Rapcabtagene autoleucel for use in treating systemic lupus erythematosus

Abstract

The invention provides methods of using a population of CD19 CAR-expressing cells (for example, rapcabtagene autoleucel) for treating autoimmune diseases or disorders. Also disclosed are methods of making a population of CD19 CAR-expressing cells.

Description

[0001] RAPCABTAGENE AUTOLEUCEL FOR USE IN TREATING SYSTEMIC LUPUS ERYTHEMATOSUS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates generally to methods of using a population of CD 19 CAR- expressing cells for treating autoimmune diseases or disorders, compositions comprising the same, and methods of making CD 19 CAR-expressing cells.

[0004] BACKGROUND OF THE INVENTION

[0005] Current therapies for severe autoimmune diseases such as systemic lupus erythematosus (SLE) include conventional immunomodulatory and anti-inflammatory agents such as antimalarials, glucocorticoids, and immunosuppressives (e.g. methotrexate, azathioprine, mycophenolate and cyclophosphamide) and biologies (such as, belimumab and very recently, anifrolumab as well as rituximab commonly used in the severe stage of the disease). Severe refractory SLE (srSLE) patients, with or without renal involvement, after having failed immunosuppressive and biological therapies, have very limited treatment options. Autologous stem cell transplantation (ASCT) may be performed; however, it remains experimental and is associated with significant toxicities including mortality. Thus, there exists an unmet need for new treatments for severe autoimmune diseases, including srSLE.

[0006] SUMMARY OF THE INVENTION

[0007] Disclosed herein are methods of using a population of CD 19 CAR-expressing cells for treating a disease, for example, an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis in a subject. The present disclosure also pertains to methods of making CD 19 CAR-expressing cells, and compositions generated using such methods.

[0008] In one aspect, the disclosure provides a method of treating a subject having an autoimmune disease or disorder, e.g., 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, antisynthetase 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD19 CAR-expressing cells”), said population comprising:

[0009] (a) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0010] (b) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0011] (c) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0012] (d) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0013] (e) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0014] (f) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0015] (g) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0016] (h) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or

[0017] (i) an increased percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

[0018] In one aspect, the disclosure provides a method of treating a subject having an autoimmune disease or disorder, e.g., 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, antisynthetase 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD 19 CAR-expressing cells”), wherein:

[0019] (a) the median GeneSetScore (Up TEM vs. Down TSCM) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, 75, 100, or 125% from the median GeneSetScore (Up TEM vs. Down TSCM) of the same population of cells prior to being engineered to express the CAR;

[0020] (b) the median GeneSetScore (Up Treg vs. Down Teff) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, 100, 150, or 200% from the median GeneSetScore (Up Treg vs. Down Teff) of the population of cells prior to being engineered to express the CAR;

[0021] (c) the median GeneSetScore (Down sternness) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 25, 50, 100, 150, 200, or 250% from the median GeneSetScore (Down sternness) of the population of cells prior to being engineered to express the CAR; (d) the median GeneSetScore (Up hypoxia) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 125, 150, 175, or 200% from the median GeneSetScore (Up hypoxia) of the population of cells prior to being engineered to express the CAR; or

[0022] (e) the median GeneSetScore (Up autophagy) of the population of cells is about the same as or differs by no more than (for example, increased by no more than) about 180, 190, 200, or 210% from the median GeneSetScore (Up autophagy) of the population of cells prior to being engineered to express the CAR.

[0023] In one aspect, the disclosure provides a method of treating a subject having an autoimmune disease or disorder, e.g., 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, antisynthetase 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, the method comprising administering to the subject rapcabtagene autoleucel.

[0024] In one aspect, the disclosure provides a method of treating a subject having an autiommune disease, the method comprising administering to the subject a population of CD 19 CAR-expressing cells in an amount sufficient to treat the autoimmune isease. In some embodiments, the autiommune disease is a severe refractory autoimmune disease selected from systemic lupus erythematosus (SLE), lupus nephritis, rheumatoid arthritis, and Sjogren's disease. In some embodiments, the SLE is a severe refractory SLE (srSLE). In some embodiments, the SLE is a severe refractory SLE (srSLE), wherein the subject has renal involvement. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel. In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 0.5 x 106to 90 x 106viable CAR-expressing cells, optionally wherein the population of CD 19 CAR-expressing cells is administered at a dose of 5 x 106viable CAR- expressing cells, optionally wherein the population of CAR-expressing cells is administered at a dose of 2.5 x 106viable CAR-expressing cells. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel.

[0025] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 2.5 x 106to 2.5 x 108viable CAR-expressing cells, for example, about 12.5 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0026] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 12.5 x 106to 1.25 x 109viable CAR-expressing cells, for example, about 25 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0027] In some embodiments, the population of CD 19 CAR-expressing cells is administered at a dose of about 2.5 x 106to 2.5 x 108viable CAR-expressing cells, for example, about 40 x 106viable CAR-expressing cells. In some embodiments, the population of CD19 CAR-expressing cells is rapcabtagene autoleucel.

[0028] In one aspect, the disclosure provides a population of CD 19 CAR-expressing cells or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE) or lupus nephritis), said method comprising administering to the subject an effective amount of the population of CD 19 CAR-expressing cells or an effective amount of the pharmaceutical composition. In some embodiments, the population of CD 19 CAR-expressing cells is rapcabtagene autoleucel.

[0029] In one aspect, the disclosure provides a method of treating a subject having an autoimmune disease, e.g., lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE) or lupus nephritis), the method comprising administering to the subject: a population of cells that express, or comprise a nucleic acid configured to express, a CD 19 chimeric antigen receptor (CD 19 CAR), optionally wherein the population of cells is rapcabtagene autoleucel, and a second therapy chosen from an antimalarial agent or a stable immunosuppressive, wherein the second therapy and CD 19 CAR cells are present in the subject at the same time, e.g., wherein the second therapy is administered at a time when the CD 19 CAR cells are present in the subject.

[0030] In one aspect, the disclosure provides rapcabtagene autoleucel or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, said method comprising administering to the subject an effective amount of the population of rapcaptagene autoleucel or an effective amount of the pharmaceutical composition

[0031] In one aspect, the disclosure provides rapcabtagene autoleucel or a pharmaceutical composition comprising the same for use in a method of modulating an immune response in a subject having lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE) or lupus nephritis), said method comprising administering to the subject an effective amount of rapcabtagene autoleucel or an effective amount of the pharmaceutical composition. Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis.

[0032] Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having severe refractory systemic lupus erythematosus (srSLE), wherein rapcabtagene autoleucel is formulated for administration in an amount sufficient to treat the srSLE.

[0033] Rapcabtagene autoleucel for use in the manufacture of a medicament for treating a subject having lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE) or lupus nephritis), rapcabtagene autoleucel is formulated for administration at a dose of 0.5 - 50 x 106viable CAR+ T cells (e.g., 2.5-, 5-, 12.5-, 25-, 40 x 106viable CAR+ T cells).

[0034] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references (for example, sequence database reference numbers) mentioned herein are incorporated by reference in their entirety. For example, all GenBank, Unigene, and Entrez sequences referred to herein, for example, in any Table herein, are incorporated by reference. When one gene or protein references a plurality of sequence accession numbers, all of the sequence variants are encompassed. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Headings, sub-headings or numbered or lettered elements, for example, (a), (b), (i) etc., are presented merely for ease of reading. The use of headings or numbered or lettered elements in this document does not require the steps or elements be performed in alphabetical order or that the steps or elements are necessarily discrete from one another. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0035] BRIEF DESCRIPTION OF THE FIGURES

[0036] FIG. 1 is a schematic showing the clinical trial design for a phase 1 / 2 study, open-label, multi-center, to assess safety, efficacy and cellular kinetics of ARM-CD19 CAR T cells in participants with severe, refractory autoimmune disorders.

[0037] FIG. 2 is a schematic comparing the ARM process to a traditional CAR T manufacturing process.

[0038] FIG. 3 demonstrates that functional (e.g., cytolytic function and cytokine production) rapcabtagene autoleucel can be generated via the ARM process using T cells obtained from subjects diagnosed with SLE. Figure 3A is a graph showing a similar transduction efficiency of a rapcabtagene autoleucel with T cells taken from subjects with SLE or healthy donors (HD). Figure 3B is a graph showing rapcabtagene autoleucel generated from T cells taken from subjects with SLE (represented by circles) and healthy donors (HD) (represented by triangles) have a comparable cytolytic function as evaluated in an in vitro killing assay. Figure 3C is graph showing rapcabtagene autoleucel generated from T cells taken from subjects with SLE and healthy donors (HD) display similar production of cytokines IL-2 and IFN-y.

[0039] FIG. 4 demonstrates that rapcabtagene autoleucel manufactured via the ARM process using T cells from subjects with SLE capable of killing autologous B cells and releasing cytokines in vitro. FIG. 4A is a graph depicting the killing of autologuous B cells in an E:T dependent manner when using CD22 as a B cell marker following co-culture of rapcabtagene autoleucel with autologous B cells at different E:T ratios (CAR-T cells v. B cells) for 24 hours and 40 hours respectively. FIG. 4B shows the IFNy in supernatants from the co-culture at 24 hours and 40 hours. The data shown is representative of data derived from 3 SLE patients. FIG. 5 is a graph depicting sustained B cell depletion in 6 subjects with srSLE during lymphodepletion and sustained for at least 60 days following administration of rapcabtagene autoleucel.

[0040] FIG. 6 is a graph depicting substantial reductions in baseline SLEDAI-2K scores in 6 subjects with srSLE following administration of rapcabtagene autoleucel.

[0041] FIG. 7 is a table demonstrating improvements in individual SLEDAI-2K domains in 6 subjects with srSLE following administration of rapcabtagene autoleucel.

[0042] FIG. 8 is a graph depicting substantial reductions in baseline PhGA scores in 6 subjects with srSLE following administration of rapcabtagene autoleucel.

[0043] FIG. 9 is a graph demonstrating a significant and sustained decrease in anti-dsDNA antibodies in 6 subjects with srSLE following administration of rapcabtagene autoleucel.

[0044] FIG. 10 is a graph showing the Complement C3 (FIG. 10A) and Complement C4 (FIG. 10B) levels in 6 subjects with srSLE following administration of rapcabtagene autoleucel.

[0045] DETAILED DESCRIPTION

[0046] Definitions

[0047] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains.

[0048] The term “a” and “an” refers to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0049] The term “about” when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or in some instances ±10%, or in some instances ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0050] The term “Chimeric Antigen Receptor” or alternatively a “CAR” refers to a recombinant polypeptide construct comprising at least an extracellular antigen-binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as “an intracellular signaling domain”) comprising a functional signaling domain derived from a stimulatory molecule as defined below. In some embodiments, the domains in the CAR polypeptide construct are in the same polypeptide chain, for example, comprise a chimeric fusion protein.

[0051] The term “autologous” refers to any material derived from the same individual to whom it is later to be re-introduced into the individual.

[0052] The term “allogeneic” refers to any material derived from a different animal of the same species as the individual to whom the material is introduced. Two or more individuals are said to be allogeneic to one another when the genes at one or more loci are not identical. In some embodiments, allogeneic material from individuals of the same species may be sufficiently unlike genetically to interact antigenically.

[0053] The term “apheresis” as used herein refers to the art-recognized extracorporeal process by which the blood of a donor or patient is removed from the donor or patient and passed through an apparatus that separates out selected constituent(s) and returns the remainder to the circulation of the donor or patient, for example, by re- transfusion. Thus, in the context of “an apheresis sample” refers to a sample obtained using apheresis.

[0054] As used herein, “lupus” refers to all types and manifestations of lupus. Manifestations of lupus include, without limitation, systemic lupus erythematosus (including severe refractory SLE (srSLE); lupus nephritis; cutaneous manifestations (e.g., manifestations seen in cutaneous lupus erythematosus, e.g., a skin lesion or rash); CNS lupus; cardiovascular, pulmonary, hepatic, haematological, gastrointestinal and musculoskeletal manifestations; neonatal lupus erythematosus; childhood systemic lupus erythematosus; drug-induced lupus erythematosus; anti-phospholipid syndrome; and complement deficiency syndromes resulting in lupus manifestations.

[0055] “Immune effector cell,” as that term is used herein, refers to a cell that is involved in an immune response, for example, in the promotion of an immune effector response. Examples of immune effector cells include T cells, for example, alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, and myeloid-derived phagocytes.

[0056] “Immune effector function or immune effector response,” as that term is used herein, refers to function or response, for example, of an immune effector cell, that enhances or promotes an immune attack of a target cell. For example, an immune effector function or response refers a property of a T or NK cell that promotes killing or the inhibition of growth or proliferation, of a target cell. In the case of a T cell, primary stimulation and costimulation are examples of immune effector function or response.

[0057] The term “effector function” refers to a specialized function of a cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines.

[0058] The term “effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result.

[0059] The term “endogenous” refers to any material from or produced inside an organism, cell, tissue, or system.

[0060] The term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue, or system.

[0061] The term “expression” refers to the transcription and / or translation of a particular nucleotide sequence. In some embodiments, expression comprises translation of an mRNA introduced into a cell.

[0062] The term “lentivirus” refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector. HIV, SIV, and FIV are all examples of lentiviruses.

[0063] The term “lentiviral vector” refers to a vector derived from at least a portion of a lentivirus genome, including especially a self-inactivating lentiviral vector as provided in Milone et al., Mol. Ther. 17(8): 1453-1464 (2009). Other examples of lentivirus vectors that may be used in the clinic, include but are not limited to, for example, the LENTIVECTOR® gene delivery technology from Oxford BioMedica, the LENTIMAX™ vector system from Lentigen and the like. Nonclinical types of lentiviral vectors are also available and would be known to one skilled in the art.

[0064] The term “parenteral” administration of an immunogenic composition includes, for example, subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, intratumoral, or infusion techniques. As used herein, “B cell antigen” refers to an antigen associated with a B cell. Non-limiting examples of molecules associated with a B cell include proteins expressed on the surface of B cells, e.g. CD19, BCMA, CD22, CD20, CD10, CD34, CD123, FLT-3, R0R1, CD79b, CD179b, or CD79a .

[0065] As used herein, the term “CD 19” refers to the Cluster of Differentiation 19 protein. The human and murine amino acid and nucleic acid sequences can be found in a public database, such as GenBank, UniProt and Swiss-Prot. For example, the amino acid sequence of human CD19 can be found as UniProt / Swiss-Prot Accession No. Pl 5391 and the nucleic acid sequence encoding of the human CD19 can be found at Accession No. NM 001178098. It is also an early marker of B cell progenitors. See, e.g., Nicholson et al. Mol. Immun. 34 (16-17): 1157-1165 (1997). In one aspect the antigen-binding portion of the CART recognizes and binds an antigen within the extracellular domain of the CD 19 protein. In one aspect, the CD 19 protein is expressed on an autoreactive B-cell. As used herein, “CD 19” includes proteins comprising mutations, e.g., point mutations, fragments, insertions, deletions and splice variants of full length wild-type CD 19.

[0066] As used herein, the terms “treat”, “treatment” and “treating” refer to the reduction or amelioration of the progression, severity and / or duration of an autoimmune disorder, or the amelioration of one or more symptoms (preferably, one or more discernible symptoms) of an autoimmune disorder resulting from the administration of one or more therapies (for example, one or more therapeutic agents such as a CAR of the invention). In specific embodiments, the terms “treat,” “treatment,” and “treating” refer to the amelioration of at least one measurable physical parameter of an autoimmune disorder, such as the level of autoantibodies, not necessarily discernible by the patient. In other embodiments the terms “treat”, “treatment” and “treating” -refer to the inhibition of the progression of an autoimmune disorder, either physically by, for example, stabilization of a discernible symptom, physiologically by, for example, stabilization of a physical parameter, or both.

[0067] The term “subject” is intended to include living organisms in which an immune response can be elicited (for example, mammals, for example, human).

[0068] The term “therapeutic” as used herein means a treatment. A therapeutic effect is obtained by reduction, suppression, remission, or eradication of a disease state.

[0069] The term “prophylaxis” as used herein means the prevention of or protective treatment for a disease or disease state. “Refractory” as used herein refers to an autoimmune disease or disorder, for example, SLE, which does not respond to a treatment. In embodiments, a refractory autoimmune disease or disorder can be resistant to a treatment before or at the beginning of the treatment. In other embodiments, the refractory autoimmune disease or disorder can become resistant during a treatment. A refractory autoimmune disease or disorder is also called a resistant autoimmune disease or disorder.

[0070] As used herein, “severe refractory autoimmune disease” refers to a manifestation of an autoimmune disease that has failed to respond (e.g., remains charactericterized by high disease activity) following at least one standard immunosuppressive therapy or at least one biological agent. One example of a severe refractory autoimmune disease is severe refractory systemic lupus erythematosus.

[0071] As used herein, “severe refractory systemic lupus erythematosus” or “srSLE” refers to a manifestation of SLE that has failed to respond (e.g., remains characterized by high disease activity) following at least one standard immunosuppressive therapy (e.g., mycophenolate, cyclophosphamide), glucocorticoids, or at least one biological agent. In some embodiments, the srSLE comprises a manifestation of SLE that has failed to respond to two or more standard immunosuppressive therapies in combination with glucocorticoids. In some embodiments, the srSLE comprises a manifestation of SLE that has failed to respond to at least one biological agent.

[0072] “Relapsed” or “relapse” as used herein refers to the return or reappearance of a disease (for example, an autoimmune disease or disorder) or the signs and symptoms of a disease such as an autoimmune disease or disorder after a period of improvement or responsiveness, for example, after prior treatment of a therapy, for example, standard of care therapy. The initial period of responsiveness may involve the level of autoantibodies cells falling below a certain threshold. The reappearance may involve the level of autoantibodies rising above a certain threshold.

