Immune cells with paired receptors and uses in treating autoimmunity

Abstract

The present disclosure relates to the fields of adoptive cell therapy and treatments for autoimmune diseases and disorders. The present disclosure provides engineered cells, compositions, and methods for treating patients suffering from autoimmune disorders and which are poorly treated by other methods. Specifically, the engineered cells, compositions, and methods selectivity destroy cells that mediate autoimmunity, both B and T cells, yet spare cells that modulate the autoimmune inflammatory response, e.g., regulatory T cells, or are not otherwise involved in the autoimmune response. Finally, the engineered cells are designed to avoid fratricide during their manufacturing.

Description

IMMUNE CELLS WITH PAIRED RECEPTORS AND USES IN TREATING AUTOIMMUNITYREFERENCE APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 664,595 filed on June 26, 2024, the contents of which are incorporated herein by reference in their entireties.REFERENCE TO SEQUENCE LISTING

[0002] The present application is being filed with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 061250-563001WO, created on June 26, 2025, and is 1,290 kilobytes in size. The information in electronic format of the Sequence Listing is incorporated by reference in its entirety.TECHNICAL FIELD

[0003] The present disclosure relates to the fields of adoptive cell therapy and treatments for autoimmune diseases and disorders. The present disclosure provides engineered cells, compositions, and methods for treating patients suffering from autoimmune disorders. Specifically, the engineered cells, compositions, and methods may selectivity destroy cells that mediate autoimmunity, both B and T cells, yet spare cells that module and / or mitigate the autoimmune inflammatory response, e.g., regulatory T cells, or are not otherwise involved in the autoimmune response. In some embodiments, the engineered cells are designed to avoid fratricide during their manufacturing.BACKGROUND

[0004] The immune system can suffer certain derangements in which subsets of either B or T cells, or both, become reactive to self-antigens and damage or destroy normal tissues. Examples of such autoimmune diseases known to be caused by autoreactivity include systemic lupus erythematosus, multiple sclerosis, and rheumatoid arthritis. New science and new modalities have changed the equation for autoimmune patients, including those suffering from treatment-refractory multiple sclerosis (MS). Cell therapy targeting immune cells offers the prospect of resetting the immune response. Indeed, stem cell transplants, though expensive, time intensive, and risky for patients, has been shown to dramatically improve the health of some patients seriously affected by autoimmune diseases. For example, in one statement from an expert: “AHSCT has been proven to be one of the most effective treatment modalities for MS in recent studies. However, debilitating complications due to immunological outcomes of the procedure have led to increased morbidity” (Mrudula et al., 2023). These risks and other difficulties of stem cell transplantation encourageexploration of different modalities, including monoclonal antibodies (mAbs). A CD20-targeting mAb is the only approved therapy for treating progressive MS. As one expert has described: “For a long time, T cells were seen as the key effector cells in MS. However, new evidence highlights the crucial role of B cells in the pathogenesis of MS. As B cells have three main immunological functions, they are assumed to be involved in the pathogenesis of MS in different ways” (Frisch et al., Neurotherapeutics 2021). There remains room for improvement over currently-available therapeutics which rely on mAbs. As another expert commented: “Among participants with relapsing multiple sclerosis, ublituximab [CD20 mAb] resulted in lower annualized relapse rates and fewer brain lesions on MRI than teriflunomide over a period of 96 weeks but did not result in a significantly lower risk of worsening of disability” (Steinman et al., 2023).

[0005] Indeed, new data suggest that targeting B cells can be an effective means to treat or ameliorate autoimmune disorders, such as systemic lupus erythematosus (SLE) and MS. These therapies generally target B cells non-selectively. Because humans can live without B cells for a significant length of time, this approach has generally targeted antigens such as CD 19 and CD20 that are expressed on nearly all B-cell lineages and cause transient B-cell aplasia, which is an acceptable health risk given the goal of treating an illness as serious as an autoimmune disease. In the words of an expert: “In this case series, CD 19 CAR T-cell transfer appeared to be feasible, safe, and efficacious in three different autoimmune diseases, providing rationale for further controlled clinical trials: (Muller et al., N Engl J Med 2024).

[0006] However, improving and extending these therapies to other patients likely requires targeting T cells. Unlike B cells, it is very hazardous to eliminate T cells wholesale as they are the central element in the adaptive immune response; for example, T cells are the most effective mediator of anti-viral immunity. In addition, certain T cell subtypes, notably regulatory T cells (Tregs), are involved in immune tolerance of normal tissues. Therefore, targeting T cells would require an especially precise therapeutic and, perhaps, one which integrates information from multiple antigens; i.e., engineered cells (e.g., Chimeric antigen receptors (CARs)) that respond to antigen profiles, rather than to single antigens. Such engineered cells respond to antigen profiles are often described as logic-gated by analogy with computational devices that respond to multiple inputs. One such logic-gated system uses NOT gate logic, which combines activating receptors and inhibitor receptors. An example of such a NOT gate is based on the LIR-1 inhibitor receptor and is called Tmod (DiAndreth et al. 2022).

[0007] Unfortunately, such logic-gated systems are currently unavailable for treating autoimmune diseases. The present disclosure remedies this unmet need.SUMMARY

[0008] The present disclosure provides engineered T cells that both remove B cells and / or target a subset of pathologic T cells and spare T cells known to provide protection against autoimmunity. Together, the present disclosure uses a designed logic-gated engineered immune cell, which, in some embodiments, is a Tmod cell that includes a NOT gate receptor system. This Tmod cell therapy provides, on one hand, a highly focused weapon that specifically targets and kills autoreactive T cell subsets and, on the other hand, a mechanism for sparing cells that modulate and / or mitigate an immune response. Thus, the present disclosure, provides a superior cell therapy for autoimmunity.

[0009] An aspect of the present disclosure is a logic-gated engineered immune cell including an activator receptor and an inhibitor receptor, wherein binding of the activator receptor to an activator antigen on a target cell promotes a cytotoxic response by the engineered immune cell against the target cell and wherein binding of the inhibitor receptor to an inhibitor antigen on a second cell protects the second cell and / or the target cell from cytotoxicity mediated by the logic-gated engineered immune cell.

[0010] As used herein “logic-gated engineered immune cells”, “immune cells of the present disclosure”, “immune cells”, “engineered immune cells”, “engineered immune cells of the present disclosure”, and the like are synonymous and refer to the same immune cell population. Thus, for example, embodiments disclosed herein relating to “immune cells” applies, at least, to “logic-gated engineered immune cells”.

[0011] Another aspect of the present disclosure is An engineered immune cell comprising: (a) at least one activator receptor that binds to an activator antigen expressed on an autoreactive target immune cell and promotes a cytotoxic response against the target cell; and (b) at least one inhibitor receptor that binds to an inhibitor antigen expressed on a non-target cell and inhibits cytotoxicity against the non-target cell; wherein the target cell is implicated in an autoimmune disease or disorder.

[0012] Another aspect of the present disclosure is An engineered immune cell comprising an activator receptor or receptors and an inhibitor receptor or receptors, wherein the activator receptor specifically binds to an activator antigen on a target cell; wherein the inhibitor receptor specifically binds to an inhibitor antigen on a non-target cell; and wherein the target cell is an autoreactive target immune cell known to mediate or is implicated in an autoimmune disease.

[0013] In embodiments, the autoreactive target immune cell associated with an autoimmune disease or disorder is an autoreactive target B cell, an autoreactive T cell, or an autoreactive myeloid cell.

[0014] In embodiments, the activator antigen is selected from a group consisting of 4-1BB, CD 19, CD20, CD22, CD69, CD25, and / or 0X40

[0015] In embodiments, the autoreactive target B cell expresses at least one activator antigen selected from: CD19, CD20 and CD22.

[0016] In embodiments, the autoreactive target T cell expresses at least one activator antigen selected from the group consisting of: CD25, CD69, 4-1BB and 0X40.

[0017] In embodiments, the non-target cell comprises a naive T cell, a memory T cell, and / or a regulatory T cell (Treg).

[0018] In embodiments, the non-target cell is a Treg.

[0019] In embodiments, the Treg expresses at least one inhibitor antigen selected from CCR8, LRRC32 (GARP), and CCR7.

[0020] In embodiments, the activator receptor comprises an extracellular ligand binding domain specific for an activator antigen expressed by an autoreactive immune cell, and wherein the extracellular ligand binding domain of the activator receptor comprises a antibody fragment or a single chain Fv antibody fragment (scFv).

[0021] In embodiments, the engineered immune cell comprises two activator receptors wherein a first activator receptor binds to an activator antigen and wherein a second activator receptor binds to a second activator antigen that is different from the first activator antigen.

[0022] In embodiments, wherein the activator receptor is a tandem activator receptor comprising a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell wherein the first extracellular ligand binding domain of the activator receptor comprises a first antibody fragment or first scFv and second extracellular ligand binding domain specific to a second activator antigen expressed by an autoreactive immune cell wherein the second extracellular ligand binding domain of the activator receptor comprises a second antibody fragment or second scFv.

[0023] In embodiments, the first activator antigen is 0X40, wherein the first scFv of the activator receptor specifically binds to 0X40 and comprises a first heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 537, a CDR- H2 of SEQ ID NO: 538, and a CDR-H3 of SEQ ID NO: 539; and a first light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 540, a CDR-L2 of SEQ ID NO: 446, and a CDR-L3 of SEQ ID NO: 541

[0024] In embodiments, the first scFV of the activator receptor comprises a first VH comprising SEQ ID NO: 624 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 624 and / or a first VL comprising SEQ ID NO: 625 or asequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 625.

[0025] In embodiments, wherein the first scFv of the activator receptor comprises SEQ ID NO: 663 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 663.

[0026] In embodiments, the activator receptor comprises SEQ ID NO: 695 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 695.

[0027] In embodiments, wherein the second activator antigen is CD19 wherein the second scFv of the activator receptor specifically binds to CD19 and comprises a second heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 959, a CDR-H2 of SEQ ID NO: 960, and a CDR-H3 of SEQ ID NO: 961; and a second light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 962, a CDR-L2 of SEQ ID NO: 963, and a CDR-L3 of SEQ ID NO: 964.

[0028] In embodiments, the second scFv of the activator receptor comprises a second VH comprising SEQ ID NO: 1001 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1001 and / or a second VL comprising SEQ ID NO: 1002 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1002.

[0029] In embodiments, the second scFv of the activator receptor comprises SEQ ID NO: 1018 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1018.

[0030] In embodiments, the activator receptor comprises SEQ ID NO: 1027 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1027.

[0031] In embodiments, the activator receptor comprises a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

[0032] In embodiments, the activator receptor comprises a transmembrane domain isolated or derived from CD8 or CD28.

[0033] In embodiments, the activator receptor comprises an intracellular domain isolated or derived from CD28, 4- IBB or CD3z, or a combination thereof.

[0034] In embodiments, the inhibitor receptor comprises an extracellular ligand binding domain specific to an inhibitor antigen expressed by a non-target cell and wherein the extracellular ligand binding domain of the inhibitor receptor comprises a antibody fragment or a scFv (single chain Fv antibody fragment ).

[0035] In embodiments, the inhibitor receptor comprises an extracellular ligand binding domain specific to an inhibitor antigen expressed by a non-target cell and wherein the extracellular ligand binding domain of the inhibitor receptor comprises a antibody fragment or a scFv (single chain Fv antibody fragment ).

[0036] In embodiments, the inhibitor antigen is LRRC32 (GARP) wherein the scFv of the inhibitor receptor specifically binds to LRR32(GARP) and comprises a heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 340, a CDR-H2 of SEQ ID NO: 341, and a CDR-H3 of SEQ ID NO: 342; and a light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 343, a CDR-L2 of SEQ ID NO: 344, and a CDR-L3 of SEQ ID NO: 345.

[0037] In embodiments, the scFv of the inhibitor receptor comprises a VH comprising SEQ ID NO: 373 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 373 and / or a VL comprising SEQ ID NO: 374 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 374.

[0038] In embodiments, the scFv of the inhibitor receptor comprises SEQ ID NO: 394 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 394.

[0039] In embodiments, the inhibitor receptor comprises SEQ ID NO: 408 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 408.

[0040] In embodiments, the inhibitor antigen is an HLA antigen to protect the non-target cell from fratricide.

[0041] In embodiments, the HLA antigen is HLA-A*02 or HLA-A*03.

[0042] In embodiments, the inhibitor receptor comprises a leukocyte immunoglobulin like receptor Bl (LILRB1) intracellular domain or a functional variant thereof.

[0043] In embodiments, the LILRB1 intracellular domain comprises SEQ ID NOs: 31 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0044] In embodiments, the inhibitor receptor comprises LILRB1 hinge and LILRB1 transmembrane domains, or functional variants thereof.

[0045] In embodiments, the LILRB1 transmembrane domain comprises SEQ ID NO: 30 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0046] In embodiments, the LILRB1 hinge domain comprises SEQ ID NO: 29, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0047] In embodiments, the LILRB1 intracellular domain, LILRB1 hinge domain, and LILRB1 transmembrane domain together comprises SEQ ID NO: 27 or 28, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0048] In embodiments, the engineered immune is modified to reduce or eliminate expression of a B2M gene product.

[0049] In embodiments, the expression of the B2M gene product is reduced or eliminated using a short hairpin RNA (shRNA).

[0050] In embodiments, the shRNA comprises a first sequence, having from 5' to 3' end a sequence complementary to the B2M gene product; and a second sequence, having from 5' to 3' end a sequence complementary to the first sequence, wherein the first sequence and second sequence form the shRNA.

[0051] In another aspect, the present disclosure relates to a pharmaceutical composition comprising a therapeutically effective amount of the engineered immune cell and a pharmaceutically acceptable excipient, wherein the effective amount is sufficient to treat or ameliorate an autoimmune disease or disorder in a subject in need thereof.

[0052] In an aspect, the present disclosure relates to a method of treating or ameliorating an autoimmune disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the engineered immune cell

[0053] In another aspect, the present disclosure relates to a method of treating or ameliorating an autoimmune disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition.

[0054] In embodiments, the autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti-synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren1s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)),severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

[0055] In an aspect, the present disclosure relates to a polynucleotide or polynucleotide system comprising one or more polynucleotides encoding the activator receptor and the inhibitor receptor of the engineered immune cell.

[0056] In another aspect, the present disclosure relates to a method of making an immune cell therapy, the method comprising transforming immune cells with the polynucleotide or polynucleotide system.

[0057] In an aspect, the present disclosure relates to a nanocarrier comprising the polynucleotide or polynucleotide system.

[0058] In embodiments, the nanocarrier is capable of delivering the polynucleotide or polynucleotide system to an immune cell in vivo or ex vivo.

[0059] In embodiments, the nanocarrier is a lipid nanoparticle (LNP).

[0060] In embodiments, the target cell is an autoreactive target immune cell associated with or causative of an autoimmune disease or disorder. In some cases,

[0061] In embodiments, the autoreactive target immune cell associated with an autoimmune disease or disorder is an autoreactive target B cell, a T cell, or a myeloid cell.

[0062] In embodiments, the autoreactive target T cell is an activated T cell, e.g., an acutely active T cell or a chronically active T cell. In some cases, the autoreactive target T cell is a Thl7 cell.

[0063] In embodiments, the target cell is not a naive T cell, a memory T cell, or a Treg.

[0064] In embodiments, an activator antigen can be any antigen that is typically and / or substantially expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease including Thl7 cells, but not typically expressed by non- autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein, or is minimally expressed by the non-autoreactive cells. In some cases, a cell will express both an activator antigen and an inhibitor antigen.

[0065] In embodiments, the activator antigen is any antigen expressed on the surface of an autoreactive immune cell. In some cases, the activator antigen is 4-1BB, CD19, CD69, CD25, and / or 0X40.

[0066] In embodiments, binding of the activator receptor to the activator antigen on a target cell without binding or with reduced binding of the inhibitor receptor to the inhibitor antigen on the second cell eliminates autoreactive immune cells, thereby treating an autoimmune disease or disorder.

[0067] In embodiments, the second cell is not causative of an autoimmune disease or disorder. In some cases, the second cell includes a naive T cell, a memory T cell, and / or a regulatory T cell (Treg).

[0068] In embodiments, binding of the inhibitor receptor to an inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logic-gated engineered immune cell. In some cases, fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0069] In embodiments, the inhibitor antigen is any antigen expressed on the surface of the second cell and / or the inhibitor antigen is any allele-specific antigen capable of protecting the second cell.

[0070] In embodiments, an inhibitor antigen can be any antigen that is not typically and / or is minimally expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease, but is typically and / or substantially expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein. In some cases, a cell will express both an activator antigen and an inhibitor antigen.

[0071] In embodiments, the allele-specific antigen capable of protecting a second cell is a component of human leukocyte antigen (HLA). In some cases, the HLA is HLA Class I or HLA Class I. In some cases, the HLA Class I includes HLA-A, HLA-B, or HLA-C. In some cases, the HLA-A includes HLA-A*02 or HLA-A*03.

[0072] In embodiments, the inhibitor antigen is HLA-A*02, CCR8, LRRC32 (GARP), and / or CCR7.

[0073] In embodiments, binding of the inhibitor antigen on the second cell reduces surface expression or activity of the activating receptor by the logic-gated engineered immune cell, thereby decreasing the cytotoxic response by the engineered immune cell against the target cell.

[0074] In embodiments, binding of the inhibitor receptor to the inhibitor antigen on the second cell without binding or with reduced binding of the activator receptor to the activator antigenon the target cell spares the target cell and provides protection from autoimmunity, thereby treating an autoimmune disease or disorder.

[0075] In embodiments, the activator receptor and / or the inhibitor receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain specific to the antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0076] In embodiments, the engineered immune cell is a T cell. In some cases, the T cell is a CD8+ CD4- T cell or a CD8- CD4+ T cell. In some cases, the T cell is a cytotoxic T cell.

[0077] In embodiments, the engineered immune cell is a natural killer (NK) cell.

[0078] In embodiments, the logic-gated engineered immune cell is allogenic to a subject with the autoimmune disease or disorder. Alternately, the logic-gated engineered immune cell is autogenic to a subject with the autoimmune disease or disorder.

[0079] Another aspect of the present disclosure is an immune cell including an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell.

[0080] In embodiments, the second cell is not causative of an autoimmune disease or disorder. In some cases, the second cell includes a naive T cell, a memory T cell, and / or a regulatory T cell (Treg) or a cell of another blood lineage.

[0081] In embodiments, binding of the first inhibitor receptor to a first inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logic-gated engineered immune cell. In some cases, fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0082] In embodiments, the first inhibitor antigen is any antigen expressed on the surface of the second cell and / or the first inhibitor antigen is any allele-specific antigen capable of protecting the second cell. In some cases, the first inhibitor antigen is HLA-A*02, CCR8, LRRC32 (GARP), and / or CCR7.

[0083] In embodiments, the inhibitor receptor further includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell.

[0084] In embodiments, binding of the second inhibitor receptor to a second inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logic-gated engineered immune cell. In some cases, fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0085] In embodiments, the second inhibitor antigen is any antigen expressed on the surface of the second cell and / or the second inhibitor antigen is any allele-specific antigen capable ofprotecting the second cell. In some cases, the second inhibitor antigen is HLA-A*02, CCR8, LRRC32 (GARP), and / or CCR7.

[0086] In embodiments, the present disclosure relates to an immune cell, further including an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell.

[0087] In embodiments, the autoreactive immune cell is associated with or causative of an autoimmune disease or disorder.

[0088] In embodiments, the autoreactive immune cell is a B cell, a T cell, or a myeloid cell. In some cases, the T cell is an activated T cell, e.g., an acutely active T cell or a chronically active T cell. In some cases, T cell is a Thl7 cell. The T cell is not a naive T cell, a memory T cell, or a Treg.

[0089] In embodiments, when the second cell expresses the first activator antigen and expresses the first inhibitor antigen, the first inhibitor receptor reduces the expression (e.g., surface expression) or activity of the activator receptor.

[0090] In embodiments, the first activator antigen is selected from 4-1BB, CD19, CD69, CD25, and 0X40.

[0091] In embodiments, the first activator antigen is CD69 and the first inhibitor antigen is HLA-A*02.

[0092] In embodiments, the first activator antigen is CD25 and the first inhibitor antigen is HLA-A*02.

[0093] In embodiments, the first activator antigen is 0X40 and the first inhibitor antigen is HLA-A*02.

[0094] In embodiments, (a) the first activator antigen is CD25 and the first inhibitor antigen is HLA-A*02 and the second inhibitor antigen is CCR8 or (b) the first activator antigen is CD25 and the first inhibitor antigen is CCR8 and the second inhibitor antigen is HL A- A* 02.

[0095] In embodiments, the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell.

[0096] In embodiments, the second activator antigen is selected from 4-1BB, CD19, and 0X40.

[0097] In embodiments, (a) the first activator antigen is 4- IBB and the second activator antigen is CD 19 or (b) the first activator antigen is CD 19 and the second activator antigen is 4- IBB.

[0098] In embodiments, (a) the first activator antigen is 0X40 and the second activator antigen is CD 19 or (b) the first activator antigen is CD 19 and the second activator antigen is 0X40.

[0099] In embodiments, the inhibitor receptor and / or the activator receptor is a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain specific to the activator antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0100] In embodiments, the immune cell is allogenic to a subject with the autoimmune disease or disorder, or the immune cell is autogenic to a subject with the autoimmune disease or disorder.

[0101] In an aspect, the present disclosure relates to a logic-gated engineered immune cell or the immune cell for use in a method of treating an autoimmune disease or disorder.

[0102] In another aspect, the present disclosure relates to a pharmaceutical composition, including an effective amount of herein-disclosed logic-gated engineered immune cells or immune cells and a pharmaceutically-acceptable excipient.

[0103] In a further aspect, the present disclosure relates to a kit including the logic-gated engineered immune cell or the immune cell, or pharmaceutical compositions comprising the cells, and instructions for use.

[0104] In an additional aspect, the present disclosure relates to methods for treating an autoimmune disease or disorder comprising administering an effective amount of herein- disclosed logic-gated engineered immune cell, herein-disclosed immune cells, or herein- disclosed pharmaceutical compositions comprising the cells.

[0105] In embodiments, the autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti-synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren' s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis. In some cases, the autoimmune disease or disorder is multiple sclerosis (MS).

[0106] In yet another aspect, the present disclosure relates to a polynucleotide or polynucleotide system, comprising one or more polynucleotides, encoding the activator receptor and / or the inhibitor receptor of a herein-disclosed logic-gated engineered immune cell or of a herein-disclosed immune cell.

[0107] In yet a further aspect, the present disclosure relates to a nanocarrier including the polynucleotide or polynucleotide system.

[0108] In yet an additional aspect, the present disclosure relates to a nanocarrier, wherein the nanocarrier is capable of delivering the polynucleotide or polynucleotide system to an immune cell in vivo or ex vivo.

[0109] An aspect of the present disclosure relates to a nanocarrier, wherein the nanocarrier is a lipid nanoparticle (LNP).

