharmaceutical composition is administered as a single therapeutically effective ose or clinically effective dose of from about 1.0x10sto about 1.0x10' cells (such as primary T cells) per kg body weight for subjects 50 kg or less. In some embodiments, the pharmaceutical composition is administered as a single therapeutically effective dose or clinically effective dose of from about 0.5 x10sto about l. OxlO7, about 1.0x10* to about 1.0x10*', about 1,0x10sto about LOxlQ7, about 5.0x103to about 1x10’', about 1,0x10* to about 1x107 about 5.0x10* to about 1.0x10', about 1,0x10sto about 5.0x106, about 1,0x10sto about 1.0x10'’. about 1.0x10sto about 5.0x10s, about 1.0x10sto about 5.0x10*. about 2.0x10sto about 5.0x10*, about 3.0x10sto about 5.0x10s, about 4.0x10sto about 5,0x10s, about 5.0x10sto about 5. Ox Hr’, about 6. Ox 10sto about 5.0x10*, about 7.0x10sto about 5.0x10*, about 8.0x10sto about 5.0x10*, or about 9.0x10sto about 5.0x10scells per kg body weight for subjects 50 kg or less. In some embodiments, the therapeutically effective dose or clinically effective dose is 0.5 x 10 0.6 x 103, 0.7 x 103, 0.8 x 10s, 0.9 x 103, 1.0 x 10s, 1.1 x 10s, 1,2 x 10s, 1.3 x 10s, 1,4 x 10s, 1.5 x 10s, 1.6 x 10s, 1.7 x 10s, 1.8 x 10s, 1.9 x 105, 2.0 x 10s, 2.1 x IO3, 2.2 x 10s, 2,3 x 10s, 2,4 x 10s, 2.5 x 103, 2.6 x 10s, 2.7 x 10s, 2.8 x 10s, 2,9 10s, 3,0 105, 3,1 x 10s, 3.2 x 10s, 3.3 x 10s, 3.4 x 10s, 3,5 x 10s, 3.6 x 10s, 3,7 x 10s, 3,8 x 103, 3.9 x 10s, 4.0 x 103, 4.1 x 10s, 4.2 x 103, 4.3 x 10s, 4.4 x 103, 4.5 x 103, 4.6 x 10s, 4.7 x 103, 4.8 10s, 4.9 x 10s, 5.0 x 10s, 0.5 x 1070.6 x 10*, 0.7 x 10 0.8 x 1070.9 x 10* 1.0 x 10 1.1 x 10:7 i.2 107 1.3 x 10*, 1.4 x 7)7 1.5 x 107 1.6 x 10*, 1.7 x 10*, 1.8 x 10s, 1.9 x 10*, 2.0 x 10*, 2.1 x 10*, 2.2 x 10*, 2.3 x 10*, 2.4 x 10*, 2.5 x 10*, 2.6 x IO6, 2,7 x 10*.2.8 x 10s, 2.9 x 10*, 3,0 x 10* 3.1 x 10*93.2 x 10* 3.3 x 10*, 3.4 x 10*, 3.5 x 10*, 3.6 x 10*, 3.7 x 10*, 3.8 x 10*, 3,9 x 10s, 4.0 x 10*, 4.1. x 10*, 4.2 x 10*, 4.3 x 10*, 4.4 x 10*, 4.5 x 10*, 4.6 x 1074.7 x 10*, 4.8 x 10*, 4.9 x 10*, 5.0 x 10*, 5.1 x 10* 5.2 x 10*, 5.3 x 10*, 5.4 x 10*s5.5 x 10*, 5.6 x 10*, 5.7 x 10*, 5,8 x 10*, 5.9 x 10*, 6.0 x 10*, 6.1 x 10*, 6.2 x 10*, 6.3 x 10*, 6.4 x 10s, 6.5 x 10s, 6,6 x 10*, 6.7 x 10s6.8 x 10* 6.9 x 10*, 7.0 x 10* 7.1 x 10*, 7.2 x 10*, 7.3 x 1077.4 x 10*, 7.5 x 10*, 7.6 x 10*, 7.7 x 10*, 7.8 x 10*, 7.9 x 10*, 70 x 10*, 8.1 x 10*, 8.2 x 10*, 8.3 x 10s, 8.4 x 7)78,5 x 1078.6 x 19 8.7 x 10*, 8.8 x 10*, 8.9 x 10*, 9.0 x 10*, 9.1 x 10*, 9.2 x 10s, 9.3 x 10*, 9.4 x 10*, 9.5 x 10*, 9.6 x IQ*, 9.7 x 10*, 9.8 x 10*, 9.9 x 10*, 0.5 x 107, 0.6 x 107 0.7 x 10 0.8 x 1070.9 x 107 or 1.0 x 10?cells per kg body weight for subjects 50 kg or less. In some embodiments, a therapeutically effecti ve dose or clinically effective dose is from about 0.2x 10® to about 5.0 x 10® cells per kg: body weight for subjects 50 kg or less. In certain embodiments, a therapeutically effective dose or clinically effective dose is at a range that is lower than from about 0.2 x 10® to about 5.0 x 10® cells per kg body weight for subjects 50:kg or less. In some embodiments, a single therapeutically effective dose or clinically effective dose is at a volume of about 10 ml. to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose is administered intravenously. In some embodiments, a therapeutically effective dose or clinically effective dose is administered to the nervous system of a subject & some embodiments, the nervous system is the central nervous system (CNS). In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered inwhecally.(01611 In exemplary embodiments, cells arc administered in a single therapeutically effective dose of from about 1.0x106to about 5.0x108cells (such as primary T cells) for subjects above 50 kg. In some embodiments, a pharmaceutical composition Is administered as a single therapeutically effective dose or clinically effective dose of from about 0,5x 10® to about 1. Ox 10®, about 1.0x10® to about 1.0x10® about 1.0x10® to about 1.0x10®, about 5.0x10® to about 1. O lO9, about 1.0x10' to about l. OxlO9, about S. QxIO7to about 1.0x10®, about 1.0x10® to about 5.0x10’', about 1.0x10® to about l. OxlO, about 1.0x10® to about 5.0x 10”, about 1.0x10' to about 5.0x10®, about 2,0x107to about 5.0x10®, about 3.0x10-' to about S. OxlO8, about 4,0x107to about 5.0x10s, about S. OxlO7to about 5.0x10s, about 6.0x10’' to about 5.0 x10s, about 7.0x10' to about 5.0x10®, about 8.0x10' to about 5.0 10®, or about 9.0x107to about 5.0x10* cells per kg body weight for subjects 50 kg or less. In some embodiments, a therapeutically effective dose or clinically effective dose is 1.0 x 1061.1 x 10®, 1.2 x 10®, 1.3 x 10®. 1.4 x 10®, 1.5 x 10®, 1.6 x 10®, 1.7 x 10®, 1.8 10s, 1.9 x 10®, 2.0 x 10®, 2.1 x IO6, 2.2 x 106, 2.3 x 106, 2.4 x 106, 2.5 x IO6, 2.6 x 10®, 2.7 x 10®, 2.8 x 10®, 2.9 x 10®, 3.0 x 10®, 3.1 x 10®, 3,2 x 10®, 3.3 x 10®, 3.4 x 10®, 3.5 x 10®, 3,6 x.10®, 3.7 x 10®, 3,8 x 10®, 3,9 x 10®, 4.0 x 10®, 4.1 x 10®, 4.2 x 10®, 4.3 x 10®, 4,4 x 10®, 4.5 x 10®, 4.6 x 10®, 4.7 x 10®, 4.8 x 10®, 4.9 x 10®, 5.0 x 10®, 5.1 x 10®, 5.2 x 10®, 5.3 x 10®, 5.4 x 10®, 5,5 x 10®, 5.6 x 10®, 5,7 x 10®, 5,8 x 10s, 5.9 x 10®, 6.0 x 10®, 6.1 x 10®, 6,2 x 10®, 6.3 x 10®, 6.4 x 10®, 6.5 x 10®, 6.6 x 10®, 6.7 x 10®, 6.8 x 10®, 6.9 x 10®, 7.0 x 10®, 7.1 x 10®, 7,2 x 10®, 7.3 x 10®, 7.4 x 10®, 7.5 x 10®, 7.6 x 10®, 7.7 x 10®, 7.8 x 10®, 7,9 x 10®, 8.0 x 10®, 8.1 x 10®, 8,2 x 10®, 8.3 x 10®, 8.4 x 10®, 8.5 x 10®, 8.6 x 10®, 8.7 x 10®, 8.8 x 10®, 8.9 x 10®, 9.0 x 10®, 9.1 x 10s, 9.2 x 10®, 9.3 x10% 9.4 X 10% 9.5 x 10% 9.6 x 10% 9.7 x 10* 9.8 x 10 9.9 x 10*, 1.0 x W7, LI x 10% 1,2 x IO7, 1.3 x 10% L4 x IO7, 1,5 x 10% 1 6 x 10% 1.7 x 10% 1.8 x 107, 1.9 x 10% 2.0 x 10% 2.1 x 10% 2.2 x 107. 2.3 x 1O'\ 2.4 x 10% 2.5 x IO7, 2.6 x 10% 2.7 x 107,.8 x 107,.9 x IO7, 3.0 x IO7, 3.1 x IO7, 3.2 x 10% 3.3 x 10% 3.4 IO7, 3.5 x 107, 3.6 x IO7, 3.7 x IO7, 3.8 x IO7, 3.9 x IO7, 4.0 x 10% 4.1 x 107. 4.2 x IO7. 4.3 x 10% 4.4 x 10% 4.5 x IO7, 4.6 IO7, 4.7 x W7, 4.8 x 10% 4.9 x 10% 5,0 x IO7, 5.1 x 10% 5.2 x IO7, 5.3 x IO7, 5.4 x 10% 5.5 x 10% 5.6 x IO7, 5.7 x 10% 5.8 x IO7, 5.9 x 107, 6.0 x 107. 6.1 x 10% 6.2 x 10% 6.3 x 107, 6.4 x 10%.5 x 10% 6.6 x IO7, 6.7 x IO7, 6.8 107, 6.9 x 107, 7.0 x IO7, 7.1 x 10% 7.2 X IO7, 7.3 x 10% 7.4 x IO7, 7.5 x 107, 7.6 x IO7, 7.7 x IO7, 7.8 x IO7, 7.9 x IO7, 8,0 x IO7, 8.1 x IO7, 8,2 x IO7, 8, 3 x IO7, 8,4 x 107, 8.5 x IO7, 8,6 x 107, 8,7 x 10% 8.8 x 10% 8.9 x 107, 9.0 x 10% 9.1 x IO7, 9.2 x 10% 9.3 x IO7, 9.4 IO7, 9.5 x IO7, 9.6 x 10% 9.7 x 10% 9.8 x 10% 9.9 x IO7, 1,0 x 10s, 1,1 x 10s, 1,2 10s1.3 x 10s, 1.4 x 10% 1,5 x 10s. L6 x 10s, 1.7 x 10% 1.8 x 10®, 1.9 x 10®,.0 x 10®, 2.1 x 10®, 2.2 x 10®, 2.3 x 10s, 2.4 x 10s, 2.5 x 10s, 2.6 x 10s, 2,7 x 10®, 2.8 x 10s, 2.9 x 10s, 3.0 x 10s, 3.1 x 10s, 3,2 x 10s, 3.3 x 10s, 3.4 x 10s, 3.5 x 10s, 3.6 x 10s, 3.7 x 10s, 3.8 x 10s, 3.9 x 10®, 4.0 x 10®, 4.1 x 10®, 4.2 x 10s, 4.3 >; 10®, 4.4 x 10s, 4.5 x 10s, 4.6 x 10% 4.7 x 10s, 4,8 x 10s, 4,9 x 10s, or 5.0 x 10® cells, per kg body weight for subjects 50 kg or less. In certain embodiments, ceils are administered in a single therapeutically effective dose or clinically effective dose of about 1.0 x IO7to about 2.5 x 10® cells for subjects above 50 kg. In some embodiments, cells are administered in a single therapeutically effective dose or clinically effective dose of a range that is less than about 1.0 x IO7lo about 2.5 x 10® cells for subjects above 50 kg. In some embodiments, cells are administered in a single therapeutically effective dose or clinically effective dose of a range that is higher than about 1.0 x IO7to about 2.5 x 10® cells for subjects above 50 kg. In some embodiments, a dose is administered Intravenously. In some embodiments, a therapeu tically effective dose or clinically effective dose is administered to the nervous system of a subject. In some embodiments, the nervous system is the central nervous system (CNS). In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered intrathecally. In some embodiments, a single therapeutically effective dose or clinically effective dose is at a volume of about 10 mL to 50 mL. In some embodiments, a therapeutically effective dose or clinically effective dose is administered intravenously. In some embodiments, a therapeutically effective dose or clinically effective dose is administered to the nervous system of a subject. In some embodiments, the nervous sy stem is the central nervoussystem (CNS), In some embodiments, the nervous system is the peripheral nervous system (PNS). In some embodiments, a therapeutically effective dose or clinically effective dose is administered in trathecally.

[0162] In. exemplary embodiments, the therapeutically effective dose or clinically effective dose is administered intravenously at a rate of about 1 to 50 ntiL per minute, 1 to 40 naL per minute, 1 to 30 mL per minute, 1 to 20 mL per minute, 10 to 20 L per minute, 10 to 30 mL per minute, 10 to 40 mL per minute, 10 to 50 mL per mmole, 20 to 50 L per minute, 30 to 50 mL per minute, 40 to 50 mL per minute. In numerous embodiments, a pharmaceutical composition is stored in one or more infusion bags for intravenous administration. In some embodiments, a dose is administered completely in no more than 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 60 minutes, 70 inutes, 80 minutes, 90 minutes, 120 minutes, 150 minutes, 180 minutes, 240 minutes, or 300 minutes.(0163 In some embodiments, a single therapeutically effective dose or clinically effects ve dose of the pharmaceutical composition is present in a single infusion bag. In other embodiments, a single therapeutically effective dose or clinically effective dose of a pharmaceutical composition is divided into 2, 3, 4 or 5 separate infusion bags.

[0164] In some embodiments, cells described herein are administered in a plurality of doses such as 2, 3, 4, 5, 6 or more doses, wherein the plurality of doses together constitute a therapeutically effective dose or clinically effective dose regimen, In some embodiments, each dose of a plurality of doses is administered to the subj ect ranging from 1 to 24 hours apart. In some instances, a subsequent dose is administered from about 1 hour to about 24 hours (e.,, about 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23 or about 24 hours) after an initial or preceding dose. In some embodiments, each dose of a plurality of doses is administered to the subject ranging from about 1 day to 28 days apart. In some instances, a subsequent dose is administered from about 1 day to about 28 days (e.g.5about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 2, 23, 24, 25, 26, 27, or about 28 days) after an initial or preceding dose. In certain embodiments, each dose of a plurality' of doses is administered to the subject ranging from I week to about 6 weeks apart. In certain instances, a subsequent dose Is administered from about 1 week to about 6 weeks (e.g., about 1, 2, 3, 4, 5, nr6 weeks) after an initial or preceding dose. In several embodiments, each dose of a plurality of doses is administered to the subject ranging from about I month to about 12 months apart. In several instances, a subsequent dose is administered from about I month to about 1 months (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months) after an Initial or preceding dose.|0165| In some embodiments, a subject is administered a first dosage regimen at a first time point, and then subsequently administered a second dosage regimen at a second time point. In some embodiments, the first dosage regimen is the same as the second dosage regimen. In other embodiments, the first dosage regimen is different than the second dosage regimen, In some instances, the number of cells in the first dosage regimen and the second do sage regimen are the same. In some instances, the number of cells in the first dosage regimen arid the second dosage regimen are differen. In some cases, the number of doses of the first dosage regimen and the second dosage regimen are the same. In some eases, the number of doses of the first dosage regimen and the second dosage regimen are different.101661 In some embodiments, CD 19 specific (CD 19) CAR-T cells described herein are administered to a subject at a dose of about 50 10st« about 200 x 10s(e.g„ 50 x 10s, 51 x 10s, 52 x 10s, 53 x 10s, 54 x i( / 55 x 10s, 56 x 10s, 57 x 10s, 58 x 10s, 59 x 10s, 60 x 10s, 61 x 10s62 x 10s, 63 x IO6, 64 x 10s, 65 x 10s, 66 x 106, 67 x 10s, 68 x 106, 69 x IQ6, 70 x 10s, 71 x 10s, 72 x if)6. 73 x 10s, 74 x 106, 75 x 1 O'; 76 x 10s, 77 1 ()6. 78 x 10s, 79 x 10s, 80 x 10s, 81 x 10s, 82 x 10s, 83 x IO6, 84 x 10s, 85 x 10s, 86 x 10s, 87 x 10s, 88 x 10s, 89 x 10s, 90 x 10s, 91 x 10s, 92 x 10s, 93 x 10s, 94 x 10s, 95 x 10s, 96 x 10s, 97 x 10s, 98 x 10s99 x 10s, 100 x 10s, 101 x 1s, 102 x 10s, 103 x 10s, 104 x 10s, 105 x 106, 106 x 10s, 107 x 10s, 108 x 10s, 109 x 106, 110 x 10s, 111 x 10s, 112 x 10sI 13 x IO6, 114 x 10s, 115 x 10s, 116 x 10s, 117 x 106, 118 x 10s, 119 x 10s, 120 x 10s, 121 x 106, 122 x 10s, 123 x 10s, 1 4 x 10s, 125 x 10s, 126 x 10s, 127 x 10s, 128 x.106, 12.9 x IO6, 130 x 10s, 131 x 106, 132 x 10s, 133 x IO6, 134 x 10s, 135 x 106, 136 x 10s, 137 x IO6, 138 x 10s, 139 x 10s, 140 x 10s, 141 x 10s, 142 x 10s, 143 x 10s, 144 x 10s, 145 x 10s, 146 x 10s, 147 x 10s, 148 x 10s, 149 x 10s, 150 x 10s, 151 x 10s, 152 x 10s, 153 x 10s, 154 x 10s, 155 x 10s, 156 x 10s, 157 x 10s158 x 106, 159 x 10s, 160 x IO6, 161 x 10s, 162 x 10s, 163 x 10s, 164 x 10s, 165 x 10s, 166 x 10s, 167 x 10s, 168 x 10s, 169 x 10s, 170 x 10s, 17 l x 10s, 172 x 10s, 173 x 10s, 174 x 10s, 175 x 10s, 176 x 10s, 177 x 10s, 178 x 10s, 179 x 10s, 180 x 10s, 181 x 10s, 182 x 10s, 183 x 10s, 184 x 10s, 185 x 10s, 186 x 10s, 187 x 10s. 188 x 10s, 189 x 10s, 190 x 10s, 191 x10\ 192 X. 10s, 193 X 10®, 194 X 10s, 195 x 10* 196 x 10s, 197 x 10* 198 x 10s, 199 x 10*5or 200 x 10*) viable CD19 specific CAR-T cells, fa some embodiments, a dose is a therapeutically effective amount of vi able CD 19 specific CAR-T cells. In some embodiments, the dose is a clinically effective amount of viable CD 19 specific CAR-T cells. In some embodiments, viable CD19 specific CAR-T cells include CD 19 specific CAR expressing CD4+ T cells and CD 19 specific CAR expressing CD8+ T cells at a ratio of about 1:1. fa some embodiments, the CD 19 specific CAR. of the cells is lisocabtagene maraleacel (BREYANZI*), a structural equivalent thereof, or a functional equivalent thereof.

[0167] In some embodiments, a subject is administered about 50 x 10sto about 200 x 10s(«?.£., 50 x 10®, 51 x 10®, 52 x 10*, 53 x 10s, 54 x 10®, 55 x 10s, 56 x 10®, 57 x 10®, 58 x 10®, 59 x 10*, 60 x 10*, 61 x 10s, 62 x 10*, 63 x 10s, 64 x 10*, 65 x 10s, 66 x 10s, 67 x 10*, 68 x 10s, 69 x 10*, 70 x 10s, 71 x 10*, 12 x 10s, 73 x 10*, 74 x 10*, 75 x 10*, 76 x 10* 77 x 10s, 78 x 10*, 79 x10s, 100 x 10*, 101 x 10*, 102 x 10*, 103 x 10s, 104 x 10*, 105 x 10s, 106 x 10*, 107 x 10s, 108 x 10*, 109 x 10s, 110 x 10*, Ill x 10s, 112 x 10*, 113 x 10s, 114 x 10*, 115 x 10s. 116 x 10*, 117 x 10*, 118 x Hl*. HO x 10*, 120 x 10*, 121 x 10s, 122 x 10*, 123 x 10*, 124 x 10s, 125 x 10s, 126 x KF. 127 x 10s, 128 x IO6, 129 x 10s, 130 x 10*, 131 x 10s, 132 x 10*, 133 x 10s, 134 x 10* 135 x 10®, 136 x 10s, 137 x 10®, 138 10s, 139 x 10s, 140 x 10s, 141 x 10* 142 x 10s, 143 x 10s, 144 x IO6, 145 x 10s, 146 x I O6147 x 10s, 148 x 10s, 149 x 10s, 150 x 10s, 151 x 10s, 152 x 10s, 153 x 10s, 154 x 10s, 155 x 10s156 x 10s, 157 x 10s, 158 x 10s, 159 x 10s, 160 x 10s, 161 x 10s, 162 x 106, 163 x 10s, 164 x 10®, 165 x 10s, 1 6 x 10s, 167 x 10s, 1 8 x 10'7 169 x 10s, 170 x 10s, 171 x 10s, 172 x 10s, 173 x 10* 174 x 10s, 175 x 10s, 176 x 10s, 1.77 x 10s, 178 x 10s, 179 x 10s, 180 x IO6, 181 x 10s, 182 x IO6, 183 x 10s, 184 x 10s, 185 x 10s, 186 x 10\ 187 x 10s, 188 x 10s, 189 x 10s, 190 x 10s, 191 x 10s, 192 x 1.0s, 193 x 10*, 194 x 10s, 1.95 x 10*, 196 x. 10s, 197 x 10®, 198 x 10*, 199 x 10*, or 200 x 10*) viable CD 19 specific CAR-T cells described herein. In some embodiments, the dose is a therapeutically effective amount of viable CD 19 specific CAR-T cells, fa other embodiments, a dose is a clinically effective amount of viable CD 19 specific CAR-T ceils. In some instances, 50% of the viable CD 19 specific CAR-T cells are CD 19 specific CAR expressing CD4+ T cells and 50% of the viable CD 19 specific CAR-1’ cells are CD19 specific CAR expressing CD8+ T cells. In some embodiments, the CD19 specific CAR ofthe cells is lisocabtagene maraleucel (BREYANZ1 A. a structural equivalent thereof, or a functional equivalent thereof.168| In some embodiments, CD 19 specific CAR-T cells as described herein are administered to a subject at a dose of about 2 x 10* per kg of body weight. In some embodiments, a maximum dose administered is about 2 x 10sviable CD19 specific CAR-T cells. In some embodiments, the dose is a therapeutically effective amount of viable CDI 9 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CDI 9 specific CAR-T cells. In some embodiments, the CD 19 specific C AR of the cells is the same CDI 9 specific CAR as axicabtagene ciloleucel (YF. SCARTA'A. a structural equivalent thereof, or a functional equivalent thereof.