[0073] “Remission” as used herein refers to a decrease in or disappearance of signs and symptoms of a disease (for example, an autoimmune disease or disorder). Remission may be partial or complete. In partial remission, some, but not all, signs and symptons of a disease have decreased or disappeared. In complete remission, all signs and symptoms of a disease have disappeared. Remission may be determined according to a defined set of criteria established for a particular disease (for example, an autoimmune disease or disorder).

[0074] Ranges: throughout this disclosure, various embodiments 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.

[0075] Administered “in combination”, as used herein, means that two (or more) different treatments are delivered to the subject during the course of the subject's affliction with the disorder, for example, the two or more treatments are delivered after the subject has been diagnosed with the disorder and before the disorder has been cured or eliminated or treatment has ceased for other reasons. In some embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery”. In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, for example, an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered. As used herein, a “naive T cell” refers to a T cell that is antigen-inexperienced. In some embodiments, an antigen-inexperienced T cell has encountered its cognate antigen in the thymus but not in the periphery. In some embodiments, naive T cells are precursors of memory cells. In some embodiments, naive T cells express both CD45RA and CCR7, but do not express CD45RO. In some embodiments, naive T cells may be characterized by expression of CD62L, CD27, CCR7, CD45RA, CD28, and CD127, and the absence of CD95 or CD45RO isoform. In some embodiments, naive T cells express CD62L, IL-7 receptor-a, IL-6 receptor, and CD 132, but do not express CD25, CD44, CD69, or CD45RO. In some embodiments, naive T cells express CD45RA, CCR7, and CD62L and do not express CD95 or IL-2 receptor 0. In some embodiments, surface expression levels of markers are assessed using flow cytometry.

[0076] The term “central memory T cells” refers to a subset of T cells that in humans are CD45RO positive and express CCR7. In some embodiments, central memory T cells express CD95. In some embodiments, central memory T cells express IL-2R, IL-7R, and / or IL-15R. In some embodiments, central memory T cells express CD45RO, CD95, IL-2 receptor 0, CCR7, and CD62L. In some embodiments, surface expression levels of markers are assessed using flow cytometry.

[0077] The term “stem memory T cells,” “stem cell memory T cells,” “stem cell-like memory T cells,” “memory stem T cells,” “T memory stem cells,” “T stem cell memory cells,” or “TSCM cells” refers to a subset of memory T cells with stem cell-like ability, for example, the ability to self-renew and / or the multipotent capacity to reconstitute memory and / or effector T cell subsets. In some embodiments, stem memory T cells express CD45RA, CD95, IL-2 receptor 0, CCR7, and CD62L. In some embodiments, surface expression levels of markers are assessed using flow cytometry. In some embodiments, exemplary stem memory T cells are disclosed in Gattinoni et al., Nat Med. 2017 January 06; 23(1): 18-27, herein incorporated by reference in its entirety.

[0078] For clarity purposes, unless otherwise noted, classifying a cell or a population of cells as “not expressing,” or having an “absence of’ or being “negative for” a particular marker may not necessarily mean an absolute absence of the marker. The skilled artisan can readily compare the cell against a positive and / or a negative control, and / or set a predetermined threshold, and classify the cell or population of cells as not expressing or being negative for the marker when the cell has an expression level below the predetermined threshold or a population of cells has an overall expression level below the predetermined threshold using conventional detection methods, e.g., using flow cytometry.

[0079] As used herein, the term “GeneSetScore (Up TEM vs. Down TSCM)” of a cell refers to a score that reflects the degree at which the cell shows an effector memory T cell (TEM) phenotype vs. a stem cell memory T cell (TSCM) phenotype. A higher GeneSetScore (Up TEM vs. Down TSCM) indicates an increasing TEM phenotype, whereas a lower GeneSetScore (Up TEM vs. Down TSCM) indicates an increasing TSCM phenotype. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is determined by measuring the expression of one or more genes that are up-regulated in TEM cells and / or down-regulated in TSCM cells, for example, one or more genes selected from the group consisting of MXRA7, CLIC1, NAT13, TBC1D2B, GLCCI1, DUSP10, APOBEC3D, CACNB3, ANXA2P2, TPRG1, EOMES, MATK, ARHGAP10, ADAM8, MAN1A1, SLFN12L, SH2D2A, EIF2C4, CD58, MY01F, RAB27B, ERN1, NPC1, NBEAL2, APOBEC3G, SYTL2, SLC4A4, PIK3AP1, PTGDR, MAF, PLEKHA5, ADRB2, PLXND1, GNA01, THBS1, PPP2R2B, CYTH3, KLRF1, FLJ16686, AUTS2, PTPRM, GNLY, and GFPT2. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is determined for each cell using RNA-seq, for example, single-cell RNA-seq (scRNA-seq), for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39A, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up TEM vs. Down TSCM) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0080] As used herein, the term “GeneSetScore (Up Treg vs. Down Teff)” of a cell refers to a score that reflects the degree at which the cell shows a regulatory T cell (Treg) phenotype vs. an effector T cell (Teff) phenotype. A higher GeneSetScore (Up Treg vs. Down Teff) indicates an increasing Treg phenotype, whereas a lower GeneSetScore (Up Treg vs. Down Teff) indicates an increasing Teff phenotype. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is determined by measuring the expression of one or more genes that are up-regulated in Treg cells and / or down-regulated in Teff cells, for example, one or more genes selected from the group consisting of C12orf75, SELPLG, SWAP70, RGS1, PRR11, SPATS2L, SPATS2L, TSHR, C14orfl45, CASP8, SYT11, ACTN4, ANXA5, GLRX, HLA-DMB, PMCH, RAB11FIP1, IL32, FAM160B1, SHMT2, FRMD4B, CCR3, TNFRSF13B, NTNG2, CLDND1, BARD1, FCER1G, TYMS, ATP1B1, GJB6, FGL2, TK1, SLC2A8, CDKN2A, SKAP2, GPR55, CDCA7, S100A4, GDPD5, PMAIP1, AC0T9, CEP55, SGMS1, ADPRH, AKAP2, HDAC9, IKZF4, CARD17, VAV3, 0BFC2A, ITGB1, CIITA, SETD7, HLA-DMA, CCR10, KIAA0101, SLC14A1, PTTG3P, DUSP1O, FAM164A, PYHIN1, MYO1F, SLC1A4, MYBL2, PTTG1, RRM2, TP53INP1, CCR5, ST8SIA6, TOX, BFSP2, ITPRIPL1, NCAPH, HLA-DPB2, SYT4, NINJ2, FAM46C, CCR4, GBP5, C15orf53, LMCD1, MKI67, NUSAP1, PDE4A, E2F2, CD58, ARHGEF12, LOC100188949, FAS, HLA-DPB1, SELP, WEE1, HLA-DPA1, FCRL1, ICA1, CNTNAP1, OAS1, METTL7A, CCR6, HLA-DRB4, ANXA2P3, STAM, HLA-DQB2, LGALS1, ANXA2, PI 16, DUSP4, LAYN, ANXA2P2, PTPLA, ANXA2P1, ZNF365, LAIR2, LOC541471, RASGRP4, BCAS1, UTS2, MIAT, PRDM1, SEMA3G, FAM129A, HPGD, NCF4, LGALS3, CEACAM4, JAKMIP1, TIGIT, HLA-DRA, IKZF2, HLA-DRB1, FANK1, RTKN2, TRIBI, FCRL3, and FOXP3. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39B, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up Treg vs. Down Teff) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0081] As used herein, the term “GeneSetScore (Down sternness)” of a cell refers to a score that reflects the degree at which the cell shows a sternness phenotype. A lower GeneSetScore (Down sternness) indicates an increasing sternness phenotype. In some embodiments, the GeneSetScore (Down sternness) is determined by measuring the expression of one or more genes that are upregulated in a differentiating stem cell vs downregulated in a hematopoietic stem cell, for example, one or more genes selected from the group consisting of ACE, BATF, CDK6, CHD2, ERCC2, HOXB4, ME0X1, SFRP1, SP7, SRF, TALI, and XRCC5. In some embodiments, the GeneSetScore (Down sternness) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39C, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Down sternness) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0082] As used herein, the term “GeneSetScore (Up hypoxia)” of a cell refers to a score that reflects the degree at which the cell shows a hypoxia phenotype. A higher GeneSetScore (Up hypoxia) indicates an increasing hypoxia phenotype. In some embodiments, the GeneSetScore (Up hypoxia) is determined by measuring the expression of one or more genes that are up- regulated in cells undergoing hypoxia, for example, one or more genes selected from the group consisting of ABCB1, ACAT1, ADM, AD0RA2B, AK2, AK3, ALDH1A1, ALDH1A3, ALDOA, ALDOC, ANGPT2, ANGPTL4, ANXA1, ANXA2, ANXA5, ARHGAP5, ARSE, ART1, BACE2, BATF3, BCL2L1, BCL2L2, BHLHE40, BHLHE41, BIK, BIRC2, BNIP3, BNIP3L, BPI, BTG1, Cllorf2, C7orf68, CA12, CA9, CALD1, CCNG2, CCT6A, CD99, CDK1, CDKN1A, CDKN1B, CITED2, CLK1, CNOT7, COL4A5, COL5A1, COL5A2, COL5A3, CP, CTSD, CXCR4, D4S234E, DDIT3, DDIT4, 1-Dec, DKC1, DR1, EDN1, EDN2, EFNA1, EGF, EGR1, EIF4A3, ELF3, ELL2, ENG, EN01, EN03, ENPEP, EPO, ERRFI1, ETS1, F3, FABP5, FGF3, FKBP4, FLT1, FN1, FOS, FTL, GAPDH, GBE1, GLRX, GPI, GPRC5A, HAP1, HBP1, HDAC1, HDAC9, HERC3, HERPUD1, HGF, HIF1A, HK1, HK2, HLA-DQB1, HM0X1, HM0X2, HSPA5, HSPD1, HSPH1, HY0U1, ICAM1, ID2, IFI27, IGF2, IGFBP1, IGFBP2, IGFBP3, IGFBP5, IL6, IL8, INSIGI, IRF6, ITGA5, JUN, KDR, KRT14, KRT18, KRT19, LDHA, LDHB, LEP, LGALS1, LONP1, LOX, LRP1, MAP4, MET, MIF, MMP13, MMP2, MMP7, MPI, MT1L, MTL3P, MUC1, MXI1, NDRG1, NFIL3, NFKB1, NFKB2, NOS1, NOS2, NOS2P1, NOS2P2, NOS3, NR3C1, NR4A1, NT5E, ODC1, P4HA1, P4HA2, PAICS, PDGFB, PDK3, PFKFB1, PFKFB3, PFKFB4, PFKL, PGAM1, PGF, PGK1, PGK2, PGM1, PIM1, PIM2, PKM2, PLAU, PLAUR, PLIN2, PLOD2, PNN, PNP, POLM, PPARA, PPAT, PR0K1, PSMA3, PSMD9, PTGS1, PTGS2, QSOX1, RBPJ, RELA, RI0K3, RNASEL, RPL36A, RRP9, SAT1, SERPINB2, SERPINE1, SGSM2, SIAH2, SIN3A, SIRPA, SLC16A1, SLC16A2, SLC20A1, SLC2A1, SLC2A3, SLC3A2, SLC6A10P, SLC6A16, SLC6A6, SLC6A8, SORL1, SPP1, SRSF6, SSSCA1, STC2, STRA13, SYT7, TBPL1, TCEAL1, TEK, TF, TFF3, TFRC, TGFA, TGFB1, TGFB3, TGFBI, TGM2, TH, THBS1, THBS2, TIMM17A, TNFAIP3, TP53, TPBG, TPD52, TPI1, TXN, TXNIP, UMPS, VEGFA, VEGFB, VEGFC, VIM, VPS11, and XRCC6. In some embodiments, the GeneSetScore (Up hypoxia) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39D, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up hypoxia) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0083] As used herein, the term “GeneSetScore (Up autophagy)” of a cell refers to a score that reflects the degree at which the cell shows an autophagy phenotype. A higher GeneSetScore (Up autophagy) indicates an increasing autophagy phenotype. In some embodiments, the GeneSetScore (Up autophagy) is determined by measuring the expression of one or more genes that are up-regulated in cells undergoing autophagy, for example, one or more genes selected from the group consisting of ABL1, ACBD5, ACINI, ACTRT1, ADAMTS7, AKR1E2, ALKBH5, ALPK1, AMBRA1, ANXA5, ANXA7, ARSB, ASB2, ATG10, ATG12, ATG13, ATG14, ATG16L1, ATG16L2, ATG2A, ATG2B, ATG3, ATG4A, ATG4B, ATG4C, ATG4D, ATG5, ATG7, ATG9A, ATG9B, ATP13A2, ATP1B1, ATPAF1-AS1, ATPIF1, BECN1, BECN1P1, BLOC1S1, BMP2KL, BNIP1, BNIP3, BOC, Cllorf2, Cl lorf41, C12orf44, C12orf5, C14orfl33, Clorf210, C5, C6orfl06, C7orf59, C7orf68, C8orf59, C9orf72, CA7, CALCB, CALCOCO2, CAPS, CCDC36, CD163L1, CD93, CDC37, CDKN2A, CHAF1B, CHMP2A, CHMP2B, CHMP3, CHMP4A, CHMP4B, CHMP4C, CHMP6, CHST3, CISD2, CLDN7, CLEC16A, CLN3, CLVS1, C0X8A, CPA3, CRNKL1, CSPG5, CTSA, CTSB, CTSD, CXCR7, DAP, DKKL1, DNAAF2, DPF3, DRAM1, DRAM2, DYNLL1, DYNLL2, DZANK1, EI24, EIF2S1, EPG5, EPM2A, FABP1, FAM125A, FAM131B, FAM134B, FAM13B, F AMI 76 A, FAM176B, FAM48A, FANCC, FANCF, FANCL, FBX07, FCGR3B, FGF14, FGF7, FGFBP1, FIS1, FNBP1L, F0X01, FUNDCI, FUNDC2, FXR2, GAB ARAP, GABARAPL1, GABARAPL2, GABARAPL3, GABRA5, GDF5, GMIP, HAP1, HAPLN1, HBXIP, HCAR1, HDAC6, HGS, HIST1H3A, HIST1H3B, HIST1H3C, HIST1H3D, HIST1H3E, HIST1H3F, HIST1H3G, HIST1H3H, HIST1H3I, HIST1H3J, HK2, HMGB1, HPR, HSF2BP, HSP90AA1, HSPA8, IFI16, IPPK, IRGM, IST1, ITGB4, ITPKC, KCNK3, KCNQ1, KIAA0226, KIAA1324, KRCC1, KRT15, KRT73, LAMP1, LAMP2, LAMT0R1, LAMT0R2, LAMT0R3, LARP1B, LENG9, LGALS8, LIX1, LIX1L, LMCD1, LRRK2, LRSAM1, LSM4, MAP1A, MAP1LC3A, MAP1LC3B, MAP1LC3B2, MAP1LC3C, MAP1S, MAP2K1, MAP3K12, MARK2, MBD5, MDH1, MEX3C, MFN1, MFN2, MLST8, MRPS10, MRPS2, MSTN, MTERFD1, MTMR14, MTMR3, MTOR, MTSS1, MYH11, MYLK, MY0M1, NBR1, NDUFB9, NEFM, NHLRC1, NME2, NPC1, NR2C2, NRBF2, NTHL1, NUP93, OBSCN, OPTN, P2RX5, PACS2, PARK2, PARK7, PDK1, PDK4, PEX13, PEX3, PFKP, PGK2, PHF23, PHYHIP, PI4K2A, PIK3C3, PIK3CA, PIK3CB, PIK3R4, PINK1, PLEKHM1, PLOD2, PNPO, PPARGC1A, PPY, PRKAA1, PRKAA2, PRKAB1, PRKAB2, PRKAG1, PRKAG2, PRKAG3, PRKD2, PRKG1, PSEN1, PTPN22, RAB12, RAB1A, RAB1B, RAB23, RAB24, RAB33B, RAB39, RAB7A, RB1CC1, RBM18, REEP2, REP15, RFWD3, RGS19, RHEB, RIMS3, RNF185, RNF41, RPS27A, RPTOR, RRAGA, RRAGB, RRAGC, RRAGD, S100A8, S100A9, SCN1A, SERPINB10, SESN2, SFRP4, SH3GLB1, SIRT2, SLC1A3, SLC1A4, SLC22A3, SLC25A19, SLC35B3, SLC35C1, SLC37A4, SLC6A1, SLCO1A2, SMURF1, SNAP29, SNAPIN, SNF8, SNRPB, SNRPB2, SNRPD1, SNRPF, SNTG1, SNX14, SPATAI 8, SQSTM1, SRPX, STAM, STAM2, STAT2, STBD1, STK11, STK32A, STOM, STX12, STX17, SUPT3H, TBC1D17, TBC1D25, TBC1D5, TCIRG1, TEAD4, TECPR1, TECPR2, TFEB, TM9SF1, TMBIM6, TMEM203, TMEM208, TMEM39A, TMEM39B, TMEM59, TMEM74, TMEM93, TNIK, TOLLIP, TOMM20, TOMM22, TOMM40, T0MM5, T0MM6, T0MM7, TOMM70A, TP53INP1, TP53INP2, TRAPPC8, TREM1, TRIM17, TRIM5, TSG101, TXLNA, UBA52, UBB, UBC, UBQLN1, UBQLN2, UBQLN4, ULK1, ULK2, ULK3, USP10, USP13, USP30, UVRAG, VAMP7, VAMP8, VDAC1, VMP1, VPS11, VPS16, VPS18, VPS25, VPS28, VPS33A, VPS33B, VPS36, VPS37A, VPS37B, VPS37C, VPS37D, VPS39, VPS41, VPS4A, VPS4B, VTA1, VTT1A, VTI1B, WDFY3, WDR45, WDR45L, WIPI1, WIPI2, XBP1, YIPF1, ZCCHC17, ZFYVE1, ZKSCAN3, ZNF189, ZNF593, and ZNF681. In some embodiments, the GeneSetScore (Up autophagy) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 39E, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up autophagy) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0084] As used herein, the term “GeneSetScore (Up resting vs. Down activated)” of a cell refers to a score that reflects the degree at which the cell shows a resting T cell phenotype vs. an activated T cell phenotype. A higher GeneSetScore (Up resting vs. Down activated) indicates an increasing resting T cell phenotype, whereas a lower GeneSetScore (Up resting vs. Down activated) indicates an increasing activated T cell phenotype. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is determined by measuring the expression of one or more genes that are up-regulated in resting T cells and / or down- regulated in activated T cells, for example, one or more genes selected from the group consisting of ABCA7, ABCF3, ACAP2, AMT, ANKH, ATF7IP2, ATG14, ATP1A1, ATXN7, ATXN7L3B, BCL7A, BEX4, BSDC1, BTG1, BTG2, BTN3A1, Cllorf21, C19orf22, C21orf2, CAMK2G, CARS2, CCNL2, CD248, CD5, CD55, CEP164, CHKB, CLK1, CLK4, CTSL1, DBP, DCUN1D2, DENND1C, DGKD, DLG1, DUSP1, EAPP, ECE1, ECHDC2, ERBB2IP, FAM117A, FAM134B, FAM134C, F AMI 69 A, FAM190B, FAU, FLJ10038, F0XJ2, F0XJ3, F0XL1, FOXO1, FXYD5, FYB, HLA-E, HSPA1L, HYAL2, ICAM2, IFIT5, IFITM1, IKBKB, IQSEC1, IRS4, KIAA0664L3, KIAA0748, KLF3, KLF9, KRT18, LEF1, LINC00342, LIPA, LIPT1, LLGL2, LMBR1L, LPAR2, LTBP3, LYPD3, LZTFL1, MANBA, MAP2K6, MAP3K1, MARCH8, MAU2, MGEA5, MMP8, MPO, MSL1, MSL3, MYH3, MYLIP, NAGPA, NDST2, NISCH, NICER, NLRP1, NOSIP, NPIP, NUMA1, PAIP2B, PAPD7, PBXIP1, PCIF1, PI4KA, PLCL2, PLEKHA1, PLEKHF2, PNISR, PPFIBP2, PRKCA, PRKCZ, PRKD3, PRMT2, PTP4A3, PXN, RASA2, RASA3, RASGRP2, RBM38, REPIN1, RNF38, RNF44, R0R1, RPL30, RPL32, RPLP1, RPS20, RPS24, RPS27, RPS6, RPS9, RXRA, RYK, SCAND2, SEMA4C, SETD1B, SETD6, SETX, SF3B1, SH2B1, SLC2A4RG, SLC35E2B, SLC46A3, SMAGP, SMARCE1, SMPD1, SNPH, SP140L, SPATA6, SPG7, SREK1IP1, SRSF5, STAT5B, SVIL, SYF2, SYNJ2BP, TAF1C, TBC1D4, TCF20, TECTA, TES, TMEM127, TMEM159, TMEM30B, TMEM66, TMEM8B, TP53TG1, TPCN1, TRIM22, TRIM44, TSC1, TSC22D1, TSC22D3, TSPYL2, TTC9, TTN, UBE2G2, USP33, USP34, VAMP1, VILL, VIPR1, VPS13C, ZBED5, ZBTB25, ZBTB40, ZC3H3, ZFP161, ZFP36L1, ZFP36L2, ZHX2, ZMYM5, ZNF136, ZNF148, ZNF318, ZNF350, ZNF512B, ZNF609, ZNF652, ZNF83, ZNF862, and ZNF91. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is determined using RNA-seq, for example, single-cell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 38D, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up resting vs. Down activated) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0085] As used herein, the term “GeneSetScore (Progressively up in memory differentiation)” of a cell refers to a score that reflects the stage of the cell in memory differentiation. A higher GeneSetScore (Progressively up in memory differentiation) indicates an increasing late memory T cell phenotype, whereas a lower GeneSetScore (Progressively up in memory differentiation) indicates an increasing early memory T cell phenotype. In some embodiments, the GeneSetScore (Up autophagy) is determined by measuring the expression of one or more genes that are up-regulated during memory differentiation, for example, one or more genes selected from the group consisting of MTCH2, RAB6C, KIAA0195, SETD2, C2orf24, NRD1, GNA13, COP A, SELT, TNIP1, CBFA2T2, LRP10, PRKCI, BRE, ANKS1A, PNPLA6, ARL6IP1, WDFY1, MAPK1, GPR153, SHKBP1, MAP1LC3B2, PIP4K2A, HCN3, GTPBP1, TLN1, C4orf34, KIF3B, TCIRG1, PPP3CA, ATG4D, TYMP, TRAF6, C17orf76, WIPF1, FAM108A1, MYL6, NRM, SPCS2, GGT3P, GALK1, CLIP4, ARL4C, YWHAQ, LPCAT4, ATG2A, IDS, TBC1D5, DMPK, ST6GALNAC6, REEP5, ABHD6, KIAA0247, EMB, TSEN54, SPIRE2, PIWIL4, ZSCAN22, ICAM1, CHD9, LPIN2, SETD8, ZC3H12A, ULBP3, IL15RA, HLA- DQB2, LCP1, CHP, RUNX3, TMEM43, REEP4, MEF2D, ABL1, TMEM39A, PCBP4, PLCD1, CHST12, RASGRP1, Clorf58, Cl lorf63, C6orfl29, FH0D1, DKFZp434F142, PIK3CG, ITPR3, BTG3, C4orf50, CNNM3, IFI16, AK1, CDK2AP1, REL, BCL2L1, MVD, TTC39C, PLEKHA2, FKBP11, EML4, FANCA, CDCA4, FUCA2, MFSD10, TBCD, CAPN2, IQGAP1, CHST11, PIK3R1, MYO5A, KIR2DL3, DLG3, MXD4, RALGDS, S1PR5, WSB2, CCR3, TIP ARP, SP140, CD151, SOX13, KRTAP5-2, NF1, PEA15, PARP8, RNF166, UEVLD, LIMK1, CACNB1, TMX4, SLC6A6, LBA1, SV2A, LLGL2, IRF1, PPP2R5C, CD99, RAPGEF1, PPP4R1, OSBPL7, FOXP4, SLA2, TBC1D2B, ST7, JAZF1, GGA2, PI4K2A, CD68, LPGAT1, STX11, ZAK, FAM160B1, RORA, C8orf80, APOBEC3F, TGFBI, DNAJC1, GPR114, LRP8, CD69, CMIP, NAT13, TGFBI, FLJ00049, ANTXR2, NR4A3, IL12RB1, NTNG2, RDX, MLLT4, GPRIN3, ADCY9, CD300A, SCD5, ABB, PTPN22, LGALS1, SYTL3, BMPR1A, TBK1, PMAIP1, RASGEF1A, GCNT1, GABARAPL1, STOM, CALHM2, ABCA2, PPP1R16B, SYNE2, PAM, C12orf75, CLCF1, MXRA7, APOBEC3C, CLSTN3, ACOT9, HIP1, LAG3, TNFAIP3, DCBLD1, KLF6, CACNB3, RNF19A, RAB27A, FADS3, DLG5, APOBEC3D, TNFRSF1B, ACTN4, TBKBP1, ATXN1, ARAP2, ARHGEF12, FAM53B, MAN1A1, FAM38A, PLXNC1, GRLF1, SRGN, HLA-DRB5, B4GALT5, WIPI1, PTPRJ, SLFN11, DUSP2, ANXA5, AHNAK, NEO1, CLIC1, EIF2C4, MAP3K5, IL2RB, PLEKHG1, MY06, GTDC1, EDARADD, GALM, TARP, ADAM8, MSC, HNRPLL, SYT11, ATP2B4, NHSL2, MATK, ARHGAP18, SLFN12L, SPATS2L, RAB27B, PIK3R3, TP53INP1, MBOAT1, GYG1, KATNAL1, FAM46C, ZC3HAV1L, ANXA2P2, CTNNA1, NPC1, C3AR1, CRIM1, SH2D2A, ERN1, YPEL1, TBX21, SLC1A4, FASLG, PHACTR2, GALNT3, ADRB2, PIK3AP1, TLR3, PLEKHA5, DUSP10, GNAO1, PTGDR, FRMD4B, ANXA2, EOMES, CADM1, MAF, TPRG1, NBEAL2, PPP2R2B, PELO, SLC4A4, KLRF1, FOSL2, RGS2, TGFBR3, PRF1, MYO1F, GAB3, C17orf66, MICAL2, CYTH3, TOX, HLA-DRA, SYNE1, WEE1, PYHIN1, F2R, PLD1, THBS1, CD58, FAS, NETO2, CXCR6, ST6GALNAC2, DUSP4, AUTS2, Clorf21, KLRG1, TNIP3, GZMA, PRR5L, PRDM1, ST8SIA6, PLXND1, PTPRM, GFPT2, MYBL1, SLAMF7, FLJ16686, GNLY, ZEB2, CST7, IL18RAP, CCL5, KLRD1, and KLRB1. In some embodiments, the GeneSetScore (Progressively up in memory differentiation) is determined using RNA-seq, for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 40B, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Progressively up in memory differentiation) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set.