[0110] Another aspect of the present disclosure relates to a nanocarrier, wherein the polynucleotide or polynucleotide system are one or more messenger ribonucleic acids (mRNAs) or modified mRNAs (mmRNAs).

[0111] Any immune cell, composition, or method disclosed herein is applicable to any herein- disclosed immune cell, composition, or method. In other words, any aspect or embodiment described herein can be combined with any other aspect or embodiment as disclosed herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0112] FIG. 1A and FIG. IB are illustrative designs of cellular logic systems to integrate multiple-antigen inputs. FIG. 1A shows Venn diagrams depicting classes of multi-antigen receptor systems that have been constructed in immune effector cells to integrate inputs (logic gates). Teal represents the antigen combination on target cells that activates killing by the engineered immune cells. In the present disclosure, the second cells, as used herein, comprise cells that modulate and / or mitigate an immune response, e.g., a regulatory T cell (Treg). FIG. IB depicts an illustrative NOT-gate engineered receptor system, cells, receptors, and antigens involved in autoimmune cell-selective killing. The engineered receptor NOT-gate system incorporates an inhibitor receptor which inhibits T cell activation when engaged with the inhibitor antigen present on second cells (e.g., Treg) but not when the inhibitor antigen is absent on autoimmune cells.

[0113] FIG. 2 is a schematic of an illustrative algorithm to identify A and B antigens for activator and inhibitor receptors for autoimmune diseases. ON indicates the antigen is express. OFF indicates the antigen is not expressed. Illustrative initial antigen candidates are shown.

[0114] FIG. 3 are graphs depicting A antigens CD25, CD69, 4- IBB and 0X40 measured in transcripts per million (TPM) in activated T cells.

[0115] FIG. 4 is a table showing the induction ratio for 0X40, CD25, 4- IBB and CD69 in activated T cells from two different RNA Sequencing data sets. Activator antigen candidates 0X40 and CD25 are highly induced upon T cell activation.

[0116] FIG. 5 is a table showing the median and mean expression level of inhibitor antigen candidates LRRC32 and CCR8. LRRC32 and CCR8 expression was measured in activated T cells, memory T cells, TH17 cells, Treg cells, and B cells.

[0117] FIG. 6 is a set of graphs showing CD25, 0X40, GARP, CD69 expression kinetics and molecules per cell values on human peripheral blood mononuclear cells (PBMCs). Expression of CD25, 0X40, CD69 and GARP on was detectable on Days 1-7 on CD3+ T cells.

[0118] FIG. 7 are a set of flow cytometry plots showing 0X40 and GARP expression post CD3 / CD28 stimulation on CD25+FOXP3+ T regulatory cells (Tregs).

[0119] FIG. 8 is a table showing Fold activation, IC50 and Percentage Block analysis in Jurkat mRNA titration assay. K562 target cells were transfected with constant amount of 0X40 mRNA and serially diluted GARP mRNA. Jurkat NF AT Luciferase (JNL) effector cells were transfected with 0X40 CARs and GARP blockers.

[0120] FIG. 9 is a table from a cell kill assay showing ET50 and selectively windows of primary T cells expressing the indicated receptors co-cultured with HeLA target cells. The number of repeats for each group tested is indicated in the chart.

[0121] FIG. 10 are a series of graphs showing specific killing percentage of NALM / 6 target cells by primary T cells expressing the indicated receptors.

[0122] FIG. 11 is a set of graphs showing specific killing of CD 19+ primary B cells and 0X40+ B cells by primary T cells expressing the indicated receptors.DETAILED DESCRIPTION

[0123] Provided herein are engineered immune cells (e.g., CAR-T), compositions, and methods for targeting specific antigens on immune cells comprising an optimized two receptor system responsive to differences in antigens expressed on a target cell (e.g., an autoreactive immune cell) or on a second cell (e.g., a cell that modulate and / or mitigates an immune reaction; for example, a Treg cell).

[0124] An aspect of the present disclosure is a logic-gated engineered immune cell including an activator receptor or receptors and an inhibitor receptor or receptors, wherein binding of the activator receptor to an activator antigen on a target cell promotes a cytotoxic response by the engineered immune cell against the target cell and wherein binding of the inhibitor receptor to an inhibitor antigen on a second cell protects the second cell and / or the target cell from cytotoxicity mediated by the logic-gated engineered immune cell.

[0125] Another aspect of the present disclosure is an immune cell including an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell. In embodiments, the inhibitor receptor further includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell. In embodiments, the immune cell includes an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell. In embodiments, the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell.

[0126] As used herein “logic-gated engineered immune cells”, “immune cells of the present disclosure”, “immune cells”, “engineered immune cells”, “engineered immune cells of the present disclosure”, and the like are synonymous and refer to the same immune cell population. Thus, for example, embodiments disclosed herein relating to “logic-gated engineered immune cells” applies to “immune cells of the present disclosure”.

[0127] As used herein, the term “autoreactive immune cell” refers to an immune cell that expresses an antigen receptor — such as a T cell receptor (TCR) or B cell receptor (BCR) — with specificity for a self-antigen. Upon engagement with the self-antigen, the autoreactive immune cell may undergo activation, proliferation, or exert effector functions (e.g., cytokine secretion, cytotoxic activity) in the absence of exogenous or foreign antigen stimulation. Such cells are implicated in the initiation or propagation of self-directed immune responses and may contribute to the pathogenesis of autoimmune diseases or disorders. Examples of autoreactive immune cells include but are not limited to T lymphocytes and B lymphocytes.

[0128] As disclosed further herein, the engineered cells, compositions, and methods which combine the power of NOT gated T cells with target antigens that include surface antigens expressed on activated T cells and which may be associated with autoimmune disease pathology, e.g., 4- IBB, CD69, 0X40 and CD25. Furthermore, the engineered cells interact with antigens present on, at least, Tregs, which are cells that play a role in regulating the immune response to normal self-tissues. Thus, the present disclosure is unique in that it provides cell therapies target B cells and / or subsets of activated T cells and takes advantage of antigens that are expressed on Tregs and absent from inflammatory cell types, such as Thl7s. More specifically, the Tmod NOT gates used herein, use specific pairs of antigens depending on their role in autoimmunity: the activator antigens that confer selectivity on the autoreactive T cell subsets that must be eliminated and the inhibitor antigens that confer selectivity on the T cell subsets that must be protected. Notably, a majority of known T cell surface antigens do not distinguish protective Tregs from the pro-inflammatory Thl 7s. However, as described herein, the antigens CCR8 and LRRC32 are expressed on Tregs but not on Thl7s and, thus, are suitable as inhibitor antigens, as used herein, for a Tmod cell therapy product designed to effectively and specifically treat autoimmune diseases. Accordingly, the present disclosure provides a superior cell therapy for autoimmunity that removes immune cells involved in the autoimmune inflammatory response; e.g., B cells and activated T cells, while sparing cells that modulate and / or mitigate the autoimmune inflammatory response, e.g., Tregs.NOT gate logic and Tmods

[0129] Provided herein are methods to target and bind sets of activator (A) antigens and inhibitor (B) antigens for use in the treatment of autoimmune diseases or disorders. Further provided are immune cells that integrate, through selective binding, signals provided by sets of A antigens and B antigens. The immune cells of the disclosure operate as a “logic gate” wherein signals from A antigens and B antigens together trigger a desired response by the immune cell.

[0130] An activator antigen can be any antigen that is typically and / or substantially expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease including Thl7 cells, but not typically expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein, or is minimally expressed by the non-autoreactive cells. In contrast, an inhibitor antigen can be any antigen that is not typically and / or is minimally expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease including Thl7 cells, but is typically and / or substantially expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein. In some cases, a cell will express both an activator antigen and an inhibitor antigen. In this case, as discussed below, activity of an immune cell of the present disclosure may be blocked or reduced by the presence of the inhibitor antigen, even in the presence of the activator antigen; thus, reduces or preventing an undesirable cytotoxic attack on the cell expressing both antigens.

[0131] Immune cells of the present disclosure are engineered to specifically activate when contacted with autoreactive cells. The immune cells do not activate when contacted with second cells, as herein described, i.e., normal cells, e.g., inactive T cells, naive T cells, regulatory T cells (Tregs). Accordingly, adverse side effects to the second cells caused by damage by the engineered immune cell may be reduced. Sets of A antigens and B antigens, where A antigens activate the engineered immune cells and B antigens inhibit them, may beused to selectively target autoreactive cells using the synthetic signal-integration systems of the present disclosure.

[0132] FIG. 1A and FIG. IB are illustrative designs of cellular logic systems to integrate multiple-antigen inputs. FIG. 1A shows Venn diagrams depicting classes of multi-antigen receptor systems that have been constructed in immune effector cells to integrate inputs (logic gates). Teal represents the antigen combination on target cells that activates killing by the engineered immune cells. In the present disclosure, the second cells, as used herein, comprise cells that modulate and / or mitigate an immune response, e.g., inactive T cells, naive T cells, and Tregs.

[0133] The cells, compositions, and method of the present disclosure are most relevant to the “NOT” embodiment shown at the right end of FIG. IB. In this case, a target cell, i.e., an autoreactive cell, expresses an activator antigen (A-Ag) which is targeted by an activator receptor expressed by an immune cell of the present disclosure. In most cases, as shown in the top left region, the target cell does not express the inhibitor receptor (B-Ag). Thus, the immune cell of the present disclosure kills the target cell. On the other hand, in the cases of the bottom right region, the second cell (e.g., an inactive T cell, naive T cell, or Treg) expresses an inhibitor antigen (B-Ag) which is targeted by an inhibitor receptor expressed by the immune cell of the present disclosure. The inhibitor receptor recognizes the binding and inhibits a cytotoxic attack upon the second cell, thereby sparing the second cell. In some cases, the spared second cell is allowed to modulate and / or mitigate and further reduce the activity of the autoreactive cell. In the overlapping region, the target / second cell expresses both the A-Ag and the B-Ag. However, the activity of the inhibitor antigen blocks the activity and / or surface expression of the activator receptor to reduce or prevent the cytotoxic attack on the target / second cell.

[0134] In some cases, the overlapping region of the “NOT” embodiment represents a situation during manufacturing of the immune cells of the present disclosure. Although Chimeric Antigen Receptors (CAR) are a promising and effective technology for adoptive T cell therapy, and especially for treating autoimmune diseases, using CAR-T cells against autoimmune diseases is frequently challenging because of shared expression of most surface antigens between normal and autoimmune T cells. This often leads to T-cell fratricide during CAR-T cell production. T cell fratricide during T cell production can lead to challenges in harvesting the number of autologous T cells needed to treat a patient. During manufacture, one immune cell of the present disclosure may contact a sibling immune cell. Should the sibling cell express the A- Ab, the immune cell would normally kill that sibling, i.e., fratricide. However, given the dual-nature of the immune cells of the present disclosure, the presence of a B-Ag (e.g., HLA-A*02) on the sibling cell will also be recognized by the immune cell of the present disclosure; binding to the B-Ag (e.g., HLA-A*02) on the sibling will block or reduce the cytotoxicity of the immune cell; thereby preventing fratricide and sparing the sibling immune cell (of the present disclosure) and allowing its survival and increasing T cell production for allogenic treatments.

[0135] The set of A antigens may comprise a single member A, or the set of A antigens may comprise n antigens denoted Ai, A2, A„. The set of B antigens may comprise a single member B, or the set of B antigens may comprise n antigens denoted Bi, B2, . . B„.

[0136] Logic gates may include OR-NOT, AND-NOT, and other variations. For example, the disclosure contemplates the following, non-limiting, list of logic gates:

[0137] A NOT B

[0138] (Ai OR A2) NOT B

[0139] (Ai AND A2) NOT B

[0140] (Ai OR A2) NOT (BI OR B2)

[0141] (Ai OR A2) NOT (BI AND B2)

[0142] A NOT (Bi OR B2)

[0143] A NOT (Bi AND B2)

[0144] NOT logic may be provided by, for example and without limitation, use of an inhibitor receptor that specifically binds a B antigen; when activated, the inhibitor receptor provides a signal that blocks the activator receptor.

[0145] OR logic may be provided by pairs of activator receptors or pairs of inhibitor receptors. For example, an (Ai OR A2) signal may be provided by a first activator receptor that specifically binds Ai and a second activator receptor that specifically binds A2. Activation of an (Ai OR A2) activator receptor occurs upon contact with a cell expressing either antigen Ai or antigen A2, or both. Likewise, a NOT (Bi OR B2) signal may be provided by a first inhibitor receptor that specifically binds Bi and a second inhibitor receptor that specifically binds B2. Activation of a (Bi OR B2) inhibitor receptor occurs upon contact with a cell expressing either antigen Bi or antigen B2, or both.

[0146] In a variation, OR logic may be provided by a single receptor that specifically binds two antigens. For example, an (Ai OR A2) signal may be provided by an activator receptor that specifically binds Ai and specifically binds A2. Likewise, a NOT (Bi OR B2) signal may be provided by an inhibitor receptor that specifically binds Bi and specifically binds B2.

[0147] AND logic may be provided by two receptors that each individually activate in response to an antigen but provide an activator (or inhibitor) signal that is below a threshold foractivation (or inhibition) of the immune cell. For example, an (Ai AND A2) signal may be provided by a first activator receptor that specifically binds Ai and a second activator receptor that specifically binds A2, where either activator signal alone is too weak to activate the immune cell. Likewise, a NOT (Bi AND B2) signal may be provided by a first inhibitor receptor that specifically binds Bi and a second inhibitor receptor that specifically binds B2, where either inhibitor signal alone is too weak to block the activation of the immune cell.

[0148] In a variation, a single activator (or inhibitor) receptor may be used, where the single receptor is activated only when contacted by both antigens. For example, an (Ai AND A2) signal may be provided by an activator receptor that specifically binds Ai and that specifically binds A2, where the activator receptor only activates when it binds both Ai and A2. Likewise, a NOT (Bi AND B2) signal may be provided by an inhibitor receptor that specifically binds Bi and that specifically binds B2, where the inhibitor receptor only activates, to block an activator signal, when it binds both Bi and B2.

[0149] Selection of suitable A antigens and B antigens maybe according to the below algorithm to find antigens with properties necessary for a NOT gated cell therapy for autoimmune disease.

[0150] The activator antigens (A-Ags) and inhibitor antigens (B-Ags) preferably have the following profiles:A-Ags (have the following profile):1. ON in activated T cells;2. OFF in resting T cells (including memory and naive T cells); and3. OFF in other cell types; but if not, then:B-Ags (have the following profile):1. ON in those cell types where given A-Ag expressed;2. OFF in activated T cells;3. OFF in Th 17; and4. ON in Tregs

[0151] Illustrative, non-limiting, A and B antigens useful in logic gates of the present disclosure are shown in Table I. Illustrative, non-limiting, examples of A and B antigen combinations in logic gates contemplated by the present disclosure are shown in Table II. See also FIG 2.Table I: Illustrative A and B antigensTable II: Illustrative A and B antigen combinations

[0152] Any combination of a single A antigen may be combined with any single B antigen is considered. As examples, the combination may include CD69 with CCR7; CD69 with CCR8; CD69 with an HLA complex; CD69 with LRRC32; CD69 with HLA-A*02; CD 19 with CCR7; CD 19 with CCR8; CD 19 with an HLA complex; CD 19 with LRRC32; CD 19 with HLA-A*02; 4-1BB with CCR7; 4-1BB with CCR8; 4-1BB with an HLA complex; 4-1BB with LRRC32; 4- IBB with HLA-A*02; CD25 with CCR7; CD25 with CCR8; CD25 with an HLA complex; CD25 with LRRC32; CD25 with HLA-A*02; 0X40 with CCR7; 0X40 with CCR8; 0X40 with an HLA complex; 0X40 with LRRC32; and 0X40 with HLA-A*02.

[0153] Any two A antigens may be combined with any single B antigen. As examples, 4-1BB and CD19 with CCR7; 4-1BB and CD19 with CCR8; 4-1BB and CD19 with an HLA complex; 4- 1BB and CD19 with LRRC32 (GARP); 4-1BB and CD19 with HLA-A*02; CD19 and 0X40 with CCR7; CD 19 and 0X40 with CCR8; CD 19 and 0X40 with an HLA complex; CD 19 and 0X40 with LRRC32 (GARP); CD19 and 0X40 with HLA-A*02; 4-1BB and CD69 with CCR7; 4-1BB and CD69 with CCR8; 4- IBB and CD69 with an HLA complex; 4- IBB and CD69 with LRRC32 (GARP); 4-1BB and CD69 with HLA-A*02; 4-1BB and CD25 with CCR7; 4-1BB and CD25 with CCR8; 4-1BB and CD25 with an HLA complex; 4-1BB and CD25 with LRRC32 (GARP); 4-1BB and CD25 with HLA-A*02; 4-1BB and 0X40 with CCR7; 4-1BB and 0X40 with CCR8; 4-1BB and 0X40 with an HLA complex; 4- IBB and 0X40 with LRRC32 (GARP); 4- IBB and 0X40with HLA-A*02; CD25 and 0X40 with CCR7; CD25 and 0X40 with CCR8; CD25 and 0X40 with an HLA complex; CD25 and 0X40 with LRRC32 (GARP); or CD25 and 0X40 with HLA- A*02.

[0154] A single A antigen may be combined with any two B antigens. As examples, CD 19 with CCR7 and CCR8; CD 19 with CCR7 and an HLA complex; CD 19 with CCR7 and LRRC32; CD 19 with CCR7 and HLA-A*02; CD 19 with CCR8 and CCR7; CD 19 with CCR8 and an HLA complex; CD 19 with CCR8 and LRRC32; CD 19 with CCR8 and HLA-A*02; CD19 with an HLA complex and CCR7; CD19 with an HLA complex and CCR8; CD19 with an HLA complex and LRRC32; CD 19 with an HLA complex and HLA-A*02; CD 19 with LRRC32 and CCR7; CD 19 with LRRC32 and CCR8; CD 19 with LRRC32 and an HLA complex; CD 19 with LRRC32 and HLA-A*02; CD 19 with HLA-A*02 and CCR7; CD 19 with HLA-A*02 and CCR8; CD 19 with HLA-A*02 and an HLA complex; or CD 19 with HLA - *02 and LRRC32.

[0155] Additionally, any two A antigens may be combined with any two B antigens. As examples, 4-1BB and CD19 with HLA-A*02 and CCR8; 4-1BB and CD19 with HLA-A*02 and CCR7; 4-1BB and CD19 with HLA-A*02 and LRRC32; 4-1BB and CD19 with an HLA complex and CCR8; 4- IBB and CD 19 with CCR8 and CCR7; 4- IBB and CD 19 with CCR8 and LRRC32; CD 19 and 0X40 with an HLA complex and CCR8; CD 19 and 0X40 with CCR8 and CCR7; CD 19 and 0X40 with CCR8 and LRRC32; CD 19 and 0X40 with HLA-A*02 and LRRC32; CD 19 and 0X40 with HLA-A*02 and CCR7; CD 19 and 0X40 with HLA-A*02 and CCR8; CD69 and 0X40 with an HLA complex and CCR8; CD69 and 0X40 with CCR8 and CCR7; CD69 and 0X40 with CCR8 and LRRC32; CD69 and 0X40 with HLA-A*02 and LRRC32; CD69 and 0X40 with HLA-A*02 and CCR7; CD69 and 0X40 with HLA-A*02 and CCR8; CD25 and 0X40 with an HLA complex and CCR8; CD25 and 0X40 with CCR8 and CCR7; CD25 and 0X40 with CCR8 and LRRC32; CD25 and 0X40 with HLA-A*02 and LRRC32; CD25 and 0X40 with HLA-A*02 and CCR7; CD25 and 0X40 with HLA-A*02 and CCR8; 4-1BB and 0X40 with HLA-A*02 and CCR8; 4-1BB and 0X40 with HLA-A*02 and CCR7; 4-1BB and 0X40 with HLA-A*02 and LRRC32; 4-1BB and 0X40 with an HLA complex and CCR8; 4-1BB and 0X40 with CCR8 and CCR7; or 4-1BB and 0X40 with CCR8 and LRRC32.

[0156] Any combination of A and B or multiples of As and multiples of Bs is considered herein.Inhibitor Receptors and Activator Receptors

[0157] An aspect of the present disclosure is a logic-gated engineered immune cell including an activator receptor and an inhibitor receptor, wherein binding of the activator receptor to an activator antigen on a target cell promotes a cytotoxic response by the engineered immune cell against the target cell and wherein binding of the inhibitor receptor to an inhibitor antigen on a second cell protects the second cell and / or the target cell from cytotoxicity mediated by the logic-gated engineered immune cell. In embodiments, the activator receptor and / or the inhibitor receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain specific to the antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0158] Another aspect of the present disclosure is an immune cell including an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell. In embodiments, the inhibitor receptor further includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell. In embodiments, the present disclosure relates to an immune cell, further including an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell. In embodiments, the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell. In embodiments, the activator receptor and / or the inhibitor receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain(s) specific to the antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0159] The term “inhibitor chimeric antigen receptor” or “inhibitor receptor” as used herein refers to an antigen-binding domain that is fused to an intracellular signaling domain capable of transducing an inhibitory signal that inhibits or suppresses the immune activity of an immune cell. Inhibitor receptors have immune cell inhibitory potential, and are distinct and distinguishable from CARs, which are receptors with immune cell activating potential. For example, CARs having activator receptors include intracellular stimulatory and / or costimulatory domains. Inhibitor receptors contain intracellular inhibitory domains.

[0160] As used herein “inhibitory signal” refers to signal transduction or changes in protein expression in an immune cell resulting in suppression of an immune response (e.g., decrease in cytokine production or reduction of immune cell activation). Inhibition or suppression of an immune cell can be selective and / or reversible, or not selective and / or reversible. Inhibitorreceptors are responsive to antigens (e.g., CD16 or HLA-A*02) expressed by immune modulatory cell or normal (non-autoreactive) allogeneic cells. For example, when a second cell, e.g., a Treg, binds to or contacts the inhibitor receptor, the inhibitor receptor is responsive and initiates an inhibitory signal in the immune cell expressing the inhibitor receptor.

[0161] Inhibitor receptors of the disclosure may comprise an extracellular ligand binding domain that binds to an inhibitor antigen relevant to the present disclosure.

[0162] As used herein, an “activator,” “activator antigen,” or “activator ligand” refers to a first antigen that binds to an activator antigen binding domain (LBD) of an engineered receptor of the disclosure, such as a CAR or TCR, thereby mediating activation of an immune cell expressing the activator receptor. When a target activator antigen is bound to activator receptor, a signal may be generated that causes activation of an immune cell expressing the activator receptor. As used herein, “activation” of an immune cell or an immune cell that is “activated” refers to an immune cell that can carry out one or more functions characteristic of an immune response. These functions include proliferation, release of cytokines, and cytotoxicity, i.e., killing of a target cell. Activated immune cells express markers that will be apparent to persons of skill in the art. For example, activated T cells can express one or more of CD69 and CD25. An immune cell expressing an activator receptor can be activated by the activator receptor when it becomes responsive to the binding of the receptor to a target antigen expressed by the target cell. A “target antigen” can also be referred to an “activator antigen” and may be isolated or expressed by a target cell.