[0169] In some embodiments, the CDI 9 specific CAR-T cells described, herein are administered to a subject at a dose of about 2 x 106per kg of body weight. In some embodiments, a maximum dose of about 2 x 108viable CD I 9 specific CAR-T cells is administered to a patient of about 100 kg of body weight and above. In some embodiments, the dose is a therapeutically effective amount of viable CD 19 specific CAR-T cells. In other embodiments, the dose Is a clinically effective amount of viable CD 19 specific CAR-T ceils. In some embodiments, the CD19 specific CAR of the cells is the same CD 19 specific CAR as brexueabtagene autoleuc-el (TECARTUS®), a structural equivalent thereof or a functional equi valent thereof.

[0170] In some embodiments, the CD 19 specific CAR-T cells described herein are administered to a subject at a dose of up to about 2 x 10sviable CD 19 specific CAR-T cells. In some embodiments, a subject is administered from about 0.2 x 10* to about 5.0 x 10* (e.g„ about 0.2 x 10\ 0.4 x K 0.5 x 10*. 0.6 x 10*, 0.8 x IO6, 0.9 10*, 1.0 x 10*, 1,2 1(A 1,4 x I ( 1,5 x 1 1.6 x 1 A. 1,8 X 10*, 1,9 x 10'22.0 x 10*. 2,2 X HA 2.4 x = 022.5 x 10s, 2,6 x 10s, 2.8 x 10* 2.9 x 10*, 3.0 x 10* 3.2 x TO*, 3.4 x 10*, 3.5 x I06, 3.6 x 10*, 3.8 x 10*, 3. x 10s, 4.0x 10s, 4,2 x 1 (A 4.4 x 10* 4.5 x 10* 4.6 x 1024.8 x 10*, 4.9 x 10s, or 5,0 x IO6) viable CD19 specific CAR-T cells per kg of body weight for a subject with a body weight ofabout 50 kg or less. In some embodiments, a subject is administered from about 0> I x 10sto about 2,5 x 10s(e.g., about 0.1 x 10s, 0.2 x 10* 0.4 x 10* 0.5 x 10* 0.6 x 1(A 0.8 x 1 (A 0.9 x 10* 1.9 x 10*, 1.2 x 10*, 1.4 x 10*, 1.5 x 10*, 1.6 x 10*, 1.8 x I06, 1.9 x 10s, 2.0 x 10s, 2.2 x 10*, 2.4 x 10*, or 2.5 x 10*) viable CDI9specific CAR- cells for a subject with a body weight of greater than about 50 kg. In some embodiments, a subject is administered from about 0.6 x 10sto about 6,0 x 10su g., about 0.6 x 10s, 0,8 x 10'\ 0,9 x 10% 1,0 x IO8, 1.2 x 10% 1,4 x 10 1.5 x 108, 1.6 x 10% 1.8 x 10s, 1.9 x 10s, 2.0 x 10s, 2.2 x 10s, 2.4 x 10s, 2.5 x 10s, 2.6 x 10s, 2,8 x 108, 2,9 x 10s, 3,0 x 108, 3.2 x 10s, 3,4 x 10s, 3.5 x 10s, 3,6 x 10®, 3,8 x 10®, 3.9 x 10®, 4,0 x 10®, 4.2 10®, 4.4 x 10% 4.5 x 10®, 4.6 x 10®, 4.8 x 10s, 4.9 x 10s, 5.0 x 10s, 5.2 x 10s, 5.4 x 10s, 5.5 x 108, 5,6 x 10s, 5.8 x IO8, 5,9 x 10% or 6,0 x 108) viable CD 19 specific CAR-T cells. In same embodiments, the dose is a therapeutically effective amount of viable CD19 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CD19 specific CAR-T cells. In some embodiments, the CD19 specific CAR of the cells is the same C D 19 specific CAR as tisagenlecleucel (KYMRIAH^), a. structural equivalent thereof, or a functional equivalent thereof.01711 In some embodiments, a single dose of any of the CD19 specific CAR-T cells described herein includes about 50 x 19® to about 200 x IO6(e.g., 50 x 10®, 51 x I O8, 52 x 10% 53 x 10% 54 x 10®, 55 x 10% 56 x 10% 57 x 10®, 58 x 10% 59 x 10% 60 x 10% 61 x 10% 62 x 10®, 63 x 10®, 64 x 10% 65 x 10% 66 x 10% 67 x 10®, 68 x 10% 69 x 10% 70 x 10% 71 x 10% 72 x 10®, 73 x 10®, 74 x 10®, 75 x 10% 76 x 10% 77 x 10% 78 x 10®, 79 x 10®, 80 x 10®, 81 x 10®, 82 x 10®, 83 x 10% 84 x 10®, 85 x 10% 86 x 10% 87 x 10% 88 x 10% 89 x 10®, 90 x 10% 91 x 10% 92 x 10®, 93 x 10®, 94 x 10®, 95 x 10% 96 x 10®, 97 x 10% 98 10®, 99 x 10% 100 10% 101 x 10% 102 x 10®, 103 10% 104 10®, 105 10s, 106 x 10% 107 x 10% 108 x 10% 109 x 10% H0 x 10% 111 x 10% 112 x 10% 113 x 10®, 114 x 10®, 115 x 10®, 116 x 10®, 117 x 10% 118 x 10®, 119 x 10®, 120 x 10®, 121 x 10% 122 x 10®, 123 x 10% 124 x 10®, 125 x 10% 126 x 10% 127 x 10% 128 x 10% 1 9 x 10% 130 x 10% 131 x 10% 132 x 10®, 133 x 10% 134 x 10% 135 x 10% 136 x 10®, 137 x 10% 138 x 10% 139 x 10% 140 x 10®, 141 x 10% 142 x 10®, 143 x 10% 144 x 10®, 145 x 10% 146 x 10% 147 x 10% 148 x 10% 149 x 10% 150 10®, 151 x 10% 152 10®, 153 x 10% 154 x 10®, 155 x 10% 156 x 10% 157 x 10% 158 x 10®, 159 x 10% 160 x 10®, 161 x 10% 162 x 10% 163 x 10% 164 x 10% 165 x 10% 166 x 10% 167 x 10% 168 x 10% 169 x 10®, 170 x 10®, 171 x 10 172 x 10% 173 x 10% 174 x 10% 175 x 10% 176 x 10% 177 x 10®, 178 x 10®, 179 x 10 180 x 10% 181 x 10% 182 x. 10% 183 x 10®, 184 x 10®, 185 x 10®, 186 x 10% 187 x 10®, 188 x 10% 189 x 10% 190 x 10®, 191 x 10®, 192 x 10% 193 x 10 194 x 10% 195 x 10®, 196 x 10®, 197 x 10% 198 x 10®, 199 x 10% or 200 x 10®) viable CD 19 specific CAR-T cells. In some embodiments, the dose is a therapeutically effective amount of viable GDI 9 specific CAR-T cells. In other embodiments, the dose is a clinicallyeffect ve amoun of viable CD 19 specific C AR - cells. In some embodiments, the viable CD 19 specific CAR-T cells include CD1 specific CAR expressing CD4+ T cells and CD 19 specific CAR expressing CD8+ cells at a ratio of about 1:1. In some embodiments, the CD 19 specifi c CAR is the same CD19 specific CAR as lisocabtagene maraleucel (BREYANZ ), a structural equivalent thereof, ora functional equivalent thereof.|< H72] In some embodiments, a single dose of any of the CD19 specific CAR-T cells described herein includes about 2 x 'Hr viable CD19 specific CAR-T cells. In some embodiments, a single infusion bag of any of the CD1.9 specific CAR-T cells described herein includes about 2 x 10sviable CD19 specific CAR-T cells in a cell suspension of about 68 ml... In some embodiments, the CD1 specific CAR is the same CD 19 specific CAR as axicabtagene ciloleucel (YESCARTA®), a structural equivalent thereof, or a fonctional equivalent thereof. (01731 In some embodiments, a single dose of any of the CD 19 specific CAR-T cells described herein includes about 2 x 10sviable CD19 specific CAR-T cells. In some embodiments, a single infusion bag of any of the CD19 specific CAR-T cells described herein includes about 2 x 10sviable CD 19 specific CAR-T cells in a cell suspension of about 68 mL. In some embodiments, the CD19 specific CAR is the same CD 19 specific CAR as brexueabtagene autoleucel (TECAR US®), a structural equivalent thereof or a functional equivalent thereof. (0174] In some embodiments, a single dose of any of the CD19 specific CAR-T cells described herein includes about 0.2 x IO6to about 5.0 x 10® (e.g., about 0.2 x IO6, 0.3 x I06, 0.4 x 106s0.5 x 106, 0.6 I06. 0.7 x 10® 0,8 x IO6, 0.9 x 10s, 1 0 x 10®, 1.1 x 10s, 1.2 x I Of. 1.3 x 10s.1.4 x 10s, 1.5 x 10s, 1.6 IO6, 1.7 x 10s, 1.8 x 10s1.9 x IO® 2.0 x IO6, 2.1 x 10H2.2 x 10s, 2.3 x IO6,,4 x 10s, 2.5 x 1062.6 x 10®,,7 x IO6, 2.8 x 10®, 2.9 x 10®, 3.0 x 10®, 3.1 x 10®, 3.2 x 10®, 3.3 x 10s, 3.4 x 10s, 3.5 x 10s, 3.6 x 10s, 3.7 x IO6, 3.8 x 10®, 3.9 1064,0 x IO6, 4,1 x IO6, 4,2 x 10s, 4.3 x 10s, 4,4 x IO6, 4.5 x 10s4.6 x 10s, 4.7 x 10s, 4.8 x 10s4.9 x 10s, or 5.0 x 10s) viable CD! 9 specific CAR-T cells per kg of body weight for a subject with a body weight of 50 kg or less. In some embodiments, a single dose of any of the CD19 specific CAR-T cells described herein includes about 0.1 x 10sto about 2.5 x 10s(e.g., about 0.1 x 10®, 0.2 x 10s, 0.3 x 10s, 0.4 x 10s, 0.5 x 10s0,6 x I / O6, 0,7 x 10s0.8 x 10s, 0.9 x 10s, LOx 10s, 1.1 x 10®, 1.2 x 10s, 1.3 x 10s, 1.4 x 10®, 1.5 x.10®, 1.6 x 10s, 1.7 x 10®, 1.8 x 10s, 1.9 x 10s, 2.0 x 10s2.1 x 10s, 2.2 x 10s, 2.3 x 10s, 2.4 x 10®, or 2.5 x 10s) viable CD 19 specific CAR-T cells per kg of body weight for asubject with a body weight of more than 50 kg. In some embodiments, a single dose of any of the CD 19 specific CAR-T cells described herein includes about 0.6 x 10sto about 6.0 x 10s(e.g., about 0.6 x IO8, 0.7 x 108, 0.8 x 108, 0.9 x IO8, 1.0x 10s, 1.1 x IO8, 1.2 x 10s, 1.3 x 10s, 1.4x 10s, 1.5 x 10s, 1.6 x 10s, 1.7 x 10s, 1.8 x 10s, 1.9 x 10s, 2.0 x 108, 2.1 x 10s, 2,2 x 108, 2.3 x 10s, 2.4 x 10s, 2.5 x 10s, 2.6 x 10s, 2.7 x 10s, 2.8 x 10s, 2.9 x 10s, 3,0 x 10s, 3.1 x. 10s, 3.2 x 10s, 3.3 x 10s, 3.4 x 10s, 3.5 x 10s, 3.6 x 10s, 3.7 x 10s, 3.8 x 10s, 3.9 x 10s, 4.0 x 10s, 4.1 x IO8, 4.2 x 10s, 4.3 x 10s, 4.4 x 10s, 4.5 x 10s, 4.6 10s, 4.7 10s, 4.8 108, 4.9 x 10s, 5.0 x 10s, 5.1 x 10s, 5.2 x.10, 5.3 x 10s, 5.4 x 10s, 5.5 x 10s, 5.6 x 10s5.7 x 10s, 5.8 x 10s, 5.9 x 10s, or 6.0 x 10s) viable CDI9 specific CAR-T cells. In some embodiments, a single infusion bag of any of the CD19 specific CAR-T cells described herein includes about 0.6 x 10sto about 6.0about 0.6 x 10s, 0,7 x 10s, 0,8 x 10s, 0,9 x 10s, 1.0 10s, Li x 10s, 1,2 x 10s, 1.3 10s, 1.4 x W8, 1.5 x 10s, i.6 x 10s, 1.7 x 10s, 1.8 x 10s, 1.9 x 10s, 2.0 x 10s, 2.1 x 10s, 2.2 x 10s, 2.3 x 10®, 2.4 x 10®, 2.5 x 10®, 2.6 x 10s, 2.7 x 10s, 2.8 x 10s, 2.9 x 10s, 3.0 x 10s, 3.1 x 108, 3.2 x 10s, 3.3 x 108, 3.4 x 10s, 3.5 x 10s, 3.6 x 10s, 3.7 x 10s, 3.8 x 108, 3.9 x 10s, 4.0 x 108, 4.1 x 10s, 4.2 x 10®, 4.3 x 10®, 4.4 x 10®, 4.5 x 10s, 4.6 x 10s, 4.7 x 10s, 4.8 x 10s, 4.9 x 10s, 5.0 x 10s, 5.1 x 10s, 5.2 x 10®, 5.3 x 10s, 5.4 x 10s, 5.5 x 10s, 5.6 x 10s, 5.7 x 10s, 5.8 x 10s, 5.9 x 108, or 6.0 x 10s) viable CD19 specific CAR-T cells in a cell suspension of from about 10 mL to about 50 mL. In some embodiments, the dose is a therapeutically effective amount of viable CD 19 specific CAR-T cells. In other embodiments, the dose is a clinically effective amount of viable CD 19 specific CAR-T cells. In some embodiment, the CD19 specific CAR of the cells is the same CD19 specific CAR as tisagealedeucel (KYMRIAH'®), a structural equivalent thereof, or a functional equivalent thereof.2. Pharmaeeutical Compositions(01751 for therapeutic application, cells prepared according to the disclosed methods can typically be supplied in the form of a pharmaceutica l composition comprising an isotonic excipient and are prepared under conditions that are sufficiently sterile for human administration. For general principles in medicinal formulation of cell compositions, see ’’Cell Therapy: Stem Cell Transplantation. Gene Therapy, and Cellular Immunotherapy," by Morstyn & Sheridan eds, Cambridge University Press, 1996; and "Hematopoietic Stem Cell Therapy," E. D. Ball, J. Lister& P. Law, Churchill Livingstone, 2000. The cells can be packaged in a device or container suitable for distribution or clinical use.[Ill 76"| In some embodiments, pharmaceutical compositions of title present disclosure comprise a population of engineered bypoim munogenic T cells comprising: (a) reduced expression of Beta-2~Microglobu1in (B2M), Class II Transactivator (ClITA), and T cell receptor alpha (TRAC) relative to a control T cell, (b) increased expression of CD47 encoded by a first exogenous polynucleotide relati ve to the control T cell, and (c) expression of a CD19-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the population of engineered hypoimmunogenic T cells are allogeneic to the subject receiving the pharmaceutical composition.a. Pharmaceutically acceptable carriers101771 In some embodiments, a pharmaceutical composition provided herein further include a pharmaceutically acceptable carrier. Acceptable carriers, exci ients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic adds; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenxyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-peutanoI; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serurn albumin, gelatin, or immunoglobulins; hydrophilic polymers such as poly vmylpyxtoli done; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; sah-fonnmg counter-ions such as sodium; metal complexes (&g., Zu-protein complexes); salts such as sodium chloride; and / or non-ionic surfactants such as polysorbates (TWEEN™), poloxamens (PLURONICS™) or polyethylene glycol (PEG). In some embodiments, the pharmaceutical composition includes a pharmaceutically acceptable buffer (e.g., neutral buffer saline or phosphate buffered saline).|01781 In some embodiments, a pharmaceutical composition includes one or more electrolyte base solutions selected from the group consisting of lactated CryoSton®, Ringer’ssolution, PlasmaLyte-ArM, Iscove’s Modified Dulbecco’s Medium, Normosol-R™:, Veen-O™, Polysal® and Hank’s Balanced Salt Solution (containing no phenol red). These base solutions closely approximate the composition of extracellular mammalian physiological fluids, 10179] In some embodiments, a pharmaceutical composition includes one or more cryoprotcctive agents selected from the group consisting of arabinogalactan, glycerol, polyvinylpyrrolidone (PVP), dextrose, dextran, trehalose, sucrose, raffinose, hydroxyethyl starch (HES), propylene glycol, human serum albumin (USA), and dimethylsulfoxide (DMSO). In some embodiments, the pharmaceutically acceptable buffer is neutral buffer saline or phosphate buffered saline. In some embodiments, pharmaceutical compositions provided herein include one or more of CryoStor® CSB, P1asma~Lyte~Aw, HSA, DMSO, and trehalose.|0180] CryoStor® is an intracellular-like optimized solution containing osmotic / oncotic agents, free radical scavengers, and energy sources to minimize apoptosis, minimize ischemiafreperfusion injury and maximize the post-thaw reco very of the greatest numbers of viable, functional cells. CryoStor® is serum- and protein-free, and non-immunogenio.CryoStor® is cGMP^anu factored from raw materials of USP grade or higher. CryoStor# is a family of solutions pre-formulated with 0%, 2%, 5% or 10% DMSO. CryoStor# CSB is a DMSO-free version of CryoStor#’. In some embodiments, a pharmaceutical composition includes abase solution of CryoStor# CSB at a concentration of about 0-100%. 5-95%, 10-90%, 15-85%, 20-80%, 30-80%, 40-80%, 50-80%, 60-80%, 70-80%, 25-75%, 30-70%. 35-65%, 40-60%, or 45-55% w / w. In some embodiments, a pharmaceutical composition includes a base soluti on of CryoStor® CSB at a concentration of about 0%, 5%, W%, 15%, 20%, 2556, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% w / w, (0181] PlasmaLyte-A™ is a non-polymeric plasma expander and contains essential salts and nutrients similar to those found in culture medium but does not contain additional constituents found In tissue culture medium which are not approved for human infusion, &,g.. phenol red, or are unavailable in U. S. P. grade. PlasmaLyte-A™ contains about 140 mEq / hter of sodium (Na), about 5 mEq / Iiter of potassium (K), about 3 mEq / liter of magnesium (Mg), about 98 mEq / liter of chloride (Cl), about 27 mEq / liter of acetate, and about 23 mEq / liter of gluconate. (PlasmaLyle-A™ Is commercially available from Baxter, Hyland Division, Glendale Calif., product No. 2B2543). In some embodiments, a pharmaceutical composition includes a basesolution of Plas aLyte-A™ at a concentration of about 0-100%, 5-95%, 10-90%, 15-85%, 15-80%, 15-75%, 15-70%, 15-65%, 15-60%, 15-55%, 15-50%, 15-45%, 15-40%, 15-35%, 15-30%, 15-25%, 20-80%, 20-75%, 20-70%, 20-65%, 20-60%, 20-55%, 20-50%, 20-45%, 20-40%, 20-35%, 20-30%, 25-75%, 30-70%, 35-65%, 40-60%, or 45-55% w / w. In some embodiments, the pharmaceutical composition includes a base solution of PlasmaLyte-A™ at a concentration of about 0%, S%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% w / w,

[0182] In some embodiments, a pharmaceutical composition includes a DMSO concentration (v / v) of at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 16%, at least about 17%, at least about 18%, at least about 19%, at least about 20%, at least about 1%, at least about 22%, at least about 23%, at least about 24%, or at least about 25%. In some embodiments, a pharmaceutical composition includes a DMSO concentration (v / v) of about 7.5%.