[0086] As used herein, the term “GeneSetScore (Up TEM vs. Down TN)” of a cell refers to a score that reflects the degree at which the cell shows an effector memory T cell (TEM) phenotype vs. a naive T cell (TN) phenotype. A higher GeneSetScore (Up TEM vs. Down TN) indicates an increasing TEM phenotype, whereas a lower GeneSetScore (Up TEM vs. Down TN) indicates an increasing TN phenotype. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is determined by measuring the expression of one or more genes that are up-regulated in TEM cells and / or down-regulated in TN cells, for example, one or more genes selected from the group consisting of MY05A, MXD4, STK3, S1PR5, GLCCI1, CCR3, SOX13, KRTAP5-2, PEA15, PARP8, RNF166, UEVLD, LIMK1, SLC6A6, SV2A, KPNA2, OSBPL7, ST7, GGA2, PI4K2A, CD68, ZAK, RORA, TGFBI, DNAJC1, JOSD1, ZFYVE28, LRP8, OSBPL3, CMIP, NAT13, TGFBI, ANTXR2, NR4A3, RDX, ADCY9, CHN1, CD300A, SCD5, PTPN22, LGALS1, RASGEF1A, GCNT1, GLUL, ABCA2, CLDND1, PAM, CLCF1, MXRA7, CLSTN3, ACOT9, METRNL, BMPR1A, LRIG1, APOBEC3G, CACNB3, RNF19A, RAB27A, FADS3, ACTN4, TBKBP1, FAM53B, MAN1A1, FAM38A, GRLF1, B4GALT5, WIPI1, DUSP2, ANXA5, AHNAK, CLIC1, MAP3K5, ST8SIA1, TARP, ADAM8, MATK, SLFN12L, PIK3R3, FAM46C, ANXA2P2, CTNNA1, NPC1, SH2D2A, ERN1, YPEL1, TBX21, STOM, PHACTR2, GBP5, ADRB2, PIK3AP1, DUSP10, PTGDR, EOMES, MAF, TPRG1, NBEAL2, NCAPH, SLC4A4, FOSL2, RGS2, TGFBR3, MYO1F, C17orf66, CYTH3, WEE1, PYHIN1, F2R, THBS1, CD58, AUTS2, FAM129A, TNIP3, GZMA, PRR5L, PRDM1, PLXND1, PTPRM, GFPT2, MYBL1, SLAMF7, ZEB2, CST7, CCL5, GZMK, and KLRB1. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is determined using RNA-seq, for example, singlecell RNA-seq (scRNA-seq) , for example, as exemplified of WO / 2020 / 047452 in Example 10 with respect to FIG. 40C, hereby incorporated by reference in its entirety. In some embodiments, the GeneSetScore (Up TEM vs. Down TN) is calculated by taking the mean log normalized gene expression value of all of the genes in the gene set. In the context of GeneSetScore values (e.g., median GeneSetScore values), when a positive GeneSetScore is reduced by 100%, the value becomes 0. When a negative GeneSetScore is increased by 100%, the value becomes 0. For example, as disclosed in WO / 2020 / 047452, the median GeneSetScore of the Dayl sample is -0.084; the median GeneSetScore of the Day9 sample is 0.035; and the median GeneSetScore of the input sample is -0.1. In WO / 2020 / 047452 in FIG. 39A, increasing the median GeneSetScore of the input sample by 100% leads to a GeneSetScore value of 0; and increasing the median GeneSetScore of the input sample by 200% leads to a GeneSetScore value of 0.1. In WO / 2020 / 047452 in FIG. 39A, decreasing the median GeneSetScore of the Day9 sample by 100% leads to a GeneSetScore value of 0; and decreasing the median GeneSetScore of the Day9 sample by 200% leads to a GeneSetScore value of -0.035.

[0087] Various embodiments of the compositions and methods herein are described in further detail below. Additional definitions are set out throughout the specification.

[0088] CD19 CAR-Expressing Cells

[0089] The present disclosure provides CAR-expressing cell compositions and their use in medicaments or methods for treating, among other diseases, autoimmune diseases (e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis involving cells or tissues which express an antigen as described herein. In some embodiments, provided herein are pharmaceutical compositions comprising a CAR-expressing cell, for example, a plurality of CD 19 CAR-expressing cells, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. In some embodiments, the CD 19 CAR-expressing cells is rapcabtag ene autoleucel.

[0090] In some embodiments, the antigen-binding domain of the CD 19 CAR has the same or a similar binding specificity as the FMC63 scFv fragment described in Nicholson et al. Mol. Immun. 34 (16-17): 1157-1165 (1997). In some embodiments, the antigen-binding domain of the CD19 CAR includes the scFv fragment described in Nicholson et al. Mol. Immun. 34 (16- 17): 1157-1165 (1997). In some embodiments, the CD19 CAR includes an antigen-binding domain (for example, a humanized antigen-binding domain) according to Table 3 of WO2014 / 153270, incorporated herein by reference. In some embodiments, the CD19 CAR comprises an amino acid sequence provided as SEQ ID NO: 12 in PCT publication WO2012 / 079000, incorporated herein by reference.

[0091] In some embodiments, the population of CAR T cells that specifically bind to CD 19 comprises rapcabtagene autoleucel. The rapcabtagene autoleucel is made using autologous T cells obtained from peripheral blood mononuclear cells from a subject (e.g., from a subject having an autoimmune disease or disorder) by leukapheresis and subsequently transduced with a self-inactivating, non-replicating lentiviral vector encoding a T cell chimeric antigen receptor targeting CD 19. The expressed transgene comprises a CD8a leader sequence, a murine anti- CD19 single chain variable fragment (scFv) derived from the mouse hybridoma FMC63, a CD8a hinge and transmembrane region, and a 4- IBB (CD 137) and CD3^ (TCRQ signaling domain, and is under control of the elongation factor 1 alpha (EFla) promoter. The construct is flanked by 5' and 3' long terminal repeats (LTRs) and also contains a *| / packaging signal, a Rev response element (RRE), a central polypurine tract (cPPT) sequence, and an optimized Woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). The leukapheresis material is enriched for CD4 / CD8 T cells by positive immunoselection, activated by CD3 and CD28 agonists and transduced with the vector. Without further cell propagation, the T cells are washed, formulated for infusion, and cryopreserved. Rapcabtagene autoleucel is composed of >80% T cells and <1% B cells, with a mixture of transgene positive (>3.4%) and negative T cells. The CD4+ and CD8+ naive T cell subsets (CD45RA+CCR7+) present in the leukapheresis material are largely retained. In some embodiments, the rapcabtagene autoleucel is made by the ARM manufacturing process described herein.

[0092] In some embodiments, the CD19 CAR-expressing cell population has:

[0093] (a) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0094] (b) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0095] (c) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;

[0096] (d) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0097] (e) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0098] (f) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR; (g) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor 0+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;

[0099] (h) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or

[0100] (i) an increased percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

[0101] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0102] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, or 50% lower), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0103] In some embodiments, the CD 19 CAR-expressing population of cells made via the ARM process described herein after being administered in vivo, persists longer or expands at a higher level (for example, at least 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90% higher), compared with cells made by an otherwise similar method which lasts, for example, more than 26 hours (for example, which lasts more than 5, 6, 7, 8, 9, 10, 11, or 12 days) or which involves expanding the population of cells in vitro for, for example, more than 3 days (for example, expanding the population of cells in vitro for 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 days).