[0163] Activator receptors of the disclosure may comprise an extracellular ligand binding domain that binds to an activator antigen relevant to the present disclosure.

[0164] Activation of an immune cell expressing an inhibitor receptor can be prevented when the inhibitor receptor becomes responsive to an inhibitor antigen on a cell, even when the activator receptor is bound to the target activator antigen.

[0165] Responsiveness of receptors, and their ability to activate or inhibit immune cells expressing the receptors, can be assayed by any means known in the art and described herein, including, but not limited to, reporter assays and cytotoxicity assays.Illustrative inhibitor antigensCCR7

[0166] In some embodiments, the inhibitor antigen is CCR7.

[0167] The term “CCR7” (also known as C-C Motif Chemokine Receptor 7, CDwl97, BLR2, CMKBR7, EBI1, Epstein-Barr Virus-Induced G-Protein Coupled Receptor 1 and MIP-3 beta receptor) refers to a member of the beta chemokine receptor family. CCR7 can be expressedby B cells, mature dendritic cells (DCs) and by several “second cell” T-cell sub-populations including naive, regulatory and central memory T cells.

[0168] CCR7 and its ligands (CCL19 and CCL21) may play a role in proper recruitment of lymphocytes and matures dendritic cells to lymphoid tissues. CCR7 was identified as a gene induced by the Epstein-Barr virus (EBV), and is thought to be a mediator of EBV effects on B lymphocytes

[0169] There are several CCR7 variants encoding distinct isoforms, all of which fall within the scope of the instant disclosure.CCR8

[0170] In some embodiments, the inhibitor antigen is CCR8.

[0171] The term “CCR8” (also known as C-C Motif Chemokine Receptor 8, TERI, GPR-CY6, CMKBRL2, CKR L1, CDwl98, CMKBR8 and CY6) refers to a member of the beta chemokine receptor family. CCR8 is found mainly on the surface of regulatory T cells and may be involved in immune suppression.

[0172] CCR8 and its ligand CCL1 may play a role in regulating the recruitment and function of Tregs within the tumor microenvironment (TME). CCR8 may be expressed on the surface of Tregs, while CCL1 is a chemokine that may be secreted by various cells within the TME. Binding of CCL1 to CCR8 on Tregs may promote the migration of regulatory immune cells to the site of CCL1, which is often the tumor site.

[0173] There are several CCR8 variants encoding distinct isoforms, all of which fall within the scope of the instant disclosure.LRRC32

[0174] In some embodiments, the inhibitor antigen is LRRC32.

[0175] The term “LRRC32” (also known as leucine rich repeat containing 32, glycoprotein A repetitions predominant, D11S833E, GARP, transforming growth factor beta activator LRRC32, garpin, CPPRDD) refers to a type I membrane protein which contains 20 leucine- rich repeats. LRRC32 consists of 662 aa and encodes an 80-kDa transmembrane protein with an extracellular region composed primarily of 20 leucine-rich repeats. LRRC32 may act as a TGFP cell surface receptor involved in the immune response.

[0176] LRRC32 may be responsible for cell-surface docking of latent TGFP before activation and release of the mature cytokine, and may have a role in maintaining peripheral tolerance and preventing inflammatory diseases.

[0177] There are several LRRC32 variants encoding distinct isoforms, all of which fall within the scope of the instant disclosure.CCR7

[0178] In some embodiments, the inhibitor antigen is CCR7.

[0179] The term “CCR7” (also known as C-C chemokine receptor type 7, BLR2, CC-CKR-7, CCR-7, CD197, CDwl97, CMKBR7, EBI1) is a seven transmembrane protein which is coupled with heterotrimeric G proteins. A main function of the CCR7 receptor is to guide immune cells to immune organs (lymph nodes, thymus, spleen) by detecting specific chemokines, which these tissues secrete. CCR7 affects not only central tolerance but also peripheral tolerance.

[0180] There are several CCR7 variants encoding distinct isoforms, all of which fall within the scope of the instant disclosure.HLA-A*02

[0181] In some embodiments, the inhibitor antigen is HLA-A *02.

[0182] The term “HLA-A*02” (also known as A*02) is a human leukocyte antigen (HLA) serotype within the HLA-A serotype group. The serotype is determined by the antibody recognition of the a2 domain of the HLA-A a-chain. For A*02, the a chain is encoded by the HLA-A*02 gene and the P chain is encoded by the B2M locus. HLA antigens are found on the surface of most cells in the body, and they play an important role in the body's immune response to foreign substances.

[0183] There are hundreds of A*02 variants encoding distinct isoforms, all of which fall within the scope of the instant disclosure.Illustrative activator antigensTNF receptor superfamily member 9 (TNFRSF9)

[0184] The activator receptors disclosed can specifically bind to a TNFRSF9 antigen.

[0185] The term “TNFRSF9” also known as TNF receptor superfamily member 9, ILA, 4- 1BB, CD137, CDwl37, IMD109 and other aliases refers to human TNFRSF9 protein and species, isoforms, and other sequence variants thereof. Thus, TNFRSF9 can be the native, full- length protein or can be a truncated fragment or a sequence variant (e.g., a naturally occurring isoform, or recombinant variant) that retains at least one biological activity of the native protein. TNFRSF9 is a member of the TNF-receptor superfamily. The TNFRSF9 receptor may contribute to the clonal expansion, survival, and development of T cells. It may also induce proliferation in peripheral monocytes, enhance T cell apoptosis induced by TCR / CD3 triggered activation, and regulate CD28 co-stimulation to promote Thl cell responses. The expression of TNFRSF9 may be induced by lymphocyte activation.

[0186] In some embodiments, the target antigen is a peptide antigen of TNFRSF9.IL2RA (CD25)

[0187] The activator receptors disclosed can specifically bind to a CD25 antigen.

[0188] The term “IL2RA” also known as interleukin 2 receptor subunit alpha, IL-2 receptor subunit alpha, TAC antigen, CD25, IDDM10, IL2R, P55, IMD41, TCGFR and other aliases refers to human IL2RA protein and species, isoforms, and other sequence variants thereof. Thus, IL2RA can be the native, full-length protein or can be a truncated fragment or a sequence variant (e.g., a naturally occurring isoform, or recombinant variant) that retains at least one biological activity of the native protein. IL2RA is a protein that may be involved in the assembly of the high-affinity Interleukin-2 receptor, consisting of alpha (IL2RA), beta (IL2RB) and the common gamma chain (IL2RG). The receptor may interact with Interleukin-2, a pleiotropic cytokine which can play an important role in immune homeostasis.

[0189] In some embodiments, the target antigen is a peptide antigen of IL2RA. CD69

[0190] The activator receptors disclosed can specifically bind to a CD69 antigen.

[0191] The term “CD69” also known as CLEC2C, early T-cell activation antigen P60, leukocyte surface antigen Leu-23, BL-AC / P26, GP32 / 28, MLR-3, AIM, EA1, C-type lectin domain family 2, activator inducer molecule (AIM / CD69) and other aliases refers to human CD69 protein and species, isoforms, and other sequence variants thereof. Thus, CD69 can be the native, full-length protein or can be a truncated fragment or a sequence variant (e.g., a naturally occurring isoform, or recombinant variant) that retains at least one biological activity of the native protein. CD69 is a human transmembrane C-Type lectin protein. CD69 may be expressed as an activation marker in hematopoietic stem cells, T cells, and many other cell types in the immune system. The activation of T lymphocytes and Natural Killer (NK) cells, both in vivo and in vitro, may induce CD69 expression. CD69, a cell surface glycoprotein expressed during lymphoid activation, may be involved in lymphocyte proliferation, and may function as a signal-transmitting receptor in lymphocytes, including natural killer (NK) cells, and platelets.

[0192] In some embodiments, the target antigen is a peptide antigen of CD69.0X40 (TNRSF4)

[0193] The activator receptors disclosed can specifically bind to a 0X40 antigen.

[0194] The term “0X40” also known as Tumor Necrosis Factor Receptor Superfamily Member 4 (TNFRSF4), TXGP1L, ACT35, CD 134, TAX transcriptionally-activated glycoprotein 1 receptor, OX40L receptor, Lymphoid Activation Antigene ACT35, IMD16, andother aliases refers to human 0X40 protein and species, isoforms, and other sequence variants thereof. Thus, 0X40 can be the native, full-length protein or can be a truncated fragment or a sequence variant (e.g., a naturally occurring isoform, or recombinant variant) that retains at least one biological activity of the native protein. 0X40, a cell surface glycoprotein and member of the tumor necrosis factor (TNF) receptor family, may be expressed by CD4 T cells and may provide a costimulatory signal for T cell activation. 0X40 has been shown to activate NF-kappaB through its interaction with adaptor proteins TRAF2 and TRAF5. OX40L may bind to 0X40 receptors on T-cells, preventing them from dying and subsequently increasing cytokine production. 0X40 may play a role in maintenance of an immune response beyond the first few days and onwards to a memory response due to its ability to enhance survival.

[0195] In some embodiments, the target antigen is a peptide antigen of 0X40.Logic-gated engineered immune cells

[0196] An aspect of the present disclosure is a logic-gated engineered immune cell including an activator receptor or receptors and an inhibitor receptor or receptors, wherein binding of the activator receptor to an activator antigen on a target cell promotes a cytotoxic response by the engineered immune cell against the target cell and wherein binding of the inhibitor receptor to an inhibitor antigen on a second cell protects the second cell and / or the target cell from cytotoxicity mediated by the logic-gated engineered immune cell.

[0197] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein a target cell is an autoreactive target immune cell associated with or causative of an autoimmune disease or disorder.

[0198] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the autoreactive target immune cell associated with an autoimmune disease or disorder is an autoreactive target B cell, a T cell, or a myeloid cell.

[0199] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the autoreactive target T cell is an activated T cell, e.g., an acutely active T cell or a chronically active T cell.

[0200] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the autoreactive target T cell is a Thl7 cell.

[0201] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the target cell is not a naive T cell, a memory T cell, or a Treg.

[0202] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the activator antigen is any antigen expressed on the surface of an autoreactive immune cell.

[0203] In embodiments, an activator antigen can be any antigen that is typically and / or substantially expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease including Thl7 cells, but not typically expressed by non- autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein, or is minimally expressed by the non-autoreactive cells. In some cases, a cell will express both an activator antigen and an inhibitor antigen.

[0204] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the activator antigen is 4- IBB, CD 19, CD20, CD22 CD69, CD25, and / or 0X40.

[0205] In embodiments, the activator receptor and / or the inhibitor receptor is a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain(s) specific to the antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0206] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the CAR includes an extracellular ligand binding domain specific to the activator antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0207] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein binding of the activator receptor to the activator antigen on a target cell without binding or with reduced binding of the inhibitor receptor to the inhibitor antigen on the second cell eliminates autoreactive immune cells, thereby treating an autoimmune disease or disorder.

[0208] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the second cell is not causative of an autoimmune disease or disorder.

[0209] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the second cell includes a naive T cell, a memory T cell, and / or a regulatory T cell (Treg).

[0210] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein binding of the inhibitor receptor to an inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logicgated engineered immune cell.

[0211] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0212] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the inhibitor antigen is any antigen expressed on the surface of the second cell.

[0213] In embodiments, an inhibitor antigen can be any antigen that is not typically and / or is minimally expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease, but is typically and / or substantially expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein. In some cases, a cell will express both an activator antigen and an inhibitor antigen.

[0214] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the inhibitor antigen is any allele-specific antigen capable of protecting the second cell.

[0215] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the allele-specific antigen capable of protecting a second cell is a component of human leukocyte antigen (HLA).

[0216] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the HLA is HLA Class I or HLA Class I.

[0217] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the HLA Class I includes HLA- A, HLA-B, or HLA-C.

[0218] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the HL A- A includes HL A- A* 02 or HL A- A* 03.

[0219] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the inhibitor antigen is HLA-A*02, CCR8, LRRC32 (GARP), and / or CCR7.

[0220] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the inhibitor receptor is TCR or a CAR.

[0221] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the CAR includes an extracellular ligand binding domain specific to the inhibitor antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0222] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein binding of the inhibitor antigen on the second cell reduces surface expression or activity of the activating receptor by the logic-gated engineered immune cell, thereby decreasing the cytotoxic response by the engineered immune cell against the target cell.

[0223] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein binding of the inhibitor receptor to the inhibitor antigen on the second cell without binding or with reduced binding of the activator receptor to the activator antigen on the targetcell spares the target cell and provides protection from autoimmunity, thereby treating an autoimmune disease or disorder.

[0224] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the engineered immune cell is a T cell.

[0225] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the T cell is a CD8+ CD4- T cell or a CD8- CD4+ T cell.

[0226] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the T cell is a cytotoxic T cell.

[0227] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the engineered immune cell is a natural killer (NK) cell.

[0228] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti- synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren' s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

[0229] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the autoimmune disease or disorder is multiple sclerosis (MS).

[0230] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the logic-gated engineered immune cell is allogenic to a subject with the autoimmune disease or disorder.

[0231] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the logic-gated engineered immune cell is autogenic to a subject with the autoimmune disease or disorder.Immune cells including inhibitor receptors.

[0232] Another aspect of the present disclosure is an immune cell including an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell. In embodiments, the inhibitor receptor includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell. In embodiments, the immune cell, further includes an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell. In embodiments, the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell. In embodiments, the activator receptor and / or the inhibitor receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In some cases, the CAR includes an extracellular ligand binding domain(s) specific to the antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0233] In embodiments, the present disclosure relates to an immune cell, wherein the second cell is not causative of an autoimmune disease or disorder.

[0234] In embodiments, the present disclosure relates to an immune cell, wherein the second cell includes a naive T cell, a memory T cell, and / or a regulatory T cell (Treg) or a cell of another blood lineage.

[0235] In embodiments, the present disclosure relates to an immune cell, wherein binding of the first inhibitor receptor to a first inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logicgated engineered immune cell.

[0236] In embodiments, the present disclosure relates to an immune cell, wherein fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0237] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is any antigen expressed on the surface of the second cell.

[0238] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is any allele-specific antigen capable of protecting the second cell.

[0239] In embodiments, the present disclosure relates to an immune cell, wherein the allelespecific antigen capable of protecting a second cell is a component of human leukocyte antigen (HLA).

[0240] In embodiments, the present disclosure relates to an immune cell, wherein the HLA is HLA Class I or HLA Class I.

[0241] In embodiments, the present disclosure relates to an immune cell, wherein the HLA Class I includes HLA-A, HLA-B, or HLA-C.

[0242] In embodiments, the present disclosure relates to an immune cell, wherein the HLA-A includes HLA-A* 02 or HLA-A* 03.

[0243] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02, CCR8, LRRC32 (GARP), and / or CCR7.

[0244] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor further includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell.

[0245] In embodiments, the present disclosure relates to an immune cell, wherein binding of the second inhibitor receptor to a second inhibitor antigen on the second cell additionally or alternately protects the second cell from fratricide, wherein the second cell is a second logicgated engineered immune cell.

[0246] In embodiments, the present disclosure relates to an immune cell, wherein fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0247] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is any antigen expressed on the surface of the second cell.

[0248] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is any allele-specific antigen capable of protecting the second cell.

[0249] In embodiments, the present disclosure relates to an immune cell, wherein the allelespecific antigen capable of protecting a second cell is a component of human leukocyte antigen (HLA).

[0250] In embodiments, the present disclosure relates to an immune cell, wherein the HLA is HLA Class I or HLA Class I.

[0251] In embodiments, the present disclosure relates to an immune cell, wherein the HLA Class I includes HLA-A, HLA-B, or HLA-C.

[0252] In embodiments, the present disclosure relates to an immune cell, wherein the HLA-A includes HLA-A* 02 or HLA-A* 03.

[0253] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is HLA-A*02 or CCR8 and wherein the first inhibitor antigen is different from the second inhibitor antigen.

[0254] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen and the second inhibitor antigen are expressed by a single second cell.

[0255] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02 and the second inhibitor antigen is CCR8.

[0256] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR8 and the second inhibitor antigen is HLA-A*02.

[0257] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain and the second extracellular ligand binding domain are separated by a linker.

[0258] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0259] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0260] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0261] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0262] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain is N-terminal to the second extracellular ligand binding domain.

[0263] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain is N-terminal to the first extracellular ligand binding domain.

[0264] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes a leukocyte immunoglobulin like receptor Bl (LILRB1) intracellular domain or a functional variant thereof.

[0265] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB1 intracellular domain includes the sequence of SEQ ID NOs: 31 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0266] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes LILRB 1 hinge and transmembrane domains, or functional variants thereof.

[0267] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB1 transmembrane domain includes the sequence of SEQ ID NO: 30 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0268] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB1 hinge domain includes the sequence of SEQ ID NO: 29, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0269] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB1 hinge domain, LILRB1 transmembrane domain, and LILRB1 intracellular domain together includes the sequence of SEQ ID NO: 27 or 28, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0270] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

[0271] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes a transmembrane domain isolated or derived from CD8 or CD28.

[0272] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes an intracellular domain isolated or derived from CD28, 4-1BB or CD3z, or a combination thereof.

[0273] In embodiments, the present disclosure relates to an immune cell, further including an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell.

[0274] In embodiments, the present disclosure relates to an immune cell, wherein the autoreactive immune cell is associated with or causative of an autoimmune disease or disorder.

[0275] In embodiments, the present disclosure relates to an immune cell, wherein the autoreactive immune cell is a B cell, a T cell, or a myeloid cell.

[0276] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is an activated T cell, e.g., an acutely active T cell or a chronically active T cell.

[0277] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is not a naive T cell, a memory T cell, or a Treg.

[0278] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a Thl7 cell.

[0279] In embodiments, the present disclosure relates to an immune cell, wherein when the second cell expresses the first activator antigen and expresses the first inhibitor antigen, theinhibitor receptor reduces the expression (e.g., surface expression) or activity of the activator receptor.

[0280] In embodiments, the present disclosure relates to an immune cell, wherein the second cell is a naive T cell, a memory T cell, or a Treg.

[0281] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is selected from 4- IBB, CD 19, CD69, CD25, and 0X40.

[0282] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD69 and the first inhibitor antigen is HLA-A*02.

[0283] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD25 and the first inhibitor antigen is HLA-A*02.

[0284] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40 and the first inhibitor antigen is HLA-A*02.

[0285] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is CD25 and the first inhibitor antigen is HLA-A*02 and the second inhibitor antigen is CCR8 or (b) the first activator antigen is CD25 and the first inhibitor antigen is CCR8 and the second inhibitor antigen is HL A- A* 02.

[0286] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell.

[0287] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is selected from 4- IBB, CD 19, and 0X40.

[0288] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is 4-1BB and the second activator antigen is CD19 or (b) the first activator antigen is CD 19 and the second activator antigen is 4- IBB.

[0289] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HL A- A* 02.

[0290] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is 0X40 and the second activator antigen is CD 19 or (b) the first activator antigen is CD 19 and the second activator antigen is 0X40.

[0291] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32.

[0292] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR).

[0293] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to the first activator antigen and the second extracellular ligand binding domain specific to the second activator antigen are separated by a linker.

[0294] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0295] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0296] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0297] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0298] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to the first activator antigen is N-terminal to the second extracellular ligand binding domain specific to the second activator antigen.

[0299] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain specific to the second activator antigen is N-terminal to the first extracellular ligand binding domain specific to the first activator antigen.

[0300] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

[0301] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes a transmembrane domain isolated or derived from CD8 or CD28.

[0302] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes an intracellular domain isolated or derived from CD28, 4- IBB or CD3z, or a combination thereof.

[0303] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is a T cell.

[0304] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a CD8+ CD4- T cell or a CD8- CD4+ T cell.

[0305] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a cytotoxic T cell.

[0306] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is a natural killer (NK) cell.

[0307] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is modified to reduce or eliminate expression of a B2M gene product.

[0308] In embodiments, the present disclosure relates to an immune cell, wherein expression of the B2M gene product is reduced or eliminated using a short hairpin RNA (shRNA).

[0309] In embodiments, the present disclosure relates to an immune cell, wherein the shRNA includes a first sequence, having from 5' to 3' end a sequence complementary to the B2M gene product; and a second sequence, having from 5' to 3' end a sequence complementary to the first sequence, wherein the first sequence and second sequence form the shRNA.

[0310] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell treats or reduces a symptom of an autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti-synthetase syndrome with ILD), vasculitis (e.g., ANCA- associated vasculitis), severe refractory Sjogren1s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

[0311] In embodiments, the present disclosure relates to an immune cell, wherein the autoimmune disease or disorder is multiple sclerosis (MS).

[0312] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is allogenic to a subject with the autoimmune disease or disorder.

[0313] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is autogenic to a subject with the autoimmune disease or disorder.

[0314] In some embodiments, the present disclosure provides chimeric antigen receptors (CARs) comprising a polypeptide. In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR) and / or wherein the activator receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). Generally, a CAR includes an extracellular ligand binding domain specific to the activator antigen, a hinge domain, a transmembrane domain, and an intracellular domain.

[0315] In some embodiments, an inhibitor or activator receptors of the present disclosure comprises an extracellular hinge region. Illustrative hinges can be isolated or derived from IgG and CD8 domains, for example IgGl. In some embodiments, the hinge is isolated or derived from CD8a or CD28. In some embodiments, the extracellular hinge region of the inhibitor receptor is isolated from leukocyte immunoglobulin like receptor Bl (LILRB1).

[0316] The inhibitor receptors of the present disclosure can be designed to comprise a transmembrane domain that is fused to the extracellular domain of the inhibitor receptors. In some instances, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex.

[0317] The transmembrane domain may be derived either from a natural or from a synthetic source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. Transmembrane regions may be isolated or derived from (i.e., comprise at least the transmembrane region(s) of) the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or from an immunoglobulin such as IgG4. In some embodiments, the transmembrane region is isolated or derived from leukocyte immunoglobulin like receptor Bl (LILRB1). Alternatively, the transmembrane domain may be synthetic, in which case it will comprise predominantly hydrophobic residues such as leucine and valine. In some embodiments, a triplet of phenylalanine, tryptophan and valine will be found at each end of a synthetic transmembrane domain. Optionally, a short oligo- or polypeptide linker, preferably between 2 and 10 amino acids in length may form the linkage between the transmembrane domain and the intracellular domain of the inhibitor receptors. A glycine-serine doublet provides a particularly suitable linker.

[0318] In some embodiments, the inhibitory intracellular domain comprises an immunoreceptor tyrosine-based inhibitory motif (ITIM). In some embodiments, the inhibitory intracellular domain comprising an ITIM can be isolated or derived from an immunecheckpoint inhibitor such as CTLA-4 and PD-1. CTLA-4 and PD-1 are immune inhibitor receptors expressed on the surface of T cells, and play a pivotal role in attenuating or terminating T cell responses.

