Treatment of autoimmune disorders with NK cells and CD38 antibody

Abstract

Provided herein are, among other things, methods for treating a patient suffering from an autoimmune disease.

Description

TREATMENT OF AUTOIMMUNE DISORDERS WITH NK CELLS AND CD38 ANTIBODYCLAIM OF PRIORITY

[0001] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 664,290, filed June 26, 2024. The entire contents of the foregoing are incorporated herein by reference.BACKGROUND

[0002] High levels of autoreactive immune cells are associated with autoimmune indications. The incomplete or reduced depletion of pathogenic B cells with antibody alone (ex. Rituximab) in autoimmune patients (for example, lupus) can result in limited efficacy or relapse of disease.

[0003] The present invention addresses these and other deficiencies in the art.SUMMARY

[0004] Disclosed herein are methods for treating disorders (e.g., autoimmune disorders) with NK cells in combination with a CD38 targeting antibody, for example, by depleting CD38 expressing B cells (e g., antibody secreting cells including plasmablasts, plasma cells, and long-lived memory cells). While other antibodies (e.g., CD19 and CD20 targeting antibodies) can target bone marrow and peripheral B cell types earlier in maturation (e.g., pre-B cells, immature B cells, and mature B cells), CD38 antibodies can also target plasma cells and plasmablasts, which express no or low CD 19 and CD20.

[0005] NK cells are immune cells that can engage tumor cells through a complex array of receptors on their cell surface, as well as through antibody-dependent cellular cytotoxicity (ADCC). To initiate ADCC, NK cells engage with antibodies via the CD16 receptor on their surface. NK cells may have an advantage over other immune cells, such as the T cells used in CAR-T cell therapy and other cell therapies. In an exemplary advantage, NK cells can be used as allogeneic therapies, meaning that NK cells from one donor can be safely used in one or many patients without the requirement for HLA matching, gene editing, or other genetic manipulations. Allogeneic NK cells with anti-tumor activity can be administered safely to patients without many of the risks associated with T cell therapies.such as severe cytokine release syndrome (CRS), and neurological toxicities or graft versus host disease (GvHD).

[0006] Allogeneic NK cells may provide an important treatment option for patients with autoimmune disorders, resulting in a deeper and more prolonged depletion of pathogenic B cells than, e.g., antibody alone, resulting in better efficacy and outcomes.

[0007] Additionally, cords with preferred characteristics for enhanced clinical activity (e.g.. high-affimty CD16 and Killer cell Immunoglobulin-like Receptor (KIR) B-haplotype) can be selected by utilizing a diverse umbilical cord blood bank as a source forNK cells.

[0008] The administration of the allogenic NK cells, as described herein, can enhance patients’ ADCC responses, e.g., when undergoing monoclonal antibody therapy.

[0009] Thus, provided herein are, among other things, methods for depleting B-cells in a patient, e.g., CD38+ B cells (e.g., plasma cells and / or plasmablasts), e.g., for treating a patient suffering from an autoimmune disorder.

[0010] Provided herein are methods for treating a patient suffering from an autoimmune disorder, the method comprising administering a population of natural killer cells (NK cells) and a CD38 targeting antibody, wherein the NK cells are allogenic to the patient. In some embodiments, an immune cell is implicated in the autoimmune disorder.

[0011] In some embodiments, the immune cell is a B cell, e.g., a CD38+ B cell (e.g., plasma cell and / or plasmablast). In some embodiments, the antibody is a B-cell depleting antibody. In some embodiments, the method further comprising administering antibodie(s) targeted to human CD19 and / or human CD20. In some embodiments, the NK cells are KIR- B haplotype and homozy gous for a CD 16 158V polymorphism.

[0012] In some embodiments, the autoimmune disease is selected from, Acromegaly, Acquired aplastic anemia, Acquired hemophilia, Primary Agammaglobulinemia. Alopecia areata, Ankylosing spondylitis (AS), Anti-NMDA receptor encephalitis, Antiphospholipid syndrome (APS) | catastrophic antiphospholipid syndrome (CAPS) / Asherson's syndrome, Arteriosclerosis, Autoimmune Addison’s disease (AAD), Autoimmune autonomic ganglionopathy (AAG) / autoimmune dysautonomia | autoimmune gastrointestinal dysmotility (AGID). Autoimmune encephalitis | acute disseminated encephalomyelitis (ADEM), Autoimmune gastritis, Autoimmune hemolytic anemia (AIHA), Autoimmune hepatitis (AIH), Autoimmune hyperlipidemia, Autoimmune hypophysitis, Autoimmune inner ear disease (AIED), Autoimmune lymphoproliferative syndrome (ALPS), Autoimmune myelofibrosis, Autoimmune myocarditis, Autoimmune oophoritis, Autoimmune pancreatitis (AIP), Autoimmune polyglandular syndromes, types I, II, & III (APS type 1, APS type 2,APS type 3, APECED), Autoimmune progesterone dermatitis, Autoimmune retinopathy (AIR), Autoimmune sudden sensorineural hearing loss (SNHL), Balo disease, Behcet’s disease. Birdshot chorioretinopathy / birdshot uveitis, Bullous pemphigoid, Castleman disease. Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy (CIDP), Chronic urticaria (CU), Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA), Cogan’s syndrome, Cold agglutinin disease, CREST syndrome | limited cutaneous systemic sclerosis, Crohn’s disease (CD). Cronkhite-Canada syndrome (CSS), Cryptogenic organizing pneumonia (COP), Dermatitis herpetiformis, Dermatomyositis, Type 1 Diabetes, Discoid lupus, Dressier’s syndrome / postmyocardial infarction / postpericardiotomy syndrome, Eczema / Atopic Dermatitis, Endometriosis, Eosinophilic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Essential mixed cryoglobulinemia, Evans syndrome, Fibrosing alveolitis / Idiopathic pulmonary fibrosis (IPF), Giant cell arteritis / temporal arteritis / Florton’s disease, Giant Cell Myocarditis, Glomerulonephritis, Goodpasture's syndrome / anti-GBM / anti-TBM disease. Granulomatosis with polyangiitis (GPA) / Wegener’s granulomatosis, Graves disease / thyroid eye disease. Guillain-Bane syndrome (GBS), Hashimoto’s thyroiditis / chronic lymphocytic thyroiditis / autoimmune thyroiditis, Henoch-Schonlein purpura / IgA vasculitis, Hidradenitis suppurativa, Hurst’s disease / acute hemorrhagic leukoencephalitis (AHLE), Hypogammaglobulinemia. IgA nephropathy / Berger's disease, Immune-mediated necrotizing myopathy (IMNM), Immune thrombocytopenia (ITP) / autoimmune thrombocytopenic purpura / autoimmune thrombocytopenia, Inclusion body myositis, IgG4-related sclerosing disease (ISD), Interstitial cystitis, Juvenile idiopathic arthritis / Adult-onset Still's disease, Juvenile polymyositis | Juvenile dermatomyositis | juvenile myositis, Kawasaki disease, Lambert-Eaton myasthenic syndrome (LEMS), Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Ligneous conjunctivitis. Linear IgA disease (LAD) | linear IgA bullous dermatosis (LABD), Lupus nephritis, Lyme disease / chronic Lyme disease / post-treatment Lyme disease syndrome (PTLDS), Ly mphocy tic colitis / microscopic colitis, Lymphocytic hypophystitis / autoimmune hypophystitis, Meniere’s disease, Microscopic polyangiitis (MPA)ZANCA-associated vasculitis, Mixed connective tissue disease (MCTD), Mooren’s ulcer, Mucha-Habermann disease, Multifocal motor neuropathy, Multiple sclerosis (MS), Myalgic encephalomyelitis (ME) / Chronic fatigue syndrome (CFS), Myasthenia gravis (MG), Narcolepsy, Neuromyelitis Optica / Devic's disease, Ocular cicatricial pemphigoid, Opsoclonus-myoclonus syndrome (OMS), Palindromic rheumatism, Paraneoplastic cerebellar degeneration. Paraneoplastic pemphigus, Parry-Romberg syndrome (PRS) / Hemifacialatrophy (HFA) / Progressive facial hemiatrophy, Paroxysmal nocturnal hemoglobinuria (PNH). Peripheral uveitis / pars planitis, PANS / PANDAS, Parsonage-Turner syndrome. Pemphigus gestationis / herpes gestationis. Pemphigus foliaceus, Pemphigus vulgaris. Pernicious anemia, POEMS syndrome, Polyarteritis nodosa, Polymyalgia rheumatica, Polymyositis, Postural orthostatic tachycardia syndrome (POTS), Primary biliary cirrhosis (PBC) / primary biliary' cholangitis, Primary sclerosing cholangitis (PSC), Psoriasis, Palmoplantar Pustulosis. Psoriatic arthritis, Pulmonary fibrosis, idiopathic (IPF). Pure red cell aplasia (PRC A), Pyoderma gangrenosum, Rasmussen's encephalitis, Raynaud’s syndrome / phenomenon, Reactive arthritis / Reiter’s syndrome, Reflex sympathetic dystrophy syndrome (RSD) / Complex regional pain syndrome (CRPS), Relapsing polychondritis, Restless leg syndrome (RLS) / Willis-Ekbom disease. Rheumatic fever, Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome / autoimmune polyendocrine syndrome type II, Scleritis, Scleroderma , Sclerosing Mesenteritis / Mesenteric Panniculitis, Serpiginous choroidopathy, Sjogren’s syndrome, Stiff person syndrome (SPS), Small fiber sensory neuropathy, Systemic lupus erythematosus (SLE), Subacute bacterial endocarditis (SBE), Subacute cutaneous lupus, Susac syndrome, Sydenham's chorea. Sympathetic ophthalmia, Takayasu’s arteritis (vasculitis). Testicular autoimmunity (vasculitis, orchitis), Tolosa-Hunt syndrome, Transverse myelitis (TM), Tubulointerstitial nephritis uveitis syndrome (TINU), Ulcerative colitis (UC), Undifferentiated connective tissue disease (UCTD), Uveitis | anterior / intermediate / posterior, Vasculitis, VEXAS Syndrome, Vitiligo, Vogt-Koyanagi- Harada syndrome (VKH), and combinations thereof.

[0013] In some embodiments, the autoimmune disorder is Systemic lupus erythematosus (SLE). In some embodiments, the patient has lupus nephritis.

[0014] In some embodiments, the CD38 targeting antibody is daratumumab.

[0015] In some embodiments, the patient is subjected to lymphodepleting chemotherapy prior to treatment. In some embodiments, the lymphodepleting chemotherapy is non-myeloablative chemotherapy.

[0016] In some embodiments, the NK cells are not genetically modified.

[0017] In some embodiments, at least 70% of the NK cells are CD56+ and CD16+.In some embodiments, at least 85% of the NK cells are CD56+ and CD3-. In some embodiments, 1 % or less of the NK cells are CD3+, 1% or less of the NK cells are CD 19+ and 1% or less of the NK cells are CD14+.

[0018] In some embodiments, each administration of NK cells is administration of 1 x 10A9 to 5 x 10A9 NK cells. In some embodiments, each administration of NK cells is administration of 1 x 10A9 to 5 x 10A9 NK cells.