[0183] In some embodiments, a pharmaceutic l composition includes human serum albumin (HSA) at a concentration of about 0-10%, 03-9.3%, 03-8.3%, 03-7.3%, 03-63%, 03-5,3%, 03-43%, 03-3.3%, 03-2.3%, 0.3- 1 %, 0.6-83%, 0,9-73%, 1.2-63%, 1.5-53%, 1.8-43%, or 2.1 -3.3% w / v. In some embodiments, a phamraceutical composition comprises human serum albumin (HSA) at a concentration (w / v) of at least about 0.1%, at least about 0.2%, at least about 03%, at least about 0.4%, at least about 0.5%, at least about 0.6%, at least about 0.7%, at least about 0.8%, at least about 0.9%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, al least about 13%, at least about 14%, at least about 15%. at least about 16%, at least about 17%, at least about 18%, at least about 1 %, at least about 20%, at least about 21 %, at least about 22%, at least about 23%, at least about 24*14, or at least about 25%. In some embodiments, a pharmaceutical composition includes HS A at a concentration of about 0%, 03%, 0.6%, 0.9%, 1.2%, 1.5%, 1.8%, 2.1%, 2.4%, 2.7%, 3.0%, 3.3%, 3.6%, 3.9%, 4.3%, 4.6%, 4.9%, 53%, 5.6%, 5.9%, 6.3%, 6.6%, 6.9%, 7.3%,7.6%, 7.9%, 8.3%, 8.6%, 8.9%, 9.3%, 9.6%, 9.9%, or 10% w / v, In some embodiments, a phar aceuticnl composition comprises US A at a concentration (w / v) of about 0.3%.10184 In some embodiments, a pharmaceutical composition includes dimethyl sulfoxide (DMSO) at a concentration of about 0- 10%, G.5-9.5%, 1-9%, 1.5-8.5%, 2-8%, 3-8%, 4-8%, 5-8%, 6-8%, 7-8%, 2.5-7.5%, 3-7%, 3.5~6.5%, 4-6%, or 4.5-5.5% v / v. In some embodiments, a pharmaceutical composition includes HSA at a concentration of about 0%, 0.25%, 0.5%, 0.75%, 1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4,75%, 5,0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8,0%s8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9,5%, 9.75%, or W.0% v / v.(01851 fa some embodiments, a pharmaceutical composition includes dimethyl sulfoxide (DMSO) ata concentration of about 0-10%, 0.5-9 5%, 1-9%, 1.5-8.5%, 2-3%, 3-8%, 4-8%, 5-8%, 6-8%, 7-8%, 2.5-7.5%, 3-7%, 3.5-6.5%, 4-6%, or 4.5-5.5% v / v. In some embodiments, a pharmaceutical composition incl udes HSA at a concentration of about 0%, 0.25%, 0.5%, 0.75%, 1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4.75%. 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0%, 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, or 10,0% w / v.(0186] In some embodiments, a pharmaceutical composition includes trehalose at a concentration of about 0-500, 50-450 mM, 100-400 mM, 150-350 M, or 200-300 mM. In some embodiments, a pharmaceutical composition includes trehalose at a concentration of about 0 M, 10 mM, 20 mM, 30 mM, 40 mM, 50 mM, 60 mM, 70 mM, 80 mM, 90 M, 100 m, 125 mM, 150 mM, 175 mM, 200 mM, 225 mM, 250 mM, 275 mM, 300 mM, 325 mM, 350 mM, 375 mM, 400 mM, 425 mM, 450 mM, 475 mM, or 500 mM,

[0187] Exemplary pharmaceutical composition components are shown in Table 1. ruble 1. Exemplary pharmaceutical eompusition components*HS in addition to PlasmaLyte.|0188] In some embodiments, a pharmacal composition comprises hypoimmunogenic cells described herein and a pharmaceuti cally acceptable carrier comprising 31.25% (v / v) Plasma-Lyte A, 31.25% (v / v) of 5% dextrose / 0.45% sodium chloride, 10% dextran 40 (LMD) / 5% dextrose, 20% (v / v) of 25% human serum albumin (HSA), and 7.5% (v / v) dimethyl sulfoxide (DMSO).(0189] In some embodiments, a pharmaceutical composition comprises hypoimmunogenic cells described herein and a pharmacenti eally acceptable carrier comprising 75% Cryostor CSW, 25% (v / v) Plasma-Lyte A, 7.5% (v / v) DMSO, and 1.2% (v / v) HSA.(0190] In some embodiments, a pharmaceutical composition comprises hypoimmunogemc cells described herein and a pharmaceutically acceptable carrier comprising 31.25% ( v / v) Plasma-Lyte A, 31.25% (v / v ) of 5% dextrose / 0.45% sodium chloride, 10% dextran 40 (LMD) / 5% dextrose, 20% (w / v) of 25% human serum albumin (HSA), and 7.5% (v / v) dimethyl, sulfoxide (DMSO),{0191] In some embodiments, a pharmaceutical composition comprises hypoimmutogenic cells described herein and a pharmaceutically acceptable carrier comprising 75% Cryostor CS10, 25% (v / v) Plasma-Lyte A, 7.5% (v / v) DMSO, and 1,2% (w / v) HSA.3, Diseases, Disorders and Conditions to be Treated|0192] B cell lymphomas encompass a group of lymphoma subtypes categorized within 2 broad categories: non-llodgkrn lymphoma (NHL) and Hodgkin lymphoma (HL), with NHLs being the majority of diagnosed cases. The therapeutic landscape varies for each distinct tumor subtype, but generally there are limited effective therapies in relapsed and / or refractory (r / r) settings across B cell lymphomas (National Comprehensive Cancer Network (NCC. NJ 2021). Despite recent a vancement, NHL remains an unmet medical need. In 20 1 American Cancer Society estimated that 82,000 eases of NHL would be diagnosed, with approximately 20,000 attributed, deaths in the US in 2021 (American Cancer Society' 2021), in 2020, there were123*000 new cases of NHL reported, with 50,000 attributed deaths in Europe (World Health Organization 2020). NHL is a heterogeneous group of cancers originating: in B lymphocytes, T lymphocytes, or natural killer (NK) cells. In the US, B cell lymphomas represent approximately 85% of all NHL oases (American Cancer Society 2019). Subtypes of NHL include large B cell lymphomas (LBCL), such as diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), and primary mediastinal B cell lymphoma (PMBCL), along with mantle lymphoma (MCL). Subtypes of indolent NHL (1NHL) include follicular lymphoma (FL) and marginal zone lymphoma (MZL).f 0193| DLBCL is the most aggressive NHL in adults, accounting for a third of all NHL cases, about 25,000 annually. In the US, the incidence of DLBCL is approximately 7 cases per 100,000 persons per year, with Caucasian- Americans having higher rates than other racial groups. There is a mate predominance, and incidence increases with age with a median age of diagnosis of 64 years (Morton 2006, Shenoy 2011). Gene expression profiling has identified 2 distinct subtypes within DLBCL that are managed with similar standard of care germinal center B cell (GCB) subtype and activated B cell (ABC) subtype. GCB subtype i associated with improved outcomes to therapy. MYC rearrangements are found in 5% to 8% of DLBCL patients and correlate with higher risk of disease progression (NCCN 2021).|(H 94| HGBCL includes DLBCLs with MFC and B cell lymphoma 2 (MX?) and / or B cell lymphoma 6 (BCL6) rearrangements (known as “double-hit” or “triple-hit lymphomas” based on the number of translocations). Generally, MYC rearrangements in DLBCLs are associated with a poor prognosis that is worsened in cases of concomitant BCL2 and / or BCL6 alterations. The majority of HGBCL are germinal center B cell like lymphomas and present with worse outcomes than DLBCL, though treatment algorithms are shared. Exceptions to HGBCL. include cases of follicular or lymphobl astic lymphoma or HGBCL, not otherwise specified (NOS), which contains blastoid -appearing LBCLs or cases that lack MFC and BCL2 QIBCLG translocations (NCCN 2021, Swerdlow 2016).|l)1 5j PMBCL has distinct clinical, pathological, and molecular characteristics compared: with DLBCL and represents approximately 2% to 4% of all patients with NHL ( Bhatt 20I5, Dabrowska-Iwanick 2014), Initial treatment for PMBCL is similar to DLBCL (i,e., anthracycline-containing regimens with rituximab with or without involved-field radiotherapy),which is curative in th© majority' of cases. Chemosensitive patients who relapse may respond to further cytotoxic therapy followed by autologous stem cell transplant (ASCT); nonresponders have limited therapeutic options that lack durability (NCCN 2021 ).

[0196] Follicular lymphoma 3B (FL3B) is the most aggressive variant of F'L, the most common subtype of indolent non~Hodgkin lymphoma. The majority of cases contain a t(I4; 18) translocation, which juxtaposes BCX2 with the IGH locus. Pathologic grading of FL according to the number of centroblasts is a clinical predictor of outcome. Grade 3 follicular lymphoma is subclassified into FL3A (centrocytes still present) or FL3B (sheets of cenlroblasts), which have similar survival outcomes but may be managed differently. FL3B is commonly treated according to treatment recommendations for DLBCL (NCCN 2021).

[0197] Treatment options for LBCL include rituximab-based chemotherapy (R-CHOP) and SCT. Most patients with DLBCL respond well to rituximab-based immuno chemother apy. However, outcome remains poor for patients with r / r DLBCL (1 -year and -year overall survival [OS] rates of 23% and 16%, respectively). While ASCT confers durable responses, approximately half of DLBCL patients are ineligible for treatment and the majority of ASCT-treated patients will experience disease progression within 3 years (Seim 2021, Crump 2014, Oisselbrecht 2010). Those patients who do not receive ASCT, either because they are not eligible even after showing response to the second-line chemotherapy or because they do not have responses to the second-line chemotherapy (refractory, have a very poor prognosis with a median OS of 4.4 months. For patients who have relapsed disease within 12 months after ASCT or primary refractory disease, only 26% of them respond io the next line of therapy (third line) with 7% of complete response and median OS of 6,2 months. Patients whose disease progresses after 2 or more prior lines of systemic therapy regimens are unlikely to benefi t further from currently available systemic therapy options, unless they have experienced a long disease-free interval. Thus, there is considerable unmet medical need for DLBCL patients who do not respond to first-line immunochemotherapy or to subsequent courses of combination chemotherapy ( NCCN 2021).

[0198] To assess the outcomes of patients with r / r LBCL, the multieenter retrospective study SCHOLAR-! analyzed data from 2 large Phase 3 studies involving 636 patients. Results showed that refractory DLBCL patients have an objective response rate (ORR) of 26%. acomplete response (CR) rate of 7%, and a median OS of 6.3 months (Crump 2017, Gisselbreeht 2012, Van Den Neste 2016). Poor outcomes were observed across all refractory subgroups (primary refractory’, refractory’ to second-line or higher therapy, or relapse < 1 year after ASCT) and regard less of disease stage, suggesting that these patients form a single population who lack effective treatment.[01.99] Recently, new therapeutic options have been approved for r / r LBCL, In 2020, FDA granted accelerated approval for tafesltamab In combination with lenalidomide for the treatment of adult patients with r / \DLBCL, NOS, including DLBCL arising from low-grade lymphoma, and who are not eligible for ASCT, Among 71 subjects with r / r DLBCL treated with tafasitamab plus lenalidomide, 37% achieved a CR and 18% achieved a partial response (PR) (MONJUVI Prescribing Information 2020). The estimated, median duration of response (DOR) was 2L7 months, EMA granted approval forpolatuzumab vedotin, a CD79b-directed antibodydrug conjugate, in combination with bendamustine and rituximab (BR) for treatment of r / r DLBCL patients who are not candidates for stem cell transplant (SCT), Polatusumab vedotin plus BR resulted in an ORR of 45% (BR alone: 18%) and a CR rate of 40% (BR alone: 18%) (POLIVY Prescribing Information 2G19, Sehn 2018). Pembrolizumab, a humanized monoclonal antibody (mAb) against programmed cell death protein I (PD- 1), received accelerated approval by FDA for the treatment of PMBCL refractory to treatment or that has relapsed after 2 or more prior lines of therapy (Armand 2019, KEYTRUD. A Prescribing Information 2019). Th e single-arm, multicenter KEYNOTE-170 study of pembrolizumab enrolled 53 patients with a median of 3 prior lines of therapy, and results showed an ORR of 45% (CR: 13%) with the median OS not reached.

[0200] Also recently, autologous CD19-directed chimeric antigen receptor (CAR) T cell therapies (tisagenlecleucel, axicabtagene ciloleucel, and lisocabtagene maraleucel ) have been approved as a second-line and / or third-line therapy for patients with r / r LBCL. These therapies offer promising options for these patients, with CR rates ranging from 39% to 60% and median DOR from 9.2 months to 16.7 months. Key toxicities include cytokine release syndrome (CRS) and neurotoxicity with rates of Grade 3 and higher events ranging from 4% to 23% and 11% to 31%, respectively (KYMRIAH® Prescribing Information 20 1; YESCARTA® Prescribing Information 2021 BREYA’NZI® Prescribing onnation 2021). Three recent randomized trialsevaluated CD19-directed:CAR T cell therapies as second-line therapy for LBCL (Locke 2022, Bishop 2021, Kamdar 2022). The ZUMA-7 stady demonstrated superiority of axicabtagene ciloleucel over standard of care (CR 65% vs 32%, event-free survival (EPS hazard ratio [HR] 0.40, p < 0.001) and TRANSFORM demonstrated superiority of lisocabtagene mural eucel over standard of care (EFS 10.1 months vs 23 months, HR 0.35). However, the BELINDA study failed to demonstrate superiority of tlsagenlecleucel versus standard of care (EPS HR 1.07, p ~ 0.69) dri ven in part by differences in trial design and treatment delays associated with manufacturing. Despite their approvals, the manufacturing and logistical challenges of autologous CAR T cell therapies make it difficult for these therapies to be widely used (Gajra 2022). Therefore, considerable unmet medical need still exists for patients with r / r LBCL, and there is urgent need to develop innovative medicines including more readily available (i.e,, off-the-shelf) CAR T cell therapies.|0201 J iNHLs include FL and M'ZL. FL is the most common indolent form of NHL, accounting for between 10% and 22% of all cases (NCCN 2021, Terns 2016). Although newly diagnosed, low-grade, limited-stage FL is often responsive to a variety of first-line treatments; the disease is characterized by a pattern of relapsing and remitting disease. Transformation from FL to DLBCL (transformed FL ftFLJ) occurs at an annual rate of approximately 3% for 15 years and is associated with poor outcomes (Montoto 2007).

[0202] Outcomes in FL are mixed. High-risk patients with > 3 Follicular Lymphoma International Prognostic Index (FLIPI) criteria have a 5-year OS rate of 53%, whereas low-risk patients with < 1 FLIPI criterion have a 5-year survival rate of over 90%. Advanced-stage FI. generally requires multiple successive lines of therapy leadi ng to progressively shorter remission periods, chemorefractory disease, transformation to DLBCL, or death due to repeated treatment-related toxicides (Cheah 2018). Thus, regardless of the disease stage at diagnosis, most patients with FL ultimately experience disease relapse, even after a long-term response to first-line therapy, representing a significant unmet need.[(1203] First-line therapies for FL include R-CHOP, R-CVP, bendamustine - CD20 mAh, and lenalidomide + rituximab. High-dose (chemo)therapy / a.uto-SCT as a consol idative therapy for patients with FL who are in second or third remission is a potential therapeutic option (NCC’N 2021). Allogeneic (allo)-SCT may offer improved relapse rates and disease controlcompared with auto-SCT; however, allo-SCT is associated with higher nonrciapse mortality rates and is not a viable treatment option for most patients with FL (Freedman 2018 )• New targeted agents have been developed to treat patients who progress following systemic chemotherapy and rituximab-containing regimens. The “enhancer of zeste homolog'” (EZH)2 histone methyl transferase inhibitor tazemetostat was granted accelerated approval by the FDA for adult subjects with EZH2 mutant r / r FL who have received > 2 prior therapies (or have no satisfactory alternative treatment options) (TAZVERIK 2020). Additionally, phosphatidylinositol 3-kina e inhibitors (Le., idelalisib, copartlisib, and duvelisib) have been approved for r / r FL after 2 prior therapies.0204] CD 19-directed CAR T cell therapy has also been utilized in the treatment of r / r FL, In a study of 21 subjects with r / r L (8 with FL and 13 with tFL) treated with an investigational autologous CD! ^directed CAR T therapy, CR rates were 88% and 46% for FL and tFL patients, respectively. All patients with FL who achieved CR remained in remission at a median follow-up of 24 months while the median duration of response for tFL was 10.2 months (Hirayama 2019). YESCARTA has received accelerated approval from the FDA for treatment of adult patients with r / r FL after 2 or more lines of systemic therapy. Approval was based on Study KTECI91O5 (ZUMA5), where treatment with YESCARTA among 81 subjects with r / r FL resulted in an ORR of 91% and a CR rate of 60% (YESCARTA 2021). Subjects had at least 9 months of potential follow-up and a median of 3 prior lines of therapy. Among the 74 responding subjects, the median DOR was not estimable with a median follow-up time of 14.5 months.| D2 5 These data highlight the need for novel treatment options for patients with r / r FL who have received 2 or more prior lines of therapy and demonstrate the promise of durable responses with CAR T cell therapy.|0206| MZL is a rare form of N HL originating in the marginal zone of B cells (Kahl 2008). The low incidence rates and heterogeneous presentation render large-scale clinical studies of MZL challenging. First-line treatment options for MZL include R-CHOP, R-CVP, rituximab, and bendamustinc + rituximab. Generally, patients respond to front-line therapy; however, patients with r / r MZL have few therapeutic options, including Bruton tyrosine kinase(BTK) inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, and lenalidomide plus rituximab (NCCN 2021).|0207| Ibrutinib, a Bruton tyrosine kinase inhibitor (BTKi), is approved for patients with MZL who have received at least 1 priiw anti-CD20-based therapy (IMBRLA'TCIA 2022).Accelerated approval of ibrutinib was granted based on a prospective, multicenter, open-label Phase 2 study of 63 patients with MZL that reported an ORR of 46% and CK rate of 3.2%. After a median follow-up time of 19.4 months, the DOR was not reached. Lenalidomide, a thalidomide analogue, is approved for the treatment of previously treated MZL in combination with a rituximab product (R. EVI. IMID 2019). In a pivotal study, among patients with MZL, the ORR by Independent Review Committee assessment was 65% (CR 29%) (Thieblemont 2019). After a median follow-up of 27.9 months, the median progression-free survival (PFS) was 20.2 months.(0208] Preliminary results with CD 19~di ented CAR T cell therapy have also demonstrated promising results in r / r MZL, Treatment with YESCARTA among 7 subjects with r / r MZL yielded an ORR of 86% and aCR rate of 71% (Jacobson 2020). To this end, effective modalities to keep patients disease-free are warranted in patients relapsing after 2 lines of therapy and CAR T therapies demonstrate the promise of durable responses. MCL comprises approximately 6% of all newly diagnosed NHL cases.(9299] MCL carries t he unfavorable characteristi cs of both indolent and aggressive NHL subtypes due to the incurability of disease with conventional chemotherapy and a generally more aggressive disease course compared to indolent NHL. The MCL International Prognostic Index (MIPI) integrates clinical variables including age, performance status, lactate dehydrogenase, and white blood cell count to stratify patien ts into low, intermediate, and high-risk categories. High proliferation index (based on KI-67), blastoid / pleomorphic histology, complex kaiyotype, and TP53 mutations predict poor outcomes for patients (Romandk 2022).(0210] Initial therapy for MCL often Involves intensive, c^tarabme-containing induction chemotherapy regimens such as RDHA, HyperCVAD, or NORDIC, followed by consolidation with an autologous stem cell transplant (ASCI) and 3 years of maintenance rituximab. Older patients and those with medical comorbidities may instead receive less intensi ve induction regimens such as bendamustme plus rituximab, R-CHOP, or lenalidomide plus rituximab.Regardless of initial therapy, most patients will eventually relapse and require multiple lines of subsequent therapies (NCCN 2021),[0211 Targeted agents such as the BTK inhibitors, bortezomib, lenalidomide, arid venetoclax are currently available to treat second-line r / r MCL. Three irreversible BTK inhibitors (ibmtinib, acalabrutinib, and zanuhrutimb) are FDA approved for the treatment of r / r MCL. Ibrutinib was the first BTK inhibitor approval based on a pivotal Phase 2 in r / r MCL demonstrating 68% ORR, 21% CR, median DOR 17.5 months, and median 2-year OS of 47% (Wang 2013). Subsequently, more selective, second-generation BTK inhibitors (acalabrotinib and zanubrutiuib) were approved. Acalabrutmib’s approval was based on the Phase 2 A. CE-LY-2004 trial with 124 r / r MCL patients demonstrating 81% ORR, 40% CR, DO 26 months and median PFS 19.5 months (Wang 2018). Zanubnitinib’s approval was based on the pivotal BGB-3111-206 and BGB3111 - AU-003 trials demonstrating an 84% ORR and 25% to 69% CR. (Tam 2021). Lenalidomide is also FDA approved for the treatment of r / r MCL. When sequenced after bortezomib, lenalidomide monotherapy is associated with an 28% ORR, 7.5% CR, and 16.6 month DOR (Goy 2013).[0212| Brexucahtagene autoleueel (formerly KTE-XI 9) is a CD 19 autologous CAR T product with a CD28 co-stimulatory domain that is now FDA approved for the treatment of r / r MCL based on the results of the ZUMA-2 trial (Wang 2020). In this study, the ORR was 85% (59% CR in the intention-to-treal population, with 12-month PFS and OS rates of 61% and 83%, respectively. Notably, responses were observed m patients with high-risk disease characteristics, including pleomoTphic / blastoid morphology, TP53 aberrations, and high Ki-67. Lisocabtagene maraleucel (1 iso-cel; formerly JCAR017) is an autologous CD1 -directed CAR T product that contains a 4- IBB signaling domain and is administered by sequential iuftjsions of the CD8-> and CD4-t CAR T cells. Lisocel is currently FDA approved for the treatment of r / r large B cell lymphomas based on the results of the TRANSCEND NHL 001 trial, While this product is not yet approved for patients with MCL, the Phase 1 portion of the TRANSCEND trial included 32 patients with r / r MCL and reported an ORR of 84% (59% CR) In this population across all dose levels (Abramson 2020).[0213| Despite the approval of brexticabtageae autoleucel, the product safety profile, manufacturing, and logistical challenges make it difficult for CT) 19-directed CAR T therapies tobe widely used in MCI.. Therefore, considerable unmet medical need still exists for patients with r / r MCL, and there is urgent need to develop innovative medicines including mere readily available (i.e., off-the-shelf) CAR T cell therapies.

[0214] C XL is the most common leukemia in the USA and among the most common globally. CLL can vary from an. indolent disease to very aggressive with potential for transformation to high-grade lymphoma (i.e., Richter’s transformation* NCCN 2022). Previous treatments for CLL included alkylating agents such as chlorambucil and cyclophosphamide and purine nucleosides such as fludarabine, pentostatm, and cladribine. Historically, chemoimmunotherapy regimens like fhid foiue, cyclophosphamide and rituximab (FCR), bendamustine plus rituximab (BR), chlorambucil, alemtuzumab, and lenalidomide were the mainstays of CLL treatment in both the -front-line and relapsed settings. Over the past decade, BTK inhibitors (ibrutinib, acalabnrtinib, zanubruttnib, and pirtobmtinib) BCL2 antagonists (venetoclax), and Pl 3K inhibitors (idelalisib and duvelisib), given as single agent or in combination with anti-CD20 antibodies (e.g., obinutuzumab), have emerged as new standards of care for front-line CLL. Front-line therapy often leads to durable responses, but despite significant progress, CLL remains incurable. Relapses frequently occur in groups with high-risk features, such as those with TP53 mutation, unmutated immunoglobulin heavy-chain variable (IGHV), deletion 11 q or deletion 17p, or presence of complex karyotype. For this reason, chemoimmunotherapies are not recommended for patients with del( 17p) / TP53 mutation due to low response rates (Rainone 2022).