[0104] Any known CD 19 CAR, for example, the CD 19 antigen-binding domain of any known CD 19 CAR, in the art can be used in accordance with the present disclosure. Examples include tisagenlecleucel, axicabtagene ciloleucel, brexucabtagene autoleucel, lisocabtagene maraleucel, relmacabtagene autoleucel, CC-97540, AT101, CABA-201, KYV-101, IMPT-514, or MB- CART19.1. Further exemplary CD 19 CARs include CD 19 CARs described herein or an antiCD F CAR described in Xu et al. Blood 123.24(2014):3750-9; Kochenderfer et al. Blood 122.25(2013):4129-39, Cruz et al. Blood 122.17(2013):2965-73, NCT00586391, NCT01087294, NCT02456350, NCT00840853, NCT02659943, NCT02650999, NCT02640209, NCT01747486, NCT02546739, NCT02656147, NCT02772198, NCT00709033, NCT02081937, NCT00924326, NCT02735083, NCT02794246, NCT02746952, NCT01593696, NCT02134262, NCT01853631, NCT02443831, NCT02277522, NCT02348216, NCT02614066, NCT02030834, NCT02624258, NCT02625480, NCT02030847, NCT02644655, NCT02349698, NCT02813837, NCT02050347, NCT01683279, NCT02529813, NCT02537977, NCT02799550, NCT02672501, NCT02819583, NCT02028455, NCT01840566, NCT01318317, NCT01864889, NCT02706405, NCT01475058, NCT01430390, NCT02146924, NCT02051257, NCT02431988, NCT01815749, NCT02153580, NCT01865617, NCT02208362, NCT02685670, NCT02535364, NCT02631044, NCT02728882, NCT02735291, NCT01860937, NCT02822326, NCT02737085, NCT02465983, NCT02132624, NCT02782351, NCT01493453, NCT02652910, NCT02247609, NCT01029366, NCT01626495, NCT02721407, NCT01044069, NCT00422383, NCT01680991, NCT02794961, or NCT02456207, NCT05338931, NCT05869955, NCT06056921, NCT06106906, NCT06121297, NCT06152172, NCT06189157, NCT05459870, NCT06153095, each of which is incorporated herein by reference in its entirety.

[0105] Methods of Treating

[0106] The present application discloses methods of treating method of treating a subject having an autoimmune disease or disorder, e.g., 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, antisynthetase 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, the method comprising administering to the subject a population of cells engineered to express a CD 19 CAR (“a population of CD19 CAR-expressing cells”). In some embodiments, the method further comprises identifying the subject as a responder or a non- responder, and / or one who has achieved remission, based on a measure of one or more of the parameters of efficacy disclosed herein. In some embodiments, the measure of one or more of the parameters disclosed herein is obtained from a sample acquired from the subject. In some embodiments of any of the methods disclosed herein, the subject is evaluated prior to receiving, during, or after receiving, the CD 19 CAR-expressing cell therapy. In some embodiments, the CD 19 CAR-expressing cell therapy is rapcabtagene autoleucel.

[0107] In some embodiments, the identification of a subject as a responder, non- responder and / or one who has achieved remission, following administration of the CD 19 CAR-expressing cells according to the methods described herein can be determined by evaluating the subject according to clinical criteria. Efficacy may be evaluated, for example, using the lupus low disease activity state (LLDAS) (as described in Franklyn et al, Ann Rheum Dis. 2016), e.g., as informed by SLEDAI-2K (described in Gladman et al, J Rheumatol. 2000), and physician's global assessment; SLE responder index (SRI-4) (as described in Furie et al, Arthritis Rheumatol. 2017); British Isles Lupus Assessment Group-based Composite Lupus Assessment (BICLA) (as described in Wallace et al. Arthritis Rheum. 2011); British Isles Lupus Activity Group score (BILAG) (as described in Isenberg et al Ann Rheum Dis 2005); urinary protein creatinine ratio (UPCR) at various time points; and / or complete renal response (CRR) at various time points. In some embodiments, remission may be evaluated using the DORIS definition (as described in van Vollenhoven et al, Lupus Sci Med. 2021). The above criteria are explained in further detail below.

[0108] Lupus Low Disease Activity State (LLDAS) is defined by the following criteria: (1) SLEDAI-2K < 4, with no activity in major organ systems (renal, CNS, cardiopulmonary, vasculitis, fever) and no hemolytic anemia or gastrointestinal activity; (2) no new features of lupus disease activity compared with the previous assessment (SLEDAI-2K); (3) physician global assessment (PhGA, scale 0-3) < 1; (4) current corticosteroid (prednisolone or equivalent) dose < 7.5 mg / day; (5) well tolerated standard maintenance doses of immunosuppressive drugs and approved biological agents (Franklyn et al 2016). LLDAS is derived at assessment visits when SLEDAI-2K and PhGA are assessed.

[0109] In some embodiments, a subject is identified as having a LLDAS defined by the above criteria following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the LLDAS is achieved within 3 months of administration. In some embodiments, the LLDAS is achieved within 6 months of administration. In some embodiments, the LLDAS is achieved within 9 months of administration. In some embodiments, the LLDAS is achieved within 12 months of administration.

[0110] In some embodiments, the LLDAS achieved by a subject following administration of the CD 19 CAR-expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, the LLDAS is sustained for 6 months or more. In some embodiments the LLDAS is sustained for 9 months or more. In some embodiments, the LLDAS is sustained for 12 months or more. In some embodiments, the LLDAS is sustained for 18 months or more. In some embodiments, the LLDAS is sustained indefinitely.

[0111] For the remission evaluations, the Definition of Remission in SLE (DORIS) definition (van Vollenhoven et al 2021) is: (1) clinical SLEDAI-2K=0; (2) Physician Global Assessment <0.5 (0-3). The participant may be on antimalarials, low-dose glucocorticoids (prednisolone < 5 mg / day), and / or stable immunosuppressives including biologies. Clinical SLEDAI-2K is a subindex of SLEDAI-2K which only includes the clinical descriptors and excludes the laboratory ones (i.e., low complement and increased DNA binding domains). In some embodiments, a subject is identified as in remission according to the DORIS definition following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to the subject according to the methods described herein. In some embodiments, the DORIS defined remission is achieved within 3 months of administration. In some embodiments, the DORIS defined remission is achieved within 6 months of administration. In some embodiments, the DORIS defined remission is achieved within 9 months of administration. In some embodiments, the DORIS defined remission is achieved within 12 months of administration.

[0112] In some embodiments, the DORIS defined remission achieved by a subject following administration of the CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) is sustained for 3 months or more. In some embodiments, the DORIS defined remission is sustained for 6 months or more. In some embodiments the LLDAS is sustained for 9 months or more. In some embodiments, the DORIS defined remission is sustained for 12 months or more. In some embodiments, the DORIS defined remission is sustained for 18 months or more. In some embodiments, the DORIS defined remission is sustained indefinitely.

[0113] The Systemic Lupus Erythematosus Disease Activity Index (SLED Al) is a validated model of experienced clinicians' global assessments of disease activity in systemic lupus erythematosus based on the consensus of a group of experts in the field of lupus research (Bombardier et al 1992). SLED Al is calculated as a total score derived from a weighted index of 9 organ systems for disease activity in SLE, as follows: 8 for central nervous system and vascular descriptors, 4 for renal and musculoskeletal descriptors, 2 for serosal, dermal, and immunologic descriptors, and 1 for constitutional and hematologic descriptors. Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) is a modified version of SLED Al to reflect persistent, active disease in those descriptors that had previously only considered new or recurrent occurrences (Gladman et al 2002). The range of SLEDAI-2K score is 0 to 105; a higher score indicating more severe disease. A score of 6 is considered clinically important and affects the decision to treat with a probability of initiating therapy in more than 50 % of cases. A score of 3 or less is considered a low disease activity state based on the presence of only one clinical manifestation (with or without positive serology results). Remission has been defined as no clinical manifestation (with or without serologic manifestations). Meaningful improvement is defined as a reduction in SLEDAI-2 K score of 4 or more points. In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods described herein results in a reduction of the SLEDAI-2K score of the subject by 4 or more points. In some embodiments, the reduction of the SLEDAI-2K score is by 5 or more points, 6 or more points, 7 or more points, 8 or more points, 9 or more points, 10 or more points, 15 or more points, 20 or more points, or 25 or more points.

[0114] In some embodiments, administration of the CD 19 CART cells (e.g., rapcabtagene autoleucel) results in a reduction of the total SLEDAI-2K score to 6 or below. In some embodiments, the total SLEDAI-2K score is reduced to 5 or below, 4 or below, 3 or below, 2 or below, or 1 or below. In some embodiments, the total SLEDAI-2K score is reduced to 0.

[0115] British Isles Lupus Activity Group score (BILAG) records disease activity occurring over the past 4 weeks. The BILAG-2004 index covers 97 items and the assessment based on the principle of the doctor’s intent to treat, which requires an assessment as 0 = not present, 1 = improving, 2 = same, 3 = worse, or 4 = new over the last month (Isenberg et al 2005, Isenberg et al 2011, Yee et al 2006, Yee et al 2010). There are nine general headings: Constitutional, Mucocutaneous, Neuropsychiatric, Musculoskeletal, Cardiorespiratory, Gastrointestinal, Ophthalmic, Renal, Haematological. Within each organ system, multiple manifestations and laboratory tests are combined into a single score for that organ. The resulting scores for each organ can be A through E, where A is very active disease, B is moderate activity, C is mild stable disease, D is resolved activity, and E indicates the organ was never involved. The BILAG-2004 system provides a disease activity measure that scores longitudinally and is clinically meaningful and easier to analyze in comparison with multiple categorical variables. This system has expected associations with change in therapy. Major clinical responses by the BILAG index are a reduction from BILAG A or B score to BILAG C score or better at 6 months with no new BILAG A or B scores and the maintenance of response with no new BILAG A or B scores between 6 and 12 months.

[0116] In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods described herein results in a reduction in the BILAG score from A to C or better. In some embodiments, the reduction in the BILAG score is from A to D. In some embodiments, reduction in the BILAG score from B to C or better. In some embodiments, the reduction in the BILAG score is from B to D. In some embodiments, the reduction in BILAG score from A to C or better is sustained for 6 months or more with no new BILAG A or B scores. In some embodiments, the reduction in BILAG score from B to C or better is sustained for 6 months or more with no new BILAG A or B scores. In some embodiments the reduction in BILAG scores described above is sustained for 9 months or more. In some embodiments, the reduction in BILAG score is sustained for 12 months or more. In some embodiments, the reduction is sustained for 18 months or more. In some embodiments, the reduction of BILAG score from A or B to C or better is maintained without any new BILAG A or B scores.

[0117] The Physician‘s global assessment (PhGA) of disease activity is performed using 100 mm visual analog scale (VAS) ranging from “0 - no disease activity” to “3 - severe disease activity”, after the question on how well the participant is doing with the disease considering all aspects affected by the disease. The distance in mm from the left edge of the scale is then measured and recorded. Values >0-5 but <1 refer to mild disease activity; values >1 but <2 refer to moderate disease activity; and values >2 up to 3 refer to severe disease activity. Meaningful improvement is defined as a change greater than or equal to 0.3 points from baseline.

[0118] In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods described herein results in reduction in PhGA score greater than or equal to 0.3 points. In some embodiments, the reduction is greater than or equal to 0.5 points. In some embodiments, the reduction is greater than or equal to 0.75 points. In some embodiments, the reduction is greater than or equal to 1 point. In some embodiments, the reduction is greater than or equal to 1.25 points. In some embodiments, the reduction is greater than or equal to 1.5 points. In some embodiments, the reduction is greater than or equal to 1.75 points. In some embodiments, the reduction is greater than or equal to 2 points. In some embodiments, the reduction is greater than or equal to 2.25 points. In some embodiments, the reduction is greater than or equal to 2.5 points. In some embodiments, the reduction is greater than or equal to 2.75 points.

[0119] In some embodiments, the methods of the present invention result in a reduction in PhGA score to 1.5 or lower. In some embodiments, the reduction in PhGA score is to 1.25 or lower. In some embodiments, the reduction in PhGA score is to 1 or lower. In some embodiments, the reduction in PhGA score is to 0.75 or lower. In some embodiments, the reduction in PhGA score is to 0.5 or lower. In some embodiments, the reduction in PhGA score is to 0.3 or lower. In some embodiments, the methods of the present invention result in a sustained reduction in PhGA from baseline to 1.5 or lower, wherein the reduction is sustained for 6 months or more. In some embodiments, the sustained reduction in PhGA from baseline is to 1.25 or lower, wherein the reduction is sustained for 6 months or more. In some embodiments, the sustained reduction in PhGA from baseline is to 1 or lower, wherein the reduction is sustained for 6 months or more. In some embodiments, the sustained reduction in PhGA from baseline is to 0.75 or lower, wherein the reduction is sustained for 6 months or more. In some embodiments, the sustained reduction in PhGA from baseline is to 0.5 or lower, wherein the reduction is sustained for 6 months or more. In some embodiments, the sustained reduction in PhGA from baseline is to 0.25 or lower, wherein the reduction is sustained for 6 months or more.

[0120] In some embodiments, the sustained reduction in PhGA as described above is sustained for 9 months or more. In some embodiments, the reduction is sustained for 12 months or more. In some embodiments, the reduction is sustained for 18 months or more. In some embodiments, the reduction is sustained for 24 months or more. In some embodiments, the sustained reduction is maintained without an increase in PhGA score to 1.0 or greater. In some embodiments, the sustained reduction is maintained without an increase in PhGA score to 1.25 or greater. In some embodiments, the sustained reduction is maintained without an increase in PhGA score to 1.5 or greater. In some embodiments, the sustained reduction is maintained without an increase in PhGA score to 1.75 or greater. In some embodiments, the sustained reduction is maintained without an increase in PhGA score to 2.0 or greater.

[0121] In some embodiments, the methods of the present invention result in no increase in PhGA score of 0.3 or more points from baseline following administration of the therapy. In some embodiments, there is no increase in PhGA of greater than 0.25 points from baseline following administration of the therapy. In some embodiments, there is no increase in PhGA score of greater than 0.2 points from baseline following administration of the therapy. In some embodiments, there is no increase in PhGA score of greater than 0.15 points from baseline following administration of the therapy. In some embodiments, there is no increase in PhGA score of greater than 0.10 points from baseline following administration of the therapy. In some embodiments, there is no increase in PhGA score of greater than 0.05 points from baseline following administration of the therapy. The SLE responder index (SRI-4) utilizes the SLEDAI-2K score to determine global improvement, BILAG-2004 domain scores to ensure no significant worsening in heretofore unaffected organ systems, and physician’s global assessment (PhGA) to ensure that improvements in disease activity are not achieved at the expense of the patient’s overall condition (Furie et al 2017). The SRI is calculated any time the SLE disease activity scores are measured in individual participants. A responder is defined as having a >4-point reduction from baseline in SLEDAI-2K score AND no new BILAG-2004 A organ domain scores or >2 new BILAG-2004 B organ domain scores compared with baseline AND no worsening in PhGA (<0.3-point increase from baseline). If all 3 criteria are met, the participant will be considered a responder at that particular point in time; otherwise, the participant will be considered a nonresponder.

[0122] In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods disclosed herein results in the subject being considered a responder according to the SRI-4 definition. In some embodiments, the response according to the SRI-4 definition is sustained for 3 or more months. In some embodiments, the response is sustained for 6 or more months. In some embodiments, the response is sustained for 9 or more months. In some embodiments, the response is sustained for 12 or more months.

[0123] BICLA (British Isles Lupus Assessment Group-based Composite Lupus Assessment, Wallace et al 2011) is a validated composite global measure of SLE disease activity that is derived from existing outcome assessments. Participants are considered as responders if they meet following criteria: (1) reduction of all baseline BILAG-2004 A to B / C / D and baseline B to C / D and no worsening in other organ systems defined as >1 new A or >2 new B items compared to baseline; (2) no worsening from baseline in SLEDAI-2K, defined as an increase from baseline of >0 points; (3) no worsening in PhGA, defined as an increase of >0.3 from baseline on a 0 to 3 visual analog scale.

[0124] In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods disclosed herein results in the subject being considered a responder according to the BICLA criteria. In some embodiments, the response according to the BICLA criteria is sustained for 3 or more months. In some embodiments, the response is sustained for 6 or more months. In some embodiments, the response is sustained for 9 or more months. In some embodiments, the response is sustained for 12 or more months. Complete Renal Response (CRR) is defined as a decrease in UPGR (urinary protein creatinine ratio) to <0.5 mg / mg in 2 consecutive, first morning void urine specimens, plus an eGFR >60 ml / min per 1.73 m2or no decrease of >20% of screening eGFR on 2 consecutive occasions (Rovin et al 2019). In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtag ene autoleucel) to a subject results in a CRR for that subject. In some embodiments, the CRR is sustained for 3 or more months. In some embodiments, the CRR is sustained for 6 or more months. In some embodiments, the CRR is sustained for 9 or more months. In some embodiments, the response is sustained for 12 or more months.

[0125] Patient reported outcomes (PRO) may also be completed at the scheduled visit before any clinical assessments are conducted. The Short Form Health Survey (SF-36 v2) is a widely used and extensively studied instrument to measure health-related quality of life among healthy participants and participants with acute and chronic conditions. It consists of eight subscales that can be scored individually: Physical Functioning, Role-Physical, Bodily Pain, General Health, Vitality, Social Functioning, Role-Emotional, and Mental Health. Two overall summary scores, the Physical Component Summary (PCS) and the Mental Component Summary (MCS) also can be computed. The SF-36 has proven useful in monitoring general and specific populations, comparing the relative burden of different diseases, differentiating the health benefits produced by different treatments, and in screening individual participants. The patient’s global assessment of disease activity is performed using a Visual Analogue Scale (VAS) of 100 mm ranging from “no disease activity” to “severe disease activity”, after the question on how well the participant is doing with the disease considering all aspects affected by the disease. The distance in mm from the left edge of the scale and the value is measured. In some embodiments, the methods of the present invention result in a decrease in the PRO as measured from baseline.