[0019] In some embodiments, the patient receives a dose of the CD38 targeted antibody before the first dose of NK cells.

[0020] In some embodiments, the expanded natural killer cells are expanded umbilical cord blood natural killer cells.

[0021] In some embodiments, the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% CD 16+ cells. In some embodiments, the population of expanded natural killer cells comprises at least 60%. e.g., at least 70%, at least 80%, at least 90% at least 95%. at least 99%, or 100% NKG2D+ cells. In some embodiments, the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp46+ cells. In some embodiments, the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp30+ cells. In some embodiments, the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% DNAM-1+ cells. In some embodiments, the population of expanded natural killer cells comprises at least 60%. e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%. or 100% NKp44+ cells. In some embodiments, the population of expanded natural killer cells comprises less than 20%, e.g., 10% or less, 5% or less, 1% or less, 0.5% or less, or 0% CD3+ cells. In some embodiments, the population of expanded natural killer cells comprises less than 20% or less, e.g., 10% or less. 5% or less. 1% or less, 0.5% or less, or 0% CD14+ cells. In some embodiments, the population of expanded natural killer cells comprises less than 20% or less, e.g., 10% or less, 5% or less, 1% or less, 0.5% or less, or 0% CD19+ cells. In some embodiments, the population of expanded natural killer cells comprises less than 20% or less, e.g., 10% or less. 5% or less. 1% or less, 0.5% or less, or 0% CD38+ cells.

[0022] In some embodiments, the natural killer cells do not comprise a CD 16 transgene. In some embodiments, the natural killer cells do not express an exogenous CD 16 protein. In some embodiments, the expanded natural killer cells are not genetically engineered. In some embodiments, the expanded natural killer cells are derived from the same umbilical cord blood donor.

[0023] In some embodiments, the population ofNK cells comprises at least 100 million expanded natural killer cells, e.g., 200 million, 250 million. 300 million, 400 million, 500 million, 600 million, 700 million, 750 million, 800 million, 900 million, 1 billion, 2 billion, 3 billion, 4 billion, 5 billion, 6 billion, 7 billion, 8 billion, 9 billion, 10 billion, 15 billion, 20 billion, 25 billion, 50 billion, 75 billion, 80 billion, 9- billion, 100 billion, 200 billion, 250 billion, 300 billion, 400 billion, 500 billion. 600 billion, 700 billion, 800 billion, 900 billion, 1 trillion, 2 trillion, 3 trillion, 4 trillion, 5 trillion. 6 trillion. 7 trillion. 8 trillion, 9 trillion, or 10 trillion expanded natural killer cells.

[0024] In some embodiments, the population of NK cells is produced by a method comprising: (a) obtaining seed cells comprising natural killer cells from umbilical cord blood; (b) depleting the seed cells of CD3+ cells; (c) expanding the natural killer cells by culturing the depleted seed cells with a first plurality of Hut78 cells engineered to express a membrane bound IL-21, a mutated TNFa, and a 4-1BBL gene to produce expanded natural killer cells, thereby producing the population of expanded natural killer cells.

[0025] In some embodiments, the population of NK cells is produced by a method comprising: (a) obtaining seed cells comprising natural killer cells from umbilical cord blood; (b) depleting the seed cells of CD3+ cells; (c) expanding the natural killer cells by culturing the depleted seed cells with a first plurality' of Hut78 cells engineered to express a membrane bound IL-21, a mutated TNFa, and a 4-1 BBL gene to produce a master cell bank population of expanded natural killer cells; and (d) expanding the master cell bank population of expanded natural killer cells by culturing with a second plurality of Hut78 cells engineered to express a membrane bound IL-21, a mutated TNFa, and a 4-1 BBL gene to produce expanded natural killer cells; thereby producing the population of expanded natural killer cells.

[0026] In some embodiments, the population of NK cells is produced by a method further comprising, after step (c), (i) freezing the master cell bank population of expanded natural killer cells in a plurality of containers; and (ii) thawing a container comprising an aliquot of the master cell bank population of expanded natural killer cells, yvherein expanding the master cell bank population of expanded natural killer cells in step (d) comprises expanding the aliquot of the master cell bank population of expanded natural killer cells.

[0027] In some embodiments, the umbilical cord blood is from a donor with the KIR- B haplotype and homozy gous for the CD16 158V polymorphism.

[0028] In some embodiments, the population of NK cells is produced by a method comprising expanding the natural killer cells from umbilical cord blood at least 10.000 fold,e.g., 15,000 fold, 20,000 fold, 25,000 fold, 30,000 fold, 35,000 fold, 40,000 fold, 45,000 fold, 50,000 fold, 55,000 fold. 60.000 fold, 65,000 fold, or 70,000 fold.

[0029] In some embodiments, the population of expanded natural killer cells is not enriched or sorted after expansion.

[0030] In some embodiments, the percentage of NK cells expressing CD 16 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood. In some embodiments, the percentage of NK cells expressing NKG2D in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood. In some embodiments, the percentage of NK cells expressing NKp30 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood. In some embodiments, the percentage of NK cells expressing NKp44 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood. In some embodiments, the percentage of NK cells expressing NKp46 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood. In some embodiments, the percentage of NK cells expressing DNAM-1 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0031] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0032] Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims.INCORPORATION BY REFERENCE

[0033] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0034] The novel features of the invention are set forth with particularity in the appended claims. The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0035] FIG. 1 is a representative flow plot showing CD38 expression on SLE donor B cells.

[0036] FIG. 2 shows % caspase positive B cells after treatment with and without daratumumab. SLE patient PBMCs were isolated from peripheral blood and combined with thawed AB-101, with or without anti-CD38 daratumumab for 4 hours. The percentage of caspase positive B-cells was determined by flow cytometry. Data are expressed as the mean ±SD of duplicate wells from n=3 donors with the indicated antibody concentration and E:T ratio (AB-101 :PBMC at either 0.2: 1 or 1 : 1). PBMC alone is shown as 0.DETAILED DESCRIPTION

[0037] Provided herein are, amongst other things, Natural Killer (NK) cells, e.g., expanded and stimulated NK cells, methods for producing the NK cells, pharmaceutical compositions comprising the NK cells, and methods of treating patients suffering, e.g.. from an autoimmune disorder, with the NK cells.I. EXPANSION AND STIMULATION OF NATURAL KILLER CELLS

[0038] In some cases, the NK cells are expanded and stimulated, e.g., as described in WO2022216813, which is hereby incorporated by reference in its entirety'.

[0039] In some cases, e.g., after having been ex vivo expanded and stimulated, e.g., as described herein, the expanded and stimulated NK cell populations not only have a number / density (e.g., as described above) that could not occur naturally in the human body, but they also differ in their phenotypic characteristics, (e.g., gene expression and / or surface protein expression) with the starting source material or other naturally occurring populations of NK cells.

[0040] In some cases, the starting NK cell source is a sample derived from a single individual, e.g., a single cord blood unit that has not been ex vivo expanded. Therefore, in some cases, the expanded and stimulated NK cells share a common lineage, i.e., they all result from expansion of the starting NK cell source, and, therefore, share a genotype via clonal expansion of a population of cells that are, themselves, from a single organism. Yet, they could not occur naturally at the density achieved with ex vivo expansion and also differ in phenotypic characteristics from the starting NK cell source.

[0041] In some cases, the population of expanded and stimulated NK cells comprises at least 100 million expanded natural killer cells, e.g., 200 million, 250 million, 300 million, 400 million, 500 million, 600 million. 700 million, 750 million. 800 million. 900 million, 1 billion, 2 billion, 3 billion, 4 billion, 5 billion, 6 billion, 7 billion, 8 billion, 9 billion, 10 billion, 15 billion, 20 billion, 25 billion, 50 billion, 75 billion, 80 billion, 9- billion, 100 billion, 200 billion, 250 billion, 300 billion, 400 billion. 500 billion, 600 billion, 700 billion, 800 billion, 900 billion, 1 trillion, 2 trillion, 3 trillion. 4 trillion. 5 trillion. 6 trillion, 7 trillion, 8 trillion, 9 trillion, or 10 trillion expanded natural killer cells.

[0042] In some embodiments, the expanded and stimulated NK cells comprise at least 80%, e.g., at least 90%, at least 95%, at least 99%, or 100% CD56+CD3- cells.

[0043] In some embodiments, the expanded and stimulated NK cells do not comprise a CD 16 transgene.

[0044] In some embodiments, the expanded and stimulated NK cells do not express an exogenous CD 16 protein.

[0045] The expanded and stimulated NK cells can be characterized, for example, by surface expression, e.g., of one or more of CD16, CD56, CD3, CD38, CD14. CD19, NKG2D, NKp46, NKp30, DNAM-1, and NKp44.

[0046] The surface protein expression levels stated herein, in some cases are achieved without positive selection on the particular surface protein referenced. For example, in some cases, the NK cell source, e.g.. a single cord unit, comprises both the KIR B allele of the KIR receptor family and the 158 V / V variant of CD 16 and is + enriched and CD3(+) depleted.e.g., by gating on CD56+CD3- expression, but no other surface protein expression selection is carried out during expansion and stimulation.

[0047] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKG2D+ cells.

[0048] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp46+ cells.

[0049] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp30+ cells.

[0050] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% DNAM-1+ cells.

[0051] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp44+ cells.

[0052] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% CD94+ (KLRD1) cells.

[0053] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprises less than or equal to 20%, e.g.. less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD3+ cells.

[0054] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprises less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD14+ cells.

[0055] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprises less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD19+ cells.

[0056] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprises less than or equal to 20%, e.g.. less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CXCR+ cells.

[0057] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e g., as described above, comprises less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD122+ (IL2RB) cells.

[0058] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e g., as described above, comprises 90% or more, e.g., 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% CD3-CD14- CD19-CD16+CD56- cells.

[0059] As described herein, the inventors have demonstrated that, surprisingly, the NK cells expanded and stimulated by the methods described herein express CD 16 at high levels throughout the expansion and stimulation process, resulting in a cell population with high CD 16 expression. The high expression of CD 16 obviates the need for engineering the expanded cells to express CD1 , which is important for initiating ADCC, and, therefore, a surprising and unexpected benefit of the expansion and stimulation methods described herein. Thus, in some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise 50% or more, e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% CD16+ NK cells.

[0060] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e g., as described above, comprises both the KIR B allele of the KIR receptor family and the 158 V / V variant of CD16 and comprise 50% or more, e.g.. 55%. 60%. 65%. 70%. 75%. 80%. 85%. 90%. or 95% CD16+ NK cells.

[0061] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing CD 16 is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0062] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing NKG2D is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0063] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing NKp30 is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0064] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing DNAM-1 is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0065] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing NKp44 is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0066] In some embodiments, the percentage of expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, expressing NKp46 is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

[0067] As described herein, the inventors have also demonstrated that, surprisingly, the NK cells expanded and stimulated by the methods described herein express CD38 at low levels. CD38 is an effective target for certain cancer therapies (e.g., multiple myeloma and acute myeloid leukemia). See. e.g., Jiao et al., CD38: Targeted Therapy in Multiple Myeloma and Therapeutic Potential for Solid Cancers,” Expert Opinion on Investigational Drugs 29(11): 1295-1308 (2020).