[0215] Targeted therapies used in second-line and subsequent settings for CLL include BTK inhibitors, BCL2 antagonists, anti-CD20 antibodies, and PI3K inhibitors. The Phase 3 DUO study of duvelisib, a PI3K. delta inhibitor, in 319 patients with r / r CLL demonstrated superior PFS to ofatumumab (13.3 months vs 9.9 months, HR ~ 0.52, p 0.001) (Flinn 2018), The Phase 3 IDELA trial in r / r CLL evaluated the PI3K inhibitor idelalisib plus rituximab vs placebo plus rituximab and demonstrated superior efficacy in PFS and OS of the ideialisib-coataimng arm (Sharman 2019). Few treatment options exist for patients who progress on these targeted therapies and therefore a significant unmet medical need still exists for CLL. Allogeneic stem cell transplantation may provide a chance of cure for seme patients but comes with high toxicities and may not be suitable for older CLL patients.[6216 Recently, CAR therapies have demonstrated encouraging durable responses for CLL patients. TRANSCEND CLL 004 is the first multicenter study of an CD19-directed CAR T cell therapy (fisocabfageae maraleucel [liso-cel]) for r / r CLL (Siddiqi 2022). Subjects had a median of 4 prior therapies and 83% had high-risk features, including mutated TP53 and del(l 7p). CRS was reported in 74% of subjects (Grade 3 in 9%), and neurological events in 3934 of subjects (Grade 3 / 4 in 22%). Of 22 efficacy-evaluable subjects, 82% and 45% achieved overall and complete responses, respectively. Of 20 minimum residual disease (MRD)-evaluable subjects, 75% and 65% achieved undetectable MRD in blood and marrow, respectively. At a median follow-up of 24 months, the duration of response was not reached and median PFS was 18 months.|0 17| In some embodiments, a subj ect of the present disclosurehas or is suspected of having a B cell malignancy. In some embodiments, a B cell malignancy is selected from the group consisting of: Non-Hodgkia’s Lymphoma (NHL),:Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), or small lymphocytic lymphoma (SLL).|021 j tu some embodiments, a subj ect of the present disclosure has or is suspected of having a CLL with high-risk features, such as those with TP53 mutation, unmutated imniunoglobulm heavy-chain variable (IGHV), deletion 1 Iq or deletion Up, or presence of complex karyotype. In some embodiments, a subject of the present disclosure who has or is suspected of having a CLL with high-risk features is treated with a composition of the present disclosure as a first-line therapy. In some embodiments, a method of the present disclosure comprises detecting one or more biomarkers in a sample from a subject of the present disclosure and identifying the subject as being a high risk CCL patient based on the one or more biomarkers, to some embodiments, the one or more biomarkers are selected from the group consisting of: del(17p), mutated TP53, unmutated immunoglobulin heavy-chain variable (IGHV) gene, and complex karyotype.10219 In some embodiments, a subject of the present disclosure who has or is suspected of having multiple myeloma is treated with a composition of the present disclosure as a first-line therapy.

[0220] B cells are thought to have multiple functions that include antigen presentetfon, cytokine secretion and autoantibody production. There is a plethora of autoantibodies associated with systemic lupus erythematosus (SEE), suggesting continued B cell stimulation and antigen processing in disease. Not all autoantibodies are known to be pathogenic, and autoantibodies are known to appear in advance of clinical disease (Arbuckle 2003). Several studies have highlighted B cell activating factor (BAFF) as having a crucial role in regulating B cell maturation, survival, and function (Vincent 2012). Elevated levels of BAFF have been reported in serum of patients suffering from autoimmune diseases, including SLE, suggesting a role in disease pathogenesis (Renaudmeai 2004). Belimumab, a folly humanized mAb which neutralizes BAFF, led to impaired B cell survival and reduction in peripheral B cell numbers in SLE patients (Vincent 2.012; Basta 2020). In the Belimumab International Study in Lupus Nephritis 2-year, randomized, placebo-controlled, Phase 3 trial, belimumab plus standard therapies (mycopheuolate mofetil [MMF] or cyclophosphamide followed by azathiop ne) were administered to adult patients with active lupus nephritis (LN). Compared with placebo, significantly more patients randomized to belimumab achieved a primary efficacy renal response (43% vs. 32%; p::::0.03) based on reductions in urinary protein to creatine ratio: (UPCR) and estimated glomerular filtration rate (eGFR), and a. complete renal response (30% vs. 20%; p-::0.02) using endpoints consistent with the proposed SC291-102 study. Renal-related events or death, were also lower in the belimumab group compared with placebo, with almost 50% lower renal-related events among patients who received belimumab than those who received standard therapies alone (Fane 2020).

[0221] Rituximab, initially approved by the FDA tn 1997 for the treatment of r / r NHL, is a mAb that selectively targets the B cell specific surface molecule CD20. The absence of CD20 antigen expression on the cell surface of hematopoietic stem cells, normal plasma cells, or other normal tissues, allows for selective depletion of B cells with rituximab (Fervenza 2008). Murine models have helped establish that CD20+ and circulating B cells are rapidly eliminated by CD20- targeting mAbs, through the reticuloendothelial system. However, B cells in somelocations, such as marginal zones, are not depleted by m Abs targeting GD20, and the reliance on the reticuloendothelial system for B cell depletion is problematic in SLE where it is known to be compromised (Gong 2005).

[0222] Two large, randomized, placebo-controlled, Phase 2 studies In non-renal lupus (EXPLORER) and LN (LUNAR) both failed to meet their primary endpoints after treatment with rituximab, although both trials demonstrated partial responses in selected patients (Merrill 2010; Rovin 2012). Complete peripheral depletion of B cells with rituximab was not observed in all participants, and even in participants wherecompleteperipheral depletion of B cells was observed, <50% achieved complete response (Gomez 2018),

[6223] As observed in murine models, the absence of a positive correlation between adequate peripheral depletion of B cells and complete response in the trial participants suggests that autoreactive B cells may persist in protected microenvironments like the lymphoid structures, kidney tubulointerstitial and bone marrow and therefore prolonged exposure to rituximab (>52 weeks) is needed (Gang 2005; Abuja 2007; Abuja 2011; Bckar 2010).Combination therapies involving rituximab and belimumab (a BAFF inhibitor) have been shown to reduce the re-emergence of autoreactive B cells following B cell depletion, enhance negative selection of autoreactive B cells, and lower anti-dsDNA antibody levels (Atisha-Fregoso 2021; van Schaik 2022). While combination treatments are effective in some patients, certain severe forms of SLE remain resistant to treatment.

[0224] Currently. there is no standard of care (SoC) treatment for achieving drag-free remission of SLE; therefore, patients often require life-long therapy. While a combination approach using antimalarials (hydroxychloroquine), systemic steroids, and conventional immunosuppressant medicines (azathioprine, MMF, and cyclophosphamide) are first-line options for SLE treatment, a significant proportion of pat ients continue to have high disease activity and recurrent relapses (Glazier 2005; Rahman 2008; Appel 2009). Despite the apparent effectiveness of these regimens, complete renal remission in 30 50% of patients within the first year, between 20-35% of patients experience renal relapse on maintenance therapy over a 3-A-year follow-up period, with one study reporting that only 38% of patients were ab le to ma intain a complete renal response over a 5-year period. A lack of response to therapy after 1 year of treatment was significantly associated with increased mortality and chronic kidney disease risk(p<0.005 ) (Anders 201 ) and cyclophosphamide carries a secondary malignancy risk, in addition to increased risk of infertility with cumulative exposure (Ginzler 2005).|l)225] After the results of the LUNAR trial demonstrated no benefit of rituximab in cornpkde renal response and a 15% improvement in partial renal response, a retrospective investigation was undertaken evaluating peripheral blood B cell depletion and occurrence of complete renal response. The data demonstrated that deeper B cell depletion was associated with improved complete renal response, and that poor peripheral blood B cell depletion was associated with non-response, This observation (Gomez 2018), coupled with data demonstrating that, obinuhizumab (a type II mAb with greater antibody dependent cellular cytotoxicity through glycoengineering) in CLL was superior to rituximab in progression-free survival and minimal residual disease negativity in a head-to-head trial of CLL (Goede 2014) prompted the initiation of a Phase 2 trial of Obinntuzumab in LN. The hypothesis driving this trial was that deeper B cell depletion should increase complete renal response in LN patients. The Phase 2 results were published in 2021 (Furie 2022) and described a 1 % improvement in complete renal response and 21% partial renal response at 52 weeks in the obinutuzumab-treated patients vs controls. The Phase 3 trial has completed enrolment and results are expected in 2024.The current SoC treatment for anti-neutrophilic cytoplasmic autoantibody (ANCA) associated vasculitis (A V) involves a combination of high dose glucocorticoids with either cyclophosphamide or rituximab, with evidence suggesting that the combination of rituximab and glucocorticoids is superior for remission induction in severe relapsing AAV compared with cyclophosphamide treatment alone (Stone 2010). Cyclophosphamide therapy is designed to target proliferating cells, resulting in blunted expansion, of pathogenic clones, while memory B and T cells are unaffected. However, due to the dose-limiting AEs induced by cyclophosphamide, coupled with severe toxicity, complete depletion of pathogenic clones is rare, resulting in treatment failure and risk of chronic relapse. Cyclophosphamide is also associated with leukopenia, cancer, and ovarian failure (Mukhtyar 2008). Severe infections are also associated with prolonged cyclophosphamide exposure and are predominant causes of ear ly death in patients (Jones 2010).|h227| Results from the RAVE trial found that rituximab was non-inferiot to cyclophosphamide for remission of disease defined as Birmingham Vasculitis Activity Scale(BVAS) / WG=0, without the use of prednisone, at 6 months (64% in rituximab group vs 53% in cyelophosphanwie group) (Stone 2010). In addition, rituximab was superior to cyclophosphamide for induction of remission for relapsing disease (67% in rituximab group, 42% in cyclophosphamide group). Finally, there were no reported significant differences in rates of adverse events between the two treatments. Despite rituximab showing superiori ty against cyclophosphamide in reducing relapse rates, remission rates are still high in rituximab-treated patients, especially in patients with a history of relapse. The RIT AZAREM trial demonstrated that repeated dosing of rituximab folio wing remission (defined, as BVAS / WG:::0) in patients was superior compared to azatbioprine at preventing relapse (HR.0.41; 95% CI 0.27 to 0.61, p<0.001) (Smith 2023). The MAINRITSAN trials demonstrated that prolonged rituximab treatment was an effective regimen in sustaining remission (defined as BVAS ==0) in patients, however, for some patients after 2 years of maintenance;, relapse risk remains high and extended therapy is advised (’Tien 2020; Charles 2018; Charles 2020; Guillevin 201 ). Despite promising results with rituximab, there remains an unmet treatment need for this group of patients as highlighted in a recent study investigating the long-term outcomes and prognostic factors for survival of patients with AAV (Sanchez 2023) The study concluded that patients with AAV had an increased risk of mortality when compared to the general population, with treatment complications and organ damage being foe main causes of limited survival and in fection the leading cause of mortality.|0228| Improved treatment options are needed as conventional therapies are not only associated with treatment failures and relapses, but also high levels of toxicity which result in Severe morbidity and lethal adverse effects (Jayne 2003). A recent randomized controlled trial (ADVOCATE) investigated the C5a receptor inhibitor, avacopan, for the treatment of AAV (Jayne 2021). Avacopan is a C5a receptor antagonist that selectively blocks tire effects of C5a preventing neutrophil chemoattraetion and activation and has demonstrated beneficial, effects on vasculitis in Phase 2 clinical trials (Jayne 2017; Merkel 2020). All patients were given either cyclophosphamide or rituximab and then assigned into two groups: one received avacopan and the other prednisone. Results showed avacopan was non-inferior but not superior to prednisone in inducing remission defined as BVAS ) of vasculitis at 26 weeks and was superior to prednisone at 52 weeks in patients who received rituximab or cyclophosphamide.

[6229] In some embodiments, a subject of the present disclosure has or is suspected of having a B cell mediated autoimmune disorder. In some embodiments, a B cell mediated autoimmune disorder is selected from the group consisting of: systemic lupus erythematosus (SEE), external systemic lupus erythematosus (ERL), lupus nephritis (LN), C S lupus, anti-neutrophilic cytoplasmic autoantibody (ANCA) associated vasculitis (AAV), granulomatous polyangiitis, microscopic polyangiitis, multiple sclerosis (relapsing and / or progressive), pemphigus vulgaris, autoimmune blistering shift diseases, membranous nephropathy (MN), anti-NMDA. receptor neuropathy, neuromyefitis optica, idiopathic thrombocytopenic purpura, autoimmune hepatitis, type 1 diabetes mellites, rheumatoid arthritis, juvenile foeomatoid arthriti s, chronic inflammatory demy elinating polyneuropathy, polymyosi ti s / dermatomy ositi, stiff persons disease, anti-NMD receptor encephalitis, anti-synthetase autoimmune syndromes, anti-phospholipid antibody syndrome, Sjogren’s syndrome, cryoglobulinemia, focal segmental glomerulosclerosis, rapidly progressive glomerulopathy, autoimmune hemolytic anemia, amyloidctsis, scleroderma, idiopathic inflammatory myositis, and immune-mediated necrotizing myopathy. In some embodiments, a subject of the present disclosure is diagnosed to have systemic lupus erythematosus (SLE), extrarenal systemic lupus erythematosus (ERL), lupus nephritis (LN), or ANCA -associated vasculitis (AAV), or membranous nephropathy MN), In some embodiments, a subject of the present disclosure has moderate to severe disease activity. In some embodiments, a subject of the present disclosure has a relapsed or refractory' autoimmune condition. In some embodiments, a subject of the present disclosure has autoantibodies. In some embodiments, depth of B ce ll depletion in a subject of the present disclosure that has or is suspected of having a B cell mediated autoimmune disorder following administration of a drug product comprising hypoirnnmne allogenic CDI9-directed CAR T cells of the present disclosure can inform treatment and may predict efficacy of the drug product, B cells drive disease pathology in many autoimmune diseases, so B cell depletion following administration of a drug product comprising hypoimmune allogenic CD19-directed CAR T cells of the present disclosure can be efficacious in many autoimmune diseases and deliver durable, long-term remission in a subject of the present disclosure that has or is suspected of having a B cell mediated autoimmune disorder.(0230] In some embodiments, a subject of the present disclosure has adequate organ function. 1ft some embodiments, a subject of ths present disclosure has adequate hepaticfunction. In some embodiments., adequate hepatic function comprises: a total bilirubin level < 1 >5x upper limit of normal (ULN); and / or alanine aminotransferase ( ALT) or aspartate aminotransferase (AST) < 3x ULN. hi some embodiments, a subject of the present disclosure has disease-related muscle involvement causing elevated ALT or AST. In some embodiments, adequate hepatic function comprises: a total bilirubin level < 3x upper limit of normal (ULN) and / or alanine aminotransferase (ALT) or aspartate aminotransferase (AST) < Sx ULN; wherein the subject has fiverin olvement; and wherein the total bilibrubin level < 3x ULN' is not attributed to Gilbert’s syndrome. In some embodiments, a subject of the present disclosure has adequate bone marrow function. In some embodiments, adequate bone marrow function comprises a hemoglobin level > 8 g / da neutrophil count > I 000 / mnri and / or a platelet count > 50,000 / mm3. In some embodiments, a subject of the present disclosure has adequate pulmonary function. In some embodiments, adequate pulmonary function comprises a baseline oxygen saturation > 92% on room air; no clinically significant pleural effusion; and / or a forced expiratory volume in the first second / Forced Vital Capacity (FEV1 / FVC) >70% as measured by spirometry. In some embodiments, a subject of the present disclosure is eligible for lymphodcpleting chemotherapy. In some embodiments, a subject of the present disclosure is diagnosed to have SLE based on 2019 EIJLAR / ACR Classification Criteria. In some embodiments, a subject of the present disclosure is confirmed to have Class III (A or A / C) or IV (A or A / C) LN based on histological analysis using ISN / RPS 2018 criteria with active or acti ve / chronic lupus nephritis performed wi thin 6 months of screening. In some embodiments, a subject of the present disclosure is confirmed to have Class V LN. In some embodiments, a subject of the present disclosure is confirmed to have Class III or Class IV LN. In some embodiments, a subject of the present disclosure has at least one elevated autoantibody. In some embodiments, at least one elevated autoantibody comprises anti-dsDNA, anti-nuclear autoantibody (ANA) or anti-Sm (Smith) autoantibody. In some embodiments, an ANA comprises a titer of at least 1: 80. In some embodiments, at least one elevated autoantibody comprises anit-PLA2R or anti-THSD7A. In some embodiments, a subject of the present disclosure has a prothrombin time-international normalized ratio < 2.5 with no disease-related coagulopathy. In some embodiments, a subject of the present disclosure has an activated partial thromboplastin ti me < 2.5 x ULN. In some embodiments, a subject of the present disclosure has an estimated GFR of > 45 mL / mln / 1.73nr’ as determined by a CKD-EPI equation. In someembodiments, a subject of the present disclosure has proteinuria with a UPCR determined by either spot urine col lection or 24-hour urine collection of> I g / dL and 7 g / dL. In some embodiments, a subject of the present disclosure has a SLEDAI-2K score > 8. In some embodiments, a subject of the present disclosure is diagnosed to have granulomatous with polyangiitis (GPA) or microscopic poly angiitis (MP A) based on the Chapel Hill Consensus Conference criteria, in some embodiments, a subject of the present disclosure is diagnosed with a disease flare with a BVAS > 2 following the achievement of remission induced by a regimen consisting of anti~CD20 B cell depletion (e.g., rituximab or obinutuxumab) and glucocorticoids*, cyclophosphamide and glucocorticoids; or anti~CD20 B cell depletion, cyclophosphamide and glucocorticoids. In some embodiments, a disease flare is organ or life threatening. In some embodirncnls, a subject of the present disclosure has had at least one GPA or MPA flare, at least one prior induction regimen with rituximab, obinutuzumab or cyclophosphamide and a baseline B V AS of > 1. In some embodiments, a subject of the present disclosure has severe disease or a history of severe disease as defined by one or more major BV AS items. In some embodiments, a subject of the present disclosure is positive for either PR3:-ANCA or MPO-ANCA, in some embodiments, a subject of the present disclosure is refractory to rituximab or has relapsed after prior rituximab therapy with evidence of reduced eGF'R.| (12311 In some embodiments, a subject of the present disclosure lias or is suspected of having a disease, disorder, or condition that is relapsed and / or refractory. In some embodiments, a B cell malignancy is relapsed and / or refractor. In some embodiments, a B cell malignancy is relapsed and / or refractory NHL. In some embodiments, a B cell malignancy is relapsed and / or refractory CLL. In some embodiments, a B cell malignancy is relapsed and / or refractory IJBCL, In some embodiments, the B cell malignancy is relapsed and / or refractory after at least 2 regimens of therapy. In some embodiments, a B cell malignancy is relapsed and / or refractory after autologous stem cell transplant (ASCT), In some embodiments, a B cell malignancy is relapsed and / or refractory, wherein relapsed and / or refractory comprises one or more oft no response to first line therapy, progressive disease as best response to most recent therapy regimen, stable disease as best response to most recent therapy with duration no longer than 6 months from last dose of therapy, disease progression or relapse within 12 months of ASCT, no response to or relapse after last line of therapy post-ASCT, and / or no response to or relapse after treatment with a first CD 19 CAR T therapy. In some embodiments, a subject of the presentdisclosure has or is suspect ed of having relapsed and / or refractory CLL and has already been treated with at least 2 regimens of therapy. In some embodi ents, a subject of the present disclosure has or is suspected of having relapsed and / or refractory multiple myeloma and has already been treated with at least I regimen of therapy, hi some embodiments, a subject of the present disclosure has or is suspected of having relapsed and / or refractory LBCL and is transplant ineligible. In some embodiments, a subject of the present disclosure has already been treated with a bispecific antibody. In some embodiments, a bispeeffic antibody is selected from the group consisting of: mosunetazumab (CD20 x CDS mAb), glofrtamab (CD2()(2) x CI)3), odronextamab (CD20 x CD3 mAb), and epcoritamab (CD20 x CD3 mAb), Tn some embodiments, a B cell mediated disorder is relapsed and / or refractory, In some embodiments, a B cel] mediated disorder is relapsed and / or refractory following a first line therapy comprising mycophenolate mofetil (MMF), cyclophosphamide, or a combination thereof, hi some embodiments, a B cell mediated disorder is relapsed and / or refractory following a second line therapy comprising rituximab, belimumab, voclosporin, obinutuzumab, or a combination thereof In some embodiments, a B cell mediated disorder is relapsed and / or refractory following at least two disease-modifying therapies selected from the group consisting of: MMF, axafhioprine, cyclophosphamide, methotrexate, ri tuximab, belimumab, intravenous immunoglobulin (Wig), and abaiacept. In some embodiments, a subject does not achieve a complete renal response to the second line therapy within 6 months of initiation of the second line therapy. In some embodiments, a subject has disease activity despite maintenance on maximally tolerated doses of drugs that block the reniu-angioteasiu system ( AS).