[0319] Inhibitory domains can be isolated from human tumor necrosis factor related apoptosis inducing ligand (TRAIL) receptor and CD200 receptor 1. In some embodiments, the TRAIL receptor comprises TR10A, TR10B or TR10D.

[0320] In some embodiments, the inhibitory domain comprises an intracellular domain, a transmembrane domain, or a combination thereof. In some embodiments, the inhibitory intracellular domain is isolated from phosphoprotein membrane anchor with glycosphingolipid microdomains 1 (PAG1). In some embodiments, the inhibitory intracellular domain is isolated from leukocyte immunoglobulin like receptor Bl (LILRB1). In some embodiments, the inhibitory domain is isolated or derived from a human protein, for example a human TRAIL receptor, CTLA-4, or PD-1, PAG1 or LILRBl protein.

[0321] In some embodiments, the engineered receptor comprises an inhibitory domain isolated or derived from killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 2 (KIR3DL2), killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 3 (KIR3DL3), leukocyte immunoglobulin like receptor Bl (LIR1, also called LIR-1 and LILRB1), programmed cell death 1 (PD-1), Fc gamma receptor IIB (FcgRIIB), killer cell lectin like receptor KI (NKG2D), CTLA-4, a domain containing a synthetic consensus ITIM, a ZAP70 SH2 domain (e.g., one or both of the N and C terminal SH2 domains), or ZAP70 KI K369A (kinase inactive ZAP70)

[0322] In some embodiments, the polypeptide comprises a ligand binding domain, such as an antigen-binding domain. Suitable antigen-binding domains include, but are not limited to, antigen-binding domains from antibodies, antibody fragments, scFv, antigen-binding domains derived from T cell receptors, and the like. All forms of antigen-binding domains known in the art are envisaged as within the scope of the disclosure.

[0323] An “extracellular domain,” as used herein, refers to the extracellular portion of a protein. For example, the TCR alpha and beta chains each comprise an extracellular domain, which comprise a constant and a variable region involved in peptide-MHC recognition. The “extracellular domain” can also comprise a fusion domain, for example of fusions between additional domains capable of binding to and targeting a specific antigen and the endogenous extracellular domain of the TCR subunit.

[0324] The term “antibody,” as used herein, refers to a protein, or polypeptide sequences derived from an immunoglobulin molecule, which specifically binds to an antigen. Antibodiescan be intact immunoglobulins of polyclonal or monoclonal origin, or fragments thereof and can be derived from natural or from recombinant sources.

[0325] The terms “antibody fragment” or “antibody binding domain” refer to at least one portion of an antibody, or recombinant variants thereof that contains the antigen-binding domain, i.e., an antigenic determining variable region of an intact antibody that is sufficient to confer recognition and specific binding of the antibody fragment to a target, such as an antigen and its defined epitope. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, single-chain (sc)Fv (“scFv”) antibody fragments, linear antibodies, single domain antibodies (abbreviated “sdAb”) (either VL or VH), camelid VHH domains, and multi-specific antibodies formed from antibody fragments.

[0326] An antibody, antibody fragment, or antibody -binding domain that “does not bind” with an antigen (e.g., CD16A) is intended to refer to an antibody, or the like portion thereof, that exhibits substantially no detectable binding above background to an antigen in a conventional immunoassay, such as ELISA or flow cytometry.

[0327] The term “scFv” refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain and at least one antibody fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked via a short flexible polypeptide linker, and capable of being expressed as a single polypeptide chain, and wherein the scFv retains the specificity of the intact antibody from which it is derived.

[0328] “Light chain variable region” or “VL” with regard to an antibody refers to the fragment of the light chain that contains three CDRs interposed between flanking stretches known as framework regions, these framework regions are generally more highly conserved than the CDRs and form a scaffold to support the CDRs.

[0329] “Heavy chain variable region” or “VH” (or, in the case of single domain antibodies, e.g., nanobodies, “VHH”) with regard to an antibody refers to the fragment of the heavy chain that contains three CDRs interposed between flanking stretches known as framework regions, these framework regions are generally more highly conserved than the CDRs and form a scaffold to support the CDRs.

[0330] Unless specified, as used herein a scFv may have the VL and VH variable regions in either order, e.g., with respect to the N-terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH-linker-VL.

[0331] The term “antibody light chain,” refers to the smaller of the two types of polypeptide chains present in antibody molecules in their naturally occurring conformations. Kappa (“K”) and lambda (“X”) light chains refer to the two major antibody light chain isotypes.

[0332] The term “recombinant antibody” refers to an antibody that is generated using recombinant DNA technology, such as, for example, an antibody expressed by a bacteriophage or yeast expression system. The term should also be construed to mean an antibody which has been generated by the synthesis of a DNA molecule encoding the antibody and which DNA molecule expresses an antibody protein, or an amino acid sequence specifying the antibody, wherein the DNA or amino acid sequence has been obtained using recombinant DNA or amino acid sequence technology which is available and well known in the art.

[0333] In embodiments, the present disclosure relates to an immune cell, wherein a first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0334] In embodiments, the present disclosure relates to an immune cell, wherein a second extracellular ligand binding domain of the inhibitor receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region.

[0335] In embodiments, the present disclosure relates to an immune cell, wherein a first extracellular ligand binding domain of the inhibitor receptor includes a first antibody fragment or a first single chain Fv antibody fragment (scFv).

[0336] In embodiments, the present disclosure relates to an immune cell, wherein a second extracellular ligand binding domain of the inhibitor receptor includes a second antibody fragment or a second single chain Fv antibody fragment (scFv).

[0337] In embodiments, the present disclosure relates to an immune cell, wherein a first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0338] In embodiments, the present disclosure relates to an immune cell, wherein a second extracellular ligand binding domain of the activator receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region.

[0339] In embodiments, the present disclosure relates to an immune cell, wherein a first extracellular ligand binding domain of the activator receptor includes a first antibody fragment or a first single chain Fv antibody fragment (scFv).

[0340] In embodiments, the present disclosure relates to an immune cell, wherein a second extracellular ligand binding domain of the activator receptor includes a second antibody fragment or a second single chain Fv antibody fragment (scFv).

[0341] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first antibody fragment or a first single chain Fv antibody fragment (scFv).

[0342] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0343] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02; and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) including an amino acid sequence including SEQ ID NO: 4; (b) a CDR-H2 including an amino acid sequence including SEQ ID NO: 5, or (c) a CDR-H3 including an amino acid sequence including SEQ ID NO: 6, and the first VL region includes: (d) a CDR-L1 including an amino acid sequence including SEQ ID NO: 1, (e) a CDR-L2 including an amino acid sequence including SEQ ID NO: 2, or (f) a CDR3-L3 including an amino acid sequence including SEQ ID NO: 3.

[0344] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02; and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) including an amino acid sequence including SEQ ID NO: 10, (b) a CDR-H2 including an amino acid sequence including SEQ ID NO: 11, or (c) a CDR-H3 including an amino acid sequence including SEQ ID NO: 12, and the first VL region includes: (d) a CDR-L1 including an amino acid sequence including SEQ ID NO: 7; (e) a CDR-L2 including an amino acid sequence including SEQ ID NO: 8; or (f) a CDR3-L3 including an amino acid sequence including SEQ ID NO: 9.

[0345] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02, and wherein the first VH region includes the sequence of any one of the SEQ IDs NOs: 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310,1312, or 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311,1313, or 1315 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0346] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02; and the scFv includes a sequence having at least 85%, at least90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 13 to 25.

[0347] Additional binding sequences for HLA-A*02 and HLA-A*03 can be found in U.S. Patent No. : 11,254,726, and PCT Application No. PCT / US24 / 40804, the contents of which are hereby incorporated by reference in their entirety.

[0348] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR8; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1180, 1186, 1192, 1197 1206, 1212, 1222, and, 1228; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1181, 1187; 1193, 1198, 1207, 1213, 1223, and 1229; or a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1182, 1188, 1194, 1195, 1199, 1203, 1208, 1214, 1218, 1224, and 1230; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1183, 1189, 1196, 1200, 1204, 1209, 1215, 1219, 1225, and 1231; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1184, 1190, 1201; 1205, 1210, 1216, 1220, 1226, and 1232; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1185, 1191, 1202, 1211, 1217, 1221, 1227, and 1233.

[0349] In embodiments, the present disclosure relates to an immune cell, wherein the first antigen is CCR8, and wherein the first VH region includes the sequence of any one of the SEQ IDs NOs: 1234, 1236, 1238, 1240, 1242, 1244, 1246, 1248, 1250, 1252, 1254, 1256, 1258 or 1260 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, or 1261 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0350] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR8; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1262-1275.

[0351] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR7; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1035, 1040, 1046, 1052, 1058, 1064, 1070, 1076, 1082,1091, 1101, and 1107; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1036, 1041, 1047, 1053, 1059, 1065, 1071, 1077, 1083, 1085, 1088;1092, 1099, and 1102; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1042, 1048, 1054, 1060, 1066, 1072, 1078, 1084, 1086,1093, 1097, 1103, and 1106; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1037, 1043, 1049, 1055, 1061, 1067, 1073; 1079, 1087, 1094, 1098, and 1104 (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1038, 1044, 1050, 1056, 1062, 1068, 1074; 1080, 1089, 1095, and 1105; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1039, 1045, 1051, 1057, 1063, 1069, 1075, 1081, 1090, 1096, and 1100.

[0352] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR7, and wherein the first VH region includes the sequence of any one of the SEQ IDs NOs: 1108, 1110, 1112, 1114, 1116, 1118, 1120, 1122, 1124, 1126, 1128, 1130, 1132, 1134, 1136, 1138, 1140, or 1142 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 1109, 1111, 1113, 1115, 1117, 1119, 1121, 1123, 1125, 1127, 1129, 1131, 1133, 1135, 1137, 1139, 1141 or 1143 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0353] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR7; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1144-1161.

[0354] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor further includes a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell.

[0355] In embodiments, the present disclosure relates to an immune cell, wherein binding of the second inhibitor receptor to a second inhibitor antigen on the second cell additionally oralternately protects the second cell from fratricide, wherein the second cell is a second logicgated engineered immune cell.

[0356] In embodiments, the present disclosure relates to an immune cell, wherein fratricide would occur during manufacture of the second logic-gated engineered immune cell.

[0357] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is any antigen expressed on the surface of the second cell.

[0358] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is any allele-specific antigen capable of protecting the second cell.

[0359] In embodiments, the present disclosure relates to an immune cell, wherein the allelespecific antigen capable of protecting a second cell is a component of human leukocyte antigen (HLA).

[0360] In embodiments, the present disclosure relates to an immune cell, wherein the HLA is HLA Class I or HLA Class I.

[0361] In embodiments, the present disclosure relates to an immune cell, wherein the HLA Class I includes HLA-A, HLA-B, or HLA-C.

[0362] In embodiments, the present disclosure relates to an immune cell, wherein the HLA-A includes HLA-A* 02 or HLA-A* 03.

[0363] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is HLA-A*02 or CCR8 and wherein the first inhibitor antigen is different from the second inhibitor antigen.

[0364] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen and the second inhibitor antigen are expressed by a single second cell.

[0365] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HLA-A*02 and the second inhibitor antigen is CCR8.

[0366] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is CCR8 and the second inhibitor antigen is HLA-A*02.

[0367] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain of the inhibitor receptor includes a second antibody fragment or a second single chain Fv antibody fragment (scFv).

[0368] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain of the inhibitor receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region.

[0369] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is HLA-A*02; and wherein the second VH region includes: (a) acomplementarity-determining region VH-1 (CDR-H1) including an amino acid sequence including SEQ ID NO: 4; (b) a CDR-H2 including an amino acid sequence including SEQ ID NO: 5, or (c) a CDR-H3 including an amino acid sequence including SEQ ID NO: 6, and the first VL region includes: (d) a CDR-L1 including an amino acid sequence including SEQ ID NO: 1, (e) a CDR-L2 including an amino acid sequence including SEQ ID NO: 2, or (f) a CDR3-L3 including an amino acid sequence including SEQ ID NO: 3.

[0370] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is HLA-A*02; and wherein the second VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) including an amino acid sequence including SEQ ID NO: 10, (b) a CDR-H2 including an amino acid sequence including SEQ ID NO: 11, or (c) a CDR-H3 including an amino acid sequence including SEQ ID NO: 12, and the first VL region includes: (d) a CDR-L1 including an amino acid sequence including SEQ ID NO: 7; (e) a CDR-L2 including an amino acid sequence including SEQ ID NO: 8; or (f) a CDR3-L3 including an amino acid sequence including SEQ ID NO: 9.

[0371] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is HLA-A*02; and the second scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 13 to 25.

[0372] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is CCR8; and wherein the second VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1180, 1186, 1192, 1197 1206, 1212, 1222, and 1228; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1181, 1187; 1193, 1198, 1207, 1213, 1223, and 1229; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1182, 1188, 1194, 1195, 1199, 1203, 1208, 1214, 1218, 1224, and 1230; and the second VL region includes: (d) a CDR- L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1183, 1189, 1196, 1200, 1204, 1209, 1215, 1219, 1225, and 1231; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1184, 1190, 1201; 1205, 1210, 1216, 1220, 1226, and 1232; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1185, 1191, 1202, 1211, 1217, 1221, 1227, and 1233.

[0373] In embodiments, the present disclosure relates to an immune cell, wherein the second inhibitor antigen is CCR8; and the second scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1262-1275.

[0374] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain and the second extracellular ligand binding domain are separated by a linker.

[0375] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0376] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0377] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0378] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0379] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain is N-terminal to the second extracellular ligand binding domain.

[0380] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain is N-terminal to the first extracellular ligand binding domain.

[0381] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 1162-1179.

[0382] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to SEQ ID NO: 1276-1289.

[0383] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 310, 323, 329, 335, 340, and 351; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 311, 324, 330, 336, 341, 346, 347, and 352; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 312, 325, 331, 337, 342, 348, and 353; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 313, 316, 320, 326, 332, 338, 343, and 354; (e) a CDR- L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 314, 318, 321, 327, 333, 339, 344, and 349; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 315, 317, 319, 322, 328, 334, 340, 345, 350, and 356. In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) comprising SEQ ID NO: 340; (b) a CDR-H2 comprising SEQ ID NO: 341(c) a CDR-H3 comprising SEQ ID NO: 342; and the first VL region includes: (d) a CDR-L1 comprising SEQ ID NOs: 343 (e) a CDR-L2 comprising SEQ ID NO: 344; and (f) a CDR3-L3 comprising SEQ ID NO: 345.

[0384] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and wherein the first VH region includes the sequence of any one of SEQ ID NOs: 369, 371, 373, 375, 377, 379, 381, 383, or 385, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 370, 372, 374, 376, 378, 380, 382, 384, or 386, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and wherein the first VH comprises SEQ ID NO: 373, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region comprises SEQ ID NOs: 374 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0385] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 387 - 400. In embodiments, thepresent disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32 (GARP); and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NO: 394.

[0386] In embodiments, the present disclosure relates to an immune cell wherein the inhibitor receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 401 - 414. In embodiments, the present disclosure relates to an immune cell wherein the inhibitor receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to SEQ ID NO: 408.

[0387] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes a leukocyte immunoglobulin like receptor Bl (LILRB1) intracellular domain or a functional variant thereof.

[0388] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB1 intracellular domain includes the sequence of SEQ ID NOs: 31 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0389] In embodiments, the present disclosure relates to an immune cell, wherein the inhibitor receptor includes LILRB 1 hinge and transmembrane domains, or functional variants thereof.

[0390] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB 1 transmembrane domain includes the sequence of SEQ ID NO: 30 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0391] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB 1 hinge domain includes the sequence of SEQ ID NO: 29, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0392] In embodiments, the present disclosure relates to an immune cell, wherein the LILRB 1 hinge domain, LILRB 1 transmembrane domain, and LILRB 1 intracellular domain together includes the sequence of SEQ ID NO: 27 or 28, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0393] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

[0394] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes a transmembrane domain isolated or derived from CD8 or CD28.

[0395] In embodiments, the present disclosure relates to an immune cell, wherein the CAR includes an intracellular domain isolated or derived from CD28, 4-1BB or CD3z, or a combination thereof.

[0396] In embodiments, the present disclosure relates to an immune cell, further including an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell.

[0397] In embodiments, the present disclosure relates to an immune cell, wherein the autoreactive immune cell is associated with or causative of an autoimmune disease or disorder.

[0398] In embodiments, the present disclosure relates to an immune cell, wherein the autoreactive immune cell is a B cell, a T cell, or a myeloid cell.

[0399] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is an activated T cell, e.g., an acutely active T cell or a chronically active T cell.

[0400] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is not a naive T cell, a memory T cell, or a Treg.

[0401] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a Thl7 cell.

[0402] In embodiments, the present disclosure relates to an immune cell, wherein when the second cell expresses the first activator antigen and expresses the first inhibitor antigen, the inhibitor receptor reduces the expression (e.g., surface expression) or activity of the activator receptor.

[0403] In embodiments, the present disclosure relates to an immune cell, wherein the second cell is a naive T cell, a memory T cell, or a Treg.

[0404] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is selected from 4- IBB, CD 19, CD69, CD25, and 0X40.

[0405] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD69, and the first inhibitor antigen is HLA-A*02.

[0406] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD25, and the first inhibitor antigen is HLA-A*02.

[0407] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40, and the first inhibitor antigen is HLA-A*02.

[0408] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is CD25, and the first inhibitor antigen is HLA-A*02 and the second inhibitor antigen is CCR8 or (b) the first activator antigen is CD25 and the first inhibitor antigen is CCR8 and the second inhibitor antigen is HL A- A* 02.

[0409] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor further includes a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell.

[0410] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is selected from 4- IBB, CD 19, and 0X40.

[0411] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is 4-1BB and the second activator antigen is CD19 or (b) the first activator antigen is CD 19, and the second activator antigen is 4- IBB.

[0412] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is HL A- A* 02.

[0413] In embodiments, the present disclosure relates to an immune cell, wherein (a) the first activator antigen is 0X40, and the second activator antigen is CD 19 or (b) the first activator antigen is CD 19 and the second activator antigen is 0X40.

[0414] In embodiments, the present disclosure relates to an immune cell, wherein the first inhibitor antigen is LRRC32.

[0415] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR).

[0416] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain of the activator receptor includes a first antibody fragment or a first single chain Fv antibody fragment (scFv).

[0417] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0418] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD69; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 702, 710, and 716; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 703, 711, and 717; or (c) a CDR- H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 704, 712, and 718; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 705, 713, and 719; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 706, 714, and 720;or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 707, 708, 709, 715, and 721.

[0419] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD69; and wherein the first VH region includes the sequence of any one of SEQ ID NOs: 722, 724, 726, 728, 730, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 723, 725, 727, 729, 731, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0420] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD69; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 732-736.

[0421] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 737-741.

[0422] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is CD25; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 52, 58, 64, 70, 76, 78, 80, 83, 91, 97, 102, 108, 114, 120, 126, 131, and 137; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs:47, 53, 59, 65, 71, 77, 79, 81, 82, 84, 87, 88, 89, 90; 92, 98, 100, 103, 109, 115, 121, 127, and 132; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 48, 54, 60, 66, 72, 85, 93, 99, 104, 110, 116, 122, 128, 133, and 139; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 49, 55, 61, 67, 73, 94, 105, 111, 117, 123, 129, and 141; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 50, 56, 62, 68, 74, 95, 106, 112; 118, 124, and 135; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 51, 57, 63, 69, 75, 86, 96, 101, 107, 113, 119, 125, 130, 136, and 140.

[0423] In embodiments, the present disclosure relates to an immune cell, wherein the first VH region of the activator receptor includes the sequence selected from the group consisting of SEQ ID NO: 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172,174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, and 224, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence selected from the group consisting of SEQ ID NO : 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, and 225 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0424] In embodiments, the present disclosure relates to an immune cell, wherein the scFv includes the sequence selected from the group consisting of SEQ ID NO : 226 - 267, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0425] In embodiments, the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 268 - 309.

[0426] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 501, 507, 513, 518, 524, 534, 539, 544, 549, 555, 561, 565, and 570; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467; 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573. In embodiments,the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40; and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) comprising SEQ ID NO: 537; (b) a CDR-H2 comprising SEQ ID NO: 538 (c) a CDR-H3 comprising SEQ ID NO: 539; and the first VL region includes: (d) a CDR-L1 comprising SEQ ID NOs: 540 (e) a CDR-L2 comprising SEQ ID NO: 446; and (f) a CDR3-L3 comprising SEQ ID NO: 541.

[0427] In embodiments, the present disclosure relates to an immune cell, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NO: 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614,616, 618, 620, 622, 624, 626, 628, 630, 632, 634, or 636, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NO : 575, 577, 579, 581, 583, 585,587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623,625, 627, 629, 631, 633, 635, or 637, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40; and wherein the first VH comprises SEQ ID NO: 624, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region comprises SEQ ID NOs: 625 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0428] In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 638 - 669. In embodiments, the present disclosure relates to an immune cell, wherein the first activator antigen is 0X40; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NO: 663.

[0429] In embodiments, the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 670 - 701. In embodiments, the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to SEQ ID NOs: 695.

[0430] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 4-1BB; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 742, 749, 755, 761, 767, 773, 779, 785, 791, 797; 803, 808, 814, 822, 827, 833, 844, 849, and 855; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 744, 750, 756, 762, 768, 774, 780, 786, 792, 798 ,809, 815, 828, 834, 839, 845, 850, and 856; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 745, 751, 757, 763, 769, 775, 781, 787, 793, 799, 804, 810, 818, 823, 829, 835, 840, 846, 851, and 857; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 746, 752, 758, 764, 770, 776, 782, 788, 794, 800, 805, 811, 816, 819, 824, 830, 836, 841, 847, 852, and 858; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 747, 753, 759, 765, 771, 777, 783, 789, 795, 801, 806, 812, 820, 825, 831, 837, 842, 853, and 859; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 748, 754, 760, 772, 778, 784, 790, 796, 802, 807, 813, 817, 821, 826, 832, 838, 843, 848, 854, and 860.

[0431] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 4-1BB; and wherein the first VH region includes the sequence of any one of SEQ ID NOs: 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881,883, 885, 887, 889, 891, 893, 895 ,897, 899, 901, 903, 905 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 862, 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 898, 900, 902, 904 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0432] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 4- IBB; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 907-929.

[0433] In embodiments, the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 1026-1034. In embodiments,the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to SEQ ID NO: 1027.

[0434] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD 19; and wherein the first VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 953, 959, 970, 976, 985, and 991; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 954, 960, 965, 971, 977, 982, 986, 992, and 997; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 955, 961, 966, 972, 978, 987, and 993; and the first VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 956, 962, 967, 973, 979, 983, 988, and 994; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 957, 963, 968, 974, 980, 989, 995, and 998; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 958, 964, 969, 975, 981, 984, 990, and 996. In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD19; and wherein the first VH region includes: (a) a complementarity-determining region VH-1 (CDR-H1) comprising SEQ ID NO: 959; (b) a CDR-H2 comprising SEQ ID NO: 960 (c) a CDR-H3 comprising SEQ ID NO: 961; and the first VL region includes: (d) a CDR- L1 comprising SEQ ID NOs: 962 (e) a CDR-L2 comprising SEQ ID NO: 963; and (f) a CDR3- L3 comprising SEQ ID NO: 964.