[0068] Thus, in some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprise less than or equal to 80% CD38+ cells, e.g., less than or equal to 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, or 20% CD38+ cells.

[0069] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e g., as described above, comprises both the KIR B allele of the KIR receptor family and the 158 V / V variant of CD16 andcomprise less than or equal to 80% CD38+ cells, e.g., less than or equal to 75%, 70%, 65%, 60%. 55%. 50%. 45%. 40%. 35%. 30%. 25%. or 20% CD38+ cells.

[0070] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e.g., as described above, comprises both the KIR B allele of the KIR receptor family and the 158 V / V variant of CD 16 and comprise less than or equal to 80% CD38+ cells, e.g., less than or equal to 75%, 70%, 65%, 60%. 55%. 50%. 45%. 40%. 35%. 30%. 25%. or 20% CD38+ cells, and 50% or more, e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% CD16+ NK cells.

[0071] In some embodiments, the expanded and stimulated NK cells, e.g., from expansion and stimulation of a single cord blood unit, e g., as described above, comprises both the KIR B allele of the KIR receptor family and the 158 V / V variant of CD16 and comprise: i) 50% or more, e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% CD16+ NK cells; and / or ii) less than or equal to 80% CD38+ cells, e.g., less than or equal to 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, or 20% CD38+ cells; and / or iii) at least 60%, e.g., at least 70%. at least 80%. at least 90% at least 95%, at least 99%, or 100% NKG2D+ cells; and / or iv) at least 60%, e.g., at least 70%, at least 80%. at least 90% at least 95%, at least 99%, or 100% NKp46+ cells; and / or v) at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp30+ cells; and / or vi) at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% DNAM-1+ cells; and / or vii) at least 60%, e.g., at least 70%, at least 80%. at least 90% at least 95%, at least 99%, or 100% NKp44+ cells; and / or viii) at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% CD94+ (KLRD1) cells; and / or ix) less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD3+ cells; and / or x) less than or equal to 20%, e.g.. less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD14+ cells; and / or xi) less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CD19+ cells; and / or xii) less than or equal to 20%, e.g., less than or equal to 10%, less than or equal to 5%, less than or equal to 1% or 0% CXCR+ cells; and / or xiii) less than or equal to 20%, e.g., less than or equal to 10%. less than or equal to 5%. less than or equal to 1% or 0% CD122+ (IL2RB) cells.

[0072] In some embodiments, the NK cell is engineered to alter, e.g., reduce, expression of one or more inhibitor receptor genes.

[0073] In some embodiments, the inhibitory receptor gene is a HLA-specific inhibitor}’ receptor. In some embodiments, the inhibitory receptor gene is a non-HLA- specific inhibitory receptor.

[0074] In some embodiments, the inhibitor receptor gene is selected from the group consisting of KIR, CD94 / NKG2A, LILRB1, PD-1, Irp60, Siglec-7, LAIR-1, and combinations thereof.

[0075] Also provided herein are pharmaceutical compositions comprising the natural killer cells described herein and dosage units of the pharmaceutical compositions described herein.

[0076] In some cases, the dosage unit comprises between 100 million and 1.5 billion cells, e.g., 100 million. 200 million, 300 million, 400 million. 500 million, 600 million, 700 million, 800 million, 900 million, 1 billion, 1.1 billion, 1.2 billion, 1.3 billion, 1.4 billion, or 1.5 billion.

[0077] In some cases, the dosage unit comprises between 100 million and 10 billion cells, e.g., 100 million. 200 million, 300 million, 400 million, 500 million, 600 million, 700 million, 800 million. 900 million, 1 billion, 1.5 billion, 2 billion, 2.5 billion, 3 billion, 3.5 billion, 4 billion, 4.5 billion, 5 billion, 5.5 billion, 6 billion, 6.5 billion, 7 billion, 7.5 billion, 8 billion, 8.5 billion, 9 billion, 9.5 billion, 10 billion cells, or thereabout.

[0078] Pharmaceutical compositions ty pically include a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0079] In some embodiments, the pharmaceutical composition comprises: a) natural killer cell(s) described herein; and b) a cryopreservation composition. Suitable cry opreservation compositions are described herein.

[0080] In some embodiments, the composition is frozen. In some embodiments, the composition has been frozen for at least three months, e.g., at least six months, at least nine months, at least 12 months, at least 15 months, at least 18 months, at least 24 months, at least 36 months, or at least 48 months.

[0081] In some embodiments, at least 60%, e.g., at least 70%, at least 80%, at least90% at least 95%, at least 99%, or 100% of the natural killer cells are viable after being thawed.

[0082] In some embodiments, the pharmaceutical composition comprises: a) a cryopreservation composition described herein; and b) therapeutic cell(s), e.g., the engineered NK cells described herein.

[0083] In some embodiments, the pharmaceutical composition further comprises: c) a buffer solution. Suitable buffer solutions are described herein, e.g., as for cry opreservation compositions.

[0084] In some embodiments, the pharmaceutical composition comprises from or from about 1x107to or to about 1x109cells / mL. In some embodiments, the pharmaceutical composition comprises IxlO8cells / mL. In some embodiments, the pharmaceutical composition comprises about IxlO8cells / mL.

[0085] In some embodiments, the pharmaceutical composition comprises from or from about IxlO8to or to about IxlO10cells / mL.

[0086] In some embodiments, the pharmaceutical composition further comprises an antibody or antigen binding fragment thereof, e.g., an antibody described herein.

[0087] Pharmaceutical compositions are ty pically formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration.

[0088] Methods of formulating suitable pharmaceutical compositions are known in the art, see. e.g., Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0089] Pharmaceutical compositions suitable for injectable use can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenousadministration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syrmgability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens. chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0090] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by fdtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile- filtered solution thereof.

[0091] Examples of suitable pharmaceutical compositions are described, for example, in WO2017 / 135631 and W02022 / 0133061, each of which is hereby incorporated by reference in its entirety.II. ANTIBODIES

[0092] The methods described herein comprise administering an antibody, e.g., an antibody that targets an immune cell, e.g., an immune cell implicated in an autoimmune reaction, e.g., a B cell, e.g., a plasmablast or plasma cell. In some cases, the antibody is a B- cell depleting antibody, e.g., a B-cell depleting monoclonal antibody (mAb). In some cases,the antibody is a CD38 targeted antibody. In some cases, the methods described herein comprise administering multiple antibodies, e.g., multiple antibodies that target immune cell(s), e.g., immune cell(s) implicated in an autoimmune reaction, e.g., a B cell. In some cases, one or more of the antibodies is a B-cell depleting antibody, e.g., a B-cell depleting monoclonal antibody (mAb). In some cases, the antibodies are selected from CD38, CD20 and / or CD19 targeted antibodies, e.g., as described herein.

[0093] The term "antibody ' refers to an immunoglobulin molecule or immunologically active portion thereof, i.e., an antigen-binding portion.

[0094] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible naturally-occurring mutations that may be present in minor amounts. An antibody can be monoclonal. An antibody can be a human or humanized antibody. The term “monoclonal antibody” encompasses intact and full-length monoclonal antibodies as well as antibody fragments (e.g., Fab, Fab’, F(ab')2, Fv), single chain antibodies (e.g., scFv), fusion proteins comprising an antibody fragment, and any other modified immunoglobulin molecule comprising at least one antigen-binding site. Furthermore, "monoclonal antibody" refers to such antibodies made by any number of techniques, including but not limited to, hybridoma production, phage library’ display, recombinant expression, and transgenic animals.

[0095] The term "chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chain is derived from a first source or species, while the remainder of the heavy and / or light chain is derived from a different source or species.

[0096] The term "humanized antibody" as used herein refers to an antibody that comprises a human heavy chain variable region and a light chain variable region wherein the native CDR residues are replaced by residues from corresponding CDRs from a nonhuman antibody (e.g., mouse, rat, rabbit, or nonhuman primate), wherein the nonhuman antibody has the desired specificity7, affinity, and / or activity. In some embodiments, one or more framework region residues of the human heavy chain or light chain variable regions are replaced by corresponding residues from nonhuman antibody. Furthermore, humanized antibodies can comprise residues that are not found in the human antibody or in the nonhuman antibody. In some embodiments, these modifications are made to further refine and / or optimize antibody characteristics. In some embodiments, the humanized antibody comprises at least a portion of an immunoglobulin constant region (e.g., CHI, CH2, CH3, Fc), typically that of a human immunoglobulin.

[0097] The term "human antibody" as used herein refers to an antibody that possesses an amino acid sequence that corresponds to an antibody produced by a human and / or an antibody that has been made using any of the techniques that are known to those of skill in the art for making human antibodies. These techniques include, but not limited to, phage display libraries, yeast display libraries, transgenic animals, recombinant protein production, and B-cell hybridoma technology.

[0098] "Antibody fragments7’ can include a portion of an intact antibody, preferably the antigen binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; singlechain antibody molecules; and multispeciftc antibodies formed from antibody fragments.

[0099] The terms "epitope" and "antigenic determinant" are used interchangeably herein and refer to that portion of an antigen or target capable of being recognized and bound by a particular antibody. When the antigen or target is a polypeptide, epitopes can be formed both from contiguous amino acids and noncontiguous amino acids juxtaposed by tertiary- folding of the protein. Epitopes formed from contiguous amino acids (also referred to as linear epitopes) are typically retained upon protein denaturing, whereas epitopes formed by tertiary folding (also referred to as conformational epitopes) are typically lost upon protein denaturing. An epitope ty pically includes at least 3, and more usually, at least 5, 6, 7, or 8-10 amino acids in a unique spatial conformation. Epitopes can be predicted using any one of a large number of software bioinformatic tools available on the internet. X-ray crystallography may be used to characterize an epitope on a target protein by analyzing the amino acid residue interactions of an antigen / antibody complex.

[0100] “Fv” includes the minimum antibody fragment which contains a complete antigen- recognition and binding site. This region consists of a dimer of one heavy- and one light- chain variable domain in tight, non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity- to the antibody. However, even a single variable domain (or half of an Fv comprising only- three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity- than the entire binding site. The Fab fragment also contains the constant domain of the light chain and the first constant domain (CHI) of the heavy- chain. Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) ofthe constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0101] Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD. IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgBl, IgG2, IgG3. IgG4, IgA. and IgA2. "‘Single-chain Fv” or “sFv” antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. In some cases, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the sFv to form the desired structure for antigen binding.

[0102] In various embodiments, the antibody or antigen binding fragment thereof comprises a human or humanized antibody. Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non- human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence. Methods for humanizing non-human antibodies are well known in the art.