[0232] In some embodiments, a subject. of the present disclosure is 18-85 years of age. In some embodiments, a subject of the present disclosure is 18-80 years of age. In some embodiments, a subject of the present: disclosure is 18-75 years of age.4. Exclusion criteria0233] In some embodiments, a subject of the present disclosure cannot orshould not be treated with a pharmaceutical composition of the present disclosure if they meet certain criteria (exclusion criteria). In some embodiments, a subject of the present disclosure has not received a prior CD19-direc ed therapy. In some embodiments, a prior CD19-directed therapy is orcomprises a CD19-riirccted CAR T cell treatment. In some embodiments, a prior C 1 Redirected therapy is or comprises a CD19-directed antibody treatment. In some embodiments, a prior CD19-directed therapy is or comprises a CD19-directed CAR NK cell treatment. In some embodiment, a subject of the present disclosure has not received a prior CD47~directed therapy. A CD47-directed therapy can include a binder of CD47 ( e.g., an anti-CD47 antibody) that specifically binds to the CD47 and interferes with the interaction of CD47 and SIRPa. Examples of binders that specifi cally bind CD47 include ruagrolimab, Trillium (TTI)-621, ALX148, AO-176, lemzopariimab, 1BI1 8, SRF231, Cf?-9(K)()2. and AK 117. A CD47-directed therapy can also include a protein and / or molecule that interferes with the interaction of CD47 and SIRPa by binding SIRPa, such as ADU-1805 and BI 765063. In some embodiments, a subject has not received a CD47~direcfed therapy wi thin 1 year of being administered the composition comprising the population of engineered hypoimniunogenic T cells. In some embodiments, a subject has not received a CD47-directed therapy within about 1 year, about 11 months, about 10 months, about 9 months, about 8 months, about 7 months, about 6 months, about 5 months, about 4 months, about 3 months, about 2 months, or about 1 month of being: administered the composition comprising thepopulation of engineered hypoi munogcnic T ceils. In some embodiments, a subject of the present disclosure having or suspected of having anti~CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until anti-CD47 antibody levels are reduced. In some embodiments, a subject of the present disclosure having or suspected of having anti-CD47 antibodies cannot or should not be treated with a pharmaceu tical composi tion of the present disclosure until a period of time known to be associated with reduced (i.e., cleared) anti-CD47 antibody levels has passed. For example, the period of time may be 1 year, abou t 11 months, about 10 months, about 9 months, about 8 months, about 7 months, about 6 months, about 5 months:, about 4 months, about 3 months, about 2 months, or about I month, As another example, the period of time may be 5, 6, 7, 8, 9, or 10 half-lives of the CD47-directed therapy. In some embodiments, a subject of the present disclosure having or suspected of having aiffi~CD47 antibodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until testing of anti-CD47 antibody levels indicates reduced levels. In some embodiments, a subject of the present disclosure having or suspected of having anti~CD47 anti bodies cannot or should not be treated with a pharmaceutical composition of the present disclosure until anti-CD47 antibody levels are atundetectable levels. Any method known in the art tor detecting antibodies in a sample from a patient can be used. For example, antibodies can be detected in a sample (e.g., serum or whole blood) using an enzyme-linked immunospot (Ell Spot) assay, ELISA, immunofluorescence, flow cytometry, Western blot, or any other methodology of determining the antibody titer of a specific antibody. In some embodiments, a subject of the present disclosure subject does not have a history of primary central nervous system (CNS) lymphoma or CNS metastases. In some embodiments, a subject of the present disclosure does not have a history of Richter’strans formation of chronic leukemic lymphoma, small lymphocytic ly mphoma, or lymphoplasmacytic lymphoma. In some embodiments, a subject of the present disclosure does not have a fungal, bacterial, viral, or other infection that is uncontrolled or requiring IV antimicrobials for management. In some embodiments, a subject of the present disclosure has not been hospitalized for treatment of an infection within 28 days of baseline of > 2 times within I year of administering the composition to the subject. In some embodiments, a subjec t of the present disclosure does not have acute symptoms of COVID-19 infection. In some embodiments, a subject of the present disclosure does not have a known history of infecti on with HIV and / or hepatitis B (HBsAg positive) and / or hepatitis C virus (anti-HC V positive). In some embodiments* a subject of the present disclosure does not have an active autoimmune disease and / or any other diseases requiring immunosuppressi e therapy or coiti costeroid therapy. In some embodiments, a subject of the present disclosure does not have primary immunodeflciency or history of concomitant genetic syndrome associated with bone marrow failure (e.g., Fanconi anemia, Kostmatm syndrome, or Shwachman- Diamond ). In some embodiments, a subject of the present disclosure has not been treated with a systemic cancer therapy within 14 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with any investigational product within 28 days of administration of the composition, fa some embodiments, a subject of the presen t disclosure has not been treated with radiotherapy within 14 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with major surgery within 28 days of administration of the composition. In some embodiments, a subject of the present disclosure has not been treated with autologous hematopoietic stem cell transplant (HSCT) within 6 weeks of administration of the composition. In some embodiments, a subject of the present disclosure has not had myocardial infarction, cardiac angioplasty or stenting, cardiac arrhythmia requiringmedication, unstable angina, New York Heart Association Class 11 or greater congestive heart failure, cardiac atrial or ventricular lymphoma involvement or clinically significant cardiac disease within one year of administration of the composition. In some embodiments, a subject of the present disclosure has not had an ejection fraction < 50%. arrythmias and any structural heart disease (including clinically significant pericardial effusion), as assessed by echocardiogram or multiple gated acquisition scan performed within 1 month of administration of the composition. In some embodiments, a subject of the present disclosure does not have a history of non-Ime associated, clinically significant deep-vein thrombosis (i.e., proximal deep vein thrombosis [DVT]) or pulmonary embolism requiring therapeutic anticoagulation within 6 months of administration of the composition. In some embodiments, a subject of the present disclosure does not have a history or presence of CNS disorder (e.g,5:seizure disorder, cerebrovascular ischemia / hemorrhage, dementia, cerebellar disease, or any autoimmune disease with CNS invol vement) within 12 months of administration of the composition. In some embodiments, a subject of the present disclosure does not have clinically significant ascites. In some embodiments, a subject of the present disclosure does not have a history of organ transp lantation and / or is not awaiting organ transplantation. In some embodiments, a subject of the present disclosure does not have an indwelling line or drain, in some embodiments, a subject of the present disclosure has not received a live vaccine within 6 weeks of administration of the composition. In some embodiments, a subject of the presen t disclosure does not have any unresolved toxicity greater than Grade 1 from a previous anticancer therapy. In some embodiments, a subject of the present disclosure does not have a history of additional malignancy other than nonmelanoma skin cancer or carcinoma in situ. In some embodiments, a subject of the present disclosure does not have a severe immediate hypersensiti vity to any of the components of a pharmaceutical composition of the present disclosure. In some embodiments, a subject of the present disclosure does not have an admission or evidence of il licit drug use, drug abuse, or alcohol abuse. In some embodiments, a subject of the present disclosure does not exhibit clinical evidence of significant unstable or uncontrolled acute or chronic disease not due to the B cell mediated disorder. In some embodiments, a significant unstable or uncontrolled acute or chronic disease comprises cardiovascular, pulmonary, hematologic, gastrointestinal, hepatic, renal, neurological, malignancy, or infectious disease. In some embodiments, a subject of the present disclosure does not have a history of an anaphylactic reaction to parenteraladministration of contrast agents, human or murine proteins, or monoclonal antibodies. In some embodiments, a subject of the present disclosure does not have a history of cancer, or the subject has been disease- fee fem a basal carcinoma of the skin, cervix, bladder, prostate or b reast for at least five yeans prior to administration of the composition. In some embodiments, a cancer comprises a hematologic malignancy, a solid tumor, or a carcinoma in situ. In some embodiments, a subject of the present disclosure is administered physiologic replacement glucocorticoids, topical glucocorticoids, or inhaled glucocorticoids. In some embodiments, a subject of the present disclosure i s not administered glucocorti coids above physiologic replacement. In some embodiments, physiologic replacement glucocorticoids comprises 5 mg prednisone dai ly or equivalent In some embodiments, a subject of the present disclosure has not received treatment with Leflunomide within 28 prior to administration of the composition. In some enibodiments, a subject of the present disclosure does not have an immunoglobulin A (IgA) deficiency (IgA level < lOmg / dL). In some embodiments, a subject of the present disclosure does not have a grade >3 laboratory abnormality, except those associated with the B cell mediated disorder. In some embodiments, a subject of the present disclosure does not have a B cell mediated disorder related flare that needs immediate treatment. In some embodiments, a subject of the present disclosure does not have a CNS lupus manifestation, a history of a CNS disorder, or a presence of a CNS disorder. In some embodiments, a CNS disorder is or comprises primary seizure disorder, cerebrovascular lschemia / hemorrhage, dementia, or cerebellar disease. In some embodiments, a subj ec t of the present disclosure has not been diagnosed with anti-phospholipid antibody syndrome that is actively treated. In some embodiments, a subject of the present disclosure has not been diagnosed with EGPA as defined by the Chapel Hill Consensus Conference criteria. In some embodiments, a subject of the present disclosure does not have an alveolar hemorrhage within 4 weeks prior to administration of the composition, In some embodi ments, a subject of the present disclosure has not been diagnosed with a cerebrovascular accident caused by vasculitis or pachymeningitis.[0:234| In some embodiments, a subject of the present disclosure having or suspected of having a B cell mediated autoimmune disorder cannot or should not be treated with a pharmaceutical composition of the present disclosure if they have an active infection. In some embodiments, a subject of the present disclosure having or suspected of having a B cell mediatedautoimmune disorder cannot or should not be treated with a pharmaceutical composition of the present disclosure if they are unable to receive a lyniphodepletion regimen.|(1235| In some embodiments, a subj ect of the present disclosure should not take corticosteroid therapy at a pharmacologic dose (>' 5 mg / day of predni sone or equi valent doses of other corticosteroids) and other immunosuppressive drugs within 5 days of administration of a lymphodepletion regimen of the present d isclosure and / or administration of hypoimrnune allogenic CD19~directed CAR T cells of the present disclosure.[02361 In some embodiments, a subject of the present disclosure shoul d not take corticosteroids and / or other immunosuppressive drugs in the 3 months after administration of hypoimrnune allogenic CDI 9-directed C AR 'I' cells of the present disclosure. In some embodiments, a subject of the present disclosure should not take nonsteroidal anti-inflammatory agents in the 3 months after administration of hypoimrnune allogenic CD19-directedC. AR T cells of the present disclosure. In some embodiments, a subject of the present disclosure should not take therapeutic doses of systemic anticoagulants, such as unfractionated heparin and low-molecular weight heparin after administration of hypoimrnune allogenic CD 19-directed CAR T cells ot the present disclosure. In some embodiments, a subject of the present disclosure does not receive treatments for underlying disease, such as surgery, chemotherapy, immunotherapy, targeted agents, radiation, and high-dose corticosteroids (other than those allowed herein) and other investigational agents after administration of hypoimrnune allogenic CDI 9-directed CAR T cells of the present disclosure.5. 1.ymphodepletiun regimen>0237] Non-limiting examples of an immunosuppressive and / or immunomodulatory agent (such as, but not. limited to a lymphodepletion agent) include cyclosporine, axathioprine, mycophenolic acid, mycophenolate moietil, corticosteroids such as prednisone, methotrexate, gold salts, sulfasalazine, antimalarials, brequinar, leflunomide, mizoribine, 15-deoxyspergualme, 6-mercaptopurine, cyclophosphamide, rapamycin, tacrolimus (FK-506), 0KT3, anti-thymocyte globulin, thymopentin, thymosin-a and similar agents. In some embodiments, the immunosuppressive and / or immunomodulatory agent Is selected from a group of immunosuppressive antibodies consisting of antibodies binding to p75 of the IL-2 receptor.antibodies binding to, for instance, MHC, CD2, CD3, CD4, CD7, CD28, B7. CD40, CD45, IFN-gamma, TNF-alpha, IL-4, IL-5, I -6R, IL-6, IGF, IGFR1, IL-7, IL -8, IL-10, CD1 la, or CD58, and antibodies binding to any of their ligands. In some embodiments, such an immunosuppressive and / or immunomodulatory agent may be selected from soluble 1L-15R, IL-10, B7 molecules (e.g„, B7-1 / B7-2, variants thereof, and fragments thereof), ICOS, and 0X40, an inhibitor of a negati ve T cell regulator (such as an antibody against CTLA-4) and simi lar agents.

[0238] In some embodiments, where an immunosuppressive and / or immunomodulatory agent is administered to the patient before or after the administration of the cells, the administation is at a lower dosage than would be required for cells with MHC I and / or MHC II expression, TCR expression and without exogenous expression of CD47. In some embodiments, where an immunosuppressive and / or irnmunomodulatory agent is administered to the patient before or after the first administration of the cells, the administration is at a lower dosage than would be required for cells with MHC I and MHC II expression, TCR expression and without exogenous expression of CD47.

[0239] In some embodiments, a subject of the present disclosure Is administered a lymphodepleting chemotherapy regimen, wherein administering the lymphodepleting chemotherapy regimen occurs prior to administering a phanuaceutical composition of the present disclosure. In some embodiments, a lymphodepleting therapy is or comprises cyclophosphamide and fludarabme. In some embodiments, cyclophosphamide is administered at a dose of 500 mgmr. In some embodiments, cyclophosphamide is administered at a dose of 500 ing / nt2daily for 3 days. Tn some embodiments, cyclophosphamide is administered at a dose of 1000 mg / m2In some embodiments, cyclophosphamide is administered at a dose of 1000 mg / rrr one, 3 days prior to administration of a composition of the present disclosure. In some embodiments, fludarabine is administered at a dose of 30 mg / m2. In some embodiments, fludarabine is administered at a dose of 25 mg / m2. In some embodiments, a subject of the present disclosure has a moderate impairment of renal function, and fludarabine is administered at a dose of about 20 mg / m2with monitoring. In some embodiments, fludarabine is administered at a dose of 30 mg / m2daily for 3 days. In some embodiments, cyclophosphamide and fludarabine are administered concurrently. In some embodiments, a lymphodepleting chemotherapy regimen Isadministered intravenously, in some embodiments, a pharmaceutical composition of the present disclosure is administered to a subject 2-7 days after completion of the lymphodepleting chemotherapy regimen.

[0240] In some embodiments, a subject of the present disclosure has not received a live vaccine within 28 days of administration of the lymphodepleting chemotherapy,

[0241] hi some embodiments, a subject of the present disclosure is weaned to discontinuation of a severe autoimmune therapy over the course of no more than 21 days prior to administration of a composition of the present disclosure to the subjec t In some embodiments, a subject is weaned to 0 mg glucocorticoids. In some embodiments, a subject is weaned to a physiologic equivalent range of < 5 mg / day of prednisone.

[0242] In some embodiments, a subject is not administered a lymphodepleting chemotherapy regimen prior to administrati on of a pharmaceutical composition of th e present disclosure. In some embodiments, the following lymphodepletion protocol is followed for a subject of the present disclosure. On Days -5 through -3, before the infusion of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure, subjects receive a non-myeloablati ve lymphodepletion (LD) regimen consisting of cyclophosphamide and fludarabine to induce lymphocyte depletion and create an optimal environment for the expansion of hypoimmune allogenic CDIV-dirccted CAR T cells of the present disclosure in vivo.Hypoimmune allogenic CD19-directed CAR T cells of the present disclosure should be administered 3 days (range 2-7 days) after completion of chemotherapy.

[0243] In some embodiments, subjects of the present disclosure receive the following 3-day chemotherapy regimen per the order identified as follows: 1) IV hydration with 1 1 of 0.9% sodium chloride (NaCl) given before cyclophosphamide on the day of infusion; 2) Cyclophosphamide 500 m IV over approximately 60 minutes on Days -5, -4, and -3; 3) Fludarabine 30 mg / m2 IV over approximately 30 minutes on Days -5, -4, and -3 and subjects with moderate impairment of renal function ( creatinine clearance 50-70 rnL / min / L73 m2) should have their fludarabine dose reduced by 2:0% and be monitored closely; 4) an additional 1 L of 0.9% NaCl at the completion of the fludarabine infusion; and 5) add Mesna (sodium 2-mercaptoethanesulfonate; a detoxifying agent used to inhibit the hemorrhagic cystitis induced bythe chemotherapy) per institutional guidelines. Subjects at risk for tumor lysis syndrome (TLS) should receive TLS prophylaxis prior to LD chemotherapy according to institutional guidelines.024 Administration of hypo immune allogenic CD19foirected CAR T cells of the present disclosure: can increase the risk of high-grade and fatal textcities in subjects with ongoing infection and / or inflammation. In some embodiments, if a subject of the present disclosure has: a temperature > 38 °C within 72 hours before LD chemotherapy; CRP > 100 mg / L any time between etirollment and the start of LD chemotherapy; and / Or a white blood cell (WBC) count or WBC differential concerning for infectious process between enrollment and the start of LD chemotherapy (e.g., WBC > 20,000 cells / pL, rapidly increasing WBC, or WBC differential with a high percentage of segments / bands), the cause of the Infection should be identified and treated before administration of an LD regimen of the present disclosure.(1245] In some embodiments, if any screening assessments or procedures are repeated between confirmation of eligibility and the start of LD chemotherapy and the results are outside of the eligibility criteria provided herein, any suspected / identified mfectious / infiammatory process condition must resolve before a subject of the present disclosure is administered an LD regimen of the present disclosure, In some embodiments, a complete medical history and physical examination of a subject of the present disclosure including the head, eyes, ears, nose, and throat (HEENT) and cardiac, vascular, respiratory, gastrointestinal, integumentary, and neurological systems does not reveal evidence of infection / infiaimnation before administration of an LD regimen of the present disclosure. In some embodiments, a subject of the present disclosure has not received systemic antimicrobials for the treatment of a known or suspected infection within the 48 hours before an LD regimen of the present disclosure (prophylactic use of antimicrobials is allowed). In some embodiments, a treatment course of any antimicrobials given for a known or suspected antecedent infection should be completed before a subject of the present disclosure is administered an LD regimen of the present disclosure, In some embodiments, if a subject of the present disclosure is confirmed to have an infectious process for which antimicrobials are not available (e.g., viral pneumonia), the infection must be clinically resolved before a subject of the present, disclosure is adminis tered an LD regimen of the present disclosure. In some embodiments, the most recently collected blood, urine, or other body fluid cultures must show no growth for at least 48 hours, and any other infectious workup performed( e.g., bacterial, PCR, or stool or imaging studies) is negative before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if there is clinical suspicion of an infection for which cultures are unlikely to be positive within 48 hours (e.g., fungal infection), then adequate time is allowed to allow the cultures to become positive before a subject of the present disclosure is administered an LD regimen of the present disclosure.6, Assessment of Subjects

[0246] In some embodiments, subjects of the present disclosure undergo safety assessments that include the collection of ad verse events (AEs) and serious adverse events (SAEs), clinical laboratory tests, imaging, physical and neurologic examinations, vital signs, cardiac function (e.g., electrocardiogram [ECG]), and ECOG performance status, as described herein. In some embodiments, subjects of the presen t disclosure asked about any AEs that might have occurred since a previous evaluation. In some embodiments, serious adverse events (SAEs), treatment-related adverse events (TRAEs), treatment-emergent adverse events (TEAEs), targeted adverse events (TAEs), adverse events of special interest ( AES Is), and / or dose-limiting toxicities (DLTs) are determined for a subject as part of an evaluation.

[0247] In some embodiments, subjects of the present disclosure undergo safety assessments that include clinical laboratory tests. Exemplary clinical laboratory tests include those found in Table 16 and Table 17 of the present disclosure. In some embodiments, subjects of the present disclosure undergo safety assessments that include physical examination. In some embodiments, a physical examination comprises measurement of, assessment of, or performance of one or more of: height, weight, vital signs (e.g., systolic / diastolic blood pressure, heart rate, respiratory rate, oxygen saturation, temperature), neurological status (e.g.,:immune effector cell-associated encephalopathy (ICE) score), Eastern Cooperative Oncology Group (ECOG) score, cardiac function, echocardiogram (ECHO), electrocardiogram (ECG), brain Magnetic Resonance Imaging (MRI), a lumbar puncture, and information about concomitant medications.

[0248] In some embodiments, subjects of the presen t disclosure are assessed for the presence of replication competent lentivirus (RCL) in peripheral blood mononuclear cells (PBMCs). In seme embodiments, subjects of the present disclosure are assessed for abnormal Tcell proliferation. In some embodiments, subjects of the present disclosure are assessed for the presence of anti"CD19-directed CAR antibodies in the subjects’ serum. In some embodiments, serum from subjects of the present disclosure is assessed for cytokine profiling.

[6249] In some embodiments, subjects of the present disclosure are assessed for disease progression and / or disease response. In some embodiments, subjects of the present disclosure with NHL will be assessed based on Lugano classification criteria. In some embodiments, subjects of the present disclosure with CLL will be assessed based on the International Workshop on Chronic Lymphocytic Leukemia (iwCLL) criteria. In some embodiments, disease assessment in subjects of the present disclosure comprises imaging. In some embodiments, imaging comprises positron emission tomography (PET)-compuied tomography (CT) and / or CT scans. In some embodiments, subjects of the present disclosure undergo PET-CT and / or CT scans within 24 months of administration of hypoimmune allogenic CD19-dircct©d CAR T cells of the present disclosure. In some embodiments, disease assessment in subjects of the present disclosure comprises bone marrow aspirate and / or biopsy.

[0250] In some embodiments, PBMC samples from subjects of the present di sclosure are assessed fbr cellular kinetics of hypoimmune allogenic C 19-directed CAR T cells of the present disclosure. In some embodiments, cellular kinetics are evaluated by droplet digital polymerase chain reaction (ddPCR) and reported as a level of hypoimmune allogenic CD 19-directed CAR T cells per pg of genomic DNA (gDNA). In some embodiments, the derived parameters ofCmax, Tjnax, fo% Gast, Tjast, and AUC are determined for hypoimmune allogenic CD1 -directed CAR T cells of the present disclosure. In some embodiments, the number of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure in a subject’s: blood will be assessed via flow cytometry. In some embodiments, cellular kinetics as described herein are used to determine dose-exposure relationships and / or correlation of persistence and expansion of hypoimmune allogenic CD19- trected CAR T cells of the present disclosure with safety and efficacy parameters.G251 In some embodiments, blood / senim samples from subjects of the present disclosure undergo pharmacodynamic assessments. In some embodiments, pharmacodynamic assessments comprise monitoring of levels of analytes in blood / semm over time. In some embodiments, analytes include one or more of: homeostatic / proliferative cytokines (e.g,,interleukin (1LJ-2, IL-7, IL-15, and granulocytecolofty-stimtilating factor (G-CSF )), intlammator^Fimmune-modulating cytokines (e.g:., interfenon-gamma (IFN-y), IFN-a. IL-ip, IL-IRa, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL43, 1L-17A, granulocyte macrophage colonystimulating factor (GM-CSF), tumor necrosis factor-alpha (TNF-a), macrophage inflammatory protein- 1 A lpha (M. IP-la), and IP-ip), correlates of acute phase response (e.g., C~reactive protein (CRP) and ferritin), chenwkines (e.g., IL-8, C-X-C motif chemokine ligand- 10 (CXL’L-10), and monocyte chemotactic protein-1 (MCP-1)), and associated response related with CD 1 -directed CAR activity (e.g., circulating CD194- B cell and lactate dehydrogenase (LDH)).|0252| In some embodiments, subjects of the present disclosure are assessed for minimal residual disease (MRD). In some embodiments, MRD is determined using a next-generation sequencing (NOS) MRD assay to measure ctDN A levels before and after administration of hypoimmune allogenic CD1 ^-directed CAR T ceils of the present disclosure. In some embodiments, subjects of the present disclosure are assessed for immune responses against the administered hypoimmtrne allogenic CD19-directed CAR I' cells of the present disclosure. In some embodiments, immune responses hypoimmune allogenic CDlfodirected CAR T cells of the present disclosure are or comprise aati-hypoimmune allogenic CD19-directed CAR T cell antibodies, antibody-dependent cellular cytotoxicity (ADCQ, ctmtplemenl-depeftdeftt cytotoxicity (CDC), mtnrune activation against hypoimmune allogenic CDlO-directe CAR T cells of the present disclosure, K cell-mediated killing of hypoimmurie allogenic CDI9-directed C AR T cells of the present disclosure.(0253] In some embodiments, methods of the presen t disclosure comprise measuring MRD in a subject to monitor treatment response or relapse of disease. In some embodiments, MRD refers to a small, number of disease cells that remain io the subject during or after treatment when the subject is in remission. In remission, the subject may no longer display obvious symptoms or signs of disease, but MRD cells may remain and cause relapse of the disease.Testing for MRD may be useful for determining whether treatment has eradicated disease cells or whether traces remain, comparing efficacy of different treatments, monitoring patient remission status as well as detecting recurrence of the disease, and adjusting treatments accordingly. The number of MRD cells in a sample from the subject may be as low as one disease cell in a million normal cells.