[0126] Anti-double stranded DNA (dsDNA) antibodies play an important role in the diagnosis, classification and management of systemic lupus erythematosus. A high level of anti-dsDNA antibodies in the blood is strongly associated with lupus disease activity. In some embodiments, administration of CD19 CAR-expressing cells (e.g., rapcabtagene autoleucel) to a subject according to the methods described herein results in a decrease in the level of anti-dsDNA antibodies for the subject as compared to baseline. Dosage regimen

[0127] In some embodiments, a dose of viable CD 19 CAR-expressing cells comprises about 0.5 x 106viable CAR-expressing cells to about 1.25 x 109viable CAR-expressing cells (for example, 0.5 x 106viable CAR-expressing cells to 1.25 x 109viable CAR-expressing cells). In some embodiments, a dose of viable CAR-expressing cells comprises about 1 x 106, about 2.5 x 106, about 5 x 106, about 1.25 x 107, about 2.5 x 107, about 5 x 107, about 5.75 x 107, or about 8 x 107viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 0.5 x 106to 90 x 106viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 5 x 106viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 2.5 x 106to 2.5 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 107viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 107to 1.25 x 109viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1.25 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 2.5 x 106to 2.5 x 108viable CAR-expressing cells. In some embodiments, a dose of viable CAR-expressing cells comprises about 1 x 107or 5 x 107viable CAR-expressing cells. In some embodiments, the viable CD 19 CAR-expressing cells are rapcabtagene autoleucel.

[0128] In some embodiments, a dose of CAR-expressing cells comprises about 1 x 106, 1.1 x

[0129] 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CAR-expressing cells comprises at least about 1 x 106, 1.1 x 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells up to about 1 x 106, 1.1 x 106, 2 x 106, 3.6 x 106, 5 x 106, 1 x 107, 1.8 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, or 5 x 108cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells comprises about 1.1 X 106- 1.8 X 107cells / kg. In some embodiments, a dose of CD 19 CAR-expressing cells comprises about 1 x 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, 5 x 108, 1 x 109, 2 x 109, or 5 x 109cells. In some embodiments, a dose of CD 19 CAR-expressing cells comprises at least about 1 x

[0130] 107, 2 x 107, 5 x 107, 1 x 108, 2 x 108, 5 x 108, 1 x 109, 2 x 109, or 5 x 109cells. In some embodiments, a dose of CD 19 CAR expressing cells comprises up to about 1 x 107, 2 x l07, 5 x 107, 1 x IO8, 2 x IO8, 5 x IO8, 1 x IO9, 2 x IO9, or 5 x IO9cells. In some embodiments, the CD 19 CAR-expressing cells are rapcabtagene autoleucel.

[0131] The level of CAR-positive cells can be determined according to the methods disclosed in Example 8 of WO / 2021 / 173985. Briefly, for CAR T cells manufactured using a continuous Activated Rapid Manufacturing (ARM) process, e.g., ARM-CD19 CAR T cells, a sentinel vial of cryopreserved cells may be thawed and cultured for up to 5 days and the CAR expression analyzed by flow cytometry. The measurement of CAR expression on, e.g., day 2 or day 3 may be used to determine the dose of viable CAR-positive T cells.

[0132] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, comprising administering to said patient CAR-expressing cells produced as described herein, at a dose of viable CAR-expressing or CAR-positive cells (for example, viable CD 19 CAR-expressing cells, viable CD 19 CAR-positive cells, or any dual CARs thereof) from about 0.5 x 106viable CAR-expressing or CAR-positive cells to about 50 x 106viable CAR-expressing or CAR-positive cells (for example, from about 0.5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 90 x 106viable CD 19 CAR-expressing or CAR-positive cells), e.g. at a dose of viable CAR-expressing or CAR-positive cells (for example, viable CD 19 CAR-expressing cells or viable CD 19 CAR-positive cells) from about 2 x 106 viable CAR-expressing or CAR-positive cells to about 40 x 106viable CAR-expressing or CARpositive cells.

[0133] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having an autoimmune disease or disorder, e.g., lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of viable CAR-expressing or CAR-positive cells from about 0.5 x 106viable CAR-expressing or CAR-positive cells to about 50 x 106viable CAR-expressing or CARpositive cells (for example, from about 0.5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 50 x 106viable CD 19 CAR-expressing or CAR-positive cells).

[0134] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 2.5 x 106viable CD 19 CAR-expressing or CARpositive cells to about 40 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0135] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 5 x 106viable CD 19 CAR-expressing or CAR-positive cells to about 12.5 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0136] In some embodiments, the disclosure provides a method of treating a patient, e.g., a patient having severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis, comprising administering to said patient CAR-expressing or CAR-positive cells produced as described herein, at a dose of from about 25 x 106viable CD 19 CAR-expressing or CARpositive cells to about 40 x 106viable CD 19 CAR-expressing or CAR-positive cells.

[0137] Patient selection

[0138] In some embodiments of any of the methods of treating a subject, or composition for use disclosed herein, the subject has an autoimmune disease or disorder, e.g., lupus (e.g., systemic lupus erythematosus (SLE), e.g., severe refractory systemic lupus erythematosus (srSLE), or lupus nephritis), systemic sclerosis (e.g., rapidlv nroeressing 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis.

[0139] In some embodiments, the subject having srSLE has previously been administered one or more of an antimalarial (e.g., hydroxychloroquine or quinacrine), a glucocorticoid (e.g., prednisone), a calcineurin inhibitor, an immunomodulatory agent (e.g., methotrexate, azathioprine, mycophenolate moefetil, cyclophosphamide, or tacrolimus), a biological agent (e.g., belimumab, rituximab, a disease-modifying antirheumatic drug (DMARD) (e.g., leflunomide). In some embodiments, the subject has been identified as not responding to treatment comprising two or more immunosuppressive therapies (e.g., mycophenolate or cyclophosphamide) in combination with a glucocorticoid) and one biological agent. In some embodiments, the subject has not previously received a therapy comprising a CD 19 CAR, an adoptive T cell therapy, or a gene therapy product.

[0140] In some embodiments, prior to administration of the CD 19 CAR therapy, the subject receives lymphodepleting therapy. In some embodiments, the subject receives a lympodepleting therapy about two weeks prior to administration of the CD 19 CAR. In some embodiments, the lympodepleting therapy comprises fludarabine (e.g., 25 mg / m2IV daily for three doses) and cyclophosphamide (e.g., 250 mg / m2IV daily for three doses). In some embodiments, the CD 19 CAR therapy is rapcabtagene autoleucel.

[0141] Evaluating CAR Safety

[0142] In some embodiments of any of the therapeutic methods disclosed herein, the method further involves evaluating the safety of the CAR-expressing cell therapy in a subject. In some embodiments, safety of the CAR-expressing cell therapy is evaluated by measuring or recording one or more of a subject’s vital signs, adverse events experienced by the subject, various laboratory parementers, and / or an electrocardiogram of the subject.

[0143] In some embodiments, the subject does not experience an adverse event of grade 4 or higher. In some embodiments, the subject does not experience cytokine release syndrome (CRS). In some embodiments, the subject does not experience CRS of grade 3 or higher. In some embodiments, the subject does not experience immune effector cell-associated neurotoxicity syndrome (ICANS).

[0144] Methods of Manufacturing

[0145] Provided herein are methods of manufacturing immune effector cells (for example, T cells or NK cells) engineered to express a CAR, for example, a CAR described herein, compositions comprising such cells, and methods of using such cells for treating a disease, such as an autoimmune disease or disorder, e.g., 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, antisynthetase 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis, in a subject. In some embodiments, the methods disclosed herein may manufacture immune effector cells engineered to express a CAR in less than 24 hours. Without wishing to be bound by theory, the methods provided herein preserve the undifferentiated phenotype of T cells, such as naive T cells, during the manufacturing process. These CAR-expressing cells with an undifferentiated phenotype may persist longer and / or expand better in vivo after infusion. In some embodiments, CART cells produced by the manufacturing methods provided herein comprise a higher percentage of stem cell memory T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 2). In some embodiments, CART cells produced by the manufacturing methods provided herein comprise a higher percentage of effector T cells, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 2). In some embodiments, CART cells produced by the manufacturing methods provided herein better preserve the sternness of T cells, compared to CART cells produced by the traditional manufacturing process. (FIG. 2). In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of hypoxia, compared to CART cells produced by the traditional manufacturing process, e.g., as measured using scRNA-seq. (FIG. 2). In some embodiments, CART cells produced by the manufacturing methods provided herein show a lower level of autophagy, compared to CART cells produced by the traditional manufacturing process. (FIG. 2).

[0146] In some embodiments, the CART cells manufactured by the methods disclosed herein may be administered to a subject with minimal ex vivo expansion, for example, less than 1 day, less than 12 hours, less than 8 hours, less than 6 hours, less than 4 hours, less than 3 hours, less than 2 hours, less than 1 hour, or no ex vivo expansion. Accordingly, the methods described herein provide a fast manufacturing process of making improved CAR-expressing cell products for use in treating a disease in a subject.

[0147] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) comprising: (i) contacting a population of cells (for example, T cells, for example, T cells isolated from a frozen or fresh leukapheresis product from a subject having an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis) with (A) an agent that stimulates a CD3 / TCR complex and / or (B) an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells; (ii) contacting the population of cells (for example, T cells) with a nucleic acid molecule (for example, a DNA or RNA molecule) encoding the CAR, thereby providing a population of cells (for example, T cells) comprising the nucleic acid molecule, and (iii) harvesting the population of cells (for example, T cells) for storage (for example, reformulating the population of cells in cryopreservation media) or administration, wherein: (a) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i), for example, no later than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 26 hours after the beginning of step (i), for example, no later than 22, 23, or 24 hours after the beginning of step (i), for example, no later than 24 hours after the beginning of step (i); (b) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i), for example, no later than 12, 13, 14, 15, 16, 17, or 18 hours after the beginning of step (i), for example, no later than 18 hours after the beginning of step (i), and step (iii) is performed no later than 30, 36, or 48 hours after the beginning of step (ii), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 hours after the beginning of step (ii); or (c) the population of cells from step (iii) are not expanded, or expanded by no more than 5, 10, 15, 20, 25, 30, 35, or 40%, for example, no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells at the beginning of step (i).

[0148] In some embodiments, the nucleic acid molecule in step (ii) is a DNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is an RNA molecule. In some embodiments, the nucleic acid molecule in step (ii) is on a viral vector, for example, a viral vector chosen from a lentivirus vector, an adenoviral vector, or a retrovirus vector. In some embodiments, the nucleic acid molecule in step (ii) is on a non-viral vector. In some embodiments, the nucleic acid molecule in step (ii) is on a plasmid. In some embodiments, the nucleic acid molecule in step (ii) is not on any vector. In some embodiments, step (ii) comprises transducing the population of cells (for example, T cells) a viral vector comprising a nucleic acid molecule encoding the CAR.

[0149] In some embodiments, the population of cells (for example, T cells) is collected from an apheresis sample (for example, a leukapheresis sample) from a subject having an autoimmune disease or disorder, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis.

[0150] In some embodiments, the apheresis sample (for example, a leukapheresis sample) is collected from the subject and shipped as a fresh product or a frozen sample (for example, a cryopreserved sample) to a cell manufacturing facility. Then the T cells (for example, CD4+ T cells and / or CD8+ T cells) are selected from the apheresis sample, for example, using a cell sorting machine (for example, a CliniMACS® Prodigy® device). The selected T cells (for example, CD4+ T cells and / or CD8+ T cells) are then seeded for CART manufacturing using the process described herein. In some embodiments, the selected T cells (for example, CD4+ T cells and / or CD8+ T cells) undergo one or more rounds of freeze-thaw before being seeded for CART manufacturing.

[0151] In some embodiments, cells (for example, T cells) are contacted with anti-CD3 and anti- CD28 antibodies for, for example, 12 hours, followed by transduction with a vector (for example, a lentiviral vector) encoding a CAR. 24 hours after culture initiation, the cells are washed and formulated for storage or administration. Without wishing to be bound by theory, brief CD3 and CD28 stimulation may promote efficient transduction of self-renewing T cells. Compared to traditional CART manufacturing approaches, the activation process provided herein does not involve prolonged ex vivo expansion. (FIG. 2). Similar to the cytokine process, the activation process provided herein also preserves undifferentiated T cells during CART manufacturing.

[0152] In some embodiments, the population of cells is contacted with (A) an agent that stimulates a CD3 / TCR complex and / or (B) an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells. In some embodiments, the agent that stimulates a CD3 / TCR complex is an agent that stimulates CD3. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28, ICOS, CD27, HVEM, LIGHT, CD40, 4-1BB, 0X40, DR3, GITR, CD30, HM1 , CD2, CD226, or any combination thereof. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28.

[0153] In some embodiments, the agent that stimulates a CD3 / TCR complex is chosen from an antibody (for example, a single-domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally existing, recombinant, or chimeric ligand). In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is chosen from an antibody (for example, a single-domain antibody (for example, a heavy chain variable domain antibody), a peptibody, a Fab fragment, or a scFv), a small molecule, or a ligand (for example, a naturally existing, recombinant, or chimeric ligand). In some embodiments, the agent that stimulates a CD3 / TCR complex does not comprise a bead. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor does not comprise a bead. In some embodiments, the agent that stimulates a CD3 / TCR complex comprises an anti-CD3 antibody. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises an anti-CD28 antibody. In some embodiments, the agent that stimulates a CD3 / TCR complex comprises an anti-CD3 antibody covalently attached to a colloidal polymeric nanomatrix. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises an anti-CD28 antibody covalently attached to a colloidal polymeric nanomatrix. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor is an agent that stimulates CD28, ICOS, CD27, CD25, 4-1BB, IL6RA, IL6RB, or CD2. In some embodiments, the agent that stimulates a costimulatory molecule and / or growth factor receptor comprises one or more of a CD28, ICOS, CD27, CD25, 4-1BB, IL6RB, and / or CD2 antigen binding domain, such as but not limited to an anti- CD28, anti-ICOS, anti-CD27, anti-CD25, anti-4-lBB, anti-IL6RA, anti-IL6RB, or anti-CD2 antibody or an antibody fragment comprising one or more CDRs, heavy chain, and / or light chain thereof - such as but not limited to an anti- CD28, anti-ICOS, anti-CD27, anti-CD25, anti-4-lBB, anti-IL6RA, anti-IL6RB, or anti-CD2 antibody provided in Table 27 of WO / 2021 / 173985, hereby incorporated by reference in its entirety. In some embodiments, the agent that stimulates a CD3 / TCR complex and the agent that stimulates a costimulatory molecule and / or growth factor receptor comprise T Cell TransAct™. In some embodiments, the agent that stimulates a CD3 / TCR complex and the agent that stimulates a costimulatory molecule and / or growth factor receptor are comprised in a multispecific binding molecule. In some embodiments, the multispecific binding molecule comprises a CD3 antigen binding domain and a CD28 or CD2 antigen-binding domain

[0154] In some embodiments, the population of cells is contacted with a nucleic acid molecule encoding a CAR. In some embodiments, the population of cells is transduced with a DNA molecule encoding a CAR. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs simultaneously with contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0.5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 20 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 19 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 17 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 16 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 15 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 14 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 13 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 12 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 11 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 10 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 9 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 8 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 7 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 6 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 5 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 4 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 3 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 2 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 1 hour after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, contacting the population of cells with the nucleic acid molecule encoding the CAR occurs no later than 30 minutes after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0155] In some embodiments, the population of cells is harvested for storage or administration. In some embodiments, the population of cells is harvested for storage or administration no later than 72, 60, 48, 36, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, or 18 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 26 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 25 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 24 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 23 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is harvested for storage or administration no later than 22 hours after the beginning of contacting the population of cells with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0156] In some embodiments, the population of cells is not expanded ex vivo. In some embodiments, the population of cells is expanded by no more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, or 60%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 5%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 10%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 15%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 20%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 25%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 30%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 35%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above. In some embodiments, the population of cells is expanded by no more than 40%, for example, as assessed by the number of living cells, compared to the population of cells before it is contacted with the agent that stimulates a CD3 / TCR complex and / or the agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells described above.

[0157] In some embodiments, the activation process is conducted in serum free cell media. In some embodiments, the activation process is conducted in cell media comprising one or more cytokines chosen from: IL-2, IL-15 (for example, hetIL-15 (IL15 / sIL-15Ra)), or IL-6 (for example, IL-6 / sIL-6Ra). In some embodiments, the activation process is conducted in cell media comprising a LSD1 inhibitor. In some embodiments, the activation process is conducted in cell media comprising a MALT1 inhibitor. In some embodiments, the serum free cell media comprises a serum replacement. In some embodiments, the serum replacement is CTS™ Immune Cell Serum Replacement (ICSR). In some embodiments, the level of ICSR can be, for example, up to 5%, for example, about 1%, 2%, 3%, 4%, or 5%.

[0158] In some embodiments, the present disclosure provides methods of making a population of cells (for example, T cells) that express a chimeric antigen receptor (CAR) comprising: (a) providing an apheresis sample (for example, a fresh or cryopreserved leukapheresis sample) collected from a subject with an autoimmune disease, e.g., 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), Sjogren's disease, neuroimmune disease (e.g. myasthenia gravis (MG), neuromyelitis optica (NMO), MOG associated disease (MOGAD), multiple sclerosis (MS)), 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, or amyotrophic lateral sclerosis; (b) selecting T cells from the apheresis sample (for example, using negative selection, positive selection, or selection without beads); (c) seeding isolated T cells at, for example, 1 x 106to 1 x 107cells / mL; (d) contacting T cells with an agent that stimulates T cells, for example, an agent that stimulates a CD3 / TCR complex and / or an agent that stimulates a costimulatory molecule and / or growth factor receptor on the surface of the cells (for example, contacting T cells with anti-CD3 and / or anti-CD28 antibody, for example, contacting T cells with TransAct); (e) contacting T cells with a nucleic acid molecule (for example, a DNA or RNA molecule) encoding the CAR (for example, contacting T cells with a virus comprising a nucleic acid molecule encoding the CAR) for, for example, 6-48 hours, for example, 20-28 hours; and (f) washing and harvesting T cells for storage (for example, reformulating T cells in cry opreservation media) or administration. In some embodiments, step (f) is performed no later than 30, 36, or 48 hours after the beginning of step (d) or (e), for example, no later than 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 hours after the beginning of step (d) or (e).