[0435] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD 19; and wherein the first VH region includes the sequence of any one of SEQ ID NOs: 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013 and 1015 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region includes the sequence of any one of SEQ ID NOs: 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014 and 1016 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD 19; and wherein the first VH comprises SEQ ID NO: 1001, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the first VL region comprises SEQ ID NOs: 1002 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0436] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD 19; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1017-1025. In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is CD 19; and the scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NOs: 1018.

[0437] In embodiments, the present disclosure relates to an immune cell wherein the activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 670-701.

[0438] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 0X40; and wherein the second VH region includes: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467, 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573, and the second VL region includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 495, 501, 507, 513, 518, 524, 528, 534, 539, 544, 549, 555, 561, 565, and 570.

[0439] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 0X40; and wherein the second VH region includes the sequence of any one of SEQ ID NOs: 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 634, or 636, or asequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and the second VL region includes the sequence of any one of SEQ ID NOs: 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, or 637, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0440] In embodiments, the present disclosure relates to an immune cell, wherein the second activator antigen is 0X40; and the second scFv includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 638 - 669.

[0441] In embodiments, the present disclosure relates to an immune cell wherein the second activator receptor includes a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to any one of SEQ ID NOs: 670 - 701.

[0442] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to the first activator antigen and the second extracellular ligand binding domain specific to the second activator antigen are separated by a linker.

[0443] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0444] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0445] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0446] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0447] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to the first activator antigen is N-terminal to the second extracellular ligand binding domain specific to the second activator antigen.

[0448] In embodiments, the present disclosure relates to an immune cell, wherein the second extracellular ligand binding domain specific to the second activator antigen is N-terminal to the first extracellular ligand binding domain specific to the first activator antigen.

[0449] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

[0450] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes a transmembrane domain isolated or derived from CD8 or CD28.

[0451] In embodiments, the present disclosure relates to an immune cell, wherein the activator receptor as a CAR includes an intracellular domain isolated or derived from CD28, 4- IBB or CD3z, or a combination thereof.

[0452] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is a T cell.

[0453] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a CD8+ CD4- T cell or a CD8- CD4+ T cell.

[0454] In embodiments, the present disclosure relates to an immune cell, wherein the T cell is a cytotoxic T cell.

[0455] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is a natural killer (NK) cell.

[0456] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is modified to reduce or eliminate expression of a B2M gene product.

[0457] In embodiments, the present disclosure relates to an immune cell, wherein expression of the B2M gene product is reduced or eliminated using a short hairpin RNA (shRNA).

[0458] In embodiments, the present disclosure relates to an immune cell, wherein the shRNA includes a first sequence, having from 5' to 3' end a sequence complementary to the B2M gene product; and a second sequence, having from 5' to 3' end a sequence complementary to the first sequence, wherein the first sequence and second sequence form the shRNA.Tandem Activator Extracellular Ligand Binding Domain

[0459] The disclosure provides an activator receptor comprising an extracellular region, the extracellular region comprising a first ligand binding domain capable of specifically binding a first ligand that activates or promotes activation of the receptor and a second ligand binding domain capable of specifically binding a second ligand that activates or promotes activation of the receptor, binding of either ligand binding domain promotes activation of effector cells expressing the tandem activator receptor.

[0460] In embodiments, the tandem activator receptor comprises two scFv domains. In embodiments the tandem activator receptor can bind any two activator antigens disclosedherein. In embodiments, the tandem activator receptor can bind two different activator antigens disclosed herein in any order (ex. first scFv binds CD 19 and the second scfv binds 0x40, or reverse order). In embodiments, the first scFv domain binds to a CD 19 antigen and the second scFV domain binds 0X40. In embodiments, the first scFv domain binds 0X40 and the second scFv domains binds CD 19. In embodiments, the first scFv domain that binds to CD 19 comprises a first VH region comprising: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 953, 959, 970, 976, 985, and 991; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 954, 960, 965, 971, 977, 982, 986, 992, and 997; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 955, 961, 966, 972, 978, 987, and 993; and a first VL region comprising: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 956, 962, 967, 973, 979, 983, 988, and 994; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 957, 963, 968, 974, 980, 989, 995, and 998; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 958, 964, 969, 975, 981, 984, 990, and 996. In embodiments, the first scFv domain that binds CD 19 antigen comprises a first VH region comprising: (a) a complementarity-determining region VH-1 (CDR-H1) comprising SEQ ID NO: 959; (b) a CDR-H2 comprising SEQ ID NO: 960 (c) a CDR-H3 comprising SEQ ID NO: 961; and a first VL region comprising: (d) a CDR-L1 comprising SEQ ID NOs: 962 (e) a CDR-L2 comprising SEQ ID NO: 963; and (f) a CDR3-L3 comprising SEQ ID NO: 964. In embodiments, the first scFv domain that binds CD 19 antigen comprises a first VH region comprising any one of SEQ ID NOs: 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013 and 1015 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and a first VL region comprising any one of SEQ ID NOs: 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014 and 1016 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the first scFV domain that binds CD19 antigen comprises a first VH comprising SEQ ID NO: 1001, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and a first VL region comprising SEQ ID NOs: 1002 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the first scFV domain that binds CD 19 antigen comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1017-1025. In embodiments, the first scFV domain that binds CD 19 antigen comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NOs: 1018. In embodiments, the second scFv of the tandem activator receptor binds 0X40. In embodiments, the second scFv domain that binds to 0X40 comprises a second VH region comprising: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 501, 507, 513, 518, 524, 534, 539, 544, 549, 555, 561, 565, and 570; and a second VL region comprising: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467; 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; (f) a CDR3- L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573. In embodiments, the second scFV domain that binds 0X40 antigen comprises a first VH region comprising: (a) a complementaritydetermining region VH-1 (CDR-H1) comprising SEQ ID NO: 537; (b) a CDR-H2 comprising SEQ ID NO: 538 (c) a CDR-H3 comprising SEQ ID NO: 539; and a second VL region comprising: (d) a CDR-L1 comprising SEQ ID NOs: 540 (e) a CDR-L2 comprising SEQ ID NO: 446; and (f) a CDR3-L3 comprising SEQ ID NO: 541. In embodiments, the second scFv domain that binds 0X40 antigen comprises a second VH comprising SEQ ID NO: 624, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and a second VL region comprising SEQ ID NOs: 625 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the second scFv domain that binds 0X40 antigen comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 638 - 669. In embodiments, the second scFv domain that binds 0X40 antigen comprises asequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NO: 663. In embodiments, the tandem activator comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NOs: 1322 or 1324.

[0461] In embodiments, the tandem activator receptor comprises a first antigen binding domain comprising an scFv that specifically binds CD 19 and a second antigen binding domain comprising the endogenous 0X40 ligand TNFS4. In embodiments, the endogenous 0X40 ligand domain comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NOs: 1325-1326. In embodiments, the In embodiments, the first scFv domain binds to a CD 19 antigen. In embodiments, the scFv domain that binds to CD19 comprises a first VH region comprising: (a) a complementaritydetermining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 953, 959, 970, 976, 985, and 991; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 954, 960, 965, 971, 977, 982, 986, 992, and 997; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 955, 961, 966, 972, 978, 987, and 993; and a VL region comprising: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 956, 962, 967, 973, 979, 983, 988, and 994; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 957, 963, 968, 974, 980, 989, 995, and 998; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 958, 964, 969, 975, 981, 984, 990, and 996. In embodiments, the scFv domain that binds CD 19 antigen comprises a VH region comprising: (a) a complementarity-determining region VH-1 (CDR-H1) comprising SEQ ID NO: 959; (b) a CDR-H2 comprising SEQ ID NO: 960 (c) a CDR-H3 comprising SEQ ID NO: 961; and a first VL region that includes: (d) a CDR-L1 comprising SEQ ID NOs: 962 (e) a CDR-L2 comprising SEQ ID NO: 963; and (f) a CDR3-L3 comprising SEQ ID NO: 964. In embodiments, the scFv domain that binds CD 19 antigen comprises a VH region comprising any one of SEQ ID NOs: 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013 and 1015 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and a VL region comprising any one of SEQ ID NOs: 1000, 1002, 1004, 1006, 1008, 1010, 1012, 1014 and 1016 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the scFv domain that binds CD19 antigen comprises a VH comprising SEQ ID NO: 1001, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or100% identical thereto; and a VL region comprising SEQ ID NOs: 1002 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto. In embodiments, the scFv domain that binds CD 19 antigen comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1017- 1025. In embodiments, the first scFv domain that binds CD 19 antigen comprises a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to SEQ ID NOs: 1018. In embodiments, the tandem activator receptor comprising a first antigen binding domain comprising a scFv that specifically binds CD 19 and a second antigen binding domain comprising the endogenous 0X40 ligand TNFS4 comprises SEQ ID NO: 1323, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identity to a sequence selected from the group consisting of any one of SEQ ID NOs: 1323.Polynucleotides

[0462] In embodiments, the present disclosure relates to a polynucleotide or polynucleotide system, including one or more polynucleotides including one or more polynucleotide sequences encoding any herein-described activator receptor or the inhibitor receptor.

[0463] The present disclosure provides nucleic acid molecules, specifically polynucleotides, and / or messenger RNAs (mRNAs) which encode one or more activator and / or inhibitor receptors. The term “nucleic acid,” in its broadest sense, includes any compound and / or substance that comprises a polymer of nucleotides. These polymers are often referred to as polynucleotides. Illustrative nucleic acids or polynucleotides of the disclosure include, but are not limited to, ribonucleic acids (RNAs), mRNAs, modified mRNAs (mmRNAs), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), and locked nucleic acids (LNAs). In some embodiments, the polynucleotide is an mRNA. In some embodiments, the mRNA which encodes one or more activator and / or inhibitor receptors is capable of being translated to produce the encoded receptors in vitro, in vivo, in situ or ex vivo. The term “polynucleotide system” refers to one or more polynucleotides intend to be used together.

[0464] The present disclosure encompasses the delivery of polynucleotides or mRNA for any of therapeutic, pharmaceutical, diagnostic or imaging by any appropriate route. Delivery may be naked or formulated.

[0465] In some embodiments, the polynucleotide encodes, in a 5' to 3' orientation, an activator receptor followed by an inhibitor receptor. In some embodiments, the polynucleotide encodes, in a 5 ' to 3 ' orientation, an inhibitor receptor followed by an activator receptor. In embodiments, the activator and inhibitor receptor are separated by a linker.Naked Delivery

[0466] The polynucleotides or mRNA of the present disclosure may be delivered to a cell naked. As used herein in, “naked” refers to delivering polynucleotides or mRNA free from agents which promote transfection. For example, the polynucleotides or mRNA delivered to the cell may contain no modifications. The naked polynucleotides or mRNA may be delivered to the cell using routes of administration described herein.

[0467] In some embodiments, the mRNA can be delivered to an immune cell through naked mRNA injection. In some embodiments, intracellular delivery of naked mRNA may be carried out in vitro, in vivo, in situ or ex vivo.Formulated Delivery

[0468] The polynucleotides or mRNA of the present disclosure may be formulated using the methods described herein. The formulations may contain polynucleotides or mRNA which may be modified and / or unmodified. The formulations may further include, but are not limited to, cell penetration agents, a pharmaceutically acceptable carrier, a delivery agent, a bioerodible or biocompatible polymer, a solvent, and a sustained-release delivery depot. The formulated polynucleotide or mRNA may be delivered to the cell using routes of administration described herein.

[0469] The compositions may also be formulated for direct delivery to an organ or tissue in any of several ways including, but not limited to, direct soaking or bathing, via a catheter, by gels, powder, ointments, creams, gels, lotions, and / or drops, and the like.Viral Vectors

[0470] A number of viral based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. A selected gene can be inserted into a plasmid and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo. A number of retroviral systems are known in the art. In some embodiments, adenovirus vectors are used. A number of adenovirus vectors are known in the art. In one embodiment, lentivirus vectors are used.

[0471] Viral vectors derived from retroviruses such as the lentivirus are suitable tools to achieve long-term gene transfer since they allow long-term, stable integration of a transgeneand its propagation in daughter cells. Lentiviral vectors have the added advantage over viral vectors derived from onco-retroviruses such as murine leukemia viruses in that they can transduce non-proliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity.Nanocarriers

[0472] In embodiments, the present disclosure relates to a nanocarrier including the polynucleotide or polynucleotide system.

[0473] In embodiments, the present disclosure relates to a nanocarrier, wherein the nanocarrier is capable of delivering the polynucleotide or polynucleotide system to an immune cell in vivo or ex vivo.

[0474] In embodiments, the present disclosure relates to a nanocarrier, wherein the nanocarrier is a lipid nanoparticle (LNP).

[0475] In embodiments, the present disclosure relates to a nanocarrier, wherein the polynucleotide or polynucleotide system are one or more messenger ribonucleic acids (mRNAs) or modified mRNAs

[0476] In embodiments, the present disclosure relates to a nanocarrier, wherein the polynucleotide or polynucleotide system are one or more messenger ribonucleic acids (mRNAs) or modified mRNAs (mmRNAs).

[0477] Nanocarriers may be used to deliver nucleic acids encoding an activator receptor (or receptors) and an inhibitor receptor (or receptors) to immune cells, either ex vivo or in vivo. For example, a nanocarrier may be used to deliver a messenger RNA (mRNA), or set of mRNAs, encoding the receptors to immune cells.

[0478] Nanocarriers include micelles, polymers, liposomes, and lipid nanoparticles (LNPs). Nanoparticles are composed of lipids, polymers, or both that encapsulate the nucleic acid. Examples of nanoparticle-type nanocarriers include ionizable lipid nanoparticles (LNP), solid lipid nanoparticles (SLN). Illustrative LNPs that may be used to deliver nucleic acids to immune cells are described in, e.g., Billingsley et al. (2020, Ionizable Lipid Nanoparticle- Mediated mRNA Delivery for Human CAR T Cell Engineering, Nano Letters). In general, suitable LNPs contain an ionizable lipid core that remains neutral in a physiologically relevant pH but builds charge in acidic environments, such as the endosome, to aid in endosomal escape and enable intracellular nucleic acid delivery, (e.g., WO 2019 / 067999 and WO 2021 / 055892).

[0479] In some embodiments, a LNP may be used to deliver an mRNA or mmRNA ex vivo.

[0480] A number of nanoparticle-based systems have been developed for gene transfer into mammalian cells. For example, lipid nanoparticles provide a convenient platform for genedelivery systems. A selected mRNA (or other nucleic acid) can be packaged in various LNP formulations. The mRNA-containing LNP can then be delivered to cells of the subject in vivo.

[0481] Nanocarriers other than LNPs may also be used to deliver nucleic acids to immune cells, both ex vivo and in vivo. For example, Parayanth et al. Nat. Commun. 11 :6080 (2020) describes an injectable nanocarrier that delivers in vitro-transcribed (IVT) receptor-encoding mRNA. Illustrative nanocarriers of the disclosure may comprise poly(beta-amino ester) (PBAE) and a coating comprising polyglutamic acid (PGA). Nanocarriers of the disclosure may comprise positively charged lipids comprising poly(beta-amino ester), poly(L-lysine), poly(ethylene imine) (PEI), poly-(amidoamine) dendrimers (PAMAMs), poly(amine-co- esters), poly(dimethylaminoethyl methacrylate) (PDMAEMA), chitosan, poly- (L-lactide-co- L-lysine), poly[a-(4-aminobutyl)-L-glycolic acid] (PAGA), or poly(4-hydroxy-L- proline ester) (PHP).

[0482] Viral vectors derived from retroviruses such as the lentivirus are suitable tools to achieve long-term gene transfer since they allow long-term, stable integration of a transgene and its propagation in daughter cells. Lentiviral vectors have the added advantage over viral vectors derived from onco-retroviruses such as murine leukemia viruses in that they can transduce non-proliferating cells, such as hepatocytes. They also have the added advantage of low immunogenicity.

[0483] The expression of natural or synthetic nucleic acids encoding engineered receptors is typically achieved by operably linking a nucleic acid encoding the fusion protein or portions thereof to a promoter, and incorporating the construct into an expression plasmid. The plasmids can be suitable for replication and integration in eukaryotes. Typical cloning plasmid contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.

[0484] The polynucleotides encoding fusion proteins can be cloned into a number of types of plasmids. For example, the polynucleotides can be cloned into a plasmid including, but not limited to a phagemid, a phage derivative, an animal virus, and a cosmid. Plasmids of particular interest include expression plasmid, replication plasmid, probe generation plasmid, and sequencing plasmid.

[0485] Further, the expression plasmid may be provided to cells, such as immune cells, in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in other virology and molecular biology manuals. Viruses, which are useful as viral vectors include, but are not limited to, retroviruses, adenoviruses,adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable viral vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01 / 96584; WO 01 / 29058; and U.S. Pat. No. 6,326,193).

[0486] A number of viral based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. A selected gene can be inserted into a plasmid and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo. A number of retroviral systems are known in the art. In some embodiments, adenovirus vectors are used. A number of adenovirus vectors are known in the art. In one embodiment, lentivirus vectors are used.

[0487] Additional promoter elements, e.g., enhancers, regulate the frequency of transcription initiation. Typically, these are located in the region 30-110 basepairs (bp) upstream of the start site, although a number of promoters have recently been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. In the thymidine kinase (tk) promoter, the spacing between promoter elements can be increased to 50 bp apart before activity begins to decline. Depending on the promoter, it appears that individual elements can function either cooperatively or independently to activate transcription.

[0488] One example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operatively linked thereto. Another example of a suitable promoter is Elongation Growth Factor-la (EF-la). Another example of a suitable promoter is the U6 promoter. Another example of a suitable promoter is the Hl promoter. However, other constitutive promoter sequences may also be used, including, but not limited to, the simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters including, but not limited to, the actin promoter, the myosin promoter, the hemoglobin promoter, and the creatine kinase promoter. Further, the invention should not be limited to the use of constitutive promoters. Inducible promoters are also contemplated as part of the invention. The use of an inducible promoter provides a molecular switch capable of turning on expression of thepolynucleotide sequence which it is operatively linked when such expression is desired, or turning off the expression when expression is not desired. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter. An illustrative sequence of the U6 promoter is set forth in SEQ ID NO: 129. An illustrative sequence of the Hl promoter is set forth in SEQ ID NO: 130.

[0489] In certain aspect, a plasmid comprises a termination sequence, for example, a T6 or T7 termination sequence.Cells

[0490] The disclosure provides cells comprising the receptors, vectors and polynucleotides described herein.

[0491] In some embodiments, the cell is an immune cell or a cell capable of differentiating into an immune cell.

[0492] As used herein, the term “immune cell” refers to a cell involved in the innate or adaptive (acquired) immune systems. Exemplary innate immune cells include phagocytic cells such as neutrophils, monocytes and macrophages, Natural Killer (NK) cells, polymorphonuclear leukocytes such as neutrophils eosinophils and basophils and mononuclear cells such as monocytes, macrophages and mast cells. Immune cells with roles in acquired immunity include lymphocytes such as T-cells and B-cells.

[0493] As used herein, a “T-cell” refers to a type of lymphocyte that originates from a bone marrow precursor that develops in the thymus gland. There are several distinct types of T-cells which develop upon migration to the thymus, which include, helper CD4+ T-cells, cytotoxic CD8+ T cells, memory T cells, regulatory CD4+ T-cells and stem memory T-cells. Different types of T- cells can be distinguished by the ordinarily skilled artisan based on their expression of markers. Methods of distinguishing between T-cell types will be readily apparent to the ordinarily skilled artisan.

[0494] In some embodiments, the activator receptor and the inhibitor receptor together specifically activate the immune cell in the presence of the target cell.

[0495] In some embodiments, the immune cell is CD4+, CD8+, a gamma delta T cell, an invariant T cells, an iNK cell, aNK cell, a macrophage, or combinations thereof. In some embodiments, the immune cell is a gamma delta (y5) T cell. In some embodiments, the immune cell is an invariant T cell. In some embodiments, the immune cell is an invariant natural killer T cell (iNKT cell). In some embodiments, the immune cell is a T cell. In some embodiments, the immune cell is aB cell.In some embodiments, the immune cell is a Natural Killer (NK) cell. In some embodiments, the immune cell is CD8-. In some embodiments, the immune cell is CD8+. In some embodiments, the immune cell is CD4+. In some embodiments, the immune cell is CD4-. In some embodiments, the immune cell is CD8- / CD4+. In some embodiments, the immune cell is a CD8+ CD4- T cell.

[0496] In some embodiments, the immune cell is non-natural. In some embodiments, the immune cell is isolated.

[0497] Methods transforming populations of immune cells, such as T cells, with the vectors of the instant disclosure will be readily apparent to the person of ordinary skill in the art. For example, CD3+ T cells can be isolated from PBMCs using a CD3+ T cell negative isolation kit (Miltenyi), according to manufacturer’s instructions. T cells can be cultured at a density of 1 x 10A6 cells / mL in X-Vivo 15 media supplemented with 5% human A / B serum and 1% Pen / strep in the presence of CD3 / 28 Dynabeads (1: 1 cell to bead ratio) and 300 Units / mL of IL-2 (Miltenyi). After 2 days, T cells can be transduced with viral vectors, such as lentiviral vectors using methods known in the art. In some embodiments, the viral vector is transduced at a multiplicity of infection (MOI) of 5. Cells can then be cultured in IL-2 or other cytokines such as combinations of IL-7 / 15 / 21 for an additional 5 days prior to enrichment. Methods of isolating and culturing other populations of immune cells, such as B cells, or other populations of T cells, will be readily apparent to the person of ordinary skill in the art. Although this method outlines a potential approach it should be noted that these methodologies are rapidly evolving. For example, excellent viral transduction of peripheral blood mononuclear cells can be achieved after 5 days of growth to generate a >99% CD3+ highly transduced cell population.

[0498] Methods of activating and culturing populations of T cells comprising the TCRs, CARs, inhibitor receptors, activator receptors, or vectors encoding same, will be readily apparent to the person of ordinary skill in the art.

[0499] In some embodiments, T cells of the instant disclosure are expanded and activated in vitro. Generally, the T cells of the instant disclosure are expanded in vitro by contact with a surface having attached thereto an agent that stimulates a CD3 / TCR complex associated signal and a ligand that stimulates a co-stimulatory molecule on the surface of the T cells. In particular, T cell populations may be stimulated as described herein, such as by contact with an anti-CD3 antibody. For co-stimulation of an accessory molecule on the surface of the T cells, a ligand that binds the accessory molecule is used. For example, a population of T cells can be contacted with an anti- CD3 antibody and an anti-CD28 antibody, under conditions appropriate for stimulating proliferation of the T cells. To stimulate proliferation of either CD4+ T cells or CD8+ T cells, an anti-CD3 antibody and an anti-CD28 antibody can be used. Examples of an anti-CD28 antibodyinclude 9.3, B-T3, XR-CD28 (Diaclone, Besangon, France) can be used as can other methods commonly known in the art (Berg et al., Transplant Proc. 30(8):3975-3977, 1998; Haanen et al., J. Exp. Med. 190(9): 13191328, 1999; Garland et al., J. Immunol Meth. 227(l-2):53-63, 1999).