[0254] In some embodiments, MRD can be measured using various methods as described in, but not limited to, US2022025 54A1, US20170335391A1, US20210238694A1, and US20200370129A1.

[0255] In some embodiments, measuring MRD comprises collecting two m more biological samples from a subject at different time points (e.g. prim to, concurrent with, or after administration of hypoimmune allogenic CD 19-directed CAR I' ceils). In some embodiments, a multiplex amplification reaction is performed on nucleic acids isolated from the biological samples to generate a set of amplicons, wherein each of the set of amplicons comprises recombined V(D)J gene segments at a gene locus of interest and / or wherein each of the set of amplicons comprises gene segments at one or more cancer specific gene loci of interest, and sequencing the set of amplicons, wherein sequences of the recombined V(D)J gene segments and / or cancer specific gene segments are indicative of presence of an immune cell and / or cancer in the biological sample.

[0256] In some embodiments, measurin g MRD comprises assigning a pair of first and second polypeptides that form a. TCR (TCRA or TCRB) or immunoglobulin (e.g., Ig heavy chain, or K or X light chain) heterodimeric a single biological source sample among the two or more biological samples. In some embodiments, the first rearranged nucleic acid sequences encoding the first polypeptides of the TC R or Ig heterodimers present in the source biological sample is determined for each of the plurality of biological samples and the first rearranged nucleic acid sequences are assigned to the source biological sample. The plurality of source biological samples are then pooled to form a combined population of cell and a plurality of cognate pairs of first and second rearranged nucleic acid sequences encoding first and second polypeptides of the TCR or Ig heterodimers is determined from the combined population of cells,-The first rearranged nucleic acid sequences determined in each of the source samples are compared to the first rearranged nucleic acid sequences determined from the plurality of cognate pairs of rearranged nucleic acid sequences to assign each first rearranged nucleic acid sequence present in the combined population to a single biological source sample. For each first rearranged nucleic acid sequence assigned to a single biological source sample, the cognate second rearranged nuclei c acid sequence of the cognate pair is assigned to the same single biological source sample.|0257j In another embodiment MRD is measured by determining a first rearranged nucleic acid sequ ence encoding the first polypeptide of the TCR or Ig heterodimer present in the biological source sample. In some embodiments, for each biological source sample, rearranged nucleic acid molecules extracted from the biological source sample are amplified in a single multiplex polymerase chain reaction (PCR) using a plurality of V-segment primersand a plurality of J-segment primers to produce a plurality of rearranged nucleic acid amplicons. The plurali ty of rearranged nucleic acid amplicons are sequenced to determine sequences of the first rearranged nucleic acid sequences in each biological source sample,|0258] In some embodime nts, MRD is measured by using a “bait set” of probes for hybridization capture. The bait set comprises a plurality of different oligonucleotide-con taming probes, where each of the oligonucleotide-containing probes comprises a sequence of at least 30 bases in length that is complementary to either; (1) a sequence of a genomic region; or (2) a sequence that varies from the sequence of (1) only by one or more transitions,(02591 In some embodiments, MRD is measured via immune repertoire sequencing. TCRs and immunoglobulins serve as a unique “barcode” for these lymphocytes. By sequencing the immune receptors, the immune response in patient samples and clonal expansion of specific lymphocyte subpopulations can be tracked and monitored,8268] In some embodiments, MRD is measured by amplifying and sequencing cell-fee nucleic acids that are released into bodily fluid from cancer cells, e.g,, circulating tumor DNA c DN A )< In some embodiments, ctDNA can be non-encapsulated tumor-derived fragmented DNA and / or may have one or more epigenetic modifications, for example, the ctDNA may be acetylated, 5~met.hylated, ubiquitylaied, phosphorylated, snmoylated, ribosylated, and / or ci trail mated,026I) In some embodiments, subjects of the present disclosure undergo tumor biopsies or liquid biopsies and the samples are evaluated for CD 19 expression. In some embodiments, subjects of foe present disetosure undergo tumor biopsies or liquid biopsies before administration of hypoimmune allogenic CD19~directed CAR T cells of the present disclosure and biopsies are assessed to determine mechanisms of cancer resistance. In some embodiments, subjects of the present disclosure undergo tumor biopsies or liquid biopsies after administration of hypoimmuneallogenic CD19-directed CAR T cells of the present disclosure and tumor cells are assessed to determine mechanisms of tumor resistance.|0262| In some embodiments, PBMCs collected from subjects of the present disclosure before and after administration of hypoirnmune allogenic CD19-directed CAR T cells of the present disclosure are assessed for immunophenotypic changes induced by the hypoimmune allogenic CD19-directed CAR T cells of the present disclosure, including expression level of CD47, broad immune cell phenotyping (e.g., lymphoid and myeloid cell phenotypes, e.g., T, B, NK and monocytes), and immune cell subsets (e.g., differentiation, activation, and exhaustion), |@263| In some embodiments, subjects of the present disclosure are assessed for their survival after administration of hypoimmune allogenic CD1 ^-directed CAR T cells of the present disclosure.|0264| In some embodiments, subjects of the present disclosure are assessed for certain safety and efficacy criteria after administration of hypoimmune allogenic CDI9-directed CAR T cells of the present disclosure. In some embodiments the certain safety criteria comprise CRS Grade 3 / 4 <10%, IOANS Grade 3 / 4 <15%, infections Grade 3 / 4< 15%, and other Grade 3 / 4 adverse events < 5%. In some embodiments the certain safety criteria comprise hypoimmune allogenic CD19~directed CAR T cells of the present disclosure being no worse than autologous CAR T cells. In some embodiments, certain efficacy criteria comprise an event-free survival (EFS) of 10 months, complete response rate (CRR) of 65%, and a progression free survival (PFS) of 15 months for subjects with large B cell lymphoma (LBCL) administered hypoimxnune allogenic CD19-directed CAR T cells of the present disclosure as a second- line treatment In some embodiments, a complete response (CR) is defined as no detectable evidence of tumor. In some embodimertte, certain efficacy criteria comprise an overall response rate 1O. RR) of 70%, CRR of 50%, durable CRR of > 40%, and median duration of response (mDOR) > 12 months for subjects with LBCL administered hypoimmune allogenic CD19-directed CAR T cells of the present disclosure as a third-line treatment. la some embodiments, certain efficacy criteria comprise an ORR of 70%, CRR of 50%, durable CRR of > 40%, and mDOR > 12 months for subjects with transplant ineligible LBCL administered hypoimmune allogenic CD19-directed CAR T cells of the present disclosure as a second-line treatment In some embodiments; certain efficacy criteria comprise an QRR of 80%, CRR of 40%, durable CRR of 30-40%, mDOR > 12months, and 70% MRD negativity for subjects with CLL administered hypoimmune allogenic CD19-directed CAR. T cells of the present disclosure as a third-line treatment. In some embodiments, certain efficacy criteria comprise >50% durable CRR and 60% best overall CRR in LBCL patients. In some embodiments, certain efficacy criteria comprise >30% best overall CRR and >3 months durabili ty in CLL patients. In some embodiments, certain efficacy criteria comprise complete response with incomplete marrow reco very (CRi) and / or nodal partial response (nPR) in C L patients. In some embodiments, CR.i criteria comprise CR per iwCLL criteria along with any of the follow mg: A C < 1.5 * Iri^ZL or requires exogenous growth factors (e.g., G-CSF) < fourteen days (for at least seven days with “standard” growth factors) before the relevant blood count assessment to maintain an A. NC > 1.5 K^ / L; platelet count < 100 x 109 / L or requires exogenous growth factors or platelet transfosfons <fourteen days before the relevant blood count assessment to maintain a platelet count > 100 x IO9 / L; and / or hemoglobin < 110 g / L (11.0 g- dL) or requires exogenous growth factors or red blood cell transfusions < fourteen days before the relevant blood count assessment to maintain a hemoglobin > 110 g / L ( 11.0 g / dL). In some embodiments, nPR criteria comprise all criteria for CR / CRi are met except for bone marrow that shows detectable lymphoid nodules. In some embrrdiments, hypoimmune^ allogenic CDffi-dkected CART cells of the present disclosure persist for at least 3 months and demonstrate HIP-mediated immune evasion in subjects of the present disclosure.7, Procedures for SubjectsIn some embodiments, subjects of the present disclosure undergo screening before administration of a lymphodepletion regiment as described herein and / or before administration of hypoimmune allogenic CD19~dlrected CAR T cells of the present disclosure, hi some embodiments, screening comprises one or more of: informed consent, demographic data, medical history, previous cancer treatment history, concomitant medications documentation, clinical examination including height and weight, vital signs, neurologic examination (ICE Score), ECOG performance status, ECHO, ECG, brain MRI, lumbar puncture (if clinically indicated), disease assessment (including imaging and bone marrow biopsy / aspirate where applicable), •tumor biopsy (if archival tissue not available), SAE reporting, local laboratory assessments (e.g.5cbcmistre panel (including creatinine clearance, as estimated by MDRD equation ) (scrum ), CBC with differential (blood), beta-human chorionic gonadotropin (p-hC’G) pregnancy test (serum or urine) for ail females of childbearing potential, serologic tests (serum) (ie., HIV, hepatitis B virus, and HCV), coagulation (plasma), urinalysis, LDH, GRP, Ferritin, beta-2 mforoglobulin, direct antiglobulin test, haptoglobin, COVID-19 test, serology (HIV, HBV, HCV), serum immunoglobulin (Ig) levels, immunohistochemistry analysis (biopsy), ctDN A, or MUD.|(1266| In some embodiments, the following lymphodepletion protocol is followed for a subject of the present disclosure. On Days -5 through -3, before the infusion of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure, subjects receive a non-myeloablative lymphodepletion (LD) regimen consisting of cyclophosphamide and fludarabine to induce lymphocyte depletion and create an optimal environment for the expansion of hypoimmune allogenic GDI 9~directed CAM T cells of the present disclosure in vivo.Hypo immune allogenic CD19-directed CAR T cells of the present disclosure should be administered 3 days (range 2-7 days) after completion of LD chemotherapy,

[0267] In some embodiments, subjects of the present disclosure receive the following 3» day chemotherapy regimen per the order identified as follows: 1) IV hydration with I L of 0.9% sodium chloride (NaCl) gi ven before cyclophosphamide on the day of infusion; 2) Cyclophosphamide 500 mg / nr IV over approximately 60 minutes on Days -5, *4, and ~3; 3) Fludarabme 30 mg / mzIV over approximately 30 minutes on Days -5, -4, and -3 and subjects with moderate impairment of renal function (creatinine clearance 50-70 mL / min / I,73 m2) should have their fludarabme dose reduced by 20% and be monitored closely; 4) an additional 1 L of 0,9% NaCl at the completion of the fludarabine infusion; and 5) add. Mesna (sodium 2-mercaptoethanesulfonate; a detoxifying agent used to inhibi t the hemorrhagic cystitis induced by the chemotherapy) per institutional guidelines. Subjects at risk for tumor lysis syndrome (TLS) should receive TLS prophylaxis prior to LD chemotherapy according to institutional guidelines.

[0268] Administration of hypoimmune allogenic GDI 9-dfrected CAR T cells of the present disclosure can increase the risk of high-grade and fatal toxicities in subjects with ongoing infection and / or inflammation. In some embodiments, if a subject of the present disclosure has: a temperature > 38i!C within 72 hours before LD chemotherapy; GRP > 100 mg / L any time between enrollment and the start of LD chemotherapy; and / or a white blood cell (WBC) count orWBC differential concernin g for infectious process between enrollment and the start of ID chemotherapy (e.g,, WBC > 20,000 cells / gL, rapidly increasing WBC, or WBC differential with a high percentage of segmeats / bands), the cause of the infection should be identified and treated before administration of an LD regimen of the present disclosure.|0269| In some embodiments, if any screening assessments or procedures are repeated between confinnation of eligibility and the start of LD chemotherapy and the results are outside of the eligibility criteria provided herein, any suspected / identified infectious / itrflarnmatory process condition must resolve before a subject of the present disclosure is administered an. LD regimen of the present disclosure, In some embodiments, a complete medical history and physical examination of a subject of the present disclosure including the head, eyes, ears, nose, and throat (HEENT) and cardiac, vascular, respiratory, gastrointestinal, integumentary, and neurological systems does not reveal evidence of infection / inflammation before administration of an ID regimen of the present disclosure. In some embodiments, a subject of the present disclosure has not received systemic antimicrobials for the treatment of a known or suspected infection within the 48 hours before an ID regimen of the present disclosure (prophylactic use of antimicrobials is allowed). In some embodiments, a treatment course of any antimicrobials given for a known or suspected antecedent infection should be completed before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if a subject of the present discl osure is confinned to have an infectious process for which antimicrobials are not available (e.g.5viral pneumonia), the infection must be clinically resolved before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, the most recently collected blood, urine, or other body fluid cultures must show no growth for at least 48 hours, and any other infectious workup performed (e.g., bacterial, PCR, or stool or imaging studies) is negative before a subject of the present disclosure is administered an LD regimen of the present disclosure. In some embodiments, if there is clinical suspicion of an infection for which cultures are unlikely to be positi ve within 48 hours (e.g., fungal infection), then adequate time is allowed to allow the cultures to become positive before a subject of the present disclosure is administered an LD regimen of the present disclosure.(0270| In some embodiments, subjects of the present disclosure are hospitalized during a treatment -period from Day 0 before infusion with hypoimmune allogenic CD19-directed CAR T cells of the present disclosure until a minimum of 7 days after infusion. In some embodiments, the volume of hypoimmune allogenic CD19-directed CAR T cells of the present disclosure infused, thaw start / stop time, and the infusion start / stop time must all be noted in a subject’s medical record. In some embodiments, hypoimmune allogenic CD W-dlrected CAR T cells of the present disclosure are not be thawed until the subject is ready for the infusion.|0271 ] In some embodiments, subjects of the presen t disclosure remain hospitalized until all CAR-T related non-hematologic toxicities resolve to Grade 1 or return to baseline. In some embodiments, subjects may be discharged for a hospital with noncritical and clinically stable or improving toxicities (e.g., renal insufficiency) even if higher than Grade 1. In some embodiments, subjects should remain hospitalized for ongoing hypoimmune allogenic CD 19’ directed CAR T cells of the present disclosure-related fever, hypotension, hypoxia, or ongoing neurotoxicity of higher than Grade I. In some embodiments, subjects of the present disclosure refrain from driving or hazardous activities for at least 8 weeks following infusion.|02:72| In some embodiments, after administration of hypoimmune allogenic CD19-direeted CAR T cells of the present disclosure and being discharged from the hospital, a subject of the present disclosure will be evaluated at the following intervals: Day 8 (± 1 day). Day 10 ± I day), Day 13 (± 1 day), Day 16 (A 1 day), Day 21 ( a. 4 days), Day 28 re 4 days), monthly in months 2-12 (* 1 week) (or until start of a new systemic therapy). In some embodiments, subjects undergo the following assessments and procedures at the aforementioned intervals: clinical examination, vital signs, weight, ECOG assessment, concomitant medications documentation, neurologic examination to assess ICE score, ECG (if clinically indicated), brain MRI (if clinically indicated), lumbar puncture (if clinically indicated), tumor biopsy, disease / overall response assessments (including imaging and bone marrow biopsy / aspirate when applicable), AE7SAE / DLT reporting, chemistry panel (serum), CBC with differential (blood), coagulation (plasma), urinalysis (if clinically indicated), LDH, CRP, ferritin, beta-2 microglobulin, direct antiglobulin test, haptoglobin, serum Ig levels, Covid-19 and other viral serology testing (as clinically indicated), -hCG pregnancy test (serum or urine) for all females of childbearing potential (as clinically indicated), analytes including cytokines (seram / plasma)and anti~CAR-T antibodies, i.e, doner specific antibody and CDlMirected CAR antibody (serum), ctDN, MRD, CD19-directed CAR T cell kinetics (VCN and FACS), CD47 expression, RCL, T cell donality, and exploratory analyses including immunophenotyping, T cell killing, NK cell killing, and scRNAseq (PBMCs), assessment of levels of analytes including cytokines (serum / plasma) and CD 1 directed C AR T cells (PBMCs) should be drawn at the first onset and first reoccurrence of any Grade 2 or higher CAR-T-related toxicity, such as a Grade 2 CRS or neurologic event, and upon resoluti on of the event, if not already collected on the day (serum / 'plasma).|0273] In some embodiments, a subject of the present disclosure does not receive subsequent anti-cancer therapy. In some embodiments, a subject of the present disclosure does receive subsequent anti-cancer therapy if their disease progression has been documented, In some embo iments, subsequent anti -cancer therapy is or comprises chemotherapy, immunotherapy, targeted agents, SCT, radiation therapy, corticosteroids (other than those used to manage CRS and ICAbiS), and other investigational agents.8. Adverse Events and Toxicity Management10274] In some embodiments, a subject of the present disclosure experiences one or more adverse events (AEs) after administration of hypoimmune allogenic CDW-directcd CAR T cells of the present disclosure. In some embodiments, an AE is or comprises any untoward medical occurrence in a subject of the present disclosure, In some embodiments, an AE is or comprises worsening of a pre-existing medical condition. Worsening indicates that the pre-existing medical condition has increased in severity, frequency, and / or duration or has an association with a worse outcome. In some embodiments, an AE does not include the following: a pre-existing condition that has not or involves an intervention such as elective cosmetic surgery or a medical procedure; interventions (and associated complications) for pre-treatment conditions (such as elective cosmetic surgery) or medical procedures that were planned before administration of hypoimmune allogenic CD19~directed CAR T cells of the present disclosure; hospitalization for infusions of hypoimmune allogenic CD ^-directed CAR T cells or related procedures, and hospitalization as a precautionary measure. In some embodiments, “disease progression” as assessed by measurement of malignant lesions on radiographs or other methods is not consideredto be an AE. In some embodiments, worsening of signs and symptoms of a B cell malignancy of the present disclosure are considered to be AEs.|0275| In some embodiments, a subject of the present disclosure experiences one or more dose limiting toxicides (DLTs) after administration of hypoimmuae allogenic CD19-directed CAR T cells of the present disclosure. In some embodiments, a DLT is or comprises one or more of the following events that are at least possibly related to treatment with hypoimmune allogenic CD19-directed CAR T cells of the present disclosure and that occur within 28 days of infusion with hypoimmune allogenic CD19~directed CAR T ceils of the present, disclosure: any Grade 5 event (death) without clinical or radiologic evidence of disease progression; CRS: all Grade 4 CRS, and Grade 3 CRS that foils to improve to < Grade 2 within 72 hours following adequate therapy; Neurotoxicity: All > Grade 3 neurotoxicity of any duration; Grade 3 or higher acute GvHD that requires oral or IV corticosteroids (> 1 mg / kg) and does not resolve within 7 days. Grade 2 acute GvHD that is steroid refractory (i.e., progression after 3 days or no response after 7 days of systemic steroid treatment [> Img / kg / day prednisone or equivalent]); Grade 4 neutropenia or thrombocytopenia, not attributable to underlying disease or lymphodepletion chemotherapy, that does not improve to < Grade 2 within 42 days. Grade 3 thrombocytopenia with clinically significant bleeding; > Grade 3 toxicity involving vital organs (e,g,, cardiac, pulmonary) of arty duration (Exceptions may be made for Grade 3 or 4 abnormal hepatic or renal function tests that improves to < Grade 2 within 7 days); all other Grade 3 toxicities:, not attributable to underlying disease or lymphodepleting chemotherapy, that do not resolve to < Grade 2 within 72 hours. In some embodiments, the following events are not considered to be DLTs: Grade 3 cytopenia or Grade 4 lymphope a / anemia, Grade 3 TLS for < 2 weeks, and / or Grade 3 infections.10276] In. some embodiments, a subject of the present dis closur e experiences one or more serious adverse events (SAEs) after administration ofhypoimmune allogenic CD -directed CAR T cells of the present disclosure. In some embodiments, an S A E is defined as an A E that meets at least one of the following serious criteria: is fatal, is life-threatening (i.e., an event that places the subject at immediate risk of death; it does not refer to an event that hypothetically might have caused death if it were more severe), requires inpatient hospitalization or prolongation of existing or planned hospitalization, an AE would meet the criterion of “requireshospitalization* ’ if the event necessitated an admission to a healthcare facility, events that require an escalation of care when the subject is already hospitalized should be recorded as an SAE (e.g., movement from routine care in the hospital to the intensive care unit or if that event resulted in a prolongation of the planned hospitalization, results in persistent or significant disability / mcapacity, is a congenital anomaly / birth defect, and / or is another medically important serious event. The terms "severe’’ and "serious” are not synonymous. Severity refers to the intensity of an AE according to NCI Common Terminology Criteria tor Ad verse Events (CTCAE); the event Itself may be of relatively minor medical significance and, therefore, may not meet the seriousness criteria listed above. Severity and seriousness need to be independently assessed for each AE. Progression of a malignancy is not considered to be a drug-related SAE signs and symptoms of disease progression can be considered to be SAEs (and documented as being due to disease progression).|0277] In some embodiments, an AE is or comprises a targeted AE. In some embodiments, an SAE is or comprises a targeted SAE Targeted AEs / SAEs include neurologic, hematologic, infection, GvHD, autoimmune disorder, and second primary' malignancy events. Adverse events of special interest (reflecting potential risks of CD lf irected CAR therapies) include: CRS, neurotoxicity (i.e., IC. NS). Macrophage Activation Syndrome, TI.. S, Grade 3 cytopenia persisting through Day 28 post-treatment. Severe (Grade 3) infection through the first 28 days post-treatment.102781 In some embodiments, AEs are determined based on abnormal clinical laboratory findings. In some embodiments, abnormal laboratory findings without clinical significance are not AEs. In some embodiments, abnormal laboratory findings that result in new or worsening clinical sequelae or that require therapy or adjustment in current therapy are considered AEs. In some embodiments, an AE is an abnormal laboratory test result that results in a medical intervention (e.g., potassium supplementation for hypokalemia or iron replacement therapy for anemia) or a change in concomitant therapy. In some embodiments, an abnormal vital sign results is an AE if it is a change from baseline and is accompanied by clinical symptoms and / or results in a medical intervention or a change in concomitant therapy.102791 In some embodiments, risks associated with CD19-directed CAR T cell treatment are or comprise cytokine release syndrome (CRS), neurotoxicity, cydopenia,hypogammaglobulinemia and infections, and TLS. In some embodiments, risks associated with allogeneic CAR T cell therapy include G HD and clonal expansion (e.g„ second primary malignancy).