[0103] An antibody that “binds to,” “specifically binds to,” or is “specific for” a particular polypeptide or an epitope on a particular polypeptide is one that binds to that particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or polypeptide epitope. The term “specifically binds” as used herein refers to a binding agent (e.g., antibody) that interacts more frequently, more rapidly, with greater duration, with greater affinity, or with some combination of the above to a particular antigen, epitope, protein, or target molecule than with alternative substances. A bindingagent (e.g. antibody) that specifically binds an antigen can be identified, for example, by immunoassays, ELISAs. Surface Plasmon Resonance (SPR) assays (e.g.. Biacore). or other techniques known to those of skill in the art. A binding agent that specifically binds an antigen binds the target antigen with a higher affinity than its affinity for a different antigen. The different antigen can be a related antigen. In some embodiments, a binding agent that specifically binds an antigen binds the target antigen with an affinity that is at least 20 times greater than its affinity for a different antigen, e.g., at least 30 times greater, at least 40 times greater, at least 50 times greater, at least 60 times greater, at least 70 times greater, at least 80 times greater, at least 90 times greater, or at least 100 times greater, than its affinity for a different antigen. In some embodiments, a binding agent that specifically binds a particular antigen binds a different antigen at such a low affinity that binding cannot be detected using an assay described herein or otherwise known in the art. In some embodiments, affinity is measured using SPR technology, e.g., in a Biacore system or other system known to those of skill in the art.

[0104] In some embodiments, the antibody or antigen-binding fragment thereof is an antibody, e.g., a full length antibody comprising an Fc domain including at least one heavy chain. In some embodiments, the antibody is a recombinant antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In some embodiments, the antibody is an IgGl antibody. In some embodiments, the antibody is an IgG2 antibody. In some embodiments, the antibody is an IgG3 antibody . In some embodiments, the antibody is an IgG4 antibody.

[0105] In some embodiments, the antibody is an antibody fragment comprising an antigen-binding site. In some embodiments, the antibody is a scFv. In some embodiments, the antibody is a disulfide-linked scFv. In some embodiments, the antibody is a bispecific antibody or a multispecific antibody . In some embodiments, the antibody is a monovalent antibody. In some embodiments, the antibody is a monospecific antibody. In some embodiments, the antibody is a bivalent antibody. In some embodiments, the antibody is isolated. In some embodiments, the antibody is substantially pure. In some embodiments, a binding agent is a polyclonal antibody. Polyclonal antibodies can be prepared by any method known to those of skill in the art.

[0106] In some embodiments, a binding agent is a monoclonal antibody. Monoclonal antibodies can be prepared by any method known to those of skill in the art. In someembodiments, a binding agent is a humanized antibody. Various methods for generating humanized antibodies are known in the art. In some embodiments, a binding agent is a human antibody. Human antibodies can be prepared using various techniques known in the art.

[0107] In some embodiments, a binding agent is a scFv antibody, a Fv, a Fab, a F(ab')2, a F(ab'), or a bispecific antibody.

[0108] In some embodiments, a bispecific antibody has decreased toxicity and / or side effects. In some embodiments, a bispecific antibody has decreased toxicity and / or side effects as compared to a mixture of the two individual antibodies or the antibodies as single agents. In some embodiments, a bispecific antibody has an increased therapeutic index. In some embodiments, a bispecific antibody has an increased therapeutic index as compared to a mixture of the two individual antibodies or the antibodies as single agents. Several techniques for making bispecific antibodies are know n by those skilled in the art. In some embodiments, the bispecific antibodies comprise heavy chain constant regions with modifications in the amino acids that are part of the interface between the two heavy chains. These modifications are made to enhance heterodimer formation and generally reduce or eliminate homodimer formation. In some embodiments, the bispecific antibodies are generated using a knobs-into-holes (KIH) strategy7. In some embodiments, the bispecific antibodies comprise variant hinge regions incapable of forming disulfide linkages betw een identical heavy chains (e.g., reduce homodimer formation). In some embodiments, the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered electrostatic interactions. In some embodiments, the bispecific antibodies comprise heavy chains with changes in amino acids that result in altered hydrophobic / hydrophilic interactions. Bispecific antibodies can be intact antibodies or antibody fragments comprising antigen-binding sites.

[0109] Binding agents with more than two valencies are also contemplated. In some embodiments, trispecific or tetraspecific antibodies are generated.

[0110] In some cases, the antibody or antigen-binding fragment thereof is an IgG, IgA, or IgE antibody or antigen-binding fragment thereof. In some cases, the antibody or antigen-binding fragment thereof is an IgG antibody or antigen-binding fragment thereof. In some cases, the antibody or antigen-binding fragment thereof is an IgGl, IgG3, or IgG4 antibody or antigen-binding fragment thereof. In some cases, the antibody or antigen-binding fragment thereof is an IgGl antibody or antigen-binding fragment thereof.

[0111] In some cases, the antibody or antigen binding fragment thereof is an antibody or combination of antibodies selected from Table 1. In some cases, the antibody or antigen binding fragment thereof is an IgG antibody selected from Table 1. In some cases, the antibody or antigen binding fragment thereof is an IgGl antibody selected from Table 1. In some cases, patient suffers from a disorder that the antibody or antigen-binding fragment thereof is approved as a therapeutic (e.g., by a regulatory agency such as the U.S. Food and Drug Administration or the European Medicines Agency), e.g.. as reflected in Table 1, and / or a formulation thereof. Formulations of daratumumab include, for example, daratumumab, daratumumab / hyaluronidase, daratumumab / hyaluronidase-fihj, darzalex, darzalex faspro, isatuximab, isatuximab-irfc, and sarclisa.

[0112] In some cases, the antibody or antigen-binding fragment thereof is engineered to enhance binding of its Fc domain to activating receptors (e.g., Fcy-receptors), relative to antibody without the engineering.Table 1. CD19, CD20, and CD38 Targeting Antibody Therapeutics Approved in US or EP. The Antibody Society. Therapeutic monoclonal antibodies approved or in review in the EU or US. (June 24. 2023); antibodysocieU.org / resources / approved-antibodies.

[0113] In some cases, the antibody or antigen-binding fragment thereof is an NK cell engager, e.g., a bispecific or trispecific antibody, that bridges NK cell activating receptor(s) (for example, CD16A, NKG2D, NKp30, or NKp46) and molecule(s) specific to disease cell (e.g., tumor cell(s)). See, e.g., Demaria et al., “Natural Killer Cell Engagers in Cancer Immunotherapy: Next Generation of Immuno-Oncology Treatments,” European Journal of Immunology 51(8):doi.org / 10.1002 / eji.202048953 (2021).III. PHARMACEUTICAL COMPOSITIONS

[0114] Provided herein are pharmaceutical compositions comprising the natural killer cells described herein and dosage units of the pharmaceutical compositions described herein.

[0115] In some cases, the dosage unit comprises between 100 million and 1.5 billion cells, e.g., 100 million. 200 million, 300 million, 400 million, 500 million, 600 million, 700 million, 800 million. 900 million, 1 billion, 1.1 billion, 1.2 billion, 1.3 billion, 1.4 billion, or 1.5 billion cells, or thereabout.

[0116] In some cases, the dosage unit comprises between 100 million and 10 billion cells, e.g., 100 million, 200 million, 300 million, 400 million, 500 million, 600 million, 700 million, 800 million. 900 million, 1 billion, 1.5 billion, 2 billion, 2.5 billion, 3 billion, 3.5 billion, 4 billion, 4.5 billion, 5 billion, 5.5 billion, 6 billion, 6.5 billion, 7 billion, 7.5 billion, 8 billion, 8.5 billion, 9 billion, 9.5 billion, 10 billion cells, or thereabout.

[0117] Pharmaceutical compositions typically include a pharmaceutically acceptable carrier. As used herein the language "pharmaceutically acceptable carrier" includes saline, solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration.

[0118] In some embodiments, the pharmaceutical composition comprises: a) natural killer cell(s) described herein; and b) a cryopreservation composition.

[0119] Suitable cryopreservation compositions are described herein.

[0120] In some embodiments, the composition is frozen. In some embodiments, the composition has been frozen for at least three months, e.g., at least six months, at least nine months, at least 12 months, at least 15 months, at least 18 months, at least 24 months, at least 36 months, or at least 48 months.

[0121] In some embodiments, at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% of the natural killer cells are viable after being thawed.

[0122] In some embodiments, the pharmaceutical composition comprises: a) a cry opreservation composition described herein; and b) therapeutic cell(s).

[0123] In some embodiments, the therapeutic cell(s) are animal cell(s). In some embodiments, the therapeutic cell(s) are human cell(s).

[0124] In some embodiments, the therapeutic cell(s) are immune cell(s). In some embodiments, the immune cell(s) are selected from basophils, eosinophils, neutrophils, mastcells, monocytes, macrophages, neutrophils, dendritic cells, natural killer cells, B cells. T cells, and combinations thereof.

[0125] In some embodiments, the immune cell(s) are natural killer (NK.) cells. In some embodiments, the natural killer cell(s) are expanded and stimulated by a method described herein.

[0126] In some embodiments, the pharmaceutical composition further comprises: c) a buffer solution. Suitable buffer solutions are described herein, e.g.. as for cry opreservation compositions.

[0127] In some embodiments, the pharmaceutical composition comprises from or from about 1x107to or to about 1x109cells / mL. In some embodiments, the pharmaceutical composition comprises 1x108cells / mL. In some embodiments, the pharmaceutical composition comprises about IxlO8cells / mL.

[0128] In some embodiments, the pharmaceutical composition comprises from or from about 1x108to or to about 1x1010cells / mL.

[0129] In some embodiments, the pharmaceutical composition further comprises an antibody or antigen binding fragment thereof, e.g., an antibody described herein.

[0130] Pharmaceutical compositions are typically formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal, and rectal administration.

[0131] Methods of formulating suitable pharmaceutical compositions are know n in the art, see, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., 2005; and the books in the series Drugs and the Pharmaceutical Sciences: a Series of Textbooks and Monographs (Dekker, NY). For example, solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted w ith acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0132] Pharmaceutical compositions suitable for injectable use can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

[0133] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the acti ve compound into a sterile vehicle, which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying, which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile- filtered solution thereof.IV. METHODS OF TREATMENT

[0134] The NK cells described herein find use for treating autoimmune disorders.

[0135] Thus, also provided herein are methods of treating a patient suffering from a disorder, e.g., a disorder associated with autoimmunity, e.g., autoreactive immune cells, e.g., autoreactive B cells, comprising administering the NK cells, e.g., the NK cells describedherein, and a CD 19, CD20, and / or CD38 targeting antibody, e.g., an antibody described herein. In some cases, the antibody is a plasma cell or plasmablast targeting antibody, e.g., a CD38-targeting antibody.

[0136] As used herein, the terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disorder associated with autoimmunity (e.g.. a disorder described herein, e.g., systemic lupus erythematosus (“SLE”)). In some embodiments, treatment may be administered after one or more symptoms have developed. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e g., in light of a history of symptoms and / or in light of genetic or other susceptibility factors). Treatment may also be continued after symptoms have resolved, for example to prevent or delay their recurrence.

[0137] As used herein, “delaying” development of a disease or disorder, or one or more symptoms thereof, means to defer, hinder, slow, retard, stabilize and / or postpone development of the disease, disorder, or symptom thereof. This delay can be of varying lengths of time, depending on the history of the disease and / or subject being treated. As is evident to one skilled in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the subject does not develop the disease, disorder, or symptom thereof. For example, a method that “delays” development of an autoimmune disorder is a method that reduces the probability of disease development in a given time frame and / or reduces extent of the disease in a given time frame, when compared to not using the method. Such comparisons may be based on clinical studies, using a statistically significant number of subjects.