[0159] In some embodiments of the aforementioned methods, the methods are performed in a closed system. In some embodiments, T cell separation, activation, transduction, incubation, and washing are all performed in a closed system. In some embodiments of the aforementioned methods, the methods are performed in separate devices. In some embodiments, T cell separation, activation and transduction, incubation, and washing are performed in separate devices.

[0160] In some embodiments of the aforementioned methods, the methods further comprise adding an adjuvant or a transduction enhancement reagent in the cell culture medium to enhance transduction efficiency. In some embodiments, the adjuvant or transduction enhancement reagent comprises a cationic polymer. In some embodiments, the adjuvant or transduction enhancement reagent is chosen from: LentiBOOST™ (Sirion Biotech), vectofusin-1, F108 (Poloxamer 338 or Pluronic® F-38), protamine sulfate, hexadimethrine bromide (Polybrene), PEA, Pluronic F68, Pluronic Fl 27, Synperonic or LentiTrans™. In some embodiments, the transduction enhancement reagent is LentiBOOST™ (Sirion Biotech). In some embodiments, the transduction enhancement reagent is Fl 08 (Poloxamer 338 or Pluronic® F-38)

[0161] In some embodiments of the aforementioned methods, the transducing the population of cells (for example, T cells) with a viral vector comprises subjecting the population of cells and viral vector to a centrifugal force under conditions such that transduction efficiency is enhanced. In an embodiment, the cells are transduced by spinoculation.

[0162] In some embodiments of the aforementioned methods, cells (e.g., T cells) are activated and transduced in a cell culture flask comprising a gas-permeable membrane at the base that supports large media volumes without substantially compromising gas exchange. In some embodiments, cell growth is achieved by providing access, e.g., substantially uninterrupted access, to nutrients through convection.

[0163] Pharmaceutical Compositions

[0164] The methods described herein can further include formulating a CD 19 CAR-expressing cell in a pharmaceutical composition. Pharmaceutical compositions may comprise a CD 19 CAR-expressing cell, for example, a plurality of CD 19 CAR-expressing cells, as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. 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; adjuvants (for example, aluminum hydroxide); and preservatives. Compositions can be formulated, for example, for intravenous administration.

[0165] In some embodiments, the pharmaceutical composition is substantially free of, for example, there are no detectable levels of a contaminant, for example, selected from the group consisting of endotoxin, mycoplasma, replication competent lentivirus (RCL), p24, VSV-G nucleic acid, HIV gag, residual anti-CD3 / anti-CD28 coated beads, mouse antibodies, pooled human serum, bovine serum albumin, bovine serum, culture media components, vector packaging cell or plasmid components, a bacterium and a fungus. In some embodiments, the bacterium is at least one selected from the group consisting of Alcaligenes faecalis, Candida albicans, Escherichia coli, Haemophilus influenza, Neisseria meningitides, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumonia, and Streptococcus pyogenes group A.

[0166] When “an immunologically effective amount” or “therapeutic amount” is indicated, the precise amount of the compositions to be administered can be determined by a physician with consideration of individual differences in age, weight, tumor size, extent of infection or metastasis, and condition of the patient (subject). It can generally be stated that a pharmaceutical composition comprising the immune effector cells (for example, T cells, NK cells) described herein may be administered at a dosage of about 0.5 x 106to 50 x 106viable CAR-expressing cells, in some instances about 12.5 x 106viable CAR-expressing cells, including all integer values within those ranges. T cell compositions may also be administered multiple times at these dosages. The cells can be administered by using infusion techniques that are commonly known in immunotherapy (see, for example, Rosenberg et al., New Eng. J. of Med. 319: 1676, 1988).

[0167] The administration of the subject compositions may be carried out in any convenient manner. The compositions described herein may be administered to a patient trans arterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection, or intraperitoneally, for example, by intradermal or subcutaneous injection. The compositions of immune effector cells (for example, T cells, NK cells) may be injected directly into a lymph node or site of disease.

[0168] EXAMPLES

[0169] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0170] Example 1: Description of the Activated Rapid Manufacturing (ARM) process

[0171] In some embodiments, CART cells are manufactured using a continuous Activated Rapid Manufacturing (ARM) process, over approximately 2 days, which will potentially allow for a greater number of less differentiated T cells (T naive and TSCM (stem central memory T) cells) to be returned to a patient for in vivo cellular expansion. The short manufacturing time period allows the early differentiated T cells profile to proliferate in the body for their desired terminal differentiated state rather that in an ex vivo culture vessel.

[0172] In some embodiments, CART cells are manufactured using cryopreserved leukapheresis source material, for example, non-mobilized autologous peripheral blood leukapheresis (LKPK) material. Cryopreserved source material undergoes processing steps for T cell enrichment on the first day of production (Day 0) by means of anti-CD4 / anti-CD8 immunomagnetic system. Positive fraction is then seeded in G-rex culture vessel, activated with an anti-CD3 / CD28 system (TransACT™) and on the same day transduced with a lentiviral vector (LV) encoding a CAR. On the following day, after 20-28 hours of transduction, the T cells are harvested, washed four times, formulated in freezing medium, and then frozen by a Controlled Rate Freezer (CRF). From the start of the process on Day 0 to the initiation of harvest on the following day, cells are cultured for 20 - 28 hours with a target of 24 hours after Day 0 seeding.

[0173] Media for Day 0 were prepared according to Table 1. Table 1: Media type and point of use during CART manufacturing

[0174] The cryopreserved leukapheresis material is thawed. The thawed cells are diluted with the Rapid Buffer (Table 21) and washed on the CliniMACS® Prodigy® device. The T cells are selected by CliniMACS® CD4 and CD8 microbeads. Once the program is finished for T cell selection (approximately 3h 40 min to 4h 40 min), the reapplication bag containing the cells suspended in Rapid Media (Table 21) are transferred in a transfer pack. A sample is taken for viability and cell count. The cell count and viability data from the positive fraction bag is used to determine the cell concentration when seeding the culture vessel for activation and vector transduction.

[0175] Following positive selection of T cells via the CliniMACS® microbeads (CD4 and CD8), the cells are seeded in the culture vessel, G-Rex. Once the cells are seeded, the activation reagent (TransACT™) is then added to the culture vessel. The cells are then transduced with a lentiviral vector encoding a CAR at a target MOI of 1.0 (0.8-1.2). Following the vector addition, the culture vessel is transported to an incubator where it is incubated for a target of 24 hours (operating range 20-28 hours) at a nominal temperature of 37 °C (operating range 36-38 °C) with nominal 5% CO2 (operating range 4.5-5.5%). Following the incubation, the cells are washed with Harvest Wash Solution (Table 1) four times to remove any non-integrated vector and residual viral particles, as well as any other process related impurities. Then, the cells are eluted and a sample for cell count and viability is taken for testing and the results are used to determine the volume required to re-suspend the cells for final formulation with CryoStor® CS10. The cells are then centrifugated to remove the Harvest Wash Solution and proceed with cryopreservation. In some embodiments, the CAR expressed in CART cells binds to CD 19. In some embodiments, the CAR made by the ARM process is rapcabtagene autoleucel. In some embodiments, IL-2 used in the Rapid Media (RM) (Table 1) can be replaced with IL-15, hetlL- 15 (IL-15 / sIL-15Ra), IL-6, or IL-6 / sIL-6Ra.

[0176] Example 2: Generation and in vitro characterization assessment of CD19-targeting CAR-T cells using T cells from Systemic Lupus Erythematosus patients

[0177] Fresh blood from five SLE patients with moderate SLE disease activity (SLED Al score of 8) and 5 age / gender-matched (± 5 years) healthy donors (HD) was collected. T cell generation and ex vivo expansion were performed according to the methods disclosed herein.

[0178] Cell characterization, including cell viability, CAR-T cells final product recovery rate post-harvesting (or post-wash), and ex vivo fold expansion was analyzed in both SLE patients and HDs.

[0179] Results: Highly pure and viable T cells were obtained from both SLE patients and HDs. The manufacturing process used allowed preservation of CD4 and CD8 naive / T scm cells in the final product. Overall, SLE patient-derived CAR-T cells displayed comparable transduction rates, T cell sternness feature (data not shown) and functional properties (cytolytic activity and cytokine production) to CAR-T cells generated from age / gender-matched HDs. (FIG. 3A-C)

[0180] The rapcabtagene autoleucel cells manufactured from SLE patients in this study were fully functional in vitro. (FIG. 4A-B). These results support the feasibility of generating cells via the methodologies described herein, using T cells from SLE patients.

[0181] Example 3: Phase 1 / 2 study, open-label, multi-center, to assess safety, efficacy and cellular kinetics of rapcabtagene autoleucel cells in participants with severe, refractory autoimmune disorders

[0182] This study evaluates the safety, efficacy, and in vivo cellular kinetics (pharmacokinetics, PK) of rapcabtagene autoleucel treatment in participants with severe refractory systemic lupus erythematosus (srSLE) and other severe forms of autoimmune diseases.

[0183] Rapcabtagene autoleucel is an autologous CD19-directed CAR-T cell therapy that is comprised of CD4+ / CD8+ T cells that have undergone ex vivo T cell activation and gene modification. Rapcabtagene autoleucel utilizes the FMC63 scFv domain for CD 19 recognition and the same lentiviral vector as tisagenlecleucel (Kymriah, CTL019) and is manufactured via the activated rapid manufacturing (ARM) process. The ARM process reduces the turnaround time compared to traditional manufacturing processes (FIG.2) and preserves T cell sternness, the ability to self-renew and mature, resulting in a product with greater proliferative potential and fewer exhausted T cells compared to traditionally manufactured CAR-T cells. With ARM, CAR- T cell expansion occurs primarily within a patient’s body (in vivo), eliminating the need for an extended culture time outside of the body (ex vivo). These unique characteristics may lead to better and more durable responses, improved long-term outcomes and a reduced risk of severe adverse events compared to CAR-T cell products manufactured via traditional manufacturing methods. Non-clinical studies show that rapcabtagene autoleucel is a product with potentially superior antitumor efficacy, a similar safety profile, and delayed expansion compared to other CD19-directed CAR-T cell therapy relying on a traditional manufacturing process (e.g. tisagenlecleucel).

[0184] Disease

[0185] Systemic lupus erythematosus (SLE) is a chronic, molecularly, pathologically and clinically heterogeneous autoimmune disease characterized by a wide range of organ damage manifestations (Fanouriakis et al 2021). Current standard of care includes conventional immunomodulatory and anti-inflammatory agents such as antimalarials, glucocorticoids and immunosuppressives (e.g. methotrexate, azathioprine, mycophenolate and cyclophosphamide) and biologies (such as, belimumab and very recently, anifrolumab as well as rituximab commonly used in this severe stage of this disease). Approximately 70% of the patients follow a relapsing-remitting disease course, the remaining divided equally between a prolonged remission and a persistently active disease despite therapy. For organ-threatening or life-threatening SLE, treatment usually includes an initial period of high-intensity immunosuppressive therapy to control disease activity, followed by a longer period of less intensive therapy to consolidate response and prevent relapses (Fanouriakis et al 2021).

[0186] Severe, refractory SLE (srSLE) patients are usually excluded from randomized clinical trials. srSLE patients, with or without renal involvement, after having failed immunosuppressive and biological therapies, have very limited treatment options. Autologous stem cell transplantation (ASCT) may be performed, however, it remains experimental and is associated with significant toxicities including mortality (Jayne et al 2004). In the present study, srSLE patients with or without lupus nephritis are included. If the initial data from srSLE patients are supportive, patients with other severe autoimmune diseases may be enrolled into a new study part (e.g. Part B, described in more detail below) after a substantial amendment.

[0187] Overall Study Design

[0188] The study starts with Part A in participants diagnosed with srSLE. Inclusion of up to 3 additional parts (Part B, C, and D) may be initiated in parallel to Part A once first clinical data is available from this study and can include one or more of the following changes: (1) to assess the effect of lymphodepletion on safety / efficacy, (2) to assess a different dose level, (3) to broaden the study population by adding other indications such as other subtypes of SLE (including other organ involvements such cutaneous or articular), or other severe, B-cell driven, autoimmune diseases, e.g. systemic sclerosis or anti-synthetase syndrome where the benefit / risk assessment of rapcabtag ene autoleucel is positive. The study design / participant journey is illustrated in FIG. 1. The total study duration for a participant is up to 27 months, after which a long-term follow-up is initiated until 15 years after rapcabtagene autoleucel administration. Participants are first evaluated for clinical eligibility (weeks -10 to -6) as detailed below. If clinically eligible, leukapheresis is scheduled (weeks -6 to -2). Once the leukapheresis product has been confirmed to be suitable for rapcabtagene autoleucel manufacturing, the manufacturing process is commenced. Before the planned apheresis and CAR-T cell injection, immunosuppressive treatments are stopped except for low-dose prednisolone (up to 10 mg / day) and antimalarials, which are allowed to continue. After the final product has been confirmed to be available, participants receive lymphodepleting therapy (up to two weeks prior to Day 1) as detailed below. Following pre-inj ection check on Day 1 and premedication, rapcabtagene autoleucel is administered as a single injection on Day 1.

[0189] Each participant is hospitalized until at least Day 14 following rapcabtagene autoleucel injection. Participants are closely monitored for any safety events for the first 2 months (twice a week visits for the first 5 weeks followed by weekly visits up to end of Month 2 (Day 60)). After that, the visit frequency is reduced to quarterly visits at Months 3, 6, 9, 12, and then twice a year (Months 18 and 24). End of study visit for a participant in this study is completed at Month 24, but participants will continue to be followed in the long-term follow-up.

[0190] Part A of this study will evaluate whether rapcabtagene autoleucel, following lymphodepletion, reduces disease activity to a low disease state for severe refractory SLE patients with at least one organ involvement. The estimand for Part A is described by the following attributes:

[0191] 1. Population: srSLE patients with at least one organ involvement.

[0192] 2. Endpoint: Achievement of Low Lupus Disease Activity State (LLDAS) at Month 6.

[0193] 3. Treatment of interest: Participants receiving lymphodepletion and a successful injection of ARM-CD19 CAR T within specified dose range, and not receiving any prohibited concomitant medication as per the protocol.

[0194] 4. Measure: Percentage of participants achieving LLDAS at Month 6.

[0195] Participant inclusion and exclusion criteria

[0196] In Part A of this Phase 1 / 2 study, approximately 12 participants diagnosed with srSLE are treated with rapcabtagene autoleucel. Of these 12 participants, at least 8 (2 / 3rds) have renal involvement, as defined below. Each subject is first evaluated for clinical eligibility during screening. Participants eligible for inclusion in this study must meet the requirements for adequate renal, hepatic, cardiac, hematological and pulmonary function as follows: (1) renal function defined as serum creatinine of < 1.5 x ULN OR eGER > 45 ml / min / 1.73m2, (2) hepatic function defined as ALT and AST <4 x ULN and total bilirubin <1.5 x ULN with the exception of participants with Gilbert syndrome who may be included if their total bilirubin is <3.0 x ULN and direct bilirubin <1.5 x ULN, and cardiac function defined as LVEL > 50% as determined by ECHO or MRA or MUGA at screening, unless cardiac impairment is clearly attributable to SLE, (3) hematologic function (regardless of transfusion) defined as absolute neutrophil count (ANC) >600 / pL (only for participants with non-historical leukapheresis), platelets >50,000 / pL, white blood cells count (WBC) >1,000 cells / pL, and absolute lymphocyte count >200 / pL, and (4) pulmonary function defined as no or mild dyspnea (< Grade 1) and oxygen saturation measured by pulse oximetry > 90% on room air. The study includes men and women with SLE, aged >18 years and <65 years at screening, fulfilling the 2019 American College of Rheumatology (ACR) classification criteria for SLE (see Table XI, Aringer et al. (2019) Arthritis Rheumatol, p. 1400- 1412, Aringer et al. (2019) Ann Rheum Dis, p. 1151-1159) at least 12 months prior to and at screening. SLE can be classified by criteria set out in Aringer et al. (2019) Ann Rheum Dis, p. 1151-1159.

[0197] Table XI. ACR 2019 definitions of SLE classification criteria

[0198] Criteria Definition

[0199] Antinuclear antibodies (ANA) Antinuclear antibodies (ANA) at a titer of >1:80 on HEp-2 cells or an equivalent positive test at least once. Testing by immunofluorescence on HEp-2 cells or a solid phase ANA screening immunoassay with at least equivalent performance is highly recommended.

[0200] Fever Temperature >38.3° Celsius.

[0201] Leukopenia White blood cell count <4,000 / mm3.

[0202] Thrombocytopenia Platelet count <100,000 / mm3.

[0203] Autoimmune hemolysis Evidence of hemolysis, such as reticulocytosis, low haptoglobin, elevated indirect bilirubin, elevated LDH AND positive Coomb’s (direct antiglobulin) test.