[0280] In some embodiments, if a subject of the disclosure experiences CRS, it is managed according to ASTCT guidelines (Lee 2019). In some embodiments, if a subject of the disclosure experiences neurotoxicity (i.e., 1CANS) it is managed according to ASTCT guidelines (Lee 2019). In some embodiments, if a subject of the disclosure experiences GvHD it is managed according to Mount. Sinai Acute GVHD International Consortium 2016 report (Harris 2016), In some embodiments, if a subject of the present disclosure experiences bemophagocytic lymphohistiocytosis (HLH), also known as Immune Effector Cell-Associated Hemophagocytic Lymphohistiocytosis-Like Syndrome (IEC-HS), it can be graded according to ASTCT IEC-HS 2023 guidelines,

[0281] Infections, hjytogammaglobulinemla, and c topenia are known side effects of CD19-directed CAR T cell therapy. Infections after CAR T cell therapy are common and have been reported in up to 70% of patients who received CAR. T cell therapy in registrational clinical trials for approved agents (KYMRIAH® (Prescribing Information, YESCARTA® Prescribing Information, BRElYANZI® Prescribing Infonnation), Bacterial, viral, and fungal infections have all been reported with use of CAR T cell therapy. Infections may occur for a number of concomitant reasons, including lymphodepleting (or antecedent) chemotherapy, CAR T cell-mediated B cell or plasma cell depletion, prolonged cytopenia, corticosteroid treatment, or as a consequence of the malignancy itself. The severity of CRS may also be associated with an increased risk of acute infections. Other potential risk factors for severe infections within the first 30 days of CAR-T treatment include (CANS, tocilizumab, and corticosteroid use. Subjects may remain at increased risk of complications for weeks to months after infusion (Logue 2021, Hill 2018). Infections are generally managed using agents that target the source of infection.Additionally, prophylaxis against vesicular stomatitis vinis / herpes simplex virus reactivation and neumocystis jirovecii pneumonia infections is generally used for patients undergoing CAR T cell therapy and for several months after. The decision to administer antibacterial or antifungal prophylaxis should be risk-adjustedbased on subject characteristics, such as prior lines of suppressive therapy and infection history (Thompson 2022).

[0282] Hypogammaglobulinemia is characterized by low antibody levels due to extremely low B cell or plasma cell counts, referred to as B cell or plasma cell aplasia, respectively. Hypogammaglobulinemia has been reported in up to 53% of patients who received CAR T cell therapy in registrational clinical trials and is associated with risk of infection (KYMRIAH® Prescribing Information, YESCARTA® Prescribing Information, BREYANZI® Prescribing Information). After ly phodepleting chemotherapy and treatment of the present disclosure, subjects can be monitored for serum immunoglobulin ( Ig ) levels and CBC with differential. Subjects with hypogammaglobulinemia (serum IgG levels < 400-600 mg / dL) and serious or recurrent infections (particularly bacterial) should be managed with intravenous immunoglobulin (IVIG) replacement therapy (400-500 rng / kg: IV1G given monthly). IV1G should be continued u til serum IgG levels normalize and infections are resolved. The optimal IgG threshold to use may depend on subject characteristics and infection frequency or severity.

[0283] Subjects who receive CAR T cell therapy are also at risk for hematologic toxicities, including prolonged cytopenia, such as neutropenia, thrombocytopenia, anemia, and / or leukopenia (KYMRIAH® Prescribing Information. YESCARTA® Prescribing Information, BREYANZI® Prescribing Information). Cytopenia may occur in the weeks to months after lymphodspleting chemotherapy and C AR T cell therapy tirfekm. Factors that may contribute to prolonged cytopenia include CRS and ICANS severity, disease burden, the number of prior therapies, baseline blood cell counts, peak CRP and ferritin levels, and CAR construct. Although lymphodepletion may be a contributing factor, the pathophysiology' of prolonged cytopenia after CAR T cell infusion remains unclear. Alternative causes of cytopenia in treated include myelodysplastic syndrome or second primary malignancy ( Jain 2020, Fried 2019). Cytopenia are generally managed with transfusion or growth factor support, if the possibility of myelodysplastic syndrome has been ruled out. Growth factors may be considered for persistent cytopenia. GM-CSF is not recommended, in the seting of C AR T cell therapy. An FDA-approved biosimilar is an appropriate substitute for filgrastim (Thompson 2022).B. Hypoimmunogenic Cells

[0284] In seme embodiments, the present disclosure is directed primary cells (such as, but not limited to, primary T cells). In some embodiments, primary cells such as primary T cells,are engineered for reduced expression or lack of expression of MHC class I and / orMHC class 11 human leukocyte antigens, and in some instances, for reduced expression or lack of expression of a T-cell receptor (TCR) complex. In some embodiments, the hypoimmune (HIP) T cells and primary T cells overexpress CD47 aad a chimeric antigen receptor (CAR) in addition to reduced expression or lack of expression of MHC class I and / or MHC class II human leukocyte antigens, and have reduced expression or lack expression of a T-cell receptor (TCR) complex. In some embodiments, the CAR is a CD I 9-specific CAR. In some embodiments, the cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or unmodified wild-type cell or control cell. In some embodiments, the wild-type cell or the control cell is a starting material. In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. In some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered cell.|'G285{ In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a chimeric antigen receptor (CAR), and include a genomic modification of the B2M gene. In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and include a genomic modification of the CIITA gene. In some ernbodiments, engineered^ and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include a genomic modification of the TRAC gene. In some embodiments, engineered and / or 'hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include one or more genomic modifications selected from the group consisting of the R2M, CUT A, and TRAC genes In some embodiments, engineered and / or hypoimmune (HIP) T cells and primary T cells overexpress CD47 and a CAR, and include genomic modifications of the B2M, CIITA, and TRAC genes. In some embodiments, the cells are JB2M- / -, CIITA- / -, TRAC- / -, CD471g: cells that also express CARs.(028^1 In certain embodiments, the cells are B2Miadel / indelsCIITAindel / indel, TRACindel / indel, CD47tg cells that also express CARs. Non-limiting examples of primary T cells include CD3+ T cells, CD4+ T cells, CD8+ T cells, naive T ceils, regulatory T ( reg) cells, non-regulatory T cells, Thl cells, Th? cells, Th9 cells, Th 17 cells, T-follicnlar helper (Tfh) cells, cytotoxic T lymphocytes (CTL), effector T (Teff) cells, central memory T (Tern) cells, effectorPage.107 of 277memory T (Tern) cells, effector memory' 1' cells express CD45R. A ( TE. MRA cells), tissueresident memory (Trnt) cells, virtual memory T cells, innate memory T cells, memory stem cell (Tse), yS T cells, and any other subtype of T cells. In some embodiments, primary T cells are selected from a group that includes cytotoxic T-cells, helper T-cells, memory T-cells, regulatory T-cells, tumor infiltrating lymphocytes, and combinations thereof. In some embodiments, cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or uumodi fied wild-type cell or control cell. In some embodiments, a wild-type cell or a control cell is a starting material. In some embodiments, starting material is a primary cell collected from a donor. In some embodiments, starting material is a primary blood cell collected from a donor, e.g., via a leukopak. In some embodiments, starting material is otherwise modified or engineered to have altered expression of one or more genes to generate an engineered cell,|0287| In some embodiments, primaiy T cells are from a pool of primary T cells from one or more donor subjects that are different than the recipient subject (e.g,, the patient administered the cells). Primary T cells can be obtained from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100 or more donor subjects and pooled together. Primary T cells can be obtained from 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10, or more 20 or more, 50 or more, or 1.00 or more donor subjects and pooled together. In some embodiments, primary T cells are harvested from one or a pl urality of indi viduals, and in some instances, primary T cells or a pool of primary T cells are cul tured in vitro. In some embodiments, primary' T cells or a pool of primary T cells are engineered to exogenously express CD47 and cultured in vitro.|9 88| fa certain embodiments, primary T cells or a pool of primary T cells are engineered to express a chimeric antigen receptor (CAR), Useful CARs include those that bind a CD 19 antigen. In some cases, a CAR is the same or equivalent to those used in FDA-approved CAR-T ceil therapies such as, but not limited to, those used in tisagenlecleucel and axicabtagcne cilolcucel, or others under investigation in clinical trials.|()289I In some embodiments, the primary T cells or the pool of primary T cells are engineered to exhibit reduced expression of an endogenous T cell receptor compared to unmodified primary T cells. Methods of genetically modifying a cell including a I' cell are described in detail, for example, in W02()20 / M8620, W02016 / 183041, PCT / US2020 / 44635,WO2021022223, W02021041316, WO2021222285, and WO2021222285, the disclosures of which are herein incorporated by reference in their entireties, including the tables, appendices, sequence listing and figures.

[0290] In some embodiments, CAR-T cells comprise a second generation CAR comprising an antigen binding domain, a transmembrane domain, and at least two signaling domains.|9291 In some embodiments, CAR-T cells comprise a CAR comprising an antigen binding domain, a transmembrane, and one or wore signaling domains. In some embodiments, the CAR also comprises a linker. In some embodiments, the CAR comprises a CD19 antigen binding domain, fa some embodiments, the CAR comprises a CD8o. transmembrane domain. In some embodiments, the CAR comprises a CD8a signal peptide. In some embodiments,: the CAR comprises a Whitlow linker GSTSGSGKPGSGEGSTK. G i'SEQ ID NO: 24). In some embodiments, the antigen binding domain of the CAR is selected from a group including, but not limited to, (a) an antigen binding domain targets an antigen characteristic of a neoplastic cell or an antigen binding domain targets an antigen characteristic of an autoimmune disorder.10292 In some embodiments, the CAR farther comprises one or more linkers. The format of an seFv is generally two variable domains linked by a flexible peptide sequence, or a “linker,” either in the orientation VH-linker-VL or VE-Iinker-VH, Any suitable linker known to those in the art in view of the specification can be used in the CARS. Examples of suitable linkers include, but are not limited, to, a GS based linker sequence, and a Whitlow linker GSTSGSGKPGSGEGSTKG (SEQ ID NO: 24).

[0293] In some embodiments, the antigen binding domain is selected, from a group that includes an antibody, an antigen-binding portion or fragment thereof, an seFv, and a Fab. In some embodiments, the antigen binding domain binds to CD 19. In some embodiments, the antigen binding domain is an anti*C 19 scFv such as but not limited to FMC63.

[9294] In some embodiments, the transmembrane domain comprises one selected from a group that includes a transmembrane region of TCRa, TCRp, TCRc, CD3c, CD3y, CD38, CD3 CD4, CDS, CD8&, CD8p, CD9, CD16, CD28, CD45, CD22, CD33, CD34, CD 7, CD40, CD40T.. / CDI 54, CD45, CD64, CD80, CD86, OX40 / CD134, 4-1 BB / CD 137, CD1 4. FceRIy,VEGFR2, FAS, FGFR2B, and functional variant thereof. In some embodiments, the transmembrane domain comprises a CD8a transmembrane domain.|(I295| In some embodiments, the signaling domainfs) of the CAR comprises a costimulatory domain(s). For instance, a signaling domain can contain a costimulatory domain. Or, a signaling domain can contain one or more costimulatory domains. In certain embodiments, the signaling domain comprises a costimulatory domain. In other embodiments, the signaling domains comprise costimulatory domains. In some cases, when the CAR comprises two or more costimulatory domains, two costimulatory domains are not the same. In some embodiments, the costimulatory domains comprise two costimulatory domains that are not the same. In some embodiments, the cosbmakitory domain enhances cytokine production, CAR-T cell proliferation, and / or CAR-T cell persistence during T cell activation, In some embodiments, the costimulatory domains enhance cytokine production, CAR-T ceil proliferation, and / or CAR-T cell persistence during T cell activation,(0296] In some embodiments, a CAR comprises a CD3 zeta (CD3Q intracellular domain or functional variant thereof. In some embodiments, a CAR comprises a 4- IBB co-sti ulatory intracellular domain or functional variant thereof. In some embodiments, the CAR comprises (i) a CD3 domain at functional variant thereof; and (ii) a 4-1 BB domain or functional variant thereof.|0297] Methods tor introducing a CAR. construct or producing CAR-T cells are well known to those skilled in the art. Detailed descriptions are found, for example, in Vormittag et at, Curt Opin Biotechnol, 2018, 53, 162-181; and Eyquem etai.. Nature, 2017, 543, 113-117. (0298] In some embodiments, the cells derived from primary T cells comprise reduced expression of an endogenous T cell receptor, for example by disruption of an endogenous T cell receptor gene (e.g., T cell receptor alpha constant region ( TRAC )),(0299] In some embodiments, a CD47 transgene is inserted into a random locus of a cell In some embodiments, a trnrisgene encoding a CAR is inserted into a random locus of a cell. In some embodiments, a transgene encoding a CAR is inserted into a random locus of a cell via viral vector transductiowlntegration. In some embodiments, a CD47 transgene and a transgene encoding a CAR are inserted into a random locus of a cell via viral vector transductifm / inlugration. In some embodiments, a vector is a self-inacti vating lenti viral vector Page.1.10 of 277pseudotyped with a vesicular stomatitis VSV-G envelope. In some embodiments, a transgene encoding a CAR is inserted into at least one allele of a cell using viral transduction. In some embodiments, an exogenous polynucleotide is inserted into at least one allele of a cell using a lentivirus based viral vector.|030O| In some embodiments, a CD47 transgene and a transgene encoding a CAR are inserted into the same loons.|(B011 In certain embodirnents, a CD47 and a CAR are controlled by a single promoter and are encoded by a single transgene. In some instances, the promoter controlling expression of any tansgene described is a constitutive promoter. In other instances, the promoter for any transgene described is an inducible promoter. In some embodiments, the promoter is an EFla promoter. In some embodiments, the promoter is C. A. G promoter. In some embodiments, a CD47 transgene and a transgene encoding a CAR are both controlled by a constitutive promoter, In some embodiments, a CD47 transgene and & transgene encoding a CAR are both controlled by an inducible promoter.(0302] Methods provided are useful for inacti vation or ablation of MI1C class I expression and / ot MHC class II expression in cells such as primary T cells. In some embodiments, genome editing technologies utilizing rare-cutting endonucleases (e.g., CRISPR / Cas) are also used to reduce or eliminate expression of genes involved in an immune response (e.g„ by deleting genomic DNA of genes invol ved in an immune response or by insertions of genomic DN A into such genes, such that gene expression is impacted) in cells, In certain embodiments, genome editing technologies or other gene modulation technologies are used to insert tolerance-inducing factors in human cells, rendering them and the differentiated cells prepared therefrom hypoimmrmogenic cells. As such, the hypoimmtmogenic eel Is have reduced or eliminated expression of HC 1 and MHC II expression, In some embodiments, the cells are »onimmunogenic (e.g., do not induce an innate and / or an adaptive immune response) in a recipient subject.|0303| In some embodiments, a cell includes a modification to increase expression of CD47.In some embodiments, a cell comprises a genomic modification of one or more target polynucleotide sequences that regulate the expression, of either MHC class I molecules,MI IC class II molecules, or MHC class I and MHC class II molecules. fa some embo iments, a genetic editing system is used to modify one or more target polynucleotide sequences. In some embodiments, the targeted polynucleotide sequence is one or more selected from the group including B2M and CIITA, In some embodiments, the cell comprises a genetic editing modification to the B2M gene. In some embodiments, the cell comprises a genetic editing modification to the CIITA gene. In some embodiments, the cell comprises genetic editing modifications to the B2M and CIITA genes. In certain embodiments, the genome of the cell has been altered to reduce or delete critical components of HLA expression. In some embodiments, the cells are modified or engineered as compared to a wild-type or control cell, including an unaltered or unmodified wild-type cell or control cell. In some embodiments, the wild-type cell or the control cell is a startingmaterial, In some embodiments, the starting material is a primary cell collected from a donor. In some embodiments, the starting material is a primary blood cell collected from a donor, e.g., via a leukopak. fa some embodiments, the starting material is otherwise modified or engineered to have altered expression of one or more genes to generate the engineered cell.|03O5| In some embodiments, the present disclosure provides a cell (e.g., primary T cell or CAR-T cell) or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of MHC class I molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell (e.g., primary T cell or CAR-T cell) or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of MHC class II molecules in the cell or population thereof In numerous embodiments, the present disclosure provides a cell (e.g,, primary T cell or CAR-T cell) or population thereof comprising a genome in which one or more genes has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of MHC class I and II molecules fa the cell or population thereof.103061 In certain embodiments, the expression of MHC I molecules and / or MHC II molecules is modulated by targeting and deleting a contiguous stretch of genomic DNA, thereby reducing or eliminating expression of a target gene selected from the group consisting of B2M and CIITA. In some embodiments, described herein are genetically edited cells (e.g., modifiedPage.112 of 277human cells) comprising exogenous CD47 proteins and inactivated or modified CHTA gene sequences, and in some instances, additional gene modifications that inactivate or modify B2M geae sequences,

[0397] Provided herein are cells exhibiting a modification of one or more targeted polynucleotide sequences that regu lates the expression of any one of the following: (a) MHC I antigens, (b) MHC II antigens, (c) TCR complexes, (d) both MHC I and II antigens, and (e) MHC I and II antigens and TCR complexes. In certain embodiments, the modification includes increasing expression of CD47. In some embodiments, the cells include an exogenous or recombinant CD47 polypeptide, In certain embodiments, the modification includes expression of a chimeric antigen receptor. In some embodiments, the cells comprise an exogenous or recombinant chimeric antigen receptor polypeptide.

[9398] In some embodiments, the present disclosure provides a cell or population thereof comprising a geuome: in which a gene has been edited to delete a contiguous stretch of genomic DMA, thereby reducing or eliminating surface expression of MHC class I molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of MHC class II molecules in the cell or population thereof. In certain embodiments, the present disclosure provides a cell or population thereof comprising a genome in which a gene has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression ofTCR. molecules in the cell or population thereof. In numerous embodiments, the present disclosure provides a cell or population thereof comprising a genome in which one or more gen.es has been edited to delete a contiguous stretch of genomic DNA, thereby reducing or eliminating surface expression of MHC class I and II molecules and TCR complex molecules in the cell or population thereof In some embodiments, the cells and methods described herein include genomically editing human ceils to cleave C1ITA gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but not limited to, B2M and TRAC, In some embodiments, the cells and methods described herein include genomically editing human cells to cleave B2M gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but Page 1.13 of 277not limited to, CUT A and TRAC. In some embodiments, the cells and methods described herein include gcnomieally editing human cells to cleave TRAC gene sequences as well as editing the genome of such cells to alter one or more additional target polynucleotide sequences such as, but not limited to, B2M and CT1TA.|031 Of In some embodiments, a population of engineered cells described evades NK cell mediated cytotoxicity upon administration to a recipient patient. In some embodiments, a population of engineered cells evades NK cell mediated cytotoxicity by one or more subpopulations of NK cells. In some embodiments, a population of engineered is protected from cell lysis by NK cells, including immature and / or mature NK cells upon administration to a recipient patient. In some embodiments, a population of engineered cells evades macrophage engtilfmcnt upon administration to a recipient patient. In some embodiments, a population of engineered cells does not induce an innate and / or an adaptive immune response to the cell upon administration to a recipient patient.|03H| In some embodiments, cells described herein are controlled by a safety switch. The term “safety switch” used herein refers to a system for controlling the expression of a gene or protein of interest that, when downregulated or upregulated, leads to clearance or death of the cell, e.g., through recognition by the host’s immune system. A safety switch can be designed to be triggered by an exogenous molecule in case of an adverse clinical event. A safety switch can be engineered by regulating the expression on the DNA, RNA and protein levels. A safety switch includes a protein rjr molecule that allows for the control of cellular activity in response to an adverse event. In some embodiments, a safety switch comprises a therapeutic agent that inhibits or blocks the interaction of CD47 and SIRPa, In some aspects, the CD47“SIRPa blockade agent is an agent that, neutralizes, blocks, antagonizes, or interferes with the cell surface expression of CD47, SIRPa, or both. In some embodiments, the CD47-SIRPa blockade agent inhibits or blocks the interaction of CD47, SIRPa or both. In some embodiments, a CD47- IRPa blockade agent (e.g., a CD47 -SIRPa blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering agent) comprises an agent selected from a group that includes an antibody or fragment thereof that binds CD47, a bispecific antibody that binds CD47, an immunocytokine fusion protein that bind CD47, a CD47 containing fusion protein, an antibody or fragmentthereof that binds SIRPa, a bispecific antibody that binds SIRPa, an imnwnocytokme fusion protein that bind SIRPa, an SIRPa containing fusion protein, and a combination thereof.|(I312| In some embodiments, the population of engineered cells described elicits a reduced level of immune activation or no immune activation upon administration to a recipient subject. In some embodiments, the cells elicit a reduced level of systemic TH 1 activation or no systemic THl activation in arecipient subject. In some embodiments, the cells elicit a reduced level of immune activation of peripheral blood mononuclear cells (PBMCs) or no immune activation of PBMCs in a recipient subject. In some embodiments, the cells elicit a reduced level of donor-specific IgC* antibodies or no donor specific IgG antibodies against the cells upon administration to a recipient subject. In some embodiments, tire cells elicit a reduced level of IgM and IgG antibody production or no IgM and IgG antibody production against the cells in a recipient subject. In some embodiments, the cells elicit a reduced level of cytotoxic T cell killing of the cells upon administration to a recipient subject.L CHTA(>313 In some embodiments, the technologies disclosed herein modulate (e.g., reduces or eliminates) the expression of MHC IIgenes by targeting and modulating (e.g., reducing or eliminating) Class II transactivator (CUT A) expression. In some embodiments, the modulation occurs using a CRISPR / Cas system. ClITA is a member of the LR or nucleotide binding domain (NBD) leucine-rich repeat (LRR) family of proteins and regulates the transcription of MHC II by associating with the MHC enhanceosome.[0314[ In some embodiments, the target polynucleotide sequence of the present disclosure is a variant of CIITA. In some embodiments, the target polynucleotide sequence is a homolog of CHIA. In some embodiments, the target polynucleotide sequence is an ortholog of CHTA.[0315| In some embodiments, reduced or eliminated expression of CHTA reduces or eliminates expression of one or more of the following MHC class II are HLA-DP, HL A- DM, HLA-DOA, HLA-DOB, HLA-DQ, and HLA-DR.