[0138] As used herein, “prevention” or “preventing” refers to a regimen that protects against the onset of the disease or disorder such that the clinical symptoms of the disease do not develop. Thus, “prevention” relates to administration of a therapy (e.g., administration of a therapeutic substance) to a subject before signs of the disease are detectable in the subject and / or before a certain stage of the disease. The subject may be an individual at risk of developing the disease or disorder, or at risk of disease progression, e.g., development of proliferative lupus nephritis. Such as an individual who has one or more risk factors known to be associated with development or onset of the disease or disorder. For example, an individual may have mutations associated with the development or progression of an autoimmune disease (e.g., SLE). Further, it is understood that prevention may not result in complete protection against onset of the disease or disorder. In some instances, preventionincludes reducing the risk of developing the disease or disorder. The reduction of the risk may not result in complete elimination of the risk of developing the disease or disorder.

[0139] In some cases, an “increased” or “enhanced” amount refers to an increase that is 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, or 50 or more times (e.g., 100, 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 2.1, 2.2, 2.3, 2.4, etc.) an amount or level described herein. It may also include an increase of at least 10%. at least 20%. at least 30%. at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 150%, at least 200%, at least 500%, or at least 1000% of an amount or level described herein.

[0140] In some cases, a “decreased” or “reduced” or “lesser” amount refers to a decrease that is about 1.1, 1.2, 1.3. 1.4, 1.5, 1.6 1.7, 1.8. 1.9, 2, 2.5. 3, 3.5, 4, 4.5. 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, or 50 or more times (e.g., 100, 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) an amount or level described herein. It may also include a decrease of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%, at least 100%, at least 150%, at least 200%. at least 500%, or at least 1000% of an amount or level described herein.

[0141] Also provided herein are methods of depleting immune cell(s), e.g., autoreactive immune cells, e.g., autoreactive B cells, comprising administering the NK cells, e.g., the NK cells described herein, and a CD19, CD20. and / or CD38 targeting antibody, e.g., an antibody described herein. In some cases, the immune cell is a CD19+ immune cell, CD20+ and / or CD38+ immune cell, e.g., a CD19+, CD20+, and / or CD38+ autoreactive immune cell, e.g., a CD19+, CD20+, and / or CD38+ autoreactive B cell.

[0142] In some cases, the immune cell(s) (e.g., the autoreactive immune cell(s)) comprise or consist of B cell(s) (e.g., autoreactive B cells). In some cases, the B cell(s) are CD19+, CD20+, or CD38+ B cell(s), or a combination thereof.

[0143] In some cases, the B cell(s) are or include plasmablasts and / or plasma cells.

[0144] In some cases, the B cell(s) are selected from B cell(s) of anorectum, B cell(s) of appendix, B cell(s) of medullary sinus of lymph node, lymph node mantle zone B cell(s), monocytoid B cell(s), CD 19-positive B cell(s), and combinations thereof.

[0145] In some cases, the CD 19-positive B cell(s) are selected from immature B cell(s), mature B cell(s), precursor B cell(s), transitional stage B cell(s), and combinations thereof.

[0146] In some cases, the immature B cell(s) are selected from CD38-negative immature B cell(s), fraction E immature B cell(s). and combinations thereof.

[0147] In some cases, the mature B cell(s) are selected from B-l B cell(s), B-2 B cell(s), Be cell(s), Peyer’s patch B cell(s), follicular B cell(s), fraction F mature B cell(s), germinal center B cell(s), marginal zone B cell(s) of lymph node, marginal zone B cell(s) of spleen, memory B cell(s), naive B cell(s), plasmablast(s), regulatory B cell(s), and combinations thereof.

[0148] In some cases, the B-l B cell(s) are selected from B-la B cell(s), B-lb B cell(s), and combinations thereof. In some cases, the B-2 B cell(s) are selected from Peyer’s patch B cell(s). follicular B cell(s), fraction F mature B cell(s), and combinations thereof. In some cases, the follicular B cell(s) are selected from Bml B cell(s). Bm2 B cell(s). and combinations thereof. In some cases, the fraction F mature B cell(s) are Bml B cell(s). In some cases, the Be cell(s) are selected from Bel cell(s), Be2 cell(s), and combinations thereof. In some cases, the germinal center B cell(s) are selected from Bm2’ B cell(s), Bm3 B cell(s), Bm3-delta B cell(s), Bm4 B cell(s), centroblast(s), centrocyte(s), tonsil germinal center B cell(s), and combinations thereof.

[0149] In some cases, the memory B cell(s) are selected from Bm5 B cell(s), IgD- negative memory B cell(s), IgM memory7B cell(s), class switched memory7B cell(s), double negative memory B cell(s), unswitched memory B cell(s), and combinations thereof.

[0150] In some cases, the IgD-negative memory B cell(s) are selected from Bm5 B cell(s), CD38-negative IgG memory B cell(s), IgD-negative CD38-positive IgG memory B cell(s), IgM memory7B cell(s), double negative memory7B cell(s), and combinations thereof. In some cases, the double negative memory B cell(s) are selected from IgG-negative double negative memory7B cell(s), IgG-positive double negative memory7B cell(s), and combinations thereof.

[0151] In some cases, the class switched memory B cell(s) are selected from IgA memory7B cell(s), IgE memory7B cell(s), IgG memory7B cell(s), IgG-negative class switched memory B cell9s), and combinations thereof. In some cases, the IgG memory B cell(s) are selected from CD38-negative IgG memory B cell(s), CD38-postiive IgG memory B cell(s), and combinations thereof. In some cases, the CD38-positive IgG memory B cell(s) are selected from IgD-negative CD38-positive IgG memory7B cell(s), IgD-positive CD38- positive IgG memory7B cell(s), and combinations thereof. In some cases, the IgG-negative class switched memory7B cell(s) ae selected from CD38-positive IgG-negative class switched memory B cell(s), CD38-positive IgG-negative class switched memory B cell(s), andcombinations thereof. In some cases, the CD38-negative IgG-negative class switched memory B cell(s) are selected from CD24-negative CD38-negative IgG-negative class switched memory B cell(s), CD24-positive CD38-negative IgG-negative class switched memory B cell(s), and combinations thereof. In some cases, the CD38-positive IgG-negative class switched memory B cell(s) are selected from B220-low CD38-positive IgG-negative class switched memory B cell(s), B220-positive CD38-positive IgG-negative class switched memory B cell(s), and combinations thereof. In some cases, the B220-positive CD38- positive IgG-negative class switched memory B cell(s) are B220-low CD38-positive IgG- negative class switched memory B cell(s). In some cases, the double negative memory7B cell(s) are selected from IgG-negative double negative memory B cell(s), IgG-positive double negative memory7B cell(s), and combinations thereof. In some cases, the unswitched memory B cell(s) are selected from CD38-negative unswitched memory B cell(s), CD38- positive unswitched memory7B cell(s), and combinations thereof. In some cases, the CD38- negative unswitched memory7B cell(s) are selected from B220-low CD38-negative unswitched memory B cell(s), B220-positive CD38-negative unswitched memory B cell(s), and combinations thereof. In some cases, the B220-positive CD38-negative unswitched memory B cell(s) are B220-low CD38-negative unswitched memory B cell(s). In some cases, the CD38-positive unswitched memory7B cell(s) are selected from B220-low CD38- positive unswitched memory B cell(s), B220-positive CD38-positive unswitched memory B cell(s), and combinations thereof. In some cases, the B220-positive CD38-positive unstitched memory B cell(s) are B220-low CD38-positive unswitched memory B cell(s). In some cases, the naive B cell(s) are selected from CD38-negative naive B cell(s), CD38-positive naive B cell(s), and combinations thereof. In some cases, the CD38-positive naive B cell(s) are B220-low CD38-positive naive B cell(s), B220-positive cd38-positive naive B cell(s), and combinations thereof. In some cases, the B220-positive CD38-positive naive B cell(s) are B220-low CD38-positive naive B cell(s). In some cases, the plasmablast(s) are selected from CD86-positive plasmablast(s), IgA plasmablast(s), IgD plasmablast(s), IgD plasmablast(s), IgE plasmablast(s), IgG plasmablast(s), IgM plasmablast(s), and combinations thereof.

[0152] In some cases, the precursor B cell(s) are selected from fraction B / C precursor B cell(s), fraction C’ precursor B cell(s), fraction D precursor B cell(s), late pro-B cell(s), pre-B-I cell(s), pre-B-II cell(s), and combinations thereof. In some cases, the pre-B-II cell(s) are selected from large pre-B-II cell(s), small pre-B-II cell(s), and combinations thereof. In some cases, the large pre-B-II cell(s) are selected from preBCR-negative large pre-B-II cell(s), preBCR-positive large pre-B-II cell(s), and combinations thereof. In some cases, thepre-BCR-positive large pre-B-II cell(s) are CD38-high pre-BCR positive cell(s). In some cases, the small pre-B-II cell(s) are CD22-positive, CD38-low small pre-B cell(s).

[0153] In some cases, the transitional stage B cell(s) are selected from T1 B cell(s), T2 B cell(s), T3 B cell(s), and combinations thereof.A. Disorders

[0154] Methods and manufactured compositions disclosed herein find use in targeting a number of disorders, such as autoimmune disorders. A benefit of the approaches herein is that allogenic cells are used in combination with exogenous antibody administration to specifically target immune cells, e.g., B cells.

[0155] In some cases, the autoimmune disorder is driven by autoantibodies and / or autoreactive B cells.