[0204] Delirium Characterized by (1) change in consciousness or level of arousal with reduced ability to focus, and (2) symptom development over hours to <2 days, and (3) symptom fluctuation throughout the day, and (4) either (4a) acute / subacute change in cognition (e.g. memory deficit or disorientation), or (4b) change in behavior, mood, or affect (e.g. restlessness, reversal of sleep / wake cycle).

[0205] Psychosis Characterized by (1) delusions and / or hallucinations without insight and (2) absence of delirium.

[0206] Seizure Primary generalized seizure or partial / focal seizure.

[0207] Non-scarring alopecia Non-scarring alopecia observed by a clinician*.

[0208] Oral ulcers Oral ulcers observed by a clinician*.

[0209] Criteria Definition

[0210] Subacute cutaneous or discoid Subacute cutaneous lupus erythematosus observed by a clinician*: lupus Annular or papulosquamous (psoriasiform) cutaneous eruption, usually photodistributed. Discoid lupus erythematosus observed by a clinician*: Erythematous-violaceous cutaneous lesions with secondary changes of atrophic scarring, dyspigmentation, often follicular hyperkeratosis / plugging (scalp), leading to scarring alopecia on the scalp. If skin biopsy is performed, typical changes must be present. Subacute cutaneous lupus: interface vacuolar dermatitis consisting of a perivascular lymphohistiocytic infiltrate, often with dermal mucin noted. Discoid lupus: interface vacuolar dermatitis consisting of a perivascular and / or periappendageal lymphohistiocytic infiltrate. In the scalp, follicular keratin plugs may be seen. In longstanding lesions, mucin deposition and basement membrane thickening may be noted.

[0211] Acute cutaneous lupus Malar rash or generalized maculopapular rash observed by a clinician*. If skin biopsy is performed, typical changes must be present (Acute cutaneous lupus: interface vacuolar dermatitis consisting of a perivascular lymphohistiocytic infiltrate, often with dermal mucin noted. Perivascular neutrophilic infiltrate may be present early in the course.

[0212] Pleural or pericardial effusion Imaging evidence (such as ultrasound, x-ray, CT scan, MRI) of pleural or pericardial effusion, or both.

[0213] Acute pericarditis >2 of (1) pericardial chest pain (typically sharp, worse with inspiration, improved by leaning forward), (2) pericardial rub, (3) EKG with new widespread ST -elevation or PR depression, (4) new or worsened pericardial effusion on imaging (such as ultrasound, x- ray, CT scan, MRI).

[0214] Joint involvement EITHER (1) synovitis involving 2 or more joints characterized by swelling or effusion OR (2) tenderness in 2 or more joints and at least 30 minutes of morning stiffness.

[0215] Proteinuria >0.5g / 24h Proteinuria >0.5g / 24h by 24 hour urine or equivalent spot urine protein-to-creatinine ratio.

[0216] Class II or V lupus nephritis on Class II: Mesangial proliferative lupus nephritis: Purely mesangial renal biopsy according to hypercellularity of any degree or mesangial matrix expansion by

[0217] ISN / RPS 2003 classification. light microscopy, with mesangial immune deposit. A few isolated subepithelial or subendothelial deposits may be visible by immune- fluorescence or electron microscopy, but not by light microscopy. Class V: Membranous lupus nephritis: Global or segmental subepithelial immune deposits or their morphologic sequelae by light microscopy and by immunofluorescence or electron microscopy, with or without mesangial alterations. Criteria Definition

[0218] Class III or IV lupus nephritis Class III: Focal lupus nephritis: Active or inactive focal, segmental on renal biopsy according to or global endo- or extracapillary glomerulonephritis involving <50%

[0219] ISN / RPS 2003. of all glomeruli, typically with focal subendothelial immune deposits, with or without mesangial alterations. Class IV: Diffuse lupus nephritis: Active or inactive diffuse, segmental or global endo- or extracapillary glomerulonephritis involving >50% of all glomeruli, typically with diffuse subendothelial immune deposits, with or without mesangial alterations. This class includes cases with diffuse wire loop deposits but with little or no glomerular proliferation.

[0220] Positive anti-phospholipid Anti-Cardiolipin antibodies (IgA, IgG, or IgM) at medium or high antibodies titer (>40 APL, GPL or MPL, or >the 99th percentile) or positive anti-P2GPl antibodies (IgA, IgG, or IgM) or positive lupus anticoagulant.

[0221] Low C3 OR low C4 C3 OR C4 below the lower limit of normal.

[0222] Low C3 AND low C4 Both C3 AND C4 below their lower limits of normal.

[0223] Anti-dsDNA antibodies OR Anti-dsDNA antibodies in an immunoassay with demonstrated >

[0224] Anti-Smith (Sm) antibodies. 90% specificity for SLE against relevant disease controls OR Anti¬

[0225] Smith (Sm) antibodies.

[0226] ‘This may include physical examination or review of a photograph. Source: Aringer et al. (2019) Ann Rheum Dis, p. 1151-1159.

[0227] Participants are positive for at least one of the following autoantibodies at screening: antinuclear antibodies (ANA) at a titer of >1 :80 (on HEp-2 cells or an equivalent positive test), or anti dsDNA (above the ULN); or anti-Sm (above the ULN). Participants also have active (severe) disease as defined by SLEDAI-2K > 8 (not including the SLEDAI-2K domains of lupus headache, cerebrovascular accident, organic brain syndrome) and at least one of the following significant SLE related organ involvements: (1) renal: histological diagnosis of proliferative lupus nephritis World Health Organization (WHO) ISN / RPS (Weening et al 2004) Class III or IV within 2 years of screening, AND at least one of the following at screening: first morning void UPCR: 0.7 to 4 mg / mg or urine sediment consistent with active proliferative lupus nephritis such as presence of cellular (granular or red blood cell) casts or hematuria (>5 red blood cells per high power field) if other causes such as menstrual bleeding are excluded; (2) endo / peri / myocarditis; (3) pleuritis or other lung involvement; or (4) vasculitis. At least 2 / 3rds of participants have lupus nephritis as defined by the above.

[0228] Patients also have failed to respond (i.e., having high disease activity as defined above despite the following therapy) to two or more standard immunosuppressive therapies (including one of mycophenolate or cyclophosphamide), unless contraindicated or having experienced documented adverse events or intolerance related to such immunosuppressive drugs not allowing their further use, in combination with glucocorticoids and failure to respond to at least one biological agent (unless contraindicated or the patient is deemed ineligible).

[0229] Exclusion criteria include: (1) prior treatment with anti-CD19 therapy, adoptive T cell therapy or any prior gene therapy product (e.g. CAR-T cell therapy), (2) any acute, severe lupus related flare during screening that needs immediate treatment and / or makes the immunosuppressive washout impossible and thus makes the patient ineligible for CD 19 CAR-T therapy, such as acute CNS lupus (e.g. psychosis, epilepsy) or catastrophic antiphospholipid syndrome, and (3) any significant, likely irreversible organ damage related to SLE, e.g. end stage renal disease, where CD 19 CAR-T cell therapy is deemed to be unlikely to benefit the patient.

[0230] Study Treatment

[0231] Eligible participants undergo the following sequence of events prior to rapcabtagene autoleucel administration: (1) leukapheresis, (2) pre-lymphodepletion evaluation, (3) lymphodepletion, (4) premedication, and (5) pre- rapcabtagene autoleucel injection check.

[0232] Lymphodepleting therapy starts within one week before rapcabtagene autoleucel injection, which means that rapcabtagene autoleucel is injected 2 to 6 days after lymphodepleting therapy is completed. Lymphodepleting therapy may be repeated in the case rapcabtagene autoleucel has been delayed by more than 2 weeks. The lymphodepleting therapy regime is as follows: (1) fludarabine administered 25 mg / m2intravenously [i.v.] daily for 3 doses (for participants with renal impairment, the dose may be reduced as per local approved labels of fludarabine), and (2) cyclophosphamide administered 250 mg / m2i.v. daily for 3 doses starting with the first dose of fludarabine.

[0233] All participants are pre-medicated with acetaminophen (paracetamol, 650 mg, orally) and diphenhydramine (25-50 mg, i.v. or orally) or another Hl antihistamine approximately 30 to 60 minutes prior to injection. These medications can be repeated every 6 hours as needed. Nonsteroidal anti-inflammatory medication may be prescribed if the participant continues to have fever not relieved with acetaminophen (paracetamol).

[0234] In Part A of this Phase 1 / 2 study, rapcabtagene autoleucel treatment consists of a single intravenous (i.v.) injection of a target dose of 12.5 x 106CAR-positive viable T cells (range 5 x 106- 12.5 x 106CAR-positive viable T cells) in cell suspension, without weight-based dose adjustment. The 12.5 x 106starting dose is selected based on data from the DLBCL arm of the Phase I oncology study, CYTB323A12101. In CYTB323A12101 DLBCL, high rates of complete tumor response by elimination of CD 19 expressing cells were shown from doses of

[0235] 12.5 x 106(63.2% CR at month 3), indicating that depletion of CD19 expressing B cells, the intended pharmacodynamic effect in srSLE, can be expected in srSLE from this dose. A dose of

[0236] 2.5 x 106in CYTB323A12101 DLBCL resulted in lower CR rates at Month 3 (25%). A study of treatment of four patients with srSLE with a traditionally manufactured CD 19 targeting CAR-T (Mougiakakos et al 2021, Schett et al 2022) reported a dose of 46 x 106CAR-T cells bein used.

[0237] Doses within the range of 2.5-40 x 106CAR-positive viable T cells can also be used, and toxicity expected to be lower than observed for participants with DLBCL in CYTB323A12101. In participants with DLBCL in CYTB323A12101, manageable safety was shown at target doses of 2.5-40 x 106, with only 3 of 45 patients experiencing a > grade 3 CRS or ICANS. There was no evidence of a dose-safety relationship for DLBCL.

[0238] Prior to leukapheresis, immunosuppressive / immunomodulatory treatments are discontinued as they may interfere with the expansion or function of the CAR-T cells and, together with the profound B cell depletion, may cause excessive immunosuppression. The washout periods for the immunosuppressive agents are relatively short to minimize the risk of disease worsening; furthermore, antimalarials and corticosteroid treatment are allowed to continue, for the latter at a maximum dose of 10 mg per day (prednisone or equivalent) during leukapheresis and after rapcabtag ene autoleucel injection. Corticosteroids are tapered to max 10 mg prednisone or equivalent at least one week before leukapheresis based on baseline dose, disease activity and clinical symptoms. After rapcabtagene autoleucel injection any remaining steroids are tapered any time post injection per the following: (a) if SLEDAI-2K reduced by 4 (or more) for at least 2 weeks: taper to 7.5mg / day or less; (b) if SLEDAI-2K below 4: taper to 5 or 0 mg / day; or (3) if in remission steroids should be discontinued completely. In the first 24 weeks after the rapcabtagene autoleucel injection, the use of all immunosuppressive / immunomodulatory treatments are prohibited with the exception of corticosteroids (max. 10 mg prednisone or equivalent per day) and / or antimalarials. It is recommended to schedule leukapheresis prior to any planned corticosteroids as an absolute T cell count (absolute lymphocyte count multiplied by the percentage of CD3 positive lymphocytes) < 300 / mm3may result in a poor T cell collection and manufacturing failure. If the patient shows signs of worsening after leukapheresis but still deemed eligible for CAR-T therapy, an additional 5 mg prednisone or equivalent (so max 15 mg prednisone per day) as bridging therapy may be administered, which should be discontinued prior to rapcabtagene autoleucel administration. Antimalarials will be allowed to be continued during the full study as needed. After 24 weeks, SLE standard of care therapy may be given. Intravenous Immunoglobulin replacement therapy can be administered using the guideline by Hill et al, 2019.

[0239] Safety, Pharmacokinetic (PK) and Efficacy Assessments Objectives and related endpoints are summarized in Table X2. Safety of rapcabtagene autoleucel in participants with srSLE and other severe forms of autoimmune diseases is evaluated using parameters including vital signs, adverse events, laboratory parameters and electrocardiogram (ECG) evaluation.

[0240] Table X2 Objectives and related endpoints

[0241] Objective(s) Endpoint(s)

[0242] Primary objective(s) Endpoint(s) for primary objective(s)

[0243] • To assess safety of rapcabtagene autoleucel in • Safety parameters include vital signs, participants with srSLE and other severe forms adverse events, laboratory parameters and of autoimmune diseases ECG evaluation

[0244] Secondary objective(s) Endpoint(s) for secondary objective(s)

[0245] • To characterize the in vivo cellular kinetics • Rapcabtagene autoleucel transgene

[0246] (pharmacokinetics, PK) of rapcabtagene concentrations by qPCR over time in autoleucel in peripheral blood by quantitative peripheral blood; cellular kinetics polymerase chain reaction (qPCR) parameters (Cmax, AUC, Tmax, Tl / 2,

[0247] Clast, Tlast)

[0248] • To characterize the incidence and prevalence of • Pre-existing and treatment induced pre-existing and treatment induced immunogenicity (cellular, humoral) of immunogenicity (cellular and humoral) of rapcabtagene autoleucel rapcabtagene autoleucel

[0249] • To evaluate feasibility of the manufacturing • Manufacture success (defined as meeting process in autoimmune disorders release specifications and at or above the planned target dose)

[0250] • Part A: To assess the effect of rapcabtagene • At various timepoints: autoleucel on the following SLE diseaseaSLEDAI-2K activity scores . BILAG-2004

[0251] • Physician's global assessment

[0252] • LLDAS

[0253] • Remission (DORIS) Objective(s) Endpoint(s)

[0254] • SRI-4

[0255] • BICLA

[0256] • Part A: To evaluate effect of rapcabtag ene • UPCR at various timepoints autoleucel for srSLE participants who also have . Complete Renal Response (CRR) at various active lupus nephritis timepoints

[0257] For pharmacokinetic (PK) analysis, serial blood samples are collected at different time points to measure rapcabtagene autoleucel transgene concentrations in peripheral blood by quantitative polymerase chain reaction (qPCR). Levels of rapcabtagene autoleucel transduced cells will be measured by flow cytometry of CD3 -positive, rapcabtagene autoleucel -positive cells. The absolute number of CD 19+ B cells in the peripheral blood is measured by flow cytometry and used as the pharmacodynamics (PD) marker. The flow cytometry analysis can be performed using a validated panel that also includes the analysis of T cells and NK cells (TBNK). Pre-existing and treatment- induced immunogenicity (cellular, humoral, neutralizing antibodies) of rapcabtagene autoleucel is assessed by one or more of the following: (1) humoral immunogenicity (anti-drug antibodies, ADA) measured by flow cytometry analysis of ADA binding to ARM-CD19 CAR-expressing cells, (2) presence of neutralizing antibodies measured by a reporter gene assay, (3) cellular immunogenicity measured by flow cytometry analysis of T cell interferon-gamma expression. Analytical methods for PK and immunogenicity assessments are listed in Table 2.

[0258] Table 2. Analytical methods associated with the PK and immunogenicity assessments

[0259] To assess the effects of rapcabtagene autoleucel on disease activity in patients with srSLE, the following assessments are performed at various time points: LLDAS, Remission (DORIS), SLEDAI-2K, BILAG-2004, Physician’s global assessment, SRI-4, and / or BICLA. Furthermore, for srSLE patients who have active lupus nephritis, in addition to the beforementioned assessments, UPCR and Complete Renal Response (CRR) at various timepoints are analyzed. These results are described below.

[0260] Results

[0261] Rapcabtagene autoleucel has been administered to 6 patients diagnosed with srSLE with or without renal involvement (LN). Patient baseline characteristics are set forth in Table 3.

[0262] Table 3: Patient Baseline Characteristics All participants received lymphodepleting therapy and were dosed with 12.5 x 106CARpositive viable T cells. Safety was assessed by evaluating vital signs, adverse events (AE), laboratory parameters, and an electrocardiogram. No deaths were reported. One Serious Adverse Event (SAE) of pneumonia (Grade 2) was reported, that improved with antibiotic treatment. All 6 participants had blood and lymphatic system disorder AEs, specifically representing cytopenias assessed as related to lymphodepleting therapy.

[0263] All 6 subjects had at least one AE suspected to be related to rapcabtagene autoleucel, with 5 participants (83.3%) reporting immune system disorders and 4 participants (66.7%) reporting blood and lymphatic system disorders. Grade 1 or 2 cytokine release (CRS) was reported in four participants and resolved within 9 days following a single dose of tocilizumab. No immune effector cell-associated neurotoxicity syndrome (ICANS) were reported. No unexpected safety signal was observed.

[0264] Preliminary results indicate rapcabtagene autoleucel is effective in treating srSLE. B cell depletion was observed in all participants during lymphodepletion and was sustained after rapcabtagene autoleucel infusion for all participants who have reached 60 days follow-up. (FIG. 5) B cell recovery was observed for Patient 4001002 starting at 90 days and Patient 4001001 at 180 days. Notably, SLEDAI-2K total scores showed substantial improvement in all participants following infusion with rapcabtagene autoleucel (FIG. 6). The improvement was sustained in Patient 4001001 who had the longest follow-up to date (183 days), with a SLEDAI-2K total score of 0 (subject is in remission) at Day 92 and 183. Improvements were also observed in the individual SLEDAI-2K domains, including pleurisy, alopecia, low complement, rash, arthritis, and vasculitis (FIG. 7). PhGA scores also showed substantial improvement over time following rapcabtagene autoleucel administration. (FIG. 8).

[0265] The observed improvements were also reflected by a corresponding change in biomarker values. More specifically, the levels of anti-dsDNA antibodies decreased (FIG. 9) following administration of rapcabtagene autoleucel whereas the levels of complement C3 and C4 increased (FIG. 10). In sum, preliminary data suggest administration of rapcabtagene autoleucel is effective in treating srSLE.