[0500] The T cells are maintained under conditions necessary to support growth, for example, an appropriate temperature (e.g., 37° C.) and atmosphere (e.g., air plus 5% CO2).

[0501] In some embodiments, the T cells comprising TCRs, CARs and inhibitor receptors of the disclosure are autologous. Prior to expansion and genetic modification, a source of T cells is obtained from a subject. Immune cells such as T cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In certain embodiments of the present disclosure, any number of T cell lines available in the art, may be used. In certain embodiments of the present disclosure, T cells can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll™ separation.

[0502] In some embodiments, cells from the circulating blood of an individual are obtained by apheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets. In some embodiments, the cells collected by apheresis may be washed to remove the plasma fraction and to place the cells in an appropriate buffer or media for subsequent processing steps. In some embodiments, the cells are washed with phosphate buffered saline (PBS). In alternative embodiments, the wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations. As those of ordinary skill in the art would readily appreciate a washing step may be accomplished by methods known to those in the art, such as by using a semi -automated “flow-through” centrifuge (for example, the Cobe 2991 cell processor, the Baxter CytoMate, or the Haemonetics Cell Saver 5) according to the manufacturer's instructions. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as, for example, Ca2+-free, Mg2+- free PBS, PlasmaLyte A, or other saline solution with or without buffer. Alternatively, the undesirable components of the apheresis sample may be removed, and the cells directly resuspended in culture media.

[0503] In some embodiments, immune cells such as T cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a PERCOLL™ gradient or by counterflow centrifugal elutriation. Specific subpopulations of immune cells, such as T cells, B cells, or CD4+ T cells can be further isolated by positive or negative selection techniques. For example, in one embodiment, T cells are isolatedby incubation with anti-CD4 -conjugated beads, for a time period sufficient for positive selection of the desired T cells.

[0504] Enrichment of an immune cell population, such as a T cell population, by negative selection can be accomplished with a combination of antibodies directed to surface markers unique to the negatively selected cells. One method is cell sorting and / or selection via negative magnetic immune-adherence or flow cytometry that uses a cocktail of monoclonal antibodies directed to cell surface markers present on the cells negatively selected. For example, to enrich for CD4+ cells by negative selection, a monoclonal antibody cocktail typically includes antibodies to CD 14, CD20, CD 1 lb, CD 16, HLA-DR, and CD8.

[0505] For isolation of a desired population of immune cells by positive or negative selection, the concentration of cells and surface (e.g., particles such as beads) can be varied. In certain embodiments, it may be desirable to significantly decrease the volume in which beads and cells are mixed together (i.e., increase the concentration of cells), to ensure maximum contact of cells and beads.

[0506] In some embodiments, the cells may be incubated on a rotator for varying lengths of time at varying speeds at either 2-10° C or at room temperature.

[0507] T cells for stimulation, or PBMCs from which immune cells such as T cells are isolated, can also be frozen after a washing step. Wishing not to be bound by theory, the freeze and subsequent thaw step provides a more uniform product by removing granulocytes and to some extent monocytes in the cell population. After the washing step that removes plasma and platelets, the cells may be suspended in a freezing solution. While many freezing solutions and parameters are known in the art and will be useful in this context, one method involves using PBS containing 20% DMSO and 8% human serum albumin, or culture media containing 10% Dextran 40 and 5% Dextrose, 20% Human Serum Albumin and 7.5% DMSO, or 31.25% Plasmalyte-A, 31.25% Dextrose 5%, 0.45% NaCl, 10% Dextran 40 and 5% Dextrose, 20% Human Serum Albumin, and 7.5% DMSO or other suitable cell freezing media containing for example, Hespan and PlasmaLyte A, the cells then are frozen to -80° C at a rate of 1° per minute and stored in the vapor phase of a liquid nitrogen storage tank. Other methods of controlled freezing may be used as well as uncontrolled freezing immediately at -20° C or in liquid nitrogen.

[0508] The disclosure provides an immune cell expressing the activator receptors and / or inhibitor receptors described herein, wherein the immune cell has reduced expression and / or function the major histocompatibility (MHC) class I complex.

[0509] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is autogenic to a subject with the autoimmune disease or disorder.

[0510] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the logic-gated engineered immune cell is autogenic to a subject with the autoimmune disease or disorder.

[0511] In some embodiments, the immune cell is autologous. For example, the immune cells are isolated or derived from same subject who will receive the cell as part of a therapeutic regimen. It can be advantageous to modify autologous immune cells to have reduced expression and / or function of MHC class I with the inhibitor receptor is specific to an MHC class I antigen. Without wishing to be bound by theory, modification of autologous immune cells to have reduced expression and / or function of MHC class I reduces binding of the inhibitor receptor by MHC class I expressed by the immune cells, either in cis or in trans. In some embodiments, the immune cells do not express any heterologous receptors other than the activator and inhibitor receptors disclosed herein. In some embodiments, the immune cells express one or more additional heterologous receptors distinct from the activator and inhibitor receptors disclosed herein. In some embodiments, the immune cells do not express an endogenous receptor specific for one or more of the antigens disclosed herein. In some embodiments, the immune cells express an endogenous receptor specific for one or more of the antigens disclosed herein.

[0512] In embodiments, the present disclosure relates to an immune cell, wherein the immune cell is allogenic to a subject with the autoimmune disease or disorder.

[0513] In embodiments, the present disclosure relates to a logic-gated engineered immune cell, wherein the logic-gated engineered immune cell is allogenic to a subject with the autoimmune disease or disorder.

[0514] In some embodiments, the immune cell is all allogeneic. Allogeneic immune cells can be derived from a donor other than the subject to which the immune cells will be administered. Allogeneic immune cells have been commonly referred to in cell therapy as “off-the-shelf’ or “universal” because of the possibility for allogeneic cells to be prepared and stored for use in subjects of a variety of genotypes.Pharmaceutical Compositions

[0515] The disclosure provides pharmaceutical compositions comprising immune cells comprising the inhibitor receptors and activator receptors of the disclosure and a pharmaceutically acceptable diluent, carrier or excipient.

[0516] Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans,mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; and preservatives.

[0517] In some embodiments, the cell expresses both the activator receptor and the inhibitor receptor. In some embodiments, at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of the cells express both the activator receptor and the inhibitor receptor. In some embodiments, at least 90% of the cells express both the activator receptor and the inhibitor receptor.Treating autoimmune diseases or disorders

[0518] In embodiments, the present disclosure relates to a method for treating an autoimmune disease or disorder. The method comprising administering to the subject an effective amount of any herein-disclosed immune cell or administering to the subject an effective amount of a pharmaceutical composition comprising any herein-disclosed immune cell.

[0519] The autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti- synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren' s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

[0520] In embodiments, the autoimmune disease or disorder is multiple sclerosis (MS).

[0521] Treating the autoimmune disease can result in a decrease in number of autoreactive cells in the subject, e.g., as measured in a sample from the subject. Preferably, after treatment, the number of metastatic lesions is reduced by 5% or greater relative to number prior to treatment; more preferably, the number of autoreactive cells is reduced by 10% or greater; more preferably, reduced by 20% or greater; more preferably, reduced by 30% or greater; more preferably, reduced by 40% or greater; even more preferably, reduced by 50% or greater; and most preferably, reduced bygreater than 75%. The number of autoreactive cells may be measured by any reproducible means of measurement.

[0522] The cells and of the present disclosure may be administered in a number of ways depending upon whether local or systemic treatment is desired.

[0523] In general, administration may be parenteral.

[0524] Methods for administration of cells for adoptive cell therapy are known and may be used in connection with the provided methods and compositions. For example, adoptive T cell therapy methods are described, e.g., in US Patent Application Publication No. 2003 / 0170238 to Gruenberg et al and U.S. Pat. No. 4,690,915 to Rosenberg.

[0525] The compositions of the disclosure are suitable for parenteral administration. As used herein, “parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue, thus generally resulting in the direct administration into the blood stream, into muscle, or into an internal organ. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, subcutaneous, intraperitoneal, intramuscular, intrastemal, intravenous, intraarterial, intrathecal, intraventricular, intraurethral, intracranial, intratumoral, intrasynovial injection or infusions, and kidney dialytic infusion techniques. In some embodiments, parenteral administration of the compositions of the present disclosure comprises intravenous or intraarterial administration.

[0526] The disclosure provides pharmaceutical compositions comprising a plurality of immune cells of the disclosure, and a pharmaceutically acceptable carrier, diluent or excipient.

[0527] Formulations of a pharmaceutical composition suitable for parenteral administration typically generally comprise of immune cells combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and the like. Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents. Parenteral formulations also include aqueous solutions which may contain excipients such as salts, carbohydrates and bufferingagents. Exemplary parenteral administration forms include solutions or suspensions in sterile aqueous solutions, for example, aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered, if desired. Formulations for parenteral administration may be formulated to be immediate and / or modified release. Modified release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.

[0528] In some embodiments, the formulated composition comprising the immune cells is suitable for administration via injection. In some embodiments, the formulated composition comprising the immune cells is suitable for administration via infusion.

[0529] The pharmaceutical compositions of the present disclosure, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the immune cells with the pharmaceutical carrier(s) or excipient(s), such as liquid carriers.

[0530] Aqueous suspensions may further contain substances that increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and / or dextran. The suspension may also contain stabilizers.

[0531] The compositions of the present disclosure may additionally contain other adjunct components conventionally found in pharmaceutical compositions. Thus, for example, the compositions may contain additional, compatible, pharmaceutically active materials such as, for example, antipruritic, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present disclosure, such as dyes, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the immune cells of the compositions of the present disclosure.

[0532] The formulation or composition may also contain more than one active ingredient useful for the particular indication, disease, or condition being treated with the immune cells, where the respective activities do not adversely affect one another. Such active ingredients are suitably present in combination in amounts that are effective for the purpose intended. Thus, in some embodiments, the pharmaceutical composition further includes other pharmaceutically active agents or drugs, such as chemotherapeutic agents.

[0533] Administration can be effected in one dose, continuously or intermittently throughout the course of treatment. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician.

[0534] The pharmaceutical composition in some embodiments contains the immune cells in amounts effective to treat, ameliorate, or prevent an autoimmune disease or disorder, such as atherapeutically effective or prophylactically effective amount. Therapeutic or prophylactic efficacy in some embodiments is monitored by periodic assessment of treated subjects. For repeated administrations over days, weeks or months, depending on the condition, the treatment can be repeated until a desired suppression of the autoimmune disease or disorder signs or symptoms occurs. However, other dosage regimens may be useful and can be determined. The desired dosage can be delivered by a single bolus administration or infusion of the composition or by multiple bolus administrations or infusions of the composition.

[0535] The cells or population of cells can be administrated in one or more doses. In some embodiments, an effective amount of cells can be administrated as a single dose. In some embodiments, an effective amount of cells can be administrated as more than one doses over a period time. Timing of administration is within the judgment of a managing physician and depends on the clinical condition of the patient.

[0536] The cells or population of cells may be obtained from any source, such as a blood bank or a donor, or the patient themselves.

[0537] An effective amount means an amount which provides a therapeutic or prophylactic benefit. The dosage administered will be dependent upon the age, health and weight of the recipient, kind of concurrent treatment, if any, frequency of treatment and the nature of the effect desired. In some embodiments, an effective amount of cells or composition comprising those cells are administrated parenterally. In some embodiments, administration can be an intravenous administration.

[0538] In some embodiments, a therapeutically effective dose of the immune cells described herein are administered. In some embodiments, the immune cells of the disclosure are administered by intravenous injection. In some embodiments, the immune cells of the disclosure are administered by intraperitoneal injection. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells, about IxlO6cells, about 2xl06cells, about 3xl06cells, 4xl06cells, about 5xl06cells, about 6xl06cells, about 7xl06cells, about 8xl06cells, about 9xl06cells, about IxlO7, about 2xl07, about 3xl07, about 4xl07, about 5xl07, about 6xl07, about 7xl07, about 8xl07, about 9xl07, about IxlO8cells, about 2xl08cells, about 3xl08cells, about 4xl08cells, about 5xl08cells, about 6xl08cells, about 7xl08cells, about 8xl08cells, about 9xl08cells, about IxlO9cells, about 2xl09cells, about 3xl09cells, about 3xl09cells, about 4xl09cells, about 5xl09cells, about 5xl09cells, about 6xl09cells, about 7xl09cells, about 8xl09cells, about 9xl09cells, about IxlO10cells, about 2xlO10cells, about 3xlO10cells, about 4xlO10cells, about 5xlO10cells, about 6xlO10cells, about 7xlO10cells, about 8xlO10cells, or about 9xlO10cells.

[0539] In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about 9xlO10cells, about IxlO6cells to about 5xlO10cells, about 2xl06cells to about 5xl09cells, about 3xl06cells to about 5xl09cells, about 4xl06cells to about 3xl09cells, about 5xl06cells to about 2xl09cells, about 6xl06cells to about IxlO9cells, 0.5xl06cells to about 6xl09cells, about IxlO6cells to about 5xl09cells, about 2xl06cells to about 5xl09cells, about 3xl06cells to about 4xl09cells, about 4xl06cells to about 3xl09cells, about 5xl06cells to about 2xl09cells, about 6xl06cells to about IxlO9cells, 0.5xl06cells to about 6xl08cells, about IxlO6cells to about 5xl08cells, about 2xl06cells to about 5xl08cells, about 3xl06cells to about 4xl08cells, about 4xl06cells to about 3xl08cells, about 5xl06cells to about 2xl08cells, about 6xl06cells to about IxlO8cells, about 7xl06cells to about 9xl08cells, about 8xl06cells to about 8xl08cells, about 9xl06cells to about 7xl08cells, about IxlO7cells to about 6xl08cells, about 2xl07cells to about 5xl08cells, about 7xl06cells to about 9xl07cells, about 8xl06cells to about 8xl07cells, about 9xl06cells to about 7xl07cells, about IxlO7cells to about 6xl07cells, or about 2xl07cells to about 5xl07cells.

[0540] In some embodiments, a therapeutically effective dose comprises about 0.5xl05cells to about 9xlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about IxlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about 5xl09cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about IxlO9cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about 6xl08cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl06cells to about 9xlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl07cells to about IxlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl07cells to about 5xl09cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl07cells to about IxlO9cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl07cells to about 6x108cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl08cells to about 9xlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl08cells to about IxlO10cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl08cells to about 5xl09cells. In some embodiments, a therapeutically effective dose comprises about 0.5xl08cells to about IxlO9cells. The term “about” as referred to in a therapeutically dose, can be, for example, ± 0.5xl06cells, ± 0.5xl07cells, or ± 0.5xl08cells.

[0541] In some embodiments, the cells are administered as part of a combination treatment, such as simultaneously with or sequentially with, in any order, another therapeutic intervention. Theimmune cells of the disclosure are in some embodiments co-administered with one or more additional therapeutic agents or in connection with another therapeutic intervention, either simultaneously or sequentially in any order. In some contexts, the immune cells are coadministered with another therapy sufficiently close in time such that the immune cell populations enhance the effect of one or more additional therapeutic agents, or vice versa. In some embodiments, the immune cells are administered prior to the one or more additional therapeutic agents. In some embodiments, the immune cells are administered after to the one or more additional therapeutic agents.

[0542] In embodiments, the autoimmune disease is treated by a herein-disclosed method along with a standard of care therapy, e.g., with anti-inflammatory drugs, corticosteroids, and / or immunosuppressant drugs.

[0543] In embodiments, the autoimmune disease is treated by a herein-disclosed method following a standard of care therapy, e.g., with anti-inflammatory drugs, corticosteroids, and / or immunosuppressant drugs.

[0544] In embodiments, the autoimmune disease is treated by a herein-disclosed method before a standard of care therapy, e.g., with anti-inflammatory drugs, corticosteroids, and / or immunosuppressant drugs.Kits and Articles of Manufacture

[0545] In embodiments, the present disclosure relates to a kit including the logic-gated engineered immune cell or the immune cell and instructions for use.

[0546] In embodiments, the present disclosure relates to a kit including the pharmaceutical composition and instructions for use.

[0547] The disclosure provides kits and articles of manufacture comprising the polynucleotides and vectors encoding the receptors described herein, and cells comprising the receptors described herein. In some embodiments, the kit comprises articles such as vials, syringes and instructions for use.

[0548] In some embodiments, the kit comprises a polynucleotide or vector comprising a sequence encoding one or more receptors of the disclosure.

[0549] In some embodiments, the kit comprises a plurality of cells comprising the receptors as described herein. In some embodiments, the plurality of cells comprises a plurality of T cells.

[0550] In some embodiments, the kit further comprises instructions for use.GvHD

[0551] There is an increasing use of cellular therapy in which modified, or unmodified cells are administered to a patient. However, the coexistence of alloreactive T cells in a donor stem cell graft may cause graft-versus-host disease (GvHD) in which the donor cells react against the recipient, which may progressively damage the skin, gut, liver, and other organs of the recipient, often with fatal consequences.

[0552] The terms “graft versus host disease” or “GvHD”, refer to a complication often associated with allogeneic bone marrow transplantation and sometimes associated with transfusions of unirradiated blood to immunocompromised patients. Graft versus host disease sometimes can occur when functional immune cells in the transplanted marrow recognize the recipient as “foreign” and mount an immunologic response. GvHD can be divided into an acute form and a chronic form. Acute GVHD (aGVHD) often is observed within the first 100 days following transplant or transfusion and can affect the liver, skin, mucosa, immune system (e.g., the hematopoietic system, bone marrow, thymus, and the like), lungs and gastrointestinal tract. Chronic GVHD (cGVHD) often begins 100 days or later post-transplant or transfusion and can attack the same organs as acute GvHD, but also can affect connective tissue and exocrine glands. Acute GvHD of the skin can result in a diffuse maculopapular rash, sometimes in a lacy pattern.

[0553] The problem of graft rejection may be overcome by a combination of appropriate conditioning and large doses of stem cells, while graft versus host disease (GvHD) may be prevented by extensive T cell-depletion of the donor graft. The immediate outcomes of such procedures have been gratifying, with engraftment rate >90% and a severe GvHD rate of <10% for both adults and children even in the absence of post-transplant immunosuppression. Unfortunately, the profound immunosuppression of the grafting procedure, coupled with the extensive T celldepletion and HLA mismatching between donor and recipient lead to an extremely high rate of post-transplant infectious complications, and contributed to high incidence of disease relapse. The engineered immune cell system described herein may help overcome these obstacles by allowing for targeting of the autoreactive T cells responsible for GvHD but sparing the other immune cell types that are critical for normal immune system function.Illustrative immune cells of the present disclosure

[0554] An aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigenexpressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD69; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0555] Another aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD69; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 4; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 5; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 2; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 3; wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 702, 710, and 716; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 703, 711, and 717; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 704, 712, and 718; and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 705, 713, and 719; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 706, 714, and 720; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 707, 708, 709, 715, and 721.

[0556] An additional aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD69; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, and 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, and 1315 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 722, 724, 726, 728, 730 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 723, 725, 727, 729, 731 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0557] A further aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD25

[0558] In an aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD25;wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 4; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 5; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 2; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 3 wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 52, 58, 64, 70, 76, 78, 80, 83, 91, 97, 102, 108, 114, 120, 126, 131, and 137; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 53, 59, 65, 71, 77, 79, 81, 82, 84, 87, 88, 89, 90; 92, 98, 100, 103, 109, 115, 121, 127, and 132 or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs48, 54, 60, 66, 72, 85, 93, 99, 104, 110, 116, 122, 128, 133, and 139; and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 49, 55, 61, 67, 73, 94, 105, 111, 117, 123, 129, and 141; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 50, 56, 62, 68, 74, 95, 106, 112; 118, 124, and 135; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 51, 57, 63, 69, 75, 86, 96, 101, 107, 113, 119, 125, 130,136, and 140.

[0559] In another aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is CD25; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the firstVH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, and 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, and 1315, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 142, 144, 146, 148, 150, 152, 154, , 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215 ,217, 219, 221, 223, 225 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0560] In a further aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes:: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is 0X40; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region,.

[0561] In an additional aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is 0X40; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the firstVH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 4; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 5; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 2; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 3. wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 501, 507, 513, 518, 524, 534, 539, 544, 549, 555, 561, 565, and 570, and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467; 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573.

[0562] In yet another aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02 and the first activator antigen is 0X40;wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, and 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, and 1315 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 574, 576, 578, 580, 582, 584, 586, 588, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632, 636 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0563] In yet a further aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell and a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02, the first activator antigen is 4-1BB, and the second activator antigen is CD19.

[0564] In yet an additional aspect, the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; an activator receptor including a first extracellular ligandbinding domain specific to a first activator antigen expressed by an autoreactive immune cell and a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02, the first activator antigen is 4- IBB, and the second activator antigen is CD 19; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 4; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 5; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 2; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 3. wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 501, 507, 513, 518, 524, 534, 539, 544, 549, 555, 561, 565, and 570, and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467; 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573; and wherein the second extracellular ligand bindingdomain of the activator receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 953, 959, 970, 976, 985, and 991; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 954, 960, 965, 971, 977, 982, 986, 992, and 997; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 955, 961, 966, 972, 978, 987, and 993; and the second VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 956, 962, 967, 973, 979, 983, 988, and 994; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs 957, 963, 968, 974, 980, 989, 995, and 998; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 958, 964, 969, 975, 981, 984, 990, and 996.

[0565] An aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; and an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell and a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02, the first activator antigen is 4- IBB, and the second activator antigen is CD 19; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1290, 1292, 1294, 1296, 1298, 1300, 1302, 1304, 1306, 1308, 1310, 1312, and 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, and 1315 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 861, 863, 865, 867, 869, 871, 873, 875, 877, 879, 881, 883, 885, 887, 889, 891, 893, 895, 897, 899, 901, 903, 905 or a sequence having at least 85%,at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 864, 866, 868, 870, 872, 874, 876, 878, 880, 882, 884, 886, 888, 890, 892, 894, 896, 898, 900, 902, 904, 906 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the second extracellular ligand binding domain of the activator receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 999, 1001, 1003, 1005,1007, 1009, 1011, 1013, 1015 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the second VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 1000, 1002, 1004,1008, 1010, 1012, 1014, 1016 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0566] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen and the second extracellular ligand binding domain specific to a second activator antigen are separated by a linker.