[0156] In some cases, the autoimmune disorder is selected from Acromegaly, Acquired aplastic anemia, Acquired hemophilia. Primary Agammaglobulinemia, Alopecia areata, Ankylosing spondylitis (AS), Anti-NMDA receptor encephalitis, Antiphospholipid syndrome (APS) | catastrophic antiphospholipid syndrome (CAPS) / Asherson's syndrome, Arteriosclerosis, Autoimmune Addison’s disease (AAD), Autoimmune autonomic ganglionopathy (AAG) / autoimmune dysautonomia | autoimmune gastrointestinal dysmotility (AGID), Autoimmune encephalitis | acute disseminated encephalomyelitis (ADEM), Autoimmune gastritis, Autoimmune hemolytic anemia (AIHA), Autoimmune hepatitis (AIH), Autoimmune hyperlipidemia. Autoimmune hypophysitis, Autoimmune inner ear disease (AIED), Autoimmune lymphoproliferative syndrome (ALPS), Autoimmune myelofibrosis, Autoimmune myocarditis, Autoimmune oophoritis, Autoimmune pancreatitis (AIP), Autoimmune polyglandular syndromes, types I, II, & III (APS type 1, APS ty pe 2, APS type 3, APECED), Autoimmune progesterone dermatitis, Autoimmune retinopathy (AIR), Autoimmune sudden sensorineural hearing loss (SNHL), Balo disease, Behcet’s disease. Birdshot chorioretinopathy / birdshot uveitis, Bullous pemphigoid, Castleman disease, Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy (CIDP), Chronic urticaria (CU), Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA) , Cogan’s syndrome. Cold agglutinin disease, CREST syndrome | limited cutaneous systemic sclerosis, Crohn’s disease (CD), Cronkhite-Canada syndrome (CSS), Cryptogenic organizing pneumonia (COP), Dermatitis herpetiformis, Dermatomyositis, Type 1 Diabetes, Discoid lupus, Dressier’s syndrome / postmyocardial infarction / postpericardiotomy syndrome. Eczema / Atopic Dermatitis, Endometriosis,Eosinophilic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Essential mixed cryoglobulinemia, Evans syndrome, Fibrosing alveolitis / Idiopathic pulmonary fibrosis (IPF), Giant cell arteritis / temporal arteritis / Horton’s disease. Giant Cell Myocarditis, Glomerulonephritis, Goodpasture’s syndrome / anti-GBM / anti-TBM disease, Granulomatosis with polyangiitis (GPA) / Wegener’s granulomatosis, Graves disease / thyroid eye disease, Guillain-Barre syndrome (GBS), Hashimoto’s thyroiditis / chronic lymphocytic thyroiditis / autoimmune thyroiditis, Henoch-Schonlein purpura / IgA vasculitis, Hidradenitis suppurativa, Hurst’s disease / acute hemorrhagic leukoencephalitis (AHLE), Hypogammaglobulinemia, IgA nephropathy / Berger's disease, Immune-mediated necrotizing myopathy (IMNM), Immune thrombocytopenia (ITP) / autoimmune thrombocytopenic purpura / autoimmune thrombocytopenia, Inclusion body myositis, IgG4-related sclerosing disease (ISD), Interstitial cystitis. Juvenile idiopathic arthritis I Adult-onset Still's disease, Juvenile polymyositis | Juvenile dermatomyositis | juvenile myositis, Kawasaki disease, Lambert-Eaton my asthenic syndrome (LEMS), Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Ligneous conjunctivitis. Linear IgA disease (LAD) | linear IgA bullous dermatosis (LABD). Lupus nephritis, Lyme disease / chronic Lyme disease / post-treatment Lyme disease syndrome (PTLDS), Lymphocytic colitis / microscopic colitis. Lymphocytic hypophystitis / autoimmune hypophystitis, Meniere’s disease, Microscopic polyangiitis (MPA) / ANCA-associated vasculitis, Mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease. Multifocal motor neuropathy, Multiple sclerosis (MS), Myalgic encephalomyelitis (ME) / Chronic fatigue syndrome (CFS), Myasthenia gravis (MG), Narcolepsy, Neuromyelitis Optica / Devic's disease, Ocular cicatricial pemphigoid, Opsoclonus-myoclonus syndrome (OMS). Palindromic rheumatism, Paraneoplastic cerebellar degeneration, Paraneoplastic pemphigus, Parry-Romberg syndrome (PRS) / Hemifacial atrophy (HFA) / Progressive facial hemiatrophy. Paroxysmal nocturnal hemoglobinuria (PNH), Peripheral uveitis / pars planitis, PANS / P ANDAS, Parsonage-Turner syndrome, Pemphigus gestationis / herpes gestationis, Pemphigus foliaceus, Pemphigus vulgaris, Pernicious anemia. POEMS syndrome, Polyarteritis nodosa, Polymyalgia rheumatica, Polymyositis, Postural orthostatic tachycardia syndrome (POTS). Primary biliary cirrhosis (PBC) / primary biliary cholangitis, Primary sclerosing cholangitis (PSC), Psoriasis, Palmoplantar Pustulosis, Psoriatic arthritis, Pulmonary fibrosis, idiopathic (IPF), Pure red cell aplasia (PRC A), Pyoderma gangrenosum, Rasmussen's encephalitis, Raynaud’s syndrome / phenomenon, Reactive arthritis / Reiter’s syndrome, Reflex sympathetic dystrophy syndrome (RSD) / Complex regional pain syndrome (CRPS), Relapsing polychondritis.Restless leg syndrome (RLS) / Willis-Ekbom disease, Rheumatic fever, Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome / autoimmune polyendocrine syndrome type II, Scleritis. Scleroderma , Sclerosing Mesenteritis I Mesenteric Panniculitis, Serpiginous choroidopathy, Sjogren’s syndrome, Stiff person syndrome (SPS), Small fiber sensory neuropathy, Systemic lupus ery thematosus (SLE), Subacute bacterial endocarditis (SBE), Subacute cutaneous lupus, Susac syndrome, Sydenham's chorea. Sympathetic ophthalmia, Takayasu's arteritis (vasculitis), Testicular autoimmunity (vasculitis, orchitis), Tolosa-Hunt syndrome.Transverse myelitis (TM), Tubulointerstitial nephritis uveitis syndrome (TINU), Ulcerative colitis (UC), Undifferentiated connective tissue disease (UCTD), Uveitis | anterior / intermediate / posterior, Vasculitis, VEXAS Syndrome, Vitiligo, Vogt-Koyanagi- Harada syndrome (VKH), and combinations thereof.

[0157] In some embodiments, treatment comprises an improvement in a patient or patient population’s symptoms (e.g., as described herein). In some cases, the improvement is in comparison to a baseline or threshold amount. In some cases, the baseline or threshold amount is a value normally considered to be within a normal (e.g., healthy) range. In some cases, the baseline or threshold amount is based on a value prior to treatment and a value after treatment (e g., a patient’s own baseline score prior to treatment or a patient population’s baseline score prior to treatment).

[0158] In some cases, the improvement is measured after administration, e.g., within a treatment cycle. In some cases, the response is measured within a week, e.g.. within 1. 2, 3, 4, 5, 6, or 7 days after administration. In some cases, the response is measured after the last administration of a first treatment cycle (e.g., as described herein). In some cases, the response is measured after the last administration of a second treatment cycle (e.g., as described herein). In some cases, the response is measured within 1, 2. 3, 4, 5, 6, 7, 8, 9. 10. or 12 months after administration, e.g., the last administration of a treatment cycle.B. Patients

[0159] Suitable patients for the compositions and methods herein include those who are suffering from, who have been diagnosed with, or who are suspected of having an autoimmune disorder, e.g.. as described herein.

[0160] In some embodiments, the methods of treatment provided herein may be used to treat a subject (e.g., human, monkey, dog, cat, mouse) who has been diagnosed with or is suspected of having an autoimmune disorder, e.g., as described herein. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0161] In some embodiments, the patient is or has been diagnosed with an autoimmune disorder, e.g.. and autoimmune disorder described herein. In some cases, the patient is resistant to, relapsed, or refractory after initial treatment for the disorder. In some cases, the patient has failed at least two lines of previous treatments in alignment with the Standard of Care (SOC).C. Lymphodepletion

[0162] In some embodiments, the patient is lymphodepleted before treatment.

[0163] Illustrative lymphodepleting chemotherapy regimens, along with correlative beneficial biomarkers, are described in WO 2016 / 191756 and WO 2019 / 079564, hereby incorporated by reference in their entirety. In certain embodiments, the lymphodepleting chemotherapy regimen comprises administering to the patient doses of cyclophosphamide (between 200 mg / m2 / day and 2000 mg / m2 / day) and doses of fludarabine (between 20 mg / m2 / day and 900 mg / m2 / day).

[0164] In some embodiments, lymphodepletion comprises administration of or of about 100 to or to about 1500 mg / m2of cyclophosphamide, e g., of or of about 250 to or to about 500 mg / m2of cyclophosphamide, e.g., from or from about 250 to or to about 500, 250, 400, 500, about 250, about 400, or about 500 mg / m2of cyclophosphamide. In some embodiments, lymphodepletion comprises administration of or of about 500 mg / m2of cyclophosphamide. In some embodiments, lymphodepletion comprises administration of or of about 100 mg / m2of cyclophosphamide.

[0165] In some embodiments, lymphodepletion comprises administration of or of about 20 mg / m2 / day to or to about 40 mg / m2 / day fludarabine, e.g., 30 or about 30 mg / m2 / day.

[0166] In some embodiments, lymphodepletion comprises administration of both cyclophosmamide and fludarabine.

[0167] In some embodiments, the patient is lymphodepleted by intravenous administration of cyclophosphamide (250 mg / m2 / day) and fludarabine (30 mg / m2 / day).

[0168] In some embodiments, the patient is lymphodepleted by intravenous administration of cyclophosphamide (300 / mg / m2 / day) and fludarabine (30 mg / m2 / day).

[0169] In some embodiments, the patient is lymphodepleted by intravenous administration of cyclophosphamide (500 mg / m2 / day) and fludarabine (30 mg / m2 / day).

[0170] In some embodiments, the lymphodepletion occurs no more than 5 days prior to the first dose of NK cells. In some embodiments, the lymphodepletion occurs no more than 7 days prior to the first dose of NK cells.

[0171] In some embodiments, lymphodepletion occurs daily for 3 consecutive days, starting 5 days before the first dose of NK cells.

[0172] In some embodiments, the first dose of NK cells is given on day 6 and lymphodepletion occurs on days 1, 2, and 3.D. Administration

[0173] Described herein are methods comprising administration of NK cells (e.g., as described herein). In some cases, the NK cells are administered as part of a therapy further comprising administration of one or more additional agents, including, for example, an antibody (e.g., as described herein), a cytokine (e.g., as described herein), a lymphodepleting agent (e.g., as described herein), a corticosteroid (e.g., prednisone, prednisolone, dexamethasone, methylprednisolone), an analgesic, an antipyretic, and / or an antihistamine. For example, patients may be pre-treated I pre-medicated prior to NK cell infusion, e.g., as described herein.1. NK Cells

[0174] In some embodiments, the NK cells are administered as part of a pharmaceutical composition, e.g., a pharmaceutical composition described herein. Cells are administered after thawing, in some cases without any further manipulation in cases where their cryoprotectant is compatible for immediate administration. For a given individual, a treatment regimen often comprises administration over time of multiple aliquots or doses of NK cells, which can be drawn from a common batch or donor. In some embodiments, the NK cells, e.g., the NK cells described herein are administered at or at about l x 108to or to about 8 x 109NK cells per dose. In some embodiments, the NK cells are administered at or at about 1 x 108, at or at about 1 x 109, at or at about 4 x 109, or at or at about 8 x 109NK cells per dose. In some cases, the NK cells are administered at or about 4 billion cells per dose. In some cases, the NK cells are administered at or about 2 billion cells per dose.

[0175] In some embodiments, the NK cells are administered weekly. In some embodiments, the NK cells are administered for or for about 3-8 weeks. In some embodiments, the NK cells are administered for or for about 4-8 weeks. In some embodiments, the NK cells are administered weekly for or for about 4 weeks. In some embodiments, the NK cells are administered weekly for or for about 8 weeks.

[0176] In some embodiments, the NK cells are administered on days 6, 9, 13, and 16 of a treatment cycle. In some embodiments, the NK cells are administered on days 6, 13, and20 of a treatment cycle. In some embodiments, the NK cells are administered on days 6 and 13 of a treatment cyce.

[0177] In some embodiments, administration is repeated for one or more additional treatment cycles, e.g., following an administration scheduled described above. In some embodiments, the additional treatment cycle is or is about 2-12 months after the previous treatment cycle, e.g., 2, 3, 4, 5, 6, 7, 8. 9, 10, 11, or 12 months, or thereabout, after the previous treatment cycle.