[0266] EQUIVALENTS

[0267] The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to certain embodiments, it is apparent that further embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

What is claimed is:

1. A method of treating a subject diagnosed with systemic lupus erythematosus (SLE) comprising administering rapcabtagene autoleucel to the subject.

2. The method of claim 1, wherein the SLE is severe refractory SLE (srSLE).

3. The method of claim 1 or 2, wherein the SLEDAI-2K score of the subject is reduced by at least 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 15 points, 20 points or 25 points as compared to the baseline SLEDAI-2K score.

4. The method of claim 3, wherein the reduction in the SLEDAI-2K score is sustained for at least 3 months, 6 months, 9 months, 12 months, or 18 months.

5. The method of any of claims 1-4, wherein the total SLEDAI-2K score of the patient is reduced to 6 or below, 5 or below, 4 or below, 3 or below, 2 or below, 1 or below, or 0.

6. The method of any of claims 1-5, wherein the PhGA score of the subject is reduced by at least 0.3 points, 0.5 points, 0.75 points, 1 points, 1.25 points, 1.5 points, 1.75 points, 2 points, 2.25 points, 2.5 points, or 2.75 points as compared to the baseline PhGA score.

7. The method of any of claims 1-6, wherein the total PhGA score is reduced to 1.5 or lower, 1.25 o lowers, 1 or lower, 0.75 or lower, 0.5 or lower, or 0.3 or lower.

8. The method of any of claims 1-7, wherein the reduction on PhGA score is sustained for at least 6 months or more, 9 months or more, 12 months or more, 18 months or more, or 24 months or more.

9. The method of claim 7, wherein the sustained reduction is maintained without an increase in PhGA score to l.O or greater, 1.25 or greater, 1.5 or greater, 1.75 or greater, or 2 or greater.

10. The method of any of claims 1 -9, wherein there is no increase in PhGA score of greater than 0.3 points, 0.25 points, 0.2 points, 0.15 points, 0.1 points, or 0.5 points from baseline following administration11. The method of any of claims 1-10, wherein the subject achieves a lupus low disease activity state (LLDAS) within 6 months, 9 months, or 12 months following administration.

12. The method of claim 11, wherein LLDAS is sustained for at least 3 months, 6 months, 12 months, or 18 months.

13. The method of any of claims 1-12, wherein the subject is in remission according to the DORIS definition within 3 months, 6 months, 9 months, or 12 months following administration.

14. The method of claim 13, wherein remission is sustained for at least 3 months, 6 months, 12 months, or 18 months.

15. The method of any of claims 1-14, wherein the subject has a reduction in BILAG score from A to C or better, A to D, B to C or better, or B to D within 6 months following administration.

16. The method of any of claims 1-15, wherein the reduction in BILAG score is sustained for at least 3 months, 6 months, 12 months, or 18 months.

17. The method of any of claims 1-16, wherein the subject is a responder according to the SRI-4 definition.

18. The method of claim 17, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

19. The method of any of claims 1-18, wherein the subject is a responder according to the BICLA definition.

20. The method of claim 19, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

21. The method of any of claims 1-20, wherein the subject has a complete renal response (CRR) following administration.

22. The method of claim 21, wherein the CRR is sustained for at least 3 months, 6 months, 9 months, 12 months, or 18 months.

23. The method of any of claims 1-22, wherein the level of anti-dsDNA antibodies in the subject decreases as compared to baseline.

24. The method of any of claims 1-23, wherein the rapcabtagene autoleucel is administered at a dose of about 0.5 x 106- 1.25 x 109viable CAR-expressing cells.

25. The method of claim 24, wherein the rapcabtagene autoleucel is administered at a dose of 2.5 x 106, 5 x 106, 12.5 x 106, 25 x 106, or 40 x 106viable CAR- expressing cells.

26. The method of claim 25, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 106viable CAR-expressing cells.

27. The method of any of claims 1-26, wherein the subject has been previously treated with, or is concurrently treated with, one or more of an antimalarial, aglucocorticoid, a calcineurin inhibitor, an immunomodulatory agent, a biological agent, rituximab, or a disease-modifying antirheumatic drug (DMARD).

28. The method of any of claims 1-27, wherein the subject is administered a second therapy chosen from an antimalarial agent or a stable immunosuppressive.

29. The method of claim 28, wherein the rapcabtagene autoleucel and the second therapy are present in the subject at the same time.

30. A method of treating a subject diagnosed with systemic lupus erythematosus (SLE) comprising administering to the subject a population of cells engineered to express a CD 19 CAR, said population comprising:(i) about the same percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(ii) a change within about 5% to about 10% of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(iii) an increased percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, for example, increased by at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the same population of cells prior to being engineered to express the CAR;(iv) about the same percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memoryT cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(v) a change within about 5% to about 10% of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(vi) a decreased percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, for example, decreased by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CD45RA- CCR7+ CD45RO+ T cells, in the same population of cells prior to being engineered to express the CAR;(vii) about the same percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR;(viii) a change within about 5% to about 10% of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR; or(ix) an increased percentage of stem memory T cells, for example,CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the same population of cells prior to being engineered to express the CAR.

31. The method of claim 30, wherein the SLE is severe refractory SLE(srSLE).

32. The method of claim 30 or 31, wherein the SLEDAI-2K score of the subject is reduced by at least 4 points, 5 points, 6 points, 7 points, 8 points, 9 points, 10 points, 15 points, 20 points or 25 points as compared to the baseline SLEDAI-2K score.

33. The method of claim 32, wherein the reduction in the SLEDAI-2K score is sustained for at least 6 months, 9 months, or 12 months.

34. The method of any of claims 30-33, wherein the total SLEDAI-2K score of the patient is reduced to 6 or below, 5 or below, 4 or below, 3 or below, 2 or below, 1 or below, or 0.

35. The method of any of claims 30-34, wherein the PhGA score of the subject is reduced by at least 0.3 points, 0.5 points, 0.75 points, 1 points, 1.25 points, 1.5 points, 1.75 points, 2 points, 2.25 points, 2.5 points, or 2.75 points as compared to the baseline PhGA score.

36. The method of any of claims 30-35, wherein the total PhGA score is reduced to 1.5 or lower, 1.25 o lowers, 1 or lower, 0.75 or lower, 0.5 or lower, or 0.3 or lower.

37. The method of any of claims 30-36, wherein the reduction on PhGA score is sustained for at least 6 months or more, 9 months or more, 12 months or more, 18 months or more, or 24 months or more.

38. The method of claim 37, wherein the sustained reduction is maintained without an increase in PhGA score to l.O or greater, 1.25 or greater, 1.5 or greater, 1.75 or greater, or 2 or greater.

39. The method of any of claims 30-38, wherein there is no increase in PhGA score of greater than 0.3 points, 0.25 points, 0.2 points, 0.15 points, 0.1 points, or 0.5 points from baseline following administration40. The method of any of claims 30-39, wherein the subject achieves a lupus low disease activity state (LLDAS) within 6 months, 9 months, or 12 months following administration.

41. The method of claim 40, wherein LLDAS is sustained for at least 3 months, 6 months, 12 months, or 18 months.

42. The method of any of claims 30-41, wherein the subject is in remission according to the DORIS definition within 3 months, 6 months, 9 months, or 12 months following administration.

43. The method of claim 42, wherein remission is sustained for at least 3 months, 6 months, 12 months, or 18 months.

44. The method of any of claims 30-43, wherein the subject has a reduction in BILAG score from A to C or better, A to D, B to C or better, or B to D within 6 months following administration.

45. The method of any of claims 30-44, wherein the reduction in BILAG score is sustained for at least 3 months, 6 months, 12 months, or 18 months.

46. The method of any of claims 30-45, wherein the subject is a responder according to the SRI-4 definition.

47. The method of claim 46, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

48. The method of any of claims 30-47, wherein the subject is a responder according to the BICLA definition.

49. The method of claim 48, wherein the response is sustained for at least 3 months, 6 months, 12 months, or 18 months.

50. The method of any of claims 30-49, wherein the subject has a complete renal response (CRR) following administration.

51. The method of claim 50, wherein the CRR is sustained for at least 3 months, 6 months, 9 months, 12 months, or 18 months.

52. The method of any of claims 30-51, wherein the level of anti-dsDNA antibodies in the subject decreases as compared to baseline.

53. The method of any of claims 30-52, wherein the population of cell is administered at a dose of about 0.5 x 106- 1.25 x 109viable CAR-expressing cells.

54. A method of reducing a SLEDAI-2K score in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject, wherein the SLEDAI-2K score is reduced by at least 4 points as compared to baseline.

55. The method of claim 54, wherein the total SLEDAI-2K score is reduced to 6 or below, 5 or below, 4 or below, 3 or below, 2 or below, 1 or below, or 0.

56. The method of claim 54 or 55, wherein the reduction in SLEDAI-2K score is sustained for at least 3 months, 6 months, 9 months, 12 months, or 18 months.

57. The method of any of claims 54-56, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 10A6 viable CAR-expressing cells.

58. A method of reducing a PhGA score in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject, wherein the PhGA score is reduced by at least 0.3 points as compared to baseline.

59. The method of claim 58, wherein the PhGA score is reduced to 1.5 or lower, 1.25 or lower, 1 or lower, 0.75 or lower, 0.5 or lower, or 0.3 or lower.

60. The method of claim 58 or 59, wherein the reduction in PhGA score is sustained for at least 9 months, 12 months, 18 months, or 24 months.

61. The method of any of claims 58-60, wherein the reduction PhGA is sustained without an increase in PhGA score to 1.0 or greater, 1.25 or greater, 1.5 or greater, 1.75 or greater, or 2.0 or greater.

62. The method of any of claims 58-61, wherein the PhGA score does not increase by more than 0.3 points, 0.25 points, 0.2 points, 0.15 points, 0.10 points, or 0.05 points as compared to baseline.

63. The method of any of claims 58-62, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 10A6 viable CAR-expressing cells.

64. A method of achieving a low lupus disease activity state (LLDAS) in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject, wherein the LLDAS is achieved within 3 months, 6 months, 9 months, or 12 months following administration.

65. The method of claim 64, wherein the LLDAS is sustained for at least 3 months, 6 months, 9 months, 12 months, or 18 months.

66. The method of claim 64 or 65, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 10A6 viable CAR-expressing cells.

67. A method of causing a subject with systemic lupus erythematosus (SLE) to be in remission according to the DORIS definition, the method comprising administering rapcabtagene autoleucel to the subject, wherein the subject is in remission according to the DORIS definition.

68. The method of claim 67, wherein the subject is in remission within 3 months, 6 months, 9 months, or 12 months following administration.

69. The method of claim 67 or 68, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 10A6 viable CAR-expressing cells.

70. A method of reducing a British Isles Lupus Activity Group (BILAG) score in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject, wherein the BILAG score of the subject is reduced from A or B to C or better.

71. The method of claim 70, wherein the reduction in score is within 6 months with no new BILAG A or B scores.

72. The method of claim 70 or 71, wherein the reduction is sustained for at least 6 months, 9 months, 12 months or 18 months without any new A or B scores.

73. The method of any of claims 70-72, wherein the rapcabtagene autoleucel is administered at a dose of 12.5 x 10A6 viable CAR-expressing cells.

74. A method of identifying a subject with systemic lupus erythematosus (SLE) as a responder according to the SRI-4 index, the method comprising administeringrapcabtag ene autoleucel to the subject at a dose of 12.5 x 10A6 viable CAR-expressing cells, wherein the subject is a responder according to the SRI-4 index.

75. A method of identifying a subject with systemic lupus erythematosus (SLE) as a responder according to the British Isles Lupus Assessment Group-based Composite Lupus Assessment (BILCA) index, the method comprising administering rapcabtag ene autoleucel to the subject at a dose of 12.5 x 10A6 viable CAR-expressing cells, wherein the subject is a responder according to the BICLA index.

76. A method of achieving a complete renal response in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject at a dose of 12.5 x 10A6 viable CAR-expressing cells, wherein the subject achieves a complete renal response.

77. A method of decreasing the level of anti-dsDNA antibodies in a subject with systemic lupus erythematosus (SLE), the method comprising administering rapcabtagene autoleucel to the subject at a dose of 12.5 x 10A6 viable CAR-expressing cells, wherein the level of anti-dsDNA antibodies in the subject is decreased as compared to baseline.

78. Rapcabtagene autoleucel for use in a method of treating a subject having systemic lupus erythematosus (SLE), said method comprising administering to the subject rapcabtagene autoleucel at a dose of 12.5 x 10A6 cells.

79. A pharmaceutical composition comprising the rapcabtagene autoleucel of claim 78 and a pharmaceutically acceptable carrier.

80. A method of treating a subject with systemic lupus erythematosus (SLE) comprising administering to the subject rapcabtagene autoleucel, wherein the rapcabtagene autoleucel is made by a method comprising:(i) contacting a population of T-cells derived from the subject with SLE with an agent that stimulates a CD3 / TCR complex;(ii) contacting the population of T-cells with a nucleic acid molecule encoding a CD 19 CAR, thereby providing a population T cells comprising the CD 19 CAR(iii) harvesting the population of T-cells for storage or administration, wherein(a) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i) and step (iii) is performed no later than 30 hours after the beginning of step (i);(b) step (ii) is performed together with step (i) or no later than 20 hours after the beginning of step (i) and step (iii) is performed no later than 30 hours after the beginning of step (ii); or,(c) the population of cells from step (iii) are not expanded as assessed by the number of living cells compared to the population of cells at the beginning of step (i).

81. The method of claim 80, wherein:(a) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is the same as or differs by no more than 5 or 10% from the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i);(b) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is increased by, for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i);(c) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells in the population of cells increases during the duration of step (ii), for example, increases by, for example, at least 30, 35, 40, 45, 50, 55, or 60%, between 18-24 hours after the beginning of step (ii); or(d) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) does not decrease, ordecreases by no more than 5 or 10%, as compared to the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ cells, in the population of cells at the beginning of step (i).

82. The method of any one of claims 80 or 81, wherein:(a) the population of cells from step (iii) shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 10, 20, 30, or 40% higher), compared with cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(b) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold higher) than the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(c) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 4, 6, 8, 10, or 12-fold higher) than the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(d) the population of cells from step (iii) shows a higher percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells (for example, at least 10, 20, 30, or 40% higher), compared with cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days;(e) the percentage of naive cells, for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 1.2, 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, or 3-fold higher) than the percentage of naive cells,for example, naive T cells, for example, CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in the population of cells from step (iii) is higher (for example, at least 4, 6, 8, 10, or 12-fold higher) than the percentage of CAR-expressing naive T cells, for example, CAR-expressing CD45RA+ CD45RO- CCR7+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

83. The method of any one of claims 80-82, wherein:(a) the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells from step (iii) is the same as or differs by no more than 5 or 10% from the percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells, in the population of cells at the beginning of step (i);(b) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells from step (iii) is reduced by at least 20, 25, 30, 35, 40, 45, or 50%, as compared to the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells at the beginning of step (i);(c) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ cells, decreases during the duration of step (ii), for example, decreases by, for example, at least 8, 10, 12, 14, 16, 18, or 20%, between 18-24 hours after the beginning of step (ii); or(d) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells from step (iii) does not increase, or increases by no more than 5 or 10%, as compared to the percentage of central memory cells, forexample, central memory T cells, for example, CCR7+CD45RO+ T cells, in the population of cells at the beginning of step (i).

84. The method of any one of claims 80-83, wherein:(a) the population of cells from step (iii) shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 10, 20, 30, or 40% lower), compared with cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(b) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 20, 30, 40, or 50% lower) than the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(c) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 10, 20, 30, or 40% lower) than the percentage of CAR-expressing central memory T cells, for example, CAR-expressing CCR7+CD45RO+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(d) the population of cells from step (iii) shows a lower percentage of central memory cells, for example, central memory T cells, for example, CD95+ central memory T cells (for example, at least 10, 20, 30, or 40% lower), compared with cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days;(e) the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 20, 30, 40, or 50% lower) than the percentage of central memory cells, for example, central memory T cells, for example, CCR7+CD45RO+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expandingthe population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing central memory T cells, for example, CAR- expressing CCR7+CD45RO+ T cells in the population of cells from step (iii) is lower (for example, at least 10, 20, 30, or 40% lower) than the percentage of CAR-expressing central memory T cells, for example, CAR-expressing CCR7+CD45RO+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

85. The method of any one of claims 80-84, wherein:(a) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is increased, as compared to the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells at the beginning of step (i);(b) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is increased, as compared to the percentage of CAR-expressing stem memory T cells, for example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells at the beginning of step (i);(c) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours after the beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i); or(d) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of CAR-expressing stem memory T cells, for example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method in which step (iii) is performed more than 26 hours afterthe beginning of step (i), for example, more than 5, 6, 7, 8, 9, 10, 11, or 12 days after the beginning of step (i);(e) the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptor 0+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of stem memory T cells, for example, CD45RA+CD95+IL-2 receptorP+CCR7+CD62L+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days; or(f) the percentage of CAR-expressing stem memory T cells, for example, CAR- expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in the population of cells from step (iii) is higher than the percentage of CAR-expressing stem memory T cells, for example, CAR-expressing CD45RA+CD95+IL-2 receptor P+CCR7+CD62L+ T cells, in cells made by an otherwise similar method which further comprises, after step (ii) and prior to step (iii), expanding the population of cells (for example, T cells) in vitro for more than 3 days, for example, for 5, 6, 7, 8 or 9 days.

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