[0316] In some embodiments, the cells described herein comprise gene modifications at the gene locus encoding the CHTA protein. In other words, the cells comprise a genetic modification at the CHTA locus. In some instances, the nucleotide sequence encoding the CHTA protein is set forth in Refoeq. No. NM MM124 andNCBI Genbank No. U 18259. In some instances, the CHTA gene locus is described in NCBI Gene ID No. 4261, In certain cases, the amino acid sequence of CHTA is depicted as NCBI GenBank No. AAA88861. I, Additional descriptions of the CHTA protein and gene locus can be found in Uniprot No. P33076, HGNC Ref No. 7067, and OMIM Ref No. 600005.

[0317] In some embodiments, the hypo immunogenic cells outlined herein comprise a genetic modification targeting the CHTA gene, In some embodiments, the genetic modification targeting the CUT A gene by the rare-cutting endonuclease comprises a Cas protein or a polynucleotide encoding a Cas protein, and at least one guide ribonucleic acid sequence for specifically targeting the CHTA gene. In some embodiments, the at least one guide ribonucleic acid sequence for specifically targeting the CHTA gene is selected from the group consisting of SEQ ID NOS:5184-36352 of Table 12 of W0...

Claims

CLAIMSI, A method of treating: a subject having or suspected of having an IgE-mediated disease or disorder, the method comprising:administering to the subject a therapeutically effective amount of a composi tion comprising a population of engineered hypcimmanogeulc T cells, wherein the engineered hypoimmunogenic T cells comprise one or more modifications that:(a) inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (jii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules,(b) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wildtype I cell that does act comprise the modification, and(c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR comprises an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of an IgE -producing cell.

2. The method of claim 1, wherein the antigen expressed on the surface of an IgE-producing cell comprises CD19, CD20, CD22, BCMA, GPRC5D, CD38, or any combination thereof3. The method of claims 1 or 2, wherein the engineered T cells comprise one or more of a CD19~specific CAR., a CD20-speeifi.c CAR, a CD22~speciflc CAR, a BCMA-specifie CAR, a GP CSD-specific CAR, a CD38-specific CAR, or any combination thereof.

4. A method of treating a subject having or suspected of having an IgE-medlated disease or disorder, the method comprising:administe rin g: to the subject a therapeutically effective amount of a composition comprising a population of engineered hypoimmunogenic T cells comprising:(a) reduced expression of b a-2 microglobulin (B2M), Class II Major Histocompatibility Complex Transactivator (CITI A). and I cell receptor alpha constant (TRAC) relative to a contra; or wild-type I cells,(b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD T9-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide,5. The method of any one of claims I -4, wherein the IgE-mediated disease or disorder is asthma, Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chrome spontaneous (idiopathic) urticaria, chronic rhinosinusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angi oedema, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or any combination thereof.

6. The method of claim 5, wherein the asthma is a steroid refractory asthma and / or a severe asthma.

7. The method of claim 5, wherein the food allergy is a severe, refiwtoty food allergies,8. The method of claim 5 or 7, wherein the food allergy is an allergy to peanut, shellfish, egg, soy, wheat, fish, or tree nut.

9. The method of any one of claims 1-8, wherein the method results in deep B cell depletion.

10. The method of any one of claims 1-9, wherein the method resul ts B cell depletion in circulation and a tissue.

11. The method of any one of cl aims 1-10, wherein the method results in B cell depletion in a test tissue sample, and the test tissue sample is or comprises tissue from a lymph node, germinal center, gastrointestinal tract, liver, spleen, gallbladder, king, kidney, and / or bone marrow.

12. The method of claim 11, wherein the test tissue sample is a tissue biopsy.

13. The method of claim 1, wherein the B cells express CD 19 and or CD20 and the CD19-expressing and / or CD20-cxprcs.smg B cells are detected in the tissue biopsy by specifically staining the CD 19-expressing and / or CD20-expressing cells.Page of 27714. The method claim 13. wherein the staining is immunohistochemical staining, 15. The method of claim 13 or 14, wherein the number of B cells detected is from 0- 20 cells / mm2, from 0-15 cel mnfe from 0-10 cel inm2, from 0-5 cells / mniz, or 0 cells / m.

16. The method of claim 15, wherein the tissue biopsy is from a lymph node.

17. The method of any one of claims 11-16, wherein the test tissue sample is collected at least 2 weeks after administration of the composition comprising a population of engineered hypoimmunogenlc T cells.

18. The method of any one of claims 11-17, wherein the test tissue sample is collected no more than 3 months after administration of the composition comprising a population of engineered hypoimmunogenic T cells.

19. The method of any one of claims 11-18, wherein the test tissue sample is collected between 2 weeks and 3 months after the administration of the composition comprising a population of engineered hypoimmunogenic T cells.

20. The method of any one of claims 11-19, wherein the test tissue sample is collected about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, and / or about 12 weeks after the administration of the composition comprising a population of engineered hypoimmunogenic T cells.

21. The method of any one of claims 11 -20, wherein the test tissue sample is collected about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks., about 7 weeks to about 8 weeks, about 8 weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, and / or about 11 weeks to about 12 weeks alter the administration of the composition comprising a population of engineered hypoimmunogenic T cells.

22. The method of any one of claims 1-21, wherein the IgE an tibodies in a test sample from the subject is reduced or depleted.

23. The method of claim 22, wherein test sample is a blood sample or serum sample.

24. The method of claim 23, wherein the test sample comprises a concentration of IgE antibodies that is decreased compared to a reference value.25 The method of claim 24, wherein the reference value is the concentration of IgE antibodies detected in a reference sample obtained from the subject prior to administering the composition comprising a population of engineered hypoimmunogenic T cells.

26. The method of claim 24 or 25, wherein the reference value is a concentration of IgE antibodies from 200 ng / 'niL to 1000 ng / mL, from 200 ng / mL to 900 ng / mL, from 200 ng / mL to 800 ng / mL, from 200 ng / mL to 700 ng / mL, from 200 ng / mL to 600 ng / mL, from 200 ng / mL to 500 ng / mL. from or 200 ng / mL to 400 ng / mL,27. The method of any one of claims 22-26, wherein the concentration of IgE antibodies in the test sample is within a range from 0 ng / mL to 200 ng / mL.

28. The method of claim 22-27, wherein the concentration of IgE antibodies in the test sample is up to 200 ng / mL, 180 ng / mL, 160 ng / mL, 140 ng / mL, 120 ng / mL, 100 ng / mL, §0 ng / mL, 60 ng / mL, 40 ng / mL, 20 ng / mL, 1 ng / mL, 0.1 ng / mL, 0.09 ngmL, 0.08 ng / mL, 0,07 ngmL, 0.06 ng / mL, 0.05 ng / mL, 0.04 ng / mL, 0.03 ng / mL, 0.02 ngmL, or 0.01 ng mL.

29. The method of any one of claims 22-28, wherein the test sample comprises no IgE antibodies or IgE antibodies are undetectable.

30. The method of any one of claims 22-29, wherein the test sample is collected between 2 days and 3 months after administration of the composition comprising a population of engineered hypoi mmu nogenic T cells.

31. The method of any one of claims 22-30, wherein the tes t sample is collec ted.from the subject about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, and / or about 12 weeks after administration of the composition comprising a population of engineered hypoiminunogenic T cells.

32. The method of any one of claims 22-31, wherein the test sample is collected from the subject about 1 week to about 2 weeks, about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 7 weeks to about 8 weeks, about 8 weeks to about 9 weeks, about 9 weeks to about 10 weeks, about 10 weeks to about 11 weeks, and / or about 11 weeks to about 12 weeks after administration of the composition comprising a population of engineered hypoimmunegeme T cells.33, The method of any one of claims 1-32, wherein the engineered hypoimmunogeitic T ceils are 82. M knock cut, CHTA knock out, and / or 'TRAC knock out cells, optionally wherein the engineered hypoimmunogenic T cells are B2M knock out, CHTA knock out, and TRAC knock out cells.

34. The method of any one of claims 1-33, wherein the engineered hypoimmunogenic T cells are B2MtOi1i:!'mc‘, CIITA^^5"^, aiid / br TRAC‘a^’wte<cells, optionally wherein the engineered hypoinmumogeme T cells arecells.

35. ’The method of any one of claims 1-34, wherein the one or more tolerogenic factors comprise or arc selected from the group consisting of CD47, SIRPalpha engagers, CD24, CD27, CD35, CD46, CD55, CD59, CD200, HLA-C, HI. A-E, HLA-E heavy chain, HLA-C, PD-LI, IDO I, CTLA4~Ig, Ci-Inhibitor, IL-10, IL-35, Fast, CCL2 L CC1.22, M / geS, A20 / WFAIP3, CD39, CRT, HLA-F, ILI5-RF, MANF, and / or Serpinb9, optionally wherein the one or more tolerogenic factors comprise CD47 or an engineered CD47.

36. The method of any one of claims 1-35, wherein the engineered T cells evade NK. cell-mediated cytotoxicity upon administration to the subject.

37. The method of any one of claims 1 -36, wherein the engineered cells are protected from cell lysis by mature NK. cells upon administration to the subject.

38. The method of any one of claims 1-37, wherein the engineered T cells evade macrophage-mediated cytotoxicity, optionally wherein the macrophage-mediated cytotoxicity involves phagocytosis and / or reactive oxygen species.

39. The method of any one of claims 1-38, wherein the subject was treated with an immunodepleting therapy prior to administering the engineered T cells.

40. The method of any one of claims 1-39, wherein the composition is administered to the subject at a dosage of about 30 x 10* CARL cells to about 1000x10® CAR-*- cells.

41. The method of any one of claims 1 -40, wherein the compositi on is administered to the subject at a dosage of at least about 3 Ox IO® CARL cells, at least about 60x10* CARL cells, at least about 90x10 CARL cells, at least about 120x10* CARL cells, at least about 200 10* CARL cells, at least about 400 xlO* CARL cells, at least about 600 xlO6CARL cells, at least about 800 xlO* CARL cells, or at least about WOO x 10* CARL cells.

42. The method of any one of claims 1- 1, wherein the control or wild-type cel l is a starting material.

43. A method of assessing B cell depletion, comprising obtaining a test sample from a subject and detecting a concentration of IgE antibodies in the test sample, wherein the subject was administered a B cell depleting composition at least 2 days priors to obtaining the test sample and a decrease in the concentration of IgE antibodies in the test sample compared to a reference value indicates depletion of B cells by the B cell depleting composition.

44. A method treating a B cell mediated disease or disorder, comprising administering to a subject an agent or therapy that is capable of treating, preventing, delaying, or atenuating development of a B cell mediated disease or disorder, wherein the subject is determined to be at risk for maintaining: or developing progressive disease by a method comprising,(a) obtaining a test sample from a subject and detecting: a concentration of IgE antibodies in the sample, wherein the subject was administered a B cell depleting composition at least 2 days priors to obtaining the test sample, and(b) comparing the concentration detected for the IgE antibodies to a reference value, wherein the subject is one in which the comparison indicates that the subject is at risk for maintaining or developing a progressive B cell mediated disease or disorder.

45. The method of claim 43 or 44, wherein the reference value is the concentration of IgE antibodies detected in a reference sample obtained prior to administering the B cell depleting composition.

46. The method of any one of claims 43-45, w herein the reference value is a concentration of IgE antibodies from 10 ng / mL to 200 ng / mL, from 50 ng / mL to 200 ng / mL, from 75 ng / L to 200 ng / mL, from 100 ng / mL to 200 ng / mL, from 50 ng / mL to 600 ng / mL, from 75 ng / mL to 600 ng / mL. from 100 ng / rtl. to 600 ng / mL, from 100 ng / mL to 500 ng / mL, from 100 ng / L to 400 ng / mL, from 200 ng / mL to 600 ng / mL, from 200 ng / L to 500 ng / mL, and / or from 200 ng / rnL to 400 ngdnL.

47. The method of any one of claims 43-46, wherein the test sample and / or the reference sample is blood or serum.

48. The method of any one of claims 43-47, wherein the B cell depicting composition comprises a therapeutically effective amount of a composition comprising a population of engineered hypoimmunogenic T cells, wherein the hypoimmunogenic T cells comprise one or more modifications that:(a) inactivate or disrupt one or more alleles of: (i) one or mare major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (ii) one or more MHC class I! molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules, and / or (lii) one or mote T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more TCR molecules,(b) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wildtype T cell that does not comprise the modification, and(c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR comprises an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of an IgE-producing cell.

49. The method of claim 48, wherein the antigen expressed on the surface of an IgE-producing eell comprises CD19. CD20, CD22. BCM A, GPRC5D. CD.

38. or any combination thereof.

50. The method of claims 48 or 49, wherein the engineered T cell s comprise one or more of a CD19-specific CAR, a CD20-specific CAR, a CD22-specific CAR, a HCMA-spedfic CAR, a GPRC5D~specific CAR, a CD38~specific CAR, or any combination thereof.

51. The method of any one of claims 43-47, wherein the B cell depleting composition comprises a therapeutically effective amount of a composition comprising a population of engineered hypoirnmunogenlc T cells comprising'.(a) reduced expression of beta-2 microglobulin (B2M), Class 11 Major Histocom atibility Complex Trausactivator (CIITA), and T cell receptor alpha constant 'TRAC) relative to a control or wild-type T cells,(b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD19-specific chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide.

52. The method of any one of claims 43 -51, wherein B cell mediated disease or disorder is aa IgE-mediated disease or disorder.

53. The method of claim 52, wherein the IgE-mcdiated disease or disorder is asthma, Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chronic spontaneous (idiopathic ) urticaria, chronic rhinosinusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angioederna, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or any combination thereof.

54. The method of any one of claims 43-51, wherein El cell mediated disease or disorder is a B cell malignancy.

55. The method of claim 54, wherein the B cell malignancy is a lymphoma or a leukemia.

56. The method of claim 54 or 55, wherein the B cell malignancy comprises or is selected from the group consisting of: Nou-Hodgkin’s Lymphoma (NHL), Chronic Lymphocytic Leukemia (CLL), large B cell lymphoma (LBCL), transplant ineligible large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinal B cell lymphoma. (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), and / or small lymphocytic lymphoma (SLL).

57. The method of any one of claims 43 -51, wherein B cell mediated disease or disorder is an autoimmune condition.

58. The method of claim 57, wherein the autoimmune condition comprises or is selected from the- group consisting of: systemic lupus erythematosus (SLE), extrarenal systemic lupus erythematosus (ERL), lupus nephritis (LN), CNS lupus, vasculitis, granulomatosis vasculitis, polyangiitis & microscopic poly angiitis vasculitis, anti -neutrophilic cytoplasmic autoantibody (ANCA) associated vasculitis (AAV), granulomatous polyangiitis, microscopic polyangiitis, multiple sclerosis (relapsing and / or progressive), pemphigus vulgaris, autoimmune blistering skin, diseases, membranous nephropathy (MIN), anti -N D receptor neuropathy, netiromyelitis optica, idiopathic thrombocytopenic purpura, autoimmime hepatitis, type 1 diabetes mellitus, rheumatoid arthritis, juvenile rheumatoid arthritis, chronic inflanimatoty demyelinating polyneuropathy, polymyositis / denuatomyositis, stiff persons disease, anti-NIMDA receptor encephalitis, anti -synthetase autoimmune syndromes, anti-phospholipid antibody syndrome, Sjogren’s syndrome, cryoglobulinemia, focal segmental glomerulosclerosis, rapidlyprogressive glomerulopathy, autoimmune hemolytic anemia, amyloidosis, scleroderma, idiopathic inflammatory myositis, and / or immune-mediated necrotizing myopathy,59. A method of assessing B cell depletion, comprising obtaining a test sample from a subject that comprises antibodies and detecting the concentration of antibodies in the test sample that bind a major histocompatibility complex (MHC) protein and comparing the concentration of antibodies that bind a MHC protein to the concentration of antibodies that bind a MHC protein in a reference sample,wherein prior to obtaining fee test sample the subject was administered a cornposition comprising a population of engineered hypoimmunogenic T cells, wherein the hypoimmunogenic T cells comprise:(a) one or more modifications that inactivate or disrupt one or more alleles of: (i) one or more major histocompatibility complex (MHC) class I molecules and / or one or more molecules that regulate expression of fee one or more MHC class I molecules, (ii) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules,(b) one or more modifications that increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to a control or wild-type cell that does not comprise the modification, and (c) express a chimeric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR comprises an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell,60. The method of claim 59, wherein the reference sample was obtained fi-om the subjec t prior to administering the composition comprising the population of engineered hypoimmunogenic T cells.

61. The method of claim 59 or 60, wherein fee test sample is obtained at least 20 days after the subject was administered the composition comprising a population of engineered hypoimmimogenic T cells.

62. The method of any one of claims 59-61, wherein the test sample and / or the reference sample is blood or scrum.

63. The method of any one of claims 59-62, wherein absence of antibodies that bind a MHC protein indicates deep B cell depletion.

64. Th© method of any on© of claims 59-63, wherein the antibodies that bind aMHC protein are IgG and / or IgM isotypes.

65. A method of treating a B cell d isease or disorder, comprising,(a) administering to a subject a therapeutically effective amount of a composition comprising a population of engineered hypoimmunogenic T cells, wherein the hypoinimunogenic T cells comprise one or more modifications that:(1) inactivate or disrupt one ormore alleles of: (i) one or more major histocompatibility complex ( HC) class I molecules and / or one or more molecules that regulate expression of the one or more MHC class I molecules, (u) one or more MHC class II molecules and / or one or more molecules that regulate expression of the one or more MHC class II molecules,(2) increase expression of a tolerogenic factor encoded by a first exogenous polynucleotide, wherein the increased expression of (2) is relative to a control or wildtype T cell that does not comprise the modification, and(3) express a ©hi meric antigen receptor (CAR) encoded by a second exogenous polynucleotide, wherein the CAR comprises an extracellular ligand-binding domain having specificity for an antigen expressed on the surface of a B cell; and (b) obtaining a test sample from the subject and detecting a concentration of antibodies that bind a MHC protein, wherein the test sample was obtained at least 20 days after the subj ect was administered the composition comprising the population of engineered h jfpoi mmogenic T cells; arid(c) comparing the concentration of antibodies that bind a MHC protein in ( b) to the concentration of anti bodies that bind a MHC protein in a reference sample obtained from the subject prior to administering the composition comprising the population of engineered hypoimmunogenic T cells; and(d) if the comparison indicates the subject has developed anti-M'HC antibodies, administering to the subject a composi tion or therapy that is capable of depleting B cells.

66. The method of claim 65, wherein the population of engineered hypoimmunogenic T cells further comprise one or more modifications that inactivate or disrupt one or more alleles of (iii) one or more T-cell receptor (TCR) molecules and / or one or more molecules that regulate expression of the one or more ‘TCR molecules.

67. The method of claim 65 or 66, wherein the antigen expressed on the surface of a B cell comprises CD 19, CD20, CD22, BCMA, GPR. C5D, CD38, or any combination thereof 68. The method of any one of claims 65-67, wherein the engineered T cells compri se one or more of a CD 19 -specific CAR, a CD2Q-specific CAR, a CD22-speciflc CAR, a BCMA-specific CAR, a GPRC5D-specific CAR, a CD38-specific CAR, or any combination thereof.

69. The method of any one of claims 65-68, wherein the B cell depleting composition comprises a therapeutically effective amount of a composition comprising a population of engineered hypoimmunogenic T cells comprising:(a) reduced expression of beta-2 microglobulin (B2 M), Class H Major Histocompatibility Complex Transactivator (C11TA), and T cell receptor alpha constant (TRAC) relative to a control or wild-type T cells,(b) increased expression of CD47 encoded by a first exogenous polynucleotide, wherein the increased expression of (b) is relative to the control or wild-type T cells, and (c) expression of a CD1 -specific chimeric antigen receptor (CAR.) encoded by a second exogenous polynucleotide.

70. The method of any one of claims 65-69, wherein B cell mediated disease or disorder is an IgE -mediated disease or disorder.

71. The method of claim 70, wherein the IgE-mediated disease or disorder is asthma. Eosinophilic Granulomatosis with Polyangiitis (E-GPA), a food allergy, a drug allergy, allergic rhinitis, hyper IgE syndrome, atopic dermatitis, chronic spontaneous (idiopathic) urticaria, chronic rhinosmusitis, systemic mastocytosis, cutaneous mastocytosis, allergic bronchopulmonary aspergillosis, recurrent idiopathic angloedema, interstitial cystitis, or eosinophil-associated gastrointestinal disorder, or any combination thereof72. The method of any one of claims 65-69, wherein B cell mediated disease or disorder is a B cell malignancy,73. The method of claim 72, wherein the B cell malignancy is a lymphoma or a leukemia.

74. The method of claim 72 or 73, wherein the B cell malignancy comprises or is selected from the group consisting of: Mon-Hodgkin 's Lymphoma (NHL), Chronic Lymphocytic Leukemia (CEL), large B cell lymphoma (LBCL), transplant ineligible large B cell lymphoma (LBCL), diffuse LBCL (DLBCL), high-grade B cell lymphoma (HGBCL), primary mediastinalB cell lymphoma (PMBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), marginal zone lymphoma (MZL), and / or small lymphocytic lymphoma (SLL).

75. The method of any one of claims 65-69, wherein B cell mediated disease or disorder is an autoimmune condition.

76. The method of claim 75, wherein the autoimmune condi tion comprises or is selected from the group consisting of: systemic lupus erythematosus (SEE), extrarenal systemic lupus erythematosus (ERL), lupus nephritis (LN), CNS lupus, vasculitis, granulomatosis vasculitis, pol arrgiitis & microscopic polyangiitis vasculitis, anti-neutrophilic cytoplasmic autoantibody (ANC A) associated vasculitis (AAV), granulomatous polyangiitis, microscopic polyangiitis, mul tiple sclerosis (relapsing and / or progressive), pemphigus vulgaris, autoimmune blistering skin diseases, membranous nephropathy (MN), and-NMDA receptor neuropathysneuromyelitis optica, idiopathic thrombocytopenic purpura, autoimmune hepatitis, type I diabetes mellitus, rheumatoid arthritis, juvenile rheumatoid arthritis, chronic inflammatory demyelinating polyneuropathy, polymyositis / dermatomyositis, stiff persons disease, antl-NMDA receptor encephalitis, anti-synthetasc autoimmune syndromes, anti-phospholipid antibody syndrome, Sjogren’s syndrome, cryoglobulinemia, focal segmental glomerulosclerosis, rapidly progressive glomerulopathy, autoimmune hemolytic anemia, amyloidosis, scleroderma, idiopathic inflammatory myositis, and / or imrnune-mediated necrotizing myopathy.

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