[0567] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0568] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0569] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0570] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0571] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0572] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0573] Another aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell and a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is LRRC32, the first activator antigen is CD 19, and the second activator antigen is 0X40; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0574] A further aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell and a second extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is LRRC32, the first activator antigen is CD 19, and the second activator antigen is 0X40; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 310, 323, 329, 335, 340, and 351 (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 311, 324, 330, 336, 341, 346, 347, and 352; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 312, 325, 331, 337, 342, 348, and 353; and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 313, 316, 320, 326, 332, 338, 343, and 354; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 314, 318, 321, 327, 333, 339, 344, and 349; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 315, 317, 319, 322, 328, 334, 340, 345, 350, and 356. wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1)including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 953, 959, 970, 976, 985, and 991; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 954, 960, 965, 971, 977, 982, 986, 992, and 997; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 955, 961, 966, 972, 978, 987, and 993; and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 956, 962, 967, 973, 979, 983, 988, and 994; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 957, 963, 968, 974, 980, 989, 995, and 998; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 958, 964, 969, 975, 981, 984, 990, and 996.; and wherein the second extracellular ligand binding domain of the activator receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 415, 421, 427, 430, 436, 442, 448, 454, 460, 464, 470, 476, 481, 487, 493, 499, 505, 511, 516, 522, 526, 532, 537, 542, 547, 553, 559, 563, and 568; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 416, 422, 428, 431, 437, 443, 449, 455, 461, 465, 471, 477, 482, 488, 494, 500, 506, 512, 517, 523, 527, 533, 538, 543, 548, 554, 560, 564, and 569; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 417, 423, 429, 432, 438, 444, 450, 456, 462, 466, 472, 478, 483, 489, 501, 507, 513, 518, 524, 534, 539, 544, 549, 555, 561, 565, and 570, and the second VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 418, 424, 433, 439, 445, 451, 457, 467; 473, 479, 484, 490, 496, 502, 508, 514, 519, 529, 540, 545, 550, 556, 566, and 571; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 419, 425, 434, 440, 446, 452, 458, 468, 474, 485, 491, 497, 503, 509, 520, 530, 535, 551, 557, and 572; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 420, 426, 435, 441, 447, 453, 459, 463, 469, 475, 480, 486, 492, 498, 504, 510, 515, 521, 525, 531, 536, 541, 546, 552, 558, 562, 567, and 573.

[0575] An additional aspect of the present disclosure is an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell; an activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell and asecond extracellular ligand binding domain specific to a second activator antigen expressed by the autoreactive immune cell; wherein the first inhibitor antigen is LRRC32, the first activator antigen is CD 19, and the second activator antigen is 0X40; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382 ,384, 386 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 999, 1001, 1003, 1005, 1007, 1009, 1011, 1013, 1015 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 1000, 1002, 1004, 1006, 1008, 1010 , 1012, 1014, 1016 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the second extracellular ligand binding domain of the activator receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the activator receptor includes the sequence of any one of SEQ ID NOs 574, 576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606, 608, 610, 612, 614, 616, 618, 620 ,622 ,624, 626, 628, 630, 632, 634, 636 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the second VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635, 637 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0576] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen and the second extracellular ligand binding domain specific to a second activator antigen are separated by a linker.

[0577] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0578] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0579] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0580] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0581] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0582] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0583] In an aspect of the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell and a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell; and a first activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HL A- A* 02, the second inhibitor antigen is CCR8, and the first activator antigen is CD25; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region.

[0584] In an additional aspect of the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell and a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell; and a first activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HLA-A*02,the second inhibitor antigen is CCR8, and the first activator antigen is CD25; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 4; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 5; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 6, and the first VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 2; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 3 wherein the second extracellular ligand binding domain of the inhibitor receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the inhibitor receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 1180, 1186, 1192, 1197 1206, 1212, 1222, and, 1228; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1181, 1187; 1193, 1198, 1207, 1213, 1223, and 1229; or (c) a CDR-H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1182, 1188, 1194, 1195, 1199, 1203, 1208, 1214, 1218, 1224, and 1230, and the second VL region of the inhibitor receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1183, 1189, 1196, 1200, 1204, 1209, 1215, 1219, 1225, and 1231; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1184, 1190, 1201; 1205, 1210, 1216, 1220, 1226, and 1232; ; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 1185, 1191, 1202, 1211, 1217, 1221, 1227, and 1233 wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the activator receptor includes: (a) a complementarity-determining region VH-1 (CDR-H1) including any one amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 52, 58, 64, 70, 76, 78, 80, 83, 91, 97, 102, 108, 114, 120, 126, 131, and 137; (b) a CDR-H2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 47, 53, 59, 65, 71, 77, 79, 81, 82, 84, 87, 88, 89, 90; 92, 98, 100, 103, 109, 115, 121, 127, and 132; or (c) a CDR- H3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 48,54, 60, 66, 72, 85, 93, 99, 104, 110, 116, 122, 128, 133, and 139; and the first VL region of the activator receptor includes: (d) a CDR-L1 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 49, 55, 61, 67, 73, 94, 105, 111, 117, 123, 129, and 141; (e) a CDR-L2 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 50, 56, 62, 68, 74, 95, 106, 112; 118, 124, and 135; or (f) a CDR3-L3 including an amino acid sequence selected from the group consisting of SEQ ID NOs: 51, 57, 63, 69, 75, 86, 96, 101, 107, 113, 119, 125, 130,136, and 140..

[0585] In a further aspect of the present disclosure provides an immune cell, e.g., for treatment of an autoimmune disease or disorder. The immune cell includes: an inhibitor receptor which includes a first extracellular ligand binding domain specific to a first inhibitor antigen expressed by a second cell and a second extracellular ligand binding domain specific to a second inhibitor antigen expressed by the second cell; and a first activator receptor including a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell; wherein the first inhibitor antigen is HL A- A* 02, the second inhibitor antigen is CCR8, and the first activator antigen is CD25; wherein the first extracellular ligand binding domain of the inhibitor receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1290, 1292, 1294, 1296, 1298, 1300, 1302,1304, 1306, 1308, 1310, 1312, and 1314 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1291, 1293, 1295, 1297, 1299, 1301, 1303, 1305, 1307, 1309, 1311, 1313, and 1315 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; wherein the second extracellular ligand binding domain of the inhibitor receptor includes a second heavy chain variable (VH) region and a second light chain variable (VL) region, wherein the second VH region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1283, 1285, 1287, 1289, 1291, 1293, 1295, 1297, 1299, 1301, 1303,1305, and 1307 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the second VL region of the inhibitor receptor includes the sequence of any one of SEQ ID NOs 1235, 1237, 1239, 1241, 1243, 1245, 1247, 1249, 1251, 1253, 1255, 1257, 1259, 1261 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto; and wherein the first extracellular ligand binding domain of the activator receptor includes a first heavy chain variable (VH) region and a first light chain variable (VL) region, wherein the first VHregion of the activator receptor includes the sequence of any one of SEQ ID NOs 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto and wherein the first VL region of the activator receptor includes the sequence of any one of SEQ ID NOs 143, 145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183, 185 ,187, 189, 191, 193, 195, 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

[0586] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen and the second extracellular ligand binding domain specific to a second activator antigen are separated by a linker.

[0587] In embodiments, the present disclosure relates to an immune cell, wherein the linker is a short oligopeptide linker or polypeptide linker.

[0588] In embodiments, the present disclosure relates to an immune cell, wherein the short oligopeptide linker or polypeptide linker includes from 2 to 20 amino acids.

[0589] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes one or more Glycines (G), one or more Serines (S), one or more Glutamines (Q), and / or one or more Glutamic Acids (E).

[0590] In embodiments, the present disclosure relates to an immune cell, wherein the linker includes GSTSGSGKPGSGEGSTKG (SEQ ID NO: 1316); GSGKPGSGEGS (SEQ ID NO: 1317); EAAAKEAAAK (SEQ ID NO: 1318); GGGGQGGGGQ (SEQ ID NO: 1319); GGGG (SEQ ID NO: 1320); or QGGGGQGGGGQQ (SEQ ID NO: 1321).

[0591] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0592] In embodiments, the present disclosure relates to an immune cell, wherein the first extracellular ligand binding domain specific to a first activator antigen is N-terminal to the second extracellular ligand binding domain specific to a second activator antigen.

[0593] In embodiments, the present disclosure relates to a pharmaceutical composition, including an effective amount of any herein-disclosed immune cell and a pharmaceutically- acceptable excipient.

[0594] In embodiments, the present disclosure relates to a kit including any herein-disclosed immune cell and instructions for use. In embodiments, the present disclosure relates to a kit including the pharmaceutical composition and instructions for use.

[0595] In embodiments, the present disclosure relates to a method for treating an autoimmune disease or disorder by administering an effective amount of any herein-disclosed immune cell or pharmaceutical composition comprising any herein-disclosed immune cell. In embodiments, the autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti -synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti-synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren' s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti- MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

[0596] In embodiments, the present disclosure relates to a polynucleotide or polynucleotide system, including one or more polynucleotides including one or more polynucleotide sequences encoding any herein-disclosed activator receptor or the inhibitor receptor.

[0597] In embodiments, the present disclosure relates to a nanocarrier including the polynucleotide or polynucleotide system. In embodiments, the present disclosure relates to a nanocarrier, wherein the nanocarrier is capable of delivering the polynucleotide or polynucleotide system to an immune cell in vivo or ex vivo. In embodiments, the present disclosure relates to a nanocarrier, wherein the nanocarrier is a lipid nanoparticle (LNP). In embodiments, the present disclosure relates to a nanocarrier, wherein the polynucleotide or polynucleotide system are one or more messenger ribonucleic acids (mRNAs) or modified mRNAs (mmRNAs).

[0598] Any immune cell, composition, or method disclosed herein is applicable to any herein- disclosed immune cell, composition, or method. In other words, any aspect or embodiment described herein can be combined with any other aspect or embodiment as disclosed herein.Definitions

[0599] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of particular embodiments, preferred embodiments of compositions, methods and materials are described herein. For the purposes of the present disclosure, the following terms are defined below. Additional definitions are set forth throughout this disclosure.

[0600] As used herein, the singular forms “a”, “an”, and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent that the terms “including”, “includes”, “having”, “has”, “with”, or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising.”

[0601] As used herein, the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “include” and “includes”) or “containing” (and any form of containing, such as “contain” and “contains”), are inclusive or open-ended and do not exclude additional, unrecited elements or process steps. As also used herein, in any instance or embodiment described herein, “comprising” may be replaced with “consisting essentially of’ and / or “consisting of’ used herein, in any instance or embodiment described.

[0602] As used herein, the term “about” or “approximately” refers to a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length that varies by as much as 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% to a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length. In one embodiment, the term “about” or “approximately” refers a range of quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, or ± 1% about a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length.

[0603] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, whenappropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. The term “about,” when immediately preceding a number or numeral, means that the number or numeral ranges plus or minus 10%.

[0604] As used herein, the phrases “at least one”, “one or more”, and “and / or” are open-ended expressions that are both conjunctive and disjunctive in operation. For example, each of the expressions “at least one of A, B and C”, “at least one of A, B, or C”, “one or more of A, B, and C”, “one or more of A, B, or C” and “A, B, and / or C” means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B and C together.

[0605] As used herein, “or” may refer to “and”, “or,” or “and / or” and may be used both exclusively and inclusively. For example, the term “A or B” may refer to “A or B”, “A but not B”, “B but not A”, and “A and B”. In some cases, context may dictate a particular meaning.

[0606] Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. As another example, a range such as 95-99% identity, includes something with 95%, 96%, 97%, 98% or 99% identity, and includes subranges such as 96-99%, 96-98%, 96-97%, 97-99%, 97-98% and 98-99% identity. This applies regardless of the breadth of the range.

[0607] As used herein, the “administering” of an agent, e.g., an engineered immune cell, to a subject or subject includes any route of introducing or delivering to a subject a compound to perform its intended function. Suitable dosage formulations and methods of administering the agents are known in the art. Route of administration can also be determined and method of determining the most effective route of administration are known to those of skill in the art and will vary with the composition used for treatment, the purpose of the treatment, the health condition or disease stage of the subject being treated and target cell or tissue. Non-limiting examples of route of administration include parenteral, enteral, and topical routes of administration. Administration includes self-administration and the administration by another. It is also to be appreciated that the various modes of treatment or prevention of medical conditions as described are intended to mean “substantial”, which includes total but also less than total treatment or prevention, and wherein some biologically or medically relevant result is achieved.

[0608] As used herein “logic-gated engineered immune cells”, “immune cells of the present disclosure”, “immune cells”, “engineered immune cells”, “engineered immune cells of the present disclosure”, and the like are synonymous and refer to the same immune cell population. Thus, for example, embodiments disclosed herein relating to “logic-gated engineered immune cells” applies to “engineered immune cells of the present disclosure”.

[0609] As used herein, the term “isolated” means material that is substantially or essentially free from components that normally accompany it in its native state.

[0610] In embodiments, the term “obtained” or “derived” is used synonymously with isolated. In embodiments, the terms “obtained” or “derived” mean selecting a known amino acid sequence and including the known amino acid sequence into an antigen binding domain, ligand binding domain, construct, receptor, polynucleotide, or polypeptide of the present disclosure. As an example, the expression a “ligand binding domain may be derived from commercially available antibodies” may be interpreted as incorporating the amino acid sequence for one or more complement determining regions (CDRs) of the commercially available antibodies into an antigen binding domain, ligand binding domain, construct, receptor, or polypeptide of the present disclosure. Additionally, a VH or VL of one or more commercially available antibodies may be incorporated into an antigen binding domain, ligand binding domain, construct, receptor, or polypeptide of the present disclosure. A known amino acid sequence comprises a non-antigen binding domain of a commercially available antibody; here the non-antigen binding domain is incorporated into a construct, receptor, or polypeptide of the present disclosure. In some embodiments, “incorporating” comprises including the nucleic acid sequence encoding the known amino acid sequence into a polynucleotide that encodes an antigen binding domain, ligand binding domain, construct, receptor, or polypeptide of the present disclosure.

[0611] The terms “subject,” “patient” and “individual” are used interchangeably herein to refer to a vertebrate, preferably a mammal, more preferably a human. Tissues, cells, and their progeny of a biological entity obtained in vivo or cultured in vitro are also encompassed. A “subject,” “patient” or “individual” as used herein, includes any animal that exhibits pain that can be treated with the vectors, compositions, and methods contemplated herein. Suitable subjects (e.g., patients) include laboratory animals (such as mouse, rat, rabbit, or guinea pig), farm animals, and domestic animals or pets (such as a cat or dog). Non-human primates and, preferably, human patients, are included.

[0612] As used herein “treatment” or “treating,” includes any beneficial or desirable effect, and may include even minimal improvement in symptoms. “Treatment” does not necessarily indicate complete eradication or cure of the disease or condition, or associated symptoms thereof.

[0613] As used herein, “prevent,” and similar words such as “prevented,” “preventing” etc., indicate an approach for preventing, inhibiting, or reducing the likelihood of a symptom of disease. It also refers to delaying the onset or recurrence of a disease or condition or delaying the occurrence or recurrence of the symptoms of a disease. As used herein, “prevention” and similar words also includes reducing the intensity, effect, symptoms and / or burden of disease prior to onset or recurrence.

[0614] As used herein, the term “amount” refers to “an amount effective” or “an effective amount” of a virus to achieve a beneficial or desired prophylactic or therapeutic result, including clinical results.

[0615] A “therapeutically effective amount” of a virus or cell may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the virus or cell to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the virus or cell are outweighed by the therapeutically beneficial effects. The term “therapeutically effective amount” includes an amount that is effective to “treat” a subject (e.g., a patient).

[0616] An “increased” or “enhanced” amount of a physiological response, e.g., electrophysiological activity or cellular activity, is typically a “statistically significant” amount, and may include an increase that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 or more times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the level of activity in an untreated cell.

[0617] A “decreased” or “reduced” amount of a physiological response, e.g., electrophysiological activity or cellular activity, is typically a “statistically significant” amount, and may include an decrease that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 ormore times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the level of activity in an untreated cell.

[0618] By “maintain,” or “preserve,” or “maintenance,” or “no change,” or “no substantial change,” or “no substantial decrease” refers generally to a physiological response that is comparable to a response caused by either vehicle, or a control molecule / composition. A comparable response is one that is not significantly different or measurable different from the reference response.

[0619] In general, “sequence identity” or “sequence homology” refers to an exact nucleotide-to- nucleotide or amino acid-to-amino acid correspondence of two polynucleotides or polypeptide sequences, respectively. Typically, techniques for determining sequence identity include determining the nucleotide sequence of a polynucleotide and / or determining the amino acidsequence encoded thereby and comparing these sequences to a second nucleotide or amino acid sequence. Two or more sequences (polynucleotide or amino acid) can be compared by determining their “percent identity.” The percent identity of two sequences, whether nucleic acid or amino acid sequences, is the number of exact matches between two aligned sequences divided by the length of the shorter sequences and multiplied by 100. Percent identity may also be determined, for example, by comparing sequence information using the advanced BLAST computer program, including version 2.2.9, available from the National Institutes of Health. The BLAST program is based on the alignment method of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990) and as discussed in Altschul, et al., J. Mol. Biol. 215:403-410 (1990); Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5877 (1993); and Altschul et al., Nucleic Acids Res. 25:3389- 3402 (1997). Briefly, the BLAST program defines identity as the number of identical aligned symbols (generally nucleotides or amino acids), divided by the total number of symbols in the shorter of the two sequences. The program may be used to determine percent identity over the entire length of the proteins being compared. Default parameters are provided to optimize searches with short query sequences in, for example, with the blastp program. The program also allows use of an SEG filter to mask-off segments of the query sequences as determined by the SEG program of Wootton and Federhen, Computers and Chemistry 17:149-163 (1993). Ranges of desired degrees of sequence identity are approximately 80% to 100% and integer values therebetween. Typically, the percent identities between a disclosed sequence and a claimed sequence are at least 80%, at least 85%, at least 90%, at least 95%, or at least 98%.

[0620] As used herein, the term “antigen” refers to a compound, composition, or substance that may be specifically bound by the products of specific humoral or cellular immunity, such as an antibody molecule or T-cell receptor. Antigens can be any type of molecule including, for example, haptens, simple intermediary metabolites, sugars (e.g., oligosaccharides), lipids, and hormones as well as macromolecules such as complex carbohydrates (e.g., polysaccharides), phospholipids, and proteins. Common categories of antigens include, but are not limited to, viral antigens, bacterial antigens, fungal antigens, protozoa and other parasitic antigens, tumor antigens, antigens involved in autoimmune disease, allergy and graft rejection, toxins, and other miscellaneous antigens.

[0621] As used herein, “binding affinity” refers to the tendency of one molecule to bind (typically non-covalently) with another molecule, such as the tendency of a member of a specific binding pair for another member of a specific binding pair. A binding affinity can be measured as a dissociation constant, which for a specific binding pair (such as an antibody / antigen pair) can be lower than I X I 05M, lower than 1x 106M, lower than 1x 107M, lower than 1X1CT8M, lower than 1x 109M, lower than 1x 10l0M, lower than Ix l 01 1M or lower than 1x 10l 2M. In one aspect, binding affinityis calculated by a modification of the Scatchard method described by Frankel et al., Mol. Immunol, 16: 101-106, 1979. In another aspect, binding affinity is measured by a binding constant. In another aspect, binding affinity is measured by an antigen / antibody dissociation rate. In yet another aspect, a high binding affinity is measured by a competition radioimmunoassay.

[0622] The term “human antibody” as used herein, is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the disclosure may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis or by somatic mutation in vivo . However, the term “human antibody” as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a rabbit, have been grafted onto human framework sequences. Thus, as used herein, the term “human antibody” refers to an antibody in which substantially every part of the protein (e.g., CDR, framework, CL, CH domains (e.g., CHI, CH2, Cm), hinge, VL, VH) is substantially non-immunogenic in humans, with only minor sequence changes or variations. Similarly, antibodies designated primate (monkey, baboon, chimpanzee, etc.), rodent (mouse, rat, rabbit, guinea pig, hamster, and the like) and other mammals designate such species, sub-genus, genus, sub-family, family specific antibodies. Further, chimeric antibodies include any combination of the above. Such changes or variations optionally and preferably retain or reduce the immunogenicity in humans or other species relative to non-modified antibodies. Thus, a human antibody is distinct from a chimeric or humanized antibody. It is pointed out that a human antibody can be produced by a non-human animal or prokaryotic or eukaryotic cell that is capable of expressing functionally rearranged human immunoglobulin (e.g., heavy chain and / or light chain) genes. Further, when a human antibody is a single chain antibody, it can comprise a linker peptide that is not found in native human antibodies. For example, an Fv can comprise a linker peptide, such as two to about eight glycine or other amino acid residues, which connects the variable region of the heavy chain and the variable region of the light chain. Such linker peptides are considered to be of human origin.

[0623] As used herein, a “target cell” refers to cell that is targeted by an adoptive cell therapy. For example, a target cell can be an autoreactive cell, which can be killed by the transplanted T cells of the adoptive cell therapy. Target cells of the disclosure express a target antigen, as described herein, and do not express a non-target antigen.

[0624] As used herein, a “non-target cell” refers to cell that is not targeted by an adoptive cell therapy. For example, in an adoptive cell targeting autoreactive cells, normal, healthy cells which modulate and / or mitigate the immune system, e.g., T regs, are non-target cells. Some, or all, non-target cells in a subject may express both the target antigen and the non-target antigen. Non-target cells in a subject may express the non-target antigen irrespective of whether or not these cells also express the target antigen.

[0625] As used herein, HLA-A*02 antibody” and “anti-HLA-A*02 antibody” are used interchangeably and refer to an antibody that specifically binds to an HLA-A*02 polypeptide. Similarly, the term “HLA-A*02”, as mentioned in “HLA-A*02 binding”, “HLA-A*02 targeting”, “HLA-A*02 specific”, “HLA-A*02 expression” or other similar terms here, refers to HLA-A*02 polypeptide. The term “A*02” is understood to refer to HLA-A*02 in any instance. HLA-A*02 antibodies of the disclosure may bind to proteins falling within the HLA-A*02 allele group (for example HLA-A*02:01, HLA-A*02:02 and the like), and may not bind, or bind with lower affinity, to proteins falling within other HL A- A allele groups (for example, HLA-A*01, HLA- A* 11 , and the like). The person of ordinary skill in the art will recognize that some cross-reactivity with other HLA antigens may exist, but that the HLA-A*02 antibodies of the disclosure will still be considered to be specific to HLA-A*02. In some cases, specificity is considered in the context of the subject to be treated with an HLA-A*02 antibody or receptor of the disclosure. When the subject has both an HLA-A*02 allele and a second HLA-A allele not recognized, or only poorly recognized, by the HLA-A*02 antibody or receptor comprising an equivalent antigen binding domain, the HLA-A*02 antibody is specific to the HLA-A*02 allele of the subject.