[0178] In some embodiments, the NK cells are cryopreserved in an infusion-ready media, e.g., a cryopreservation composition suitable for intravenous administration, e.g., as described herein.

[0179] In some embodiments, the NK cells are cryopreserved in vials containing from or from about 1 x I08to or to about 8 x IO9cells per vial. In some embodiments, the NK cells are cryopreserved in vials containing a single dose.

[0180] In some embodiments, the cells are thawed, e.g., in a 37°C water bath, prior to administration.

[0181] In some embodiments, the thawed vial(s) of NK cells are aseptically transferred to a single administration vessel, e.g., administration bag using, e.g., a vial adapter and a sterile syringe. The NK cells can be administered to the patient from the vessel through a Y-type blood / solution set filter as an IV infusion, by gravity.

[0182] In some embodiments, the NK cells are administered as soon as practical, preferably less than 90 minutes, e.g., less than 80, 70, 60, 50, 40, 30, 20, or 10 minutes after thawing. In some embodiments, the NK cells are administered within 30 minutes of thawing.

[0183] In some embodiments, the pharmaceutical composition is administered intravenously via syringe.

[0184] In some embodiments, 1 mL, 4 mL, or 10 mL of drug product is administered to the patient intravenously via syringe. In some embodiments, 30 mL or 40 mL of drug product is administered to the patient intravenously via syringe.2. Antibodies

[0185] In some embodiments, the NK cell(s) described herein, e.g.. the pharmaceutical compositions comprising NK cell(s) described herein, are administered in combination with an antibody or antibodies, e.g., an antibody or antibodies described herein (e.g., in Table 1), e.g., a B-cell depleting antibody, e.g., a CD19, CD20 and / or CD38 antibody, e.g., tafasitamab, rituximab, obinutuzumab, and / or daratumumab. In someembodiments, the antibodie(s) are administered together with the NK cells as part of a pharmaceutical composition. In some embodiments, the antibodie(s) are administered separately from the NK cells, e.g., as part of a separate pharmaceutical composition. Antibodies can be administered prior to, subsequent to, or simultaneously with administration of the NK cells.

[0186] In some embodiments, the antibody is administered before the NK cells. In some embodiments, the antibody is administered after the NK cells.

[0187] In some embodiments, the NK cells are administered at least 30 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes, 180 minutes, 210 minutes, or 240 minutes after completing administration of the antibody.

[0188] In some embodiments, the NK cells are administered the day after the antibody is administered.

[0189] In some embodiments, the NK cells are administered at each administration, while the antibody is administered at a subset of the administrations. For example, in some embodiments, the NK cells are administered once a week and the antibody is administered once a month.

[0190] In some embodiments, the antibody is administered weekly for 8 weeks. In some embodiments, the antibody is administered every two weeks for 8 weeks.

[0191] In some embodiments, a dose of antibody is given prior to the first dose of cells. In some embodiments, a debulking dose of the antibody is given prior to the first dose of cells.

[0192] In some embodiments, the antibody is administered on days 2 and 13 of a treatment cycle. In some embodiments, the antibody is administered on days 1 and 15 of a treatment cycle.

[0193] In some embodiments, the antibody is administered weekly for four weeks, e.g., on days 1, 8, 15, and 22 of a treatment cycle. In some embodiments, the antibody is administered weekly for four weeks, e.g., on days 1, 8, 15, and 22 of a treatment cycle, followed by additional doses every two weeks, e.g., weeks 5, 7. 9, and 11.

[0194] In some cases, the antibody is administered intravenously. In some cases, the antibody is administered subcutaneously. In some cases, the antibody (e.g., daratumumab) is administered intravenously, e.g., weekly, e.g., for four weeks, e.g., on days 1, 8, 15, and 22, followed by subcutaneous administration, e.g., every' two weeks, e.g., weeks 5, 7, 9, and 11.3. Cytokines

[0195] In some embodiments, a cytokine is administered to the patient.

[0196] In some embodiments, the cytokine is administered together with the NK cells as part of a pharmaceutical composition. In some embodiments, the cytokine is administered separately from the NK cells, e.g., as part of a separate pharmaceutical composition. In some embodiments, the cytokine is IL-2.

[0197] In some embodiments, a cytokine is not administered to the patient.E. Dosing

[0198] An ‘‘effective amount” is an amount sufficient to effect beneficial or desired results. For example, a therapeutic amount is one that achieves the desired therapeutic effect. This amount can be the same or different from a prophylactically effective amount, which is an amount necessary to prevent onset of disease or disease symptoms. An effective amount can be administered in one or more administrations, applications or dosages. A therapeutically effective amount of a therapeutic compound (i.e., an effective dosage) depends on the therapeutic compounds selected. The compositions can be administered one from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors may influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Moreover, treatment of a subject with a therapeutically effective amount of the therapeutic compounds described herein can include a single treatment or a series of treatments.

[0199] Dosage, toxicity and therapeutic efficacy of the therapeutic compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50 / ED50. Compounds which exhibit high therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.

[0200] The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds may bewithin a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary’ within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e.. the concentration of the test compound which achieves a half- maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography, flow cytometry', or a molecular assay.F. Treatment Cycles

[0201] In some cases, treatment comprises administration of the NK cells and antibodies (e.g., as described herein), over the course of a treatment cycle. For example, in some cases, a treatment cycle comprises lymphodepletion followed by administrations of NK cells and a B-cell depleting antibody. In some cases, the B-cell depleting antibody is administered prior to the first administration of NK cells and.V. VARIANTS

[0202] In some embodiments, the fusion protein(s) or components thereof described herein, or the NK cell genotypes described herein, are at least 80%, e.g., at least 85%, 90%, 95%, 98%, or 100% identical to the amino acid sequence of an exemplary' sequence (e.g., as provided herein), e.g., have differences at up to 1%. 2%, 5%, 10%, 15%, or 20% of the residues of the exemplary sequence replaced, e.g., with conservative mutations, e.g., including or in addition to the mutations described herein. In preferred embodiments, the variant retains desired activity of the parent.

[0203] To determine the percent identity of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non- homologous sequences can be disregarded for comparison purposes). The length of a reference sequence aligned for comparison purposes is at least 80% of the length of the reference sequence, and in some embodiments is at least 90% or 100%. The nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as usedherein nucleic acid "identity" is equivalent to nucleic acid "homology"). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0204] Percent identity between a subject polypeptide or nucleic acid sequence (i.e. a query) and a second polypeptide or nucleic acid sequence (i.e. target) is determined in various ways that are within the skill in the art, for instance, using publicly available computer software such as Smith Waterman Alignment (Smith, T. F. and M. S. Waterman (1981) J Mol Biol 147:195-7); "BestFit" (Smith and Waterman, Advances in Applied Mathematics, 482- 489 (1981)) as incorporated into GeneMatcher PlusTM. Schwarz and Dayhof (1979) Atlas of Protein Sequence and Structure, Dayhof, M.O., Ed, pp 353-358; BLAST program (Basic Local Alignment Search Tool; (AltschuL S. F., W. Gish, et al. (1990) J Mol Biol 215: 403- 10), BLAST-2, BLAST-P, BLAST-N, BLAST-X, WU-BL AST-2, ALIGN, ALIGN-2, CLUSTAL, or Megalign (DNASTAR) software. In addition, those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the length of the sequences being compared. In general, for target proteins or nucleic acids, the length of comparison can be any length, up to and including full length of the target (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100%). For the purposes of the present disclosure, percent identity is relative to the full length of the query sequence.

[0205] For purposes of the present disclosure, the comparison of sequences and determination of percent identity7between two sequences can be accomplished using a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0206] Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.VI. DEFINITIONS

[0207] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should notnecessarily be construed to represent a substantial difference over what is generally understood in the art.

[0208] Throughout this application, various embodiments may 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 disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numencal 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, 3, 4. 5, and 6. This applies regardless of the breadth of the range.

[0209] As used in the specification and claims, the singular forms “a”, “an” and ‘‘the’’ include plural references unless the context clearly dictates otherwise. For example, the term “a sample’" includes a plurality of samples, including mixtures thereof.

[0210] The terms ‘"determining,” ‘"measuring,"’ "‘evaluating.” “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement. The terms include determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative or absolute. “Detecting the presence of’ can include determining the amount of something present in addition to determining whether it is present or absent depending on the context.

[0211] The terms “subject,” “individual,” or “patient” are often used interchangeably herein.

[0212] The term in vivo is used to describe an event that takes place in a subject s body.

[0213] The term "e vivo” is used to describe an event that takes place outside of a subject’s body. An ex vivo assay is not performed on a subject. Rather, it is performed upon a sample separate from a subject. An example of an ex vivo assay performed on a sample is an "in vitro” assay.

[0214] The term "in vitro” is used to describe an event that takes place contained in a container for holding laboratory reagent such that it is separated from the biological source from which the material is obtained. In vitro assays can encompass cell-based assays in which living or dead cells are employed. In vitro assays can also encompass a cell-free assay in which no intact cells are employed.

[0215] As used herein, the term ‘‘about’' a number refers to that number plus or minus 10% of that number. The term “about” a range refers to that range minus 10% of its lowest value and plus 10% of its greatest value.

[0216] As used herein, the term "buffer solution" refers to an aqueous solution consisting of a mixture of a weak acid and its conjugate base, or vice versa.

[0217] As used herein, the term "cell culture medium" refers to a mixture for growth and proliferation of cells in vitro, which contains essential elements for growth and proliferation of cells such as sugars, amino acids, various nutrients, inorganic substances, etc.

[0218] A buffer solution, as used herein, is not a cell culture medium.

[0219] As used herein, the term “bioreactor” refers to a culture apparatus capable of continuously controlling a series of conditions that affect cell culture, such as dissolved oxygen concentration, dissolved carbon dioxide concentration, pH, and temperature.

[0220] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Some vectors are suitable for delivering the nucleic acid molecule(s) or polynucleotide(s) of the present application. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as expression vectors.

[0221] The term “operably linked” refers to two or more nucleic acid sequence or polypeptide elements that are usually physically linked and are in a functional relationship with each other. For instance, a promoter is operably linked to a coding sequence if the promoter is able to initiate or regulate the transcription or expression of a coding sequence, in which case, the coding sequence should be understood as being “under the control of’ the promoter.

[0222] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Host cells include “engineered cells,” “transformants.” and “transformed cells,” which include the primary engineered (e.g., transformed) cell and progeny derived therefrom without regard to the number of passages. Progeny may not be completely identical in nucleic acid content to a parent cell, but may contain mutations. Mutant progeny that have the same function or biological activity as screened or selected for in the originally transformed cell are included herein.

[0223] As appropriate, the host cells can be stably or transiently transfected with a polynucleotide encoding a fusion protein, as described herein.