[0626] As used herein, a “TCR”, sometimes also called a “TCR complex” or “TCR / CD3 complex” refers to a protein complex comprising a TCR alpha chain, a TCR beta chain, and one or more of the invariant CD3 chains (zeta, gamma, delta and epsilon), sometimes referred to as subunits. The TCR alpha and beta chains can be disulfide-linked to function as a heterodimer to bind to peptide- MHC complexes. Once the TCR alpha / beta heterodimer engages peptide-MHC, conformational changes in the TCR complex in the associated invariant CD3 subunits are induced, which leads to their phosphorylation and association with downstream proteins, thereby transducing a primary stimulatory signal. In an exemplary TCR complex, the TCR alpha and TCR beta polypeptides form a heterodimer, CD3 epsilon and CD3 delta form a heterodimer, CD3 epsilon and CD3 gamma for a heterodimer, and two CD3 zeta form a homodimer.

[0627] As used herein, “specific to” or “specifically binds to” when used with respect to a ligand binding domain, such as an antigen binding domain, refers to a ligand binding domain that has a high specificity for a named target. Antibody specificity can be viewed as a measure of the goodness of fit between the ligand binding domain and the corresponding ligand, or the ability of the ligand binding domain to discriminate between similar or even dissimilar ligands. In comparison with specificity, affinity is a measure of the strength of the binding between the ligandbinding domain and ligand, such that a low-affinity ligand binding domain binds weakly and high- affinity ligand binding domain binds firmly. The person of skill in the art will appreciate that a ligand binding domain can be said to be specific to a particular target, and yet still have low levels of binding to one or more additional targets that do not affect its function in the receptor systems described herein.

[0628] As used herein, a “target antigen,” whether referred to using the term antigen or the name of a specific antigen, refers to an antigen expressed by a target cell, such as a autoreactive cell. Expression of target antigen is not limited to target cells. Target antigens may be expressed by both autoreactive cells and normal, second cells, as described herein, in a subject.

[0629] As used herein, “activation” of an immune cell or an immune cell that is “activated” refers to an immune cell that can carry out one or more functions characteristic of an immune response. These functions include proliferation, release of cytokines, and cytotoxicity, i.e., killing of a target cell. Activated immune cells express markers that will be apparent to persons of skill in the art. For example, activated T cells can express one or more of CD69, CD71, CD25 and HLA-DR.

[0630] Receptor expression on an immune cell can be verified by assays that report the presence of the activator receptors and inhibitor receptors described herein. For example, a population of immune cells can be stained with a labeled molecule (e.g., a fluorophore labeled receptor-specific antibody or a fluorophore-labeled receptor-specific ligand) and quantified using fluorescence activated cell sorting (FACS) flow cytometry. This method allows a percentage of immune cells in a population of immune cells to be characterized as expressing an activator receptor, an inhibitor receptor, or both receptors. The ratio of activator receptor and inhibitor receptors expressed by the immune cells described herein can be determined by, for example, digital droplet PCR. These approaches can be used to characterize the population of cells for the production and manufacturing of the immune cells, pharmaceutical compositions, and kits described herein. For the immune cells, pharmaceutical compositions, and kits described herein, it is understood that a suitable percentage of immune cells expressing both an activator receptor and an inhibitor receptor is determined specifically for the methods described herein. For example, a suitable percentage of immune cells expressing both an activator receptor and in inhibitor receptor can be 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%, or at least 95%. As a further example, between 50% and 99%, between 60% and 95%, between 65% and 95%, between 65% and 90%, between 70% and 90%, between 75% and 90%, between 75% and 85%, between 80% and 99%, between 85% and 99%, between 90% and 99% or between 95% and 99% of immune cells can express both the activator receptor and the inhibitor receptor. For example, a suitable ratio of activator receptor and inhibitor receptor in an immune cell can be about5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, or about 1:5. It is understood that purification, enrichment, and / or depletion steps can be used on populations of immune cells to meet suitable values for the immune cells, pharmaceutical compositions, and kits described herein.

[0631] An activator antigen can be any antigen that is typically and / or substantially expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease including Thl7 cells, but not typically expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein, or is minimally expressed by the non-autoreactive cells. In contrast, an inhibitor antigen can be any antigen that is not typically and / or is minimally expressed on an activated cell and / or an autoreactive cell, e.g., a cell associated with an autoimmune disease, but is typically and / or substantially expressed by non-autoreactive cells, e.g., a naive T cell, a memory T cell, or a Treg, including any “second cell” as used herein. In some cases, a cell will express both an activator antigen and an inhibitor antigen. In this case, as discussed below, activity of an immune cell of the present disclosure may be blocked or reduced by the presence of the inhibitor antigen, even in the presence of the activator antigen; thus, reduces or preventing an undesirable cytotoxic attack on the cell expressing both antigens.

[0632] A responsive receptor expressed by the immune cells described herein can be verified by assays that measure the generation of a signal expected to be generated by the intracellular domain of the receptor. Reporter cell lines, such as Jurkat-Luciferase NF AT cells (Jurkat cells), can be used to characterize a responsive receptor. Jurkat cells are derived from T cells and comprise a stably integrated nuclear factor of activated T-cells (NFAT)-inducible luciferase reporter system. NF AT is a family of transcription factors required for immune cell activation, whose activation can be used as a signaling marker for T cell activation. Jurkat cells can be transduced or transfected with the activator receptors and / or inhibitor receptors described herein. The activator receptor is responsive to the binding of a ligand if the Jurkat cell expresses a luciferase reporter gene, and the level of responsiveness can be determined by the level of reporter gene expression. The presence of luciferase can be determined using any known luciferase detection reagent, such as luciferin. An inhibitor receptor is responsive to the binding of a ligand if, when co-expressed with an activator receptor in Jurkat cells, it prevents a normally responsive immune cell from expressing luciferase in response to the activator receptor. For example, the responsiveness of an inhibitor receptor can be determined and quantified in a Jurkat cell expressing both an activator and an inhibitor by observing the following: 1) the Jurkat cell expresses luciferase in the presence of activator receptor ligand and absence of inhibitor receptor ligand; and 2) luciferase expression in the Jurkat cell isreduced or eliminated in the presence of both an activator receptor ligand and an inhibitor receptor ligand. This approach can be used to determine the sensitivity, potency, and selectivity of activator receptors and specific pairs of activator receptors and inhibitor receptors. The sensitivity, potency, and selectivity can be quantified by EC50 or IC50 values using dose-response experiments, where an activator receptor ligand and / or inhibitor receptor ligand is titrated into a culture of Jurkat cells expressing an activator receptor or a specific pair of activator and inhibitor receptors. Alternatively, the EC50 and IC50 values can be determined in a co-culture of immune cells (e.g., Jurkat cells or primary immune cells) expressing an activator receptor or a specific pair of activator and inhibitor receptors and target cells expressing an increasing amount of an activator ligand or inhibitor ligand. An increasing amount of activator ligand or inhibitor ligand can be accomplished in the target cell by, for example, titration of activator ligand or inhibitor ligand encoding mRNA into target cells, or use of target cells that naturally express different levels of the target ligands. Exemplary suitable EC50 and IC50 values for the activator and inhibitor receptors as determined used target cells expressing varying amounts of the target and non-target ligands include an EC50 of 10 transcripts per million (TPM) or less for the activator receptor, for example an EC50 of between 2-10 TPM, and an IC50 of 25 TPM or less for the inhibitor receptor, for example an IC50 of 5-21 TPM.

[0633] Activation of the immune cells described herein that express an activator receptor or specific pairs of activator and inhibitor receptors can be further determined by assays that measure the viability of a target cell following co-incubation with said immune cells. The immune cells, sometimes referred to as effector cells, are co-incubated with target cells that express an activator receptor ligand, an inhibitor receptor ligand, or both an activator and inhibitor receptor ligand. Following co-incubation, viability of the target cell is measured using any method to measure viability in a cell culture. For example, viability can be determined using a mitochondrial function assay that uses a tetrazolium salt substrate to measure active mitochondrial enzymes. Viability can also be determined using imaging-based methods. Target cells can express a fluorescent protein, such as green fluorescent protein or red fluorescent protein. Reduction in total cell fluorescence indicates a reduction in viability of the target cell. A reduction in viability of the target cell following incubation with immune cells expressing an activator receptor or a specific pair of activator and inhibitor receptors is interpreted as target cell-mediated activation of the immune cell. A measure of the selectivity of the immune cells can also be determined using this approach. The immune cell expressing a pair of activator and inhibitor receptors is selective if the following is observed: 1) viability is reduced in target cells expressing the activator receptor ligand but not the inhibitor receptor ligand; 2) viability is not reduced in target cells expressing both an activator receptor ligand and an inhibitor receptor ligand. From these measurements, a “specific killing”value can be derived that quantifies the percentage of immune cell activation based on the reduction in viability of target cell as a percentage of a negative control (immune cells that do not express an activator receptor). Further, from these measurements a “selectivity ratio” value can be derived that represents the ratio of the specific killing observed in target cells expressing an activator receptor ligand in the absence of inhibitor receptor ligand to the specific killing observed in target cells expressing both an activator receptor ligand and an inhibitor receptor ligand. This approach can be used to characterize the population of cells for the production and manufacturing of the immune cells, pharmaceutical compositions, and kits described herein. A suitable specific killing value for the immune cells, pharmaceutical compositions, and kits can be, for example, the following criteria: 1) at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or at least 99% specific killing following a 48 hour co-incubation of immune cells and target cells expressing activator receptor ligand in the absence of inhibitor receptor ligand; and 2) less than or equal to 40%, less than or equal to 35%, less than or equal to 30%, less than or equal to 25%, less than or equal to 20%, less than or equal to 15%, less than or equal to 10%, less than or equal to 5%, less than or equal to 3% or less than or equal to 1% specific killing of target cell expressing both an activator receptor ligand and an inhibitor receptor ligand.

[0634] As a further example, a suitable specific killing value for the immune cells, pharmaceutical compositions and kits can be the following criteria: 1) between 30% and 99%, between 40% and 99%, between 50% and 99%, between 55% and 95%, between 60% and 95%, between 60% and 90%, between 50% and 80%, between 50% and 70% or between 50% and 60% of target cells expressing the activator ligand but not the inhibitor ligand are killed; and 2), between 1% and 40%, between 3% and 40%, between 5% and 40%, between 5% and 30%, between 10% and 30%, between 15% and 30% or between 5% and 20% of target cells expressing the activator ligand and the inhibitor ligand are killed. As a still further example, a suitable specific killing value for the immune cells, pharmaceutical compositions, and kits can be, for example, the following criteria: 1) at least 50% specific killing following a 48 hour co-incubation of immune cells and target cells expressing activator receptor ligand in the absence of inhibitor receptor ligand; and 2) less than or equal to 20% specific killing of target cell expressing both an activator receptor ligand and an inhibitor receptor ligand. As a further example, the immune cells are capable of killing at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or at least 99% of target cells expressing the activator ligand and not the inhibitor ligand over a period of 6 hours, 12 hours, 18 hours, 24 hours, 30 hours, 36 hours, 42 hours, 48 hours, 54 hours, or 60 hours, while killing less than 40%, less than 30%, less than 20%, less than 10%, lessthan 5%, less than 3% or less than 1% of target cells expressing the activator and inhibitor ligands over the same time period.

[0635] A suitable specific killing value of the target cell expressing an activator ligand in the absence of an inhibitory ligand value for the immune cells, pharmaceutical compositions, and kits can be, for example, at least about 50% to at least about 95%. A suitable specific killing value of the target cell expressing an activator ligand in the absence of an inhibitory ligand value for the immune cells, pharmaceutical compositions, and kits can be, for example, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95%. A suitable specific killing value of the target cell expressing an activator ligand in the absence of an inhibitory ligand value for the immune cells, pharmaceutical compositions, and kits can be, for example, at most about 50%, at most about 55%, at most about 60%, at most about 65%, at most about 70%, at most about 75%, at most about 80%, at most about 85%, at most about 90%, or at most about 95%. A suitable specific killing value of target cells expressing both an activator receptor ligand and an inhibitor receptor ligand for the immune cells, pharmaceutical compositions, and kits can be less than about 50%, less than about 45%, less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5%. The suitable specific killing value for the immune cells, pharmaceutical compositions, and kits can be determined following about 6 hours, about 12 hours, about 18 hours, about 24, about 30 hours, about 36 hours, about 42 hours, about 48 hours, about 54 hours, about 60 hours, about 66 hours, or about 72 hours of co-incubation of immune cells with target cells.

[0636] A suitable specific killing value of the target cell expressing an activator ligand in the absence of an inhibitory ligand value for the immune cells, pharmaceutical compositions, and kits can be, for example, at least about 50% to at least about 95%. A suitable specifi...

Claims

CLAIMSWhat is claimed is:

1. An engineered immune cell comprising:(a) an activator receptor that binds to an activator antigen expressed on an autoreactive target immune cell and promotes a cytotoxic response against a target cell; and(b) an inhibitor receptor that binds to an inhibitor antigen expressed on a nontarget cell and inhibits cytotoxicity against the non-target cell; wherein the target cell is implicated in an autoimmune disease or disorder.

2. An engineered immune cell comprising an activator receptor or receptors and an inhibitor receptor or receptors, wherein the activator receptor specifically binds to an activator antigen on a target cell; wherein the inhibitor receptor specifically binds to an inhibitor antigen on a non-target cell; and wherein the target cell is an autoreactive target immune cell known to mediate or is implicated in an autoimmune disease.

3. The engineered immune cell of claim 1 or claim 2, wherein the autoreactive target immune cell associated with an autoimmune disease or disorder is an autoreactive target B cell, an autoreactive T cell, or an autoreactive myeloid cell.

4. The engineered immune cell of any one of claims 1-3, wherein the activator antigen is selected from a group consisting of 4- IBB, CD 19, CD20, CD22, CD69, CD25, and / or 0X40.

5. The engineered immune cell of claim 3 or claim 4, wherein the autoreactive target B cell expresses at least one activator antigen selected from: CD19, CD20 and CD22.

6. The engineered immune cell of claim 3, wherein a autoreactive target T cell expresses at least one activator antigen selected from a group consisting of: CD25, CD69, 4- IBB and 0X40.

7. The engineered immune cell of any one of claims 1-6, wherein the non-target cell comprises a naive T cell, a memory T cell, and / or a regulatory T cell (Treg).

8. The engineered immune cell of claim 7, wherein the non-target cell is a Treg.

9. The engineered immune cell of claim 8, wherein the Treg expresses at least one inhibitor antigen selected from CCR8, LRRC32 (GARP), and CCR7.

10. The engineered immune cell of any one of claims 1-9, wherein the activator receptor comprises an extracellular ligand binding domain specific for an activator antigen expressed by an autoreactive immune cell, and wherein the extracellular ligand binding domain of the activator receptor comprises a antibody fragment or a single chain Fv antibody fragment (scFv).

11. The engineered immune cell of claim 1-9, wherein the engineered immune cell comprises two activator receptors wherein a first activator receptor binds to an activator antigen and wherein a second activator receptor binds to a second activator antigen that is different from a first activator antigen.

12. The engineered immune cell of claim 11, wherein the activator receptor is a tandem activator receptor comprising a first extracellular ligand binding domain specific to a first activator antigen expressed by an autoreactive immune cell wherein the first extracellular ligand binding domain of the activator receptor comprises a first antibody fragment or first scFv and second extracellular ligand binding domain specific to a second activator antigen expressed by an autoreactive immune cell wherein the second extracellular ligand binding domain of the activator receptor comprises a second antibody fragment or second scFv.

13. The engineered immune cell of any one of claims 12, wherein the first activator antigen is 0X40, wherein the first scFv of the activator receptor specifically binds to 0X40 and comprises a first heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 537, a CDR-H2 of SEQ ID NO: 538, and a CDR-H3 of SEQ ID NO: 539; and a first light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 540, a CDR-L2 of SEQ ID NO: 446, and a CDR-L3 of SEQ ID NO: 541.

14. The engineered immune cell of claim 13, wherein the first scFV of the activator receptor comprises a first VH comprising SEQ ID NO: 624 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 624 and / or a first VL comprising SEQ ID NO: 625 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 625.

15. The engineered immune cell of claim 13, wherein the first scFv of the activator receptor comprises SEQ ID NO: 663 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 663.

16. The engineered immune cell of claim 13, wherein the activator receptor comprises SEQ ID NO: 695 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 695.

17. The engineered immune cell of any one of claims 12-16, wherein the second activator antigen is CD19 wherein the second scFv of the activator receptor specifically binds to CD19 and comprises a second heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 959, a CDR-H2 of SEQ ID NO: 960, and a CDR-H3 of SEQ ID NO: 961; and a second light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 962, a CDR-L2 of SEQ ID NO: 963, and a CDR-L3 of SEQ ID NO: 964.

18. The engineered immune cell of claim 17, wherein the second scFv of the activator receptor comprises a second VH comprising SEQ ID NO: 1001 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1001 and / or a second VL comprising SEQ ID NO: 1002 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1002.

19. The engineered immune cell of claim 17, wherein the second scFv of the activator receptor comprises SEQ ID NO: 1018 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1018.

20. The engineered immune cell of claim 17, wherein the activator receptor comprises SEQ ID NO: 1027 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 1027.

21. The engineered immune cell of any one of claims 9-20, wherein the activator receptor comprises a hinge sequence isolated or derived from CD8, CD28, IgGl, or IgG4, or a synthetic hinge.

22. The engineered immune cell of any one of claims 9-21, wherein the activator receptor comprises a transmembrane domain isolated or derived from CD8 or CD28.

23. The engineered immune cell of any one of claims 9-22, wherein the activator receptor comprises an intracellular domain isolated or derived from CD28, 4- IBB or CD3z, or a combination thereof.

24. The engineered immune cell of any one of claims 1-23, wherein the inhibitor receptor comprises an extracellular ligand binding domain specific to an inhibitor antigen expressed by a non-target cell and wherein the extracellular ligand binding domain of the inhibitor receptor comprises a antibody fragment or a scFv (single chain Fv antibody fragment).

25. The engineered immune cell of claim 24, wherein the inhibitor antigen is LRRC32 (GARP) wherein the scFv of the inhibitor receptor specifically binds to LRR32(GARP) and comprises a heavy chain variable region (VH) comprising a complementarity determining region CDR-H1 of SEQ ID NO: 340, a CDR-H2 of SEQ ID NO: 341, and a CDR-H3 of SEQ ID NO: 342; and a light chain variable region (VL) comprising a CDR-L1 of SEQ ID NO: 343, a CDR-L2 of SEQ ID NO: 344, and a CDR- L3 of SEQ ID NO: 345.

26. The engineered immune cell of claim 24, wherein the scFv of the inhibitor receptor comprises a VH comprising SEQ ID NO: 373 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 373 and / or a VL comprising SEQ ID NO: 374 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 374.

27. The engineered immune cell of claim 24, wherein the scFv of the inhibitor receptor comprises SEQ ID NO: 394 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 394.

28. The engineered immune cell of claim 24, wherein the inhibitor receptor comprises SEQ ID NO: 408 or a sequence having at least 70%, at least 85%, at least 90%, or at least 95% identity to SEQ ID NO: 408.

29. The engineered immune cell of any one of claims 1-24, wherein the inhibitor antigen is an HLA antigen to protect the non-target cell from fratricide.

30. The engineered immune cell of claim 29, wherein the HLA antigen is HLA-A*02 or HLA-A*03.

31. The engineered immune cell of any one of claims 1-30, wherein the inhibitor receptor comprises a LILRB1 intracellular domain or a functional variant thereof.

32. The engineered immune cell of claim 31, wherein the LILRB1 intracellular domain comprises SEQ ID NOs: 31 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

33. The engineered immune cell of any one of claims 1-30, wherein the inhibitor receptor comprises a LILRB1 hinge domain and a LILRB1 transmembrane domain, or functional variants thereof.

34. The engineered immune cell of claim 33, wherein the LILRB1 transmembrane domain comprises SEQ ID NO: 30 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

35. The engineered immune cell of claims 33, wherein the LILRB1 hinge domain comprises SEQ ID NO: 29, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

36. The engineered immune cell of any one of claims 31-35, wherein the LILRB1 intracellular domain, LILRB1 hinge domain, and LILRB1 transmembrane domain together comprises SEQ ID NO: 27 or 28, or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical thereto.

37. The engineered immune cell of any one of claims 1-36, wherein an engineered immune is modified to reduce or eliminate expression of a B2M gene product.

38. The engineered immune cell of claim 37, wherein the expression of the B2M gene product is reduced or eliminated using a short hairpin RNA (shRNA).

39. The engineered immune cell of claim 38, wherein the shRNA comprises a first sequence, having from 5' to 3' end a sequence complementary to the B2M gene product; and a second sequence, having from 5' to 3' end a sequence complementary to the first sequence, wherein the first sequence and second sequence form the shRNA.

40. The engineered immune cell of any one of claims 1-39, wherein the activator receptor comprises SEQ ID NO: 1322 or 1324 or a sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 99% identity, or 100% identical to SEQ ID NO: 1322 or 1324.

41. A pharmaceutical composition comprising a therapeutically effective amount of the engineered immune cell of any one of claims 1-40 and a pharmaceutically acceptable excipient, wherein the therapeutically effective amount is sufficient to treat or ameliorate an autoimmune disease or disorder in a subject in need thereof.

42. A method of treating or ameliorating an autoimmune disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the engineered immune cell of any one of claims 1-40.

43. A method of treating or ameliorating an autoimmune disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 41.

44. The method of claims 42-43, wherein the autoimmune disease or disorder is selected from lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidly progressing systemic sclerosis (SSc) with significant lung involvement (e.g., as for autoHSCT)), idiopathic inflammatory myopathies (e.g., polymyositis, dermatomyositis, anti-synthetase syndrome, immune-mediated necrotizing myopathy, inclusion body myositis, overlap myositis, cancer associated myositis, e.g., anti-synthetase syndrome with ILD), vasculitis (e.g., ANCA-associated vasculitis), severe refractory Sjogren' s, severe refractory neuroimmune disease (e.g., myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), severe refractory rheumatoid arthritis, antibody mediated neuroimmune diseases (e.g., AChR+ and MuSK+ myasthenia gravis (MG), AQP4+ neuromyelitis optica (NMO), MOGAD (anti-MOG associated disease), NMDAR+ encephalitis, or antibody-associated neurological paraneoplastic diseases), Addison's disease, Goodpasture's syndrome, thyrotoxicosis, chronic active hepatitis, relapsing polychondritis, pemphigus vulgaris, and amyotrophic lateral sclerosis.

45. A composition for use in the method of claim 42 or claim 43.

46. A polynucleotide or polynucleotide system comprising one or more polynucleotides encoding the activator receptor and the inhibitor receptor of the engineered immune cell of any one of claims 1-40.

47. A method of making an immune cell therapy, the method comprising transforming immune cells with the polynucleotide or polynucleotide system of claim 46.

48. A nanocarrier comprising the polynucleotide or polynucleotide system of claim46.

49. The nanocarrier of claim 48, wherein the nanocarrier is capable of delivering the polynucleotide or polynucleotide system to an immune cell in vivo or ex vivo.

50. The nanocarrier of claims 48-49, wherein the nanocarrier is a lipid nanoparticle (LNP).

51. Any composition or method disclosed herein.

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