[0224] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.VII. EXAMPLES

[0225] The following examples are included for illustrative purposes only and are not intended to limit the scope of the invention.Example 1: AB-101

[0226] AB- 101 is a universal, off-the-shelf, cryopreserved allogeneic cord blood derived NK cell therapy product comprising ex vivo expanded and activated effector cells designed to enhance ADCC anti -tumor responses in patients, e.g., patients treated with monoclonal antibodies or NK cell engagers. AB-101 was prepared as described, for example, in WO2022 / 133056.Example 2: CD38 Expression on SLE donor B cells

[0227] Whole blood from SLE donors (n=3) was collected in Anticoagulant Citrate Dextrose Solution, Solution A (ACD-A) tubes and shipped overnight at ambient temperature. PBMC were isolated using Lymphoprep according to the manufacturer instructions. Briefly, blood was diluted 1: 1 in wash medium (PBS containing 2% human AB serum) and then centrifuged at 800 x g for 30 minutes with brake off. The interface was collected and washed twice in wash medium. Viable cell numbers were determined using a CellacaTM MX cell counter. PBMC were either characterized by flow cytometry with cell-specific markers, including the B-cell markers CD19 and CD38 or utilized for cy totoxicity assay s.

[0228] Flow cytometry characterization. Flow cytometric data acquisition was performed using a Cytek®1Aurora (Cytek®, Fremont CA) and data acquired using SpectroFlo® Flow Cytometry Software. Data was analyzed with FlowJo (FlowJo, LLC, Ashland, OR) software version 10.8.1. CD38 expression was assessed on CD19+ B-cells. The percent CD19+CD38+ B-cells are shown in the upper right quadrant of the representative flow plot in FIG. 1 and summarized for 3 SLE donors in the table. The average percentage of B-cells that express CD38 is 72% across 3 SLE donors indicating that there are CD38-expressing B-cells in the SLE PBMC population.Table 2. CD38 Expression on SLE donor B Cells.Example 3: Antibody-Dependent Cytotoxicity Assay

[0229] PBMC were resuspended in assay medium (RPMI1640, 5% human AB serum, and 1 ng / mL human IL-15) and plated at 2xlOA5 cells / well in a 96-well U-bottom plate (Coming #3879) with or without AB-101 (4x10A4 or 2x10A5 cells / well) and anti-CD38 monoclonal antibody (0.1 or 1 pg / mL of daratumumab) in a total volume of 200 pL / well. Plates were incubated for 3.5 hours in a 37°C incubator with 5% CO2. Caspase 3 / 7 green (ThermoFisher) was diluted 1 : 100 in assay medium then added at lOpL / well (IpM final concentration) and incubated for an additional 30 minutes at 37°C. Plates were centrifuged at 800 x g for 3 minutes, supernatant was removed and cells were incubated for 25 minutes at 4°C with antibodies (Table 4) and human Fc block (human TruStain FcX) diluted in brilliant stain buffer. Plates were washed with BD stain buffer (BD Biosciences) and centrifuged at 800 x g for 3 minutes. Samples were resuspended in BD stain buffer for analysis by flow cytometry.

[0230] Flow cytometric data acquisition was performed using a Cytek® Aurora (Cytek®, Fremont CA) and data acquired using SpectroFlo® Flow Cytometry Software. Caspase 3 / 7 green was detected in the FITC channel. Data was analyzed with FlowJo (FlowJo, LLC, Ashland, OR) software version 10.8.1. PBMCs were first gated for single cells using forward scatter (FSC) and side scatter (SSC) followed by CD45 expression. Caspase expression in B cells was gated as follows: B-cells were first gated as CD14 negative and CD19 or CD20 positive followed by gating out of CD3, CD56. and CD16 positive cells. Data is expressed as the percentage of caspase-positive B cells.

[0231] As shown in FIG. 2, antibody-dependent cellular cytotoxicity (ADCC) experiments utilizing human SLE donor PBMCs co-cultured with AB-101 demonstrated AB- 101-mediated B-cell killing at different effector to target (E:T) ratios in the presence of the anti-CD38 monoclonal antibody, daratumumab. Enhanced AB-101-mediated B-cell killing was observed in an E:T ratio-dependent manner.OTHER EMBODIMENTSIt is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are w ithin the scope of the following claims.

Claims

CLAIMS1. A method for treating a patient suffering from an autoimmune disorder, the method comprising administering a population of natural killer cells (NK cells) and a CD38 targeting antibody, wherein the NK cells are allogenic to the patient.

2. The method of claim 1, wherein the CD38 antibody targets an immune cell implicated in the autoimmune disorder.

3. The method of claim 1 or claim 2, wherein the immune cell is a B cell, optionally a plasma cell and / or plasmablast.

4. The method of any one of claims 1 to 3, wherein the antibody is a B-cell depleting antibody.

5. The method of any one of claims 1 to 4, further comprising administering antibodie(s) targeted to human CD 19 and / or human CD20.

6. The method of any one of claims 1 to 5, wherein the NK cells are KIR-B haplotype and homozygous for a CD 16 158V polymorphism.

7. The method of any one of claims 1 to 6, wherein the autoimmune disorder is selected from, Acromegaly, Acquired aplastic anemia, Acquired hemophilia, Primary Agammaglobulinemia. Alopecia areata, Ankylosing spondylitis (AS), Anti-NMDA receptor encephalitis. Antiphospholipid syndrome (APS) | catastrophic antiphospholipid syndrome (CAPS) / Asherson's syndrome, Arteriosclerosis, Autoimmune Addison’s disease (AAD), Autoimmune autonomic ganglionopathy (AAG) / autoimmune dysautonomia | autoimmune gastrointestinal dysmotility (AGID). Autoimmune encephalitis | acute disseminated encephalomyelitis (ADEM). Autoimmune gastritis. Autoimmune hemolytic anemia (AIHA), Autoimmune hepatitis (AIH), Autoimmune hyperlipidemia. Autoimmune hypophysitis, Autoimmune inner ear disease (AIED), Autoimmune lymphoproliferative syndrome (ALPS), Autoimmune myelofibrosis, Autoimmune myocarditis, Autoimmune oophoritis, Autoimmune pancreatitis (AIP), Autoimmune polyglandular syndromes, types I, II, & III (APS type 1, APS type 2, APS type 3, APECED), Autoimmune progesterone dermatitis,Autoimmune retinopathy (AIR), Autoimmune sudden sensorineural hearing loss (SNHL), Balo disease. Behcets disease, Birdshot chorioretinopathy / birdshot uveitis, Bullous pemphigoid, Castleman disease, Celiac disease, Chagas disease. Chronic inflammatory demyelinating polyneuropathy (CIDP), Chronic urticaria (CU), Churg-Strauss syndrome / eosinophilic granulomatosis with polyangiitis (EGPA), Cogan’s syndrome, Cold agglutinin disease, CREST syndrome | limited cutaneous systemic sclerosis, Crohn’s disease (CD), Cronkhite-Canada syndrome (CSS). Cryptogenic organizing pneumonia (COP), Dermatitis herpetiformis, Dermatomyositis, Type 1 Diabetes, Discoid lupus, Dressier’s syndrome / postmyocardial infarction / postpericardiotomy syndrome, Eczema / Atopic Dermatitis, Endometriosis, Eosinophilic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Essential mixed cryoglobulinemia, Evans syndrome, Fibrosing alveolitis / Idiopathic pulmonary’ fibrosis (IPF), Giant cell arteritis I temporal arteritis I Horton’s disease, Giant Cell Myocarditis, Glomerulonephritis, Goodpasture’s syndrome / anti-GBM / anti-TBM disease, Granulomatosis with polyangiitis (GPA) / Wegener’s granulomatosis, Graves disease / thyroid eye disease, Guillain-Bane syndrome (GBS), Hashimoto’s thyroiditis / chronic lymphocytic thyroiditis / autoimmune thyroiditis, Henoch-Schonlein purpura / IgA vasculitis, Hidradenitis suppurativa, Hurst’s disease / acute hemorrhagic leukoencephalitis (AHLE), Hypogammaglobulinemia, IgA nephropathy / Berger's disease, Immune-mediated necrotizing myopathy (IMNM), Immune thrombocytopenia (ITP) / autoimmune thrombocytopenic purpura / autoimmune thrombocytopenia, Inclusion body myositis, IgG4-related sclerosing disease (ISD), Interstitial cystitis. Juvenile idiopathic arthritis / Adult-onset Still's disease, Juvenile polymyositis | Juvenile dermatomyositis | juvenile myositis, Kawasaki disease, Lambert-Eaton myasthenic syndrome (LEMS), Leukocytoclastic vasculitis, Lichen planus, Lichen sclerosus, Ligneous conjunctivitis. Linear IgA disease (LAD) | linear IgA bullous dermatosis (LABD), Lupus nephritis, Lyme disease / chronic Lyme disease / post-treatment Lyme disease syndrome (PTLDS), Lymphocytic colitis / microscopic colitis. Lymphocytic hypophystitis / autoimmune hypophystitis, Meniere’s disease, Microscopic polyangiitis (MPA) / ANCA-associated vasculitis, Mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease. Multifocal motor neuropathy, Multiple sclerosis (MS), Myalgic encephalomyelitis (ME) / Chronic fatigue syndrome (CFS), Myasthenia gravis (MG), Narcolepsy, Neuromyelitis Optica / Devic's disease, Ocular cicatricial pemphigoid, Opsoclonus-myoclonus syndrome (OMS), Palindromic rheumatism, Paraneoplastic cerebellar degeneration, Paraneoplastic pemphigus, Parry-Romberg syndrome (PRS) / Hemifacial atrophy (HFA)ZProgressive facial hemiatrophy, Paroxysmal nocturnal hemoglobinuria(PNH), Peripheral uveitis / pars planitis, PANS / PANDAS, Parsonage-Turner syndrome. Pemphigus gestationis / herpes gestationis, Pemphigus foliaceus, Pemphigus vulgaris, Pernicious anemia, POEMS syndrome. Polyarteritis nodosa. Polymyalgia rheumatica. Polymyositis, Postural orthostatic tachycardia syndrome (POTS), Primary biliary' cirrhosis (PBC) / primary7biliary' cholangitis, Primary sclerosing cholangitis (PSC), Psoriasis, Palmoplantar Pustulosis. Psoriatic arthritis, Pulmonary fibrosis, idiopathic (IPF), Pure red cell aplasia (PRC A), Pyoderma gangrenosum. Rasmussen's encephalitis, Raynaud’s syndrome / phenomenon, Reactive arthritis / Reiter’s syndrome, Reflex sympathetic dystrophy syndrome (RSD) / Complex regional pain syndrome (CRPS), Relapsing polychondritis, Restless leg syndrome (RLS) / Willis-Ekbom disease, Rheumatic fever, Rheumatoid arthritis, Sarcoidosis, Schmidt syndrome / autoimmune polyendocrine syndrome type II, Scleritis. Scleroderma , Sclerosing Mesenteritis I Mesenteric Panniculitis, Serpiginous choroidopathy, Sjogren’s syndrome, Stiff person syndrome (SPS), Small fiber sensory neuropathy, Systemic lupus erythematosus (SLE), Subacute bacterial endocarditis (SBE), Subacute cutaneous lupus, Susac syndrome, Sydenham's chorea. Sympathetic ophthalmia, Takayasu’s arteritis (vasculitis), Testicular autoimmunity (vasculitis, orchitis), Tolosa-Hunt syndrome.Transverse myelitis (TM), Tubulointerstitial nephritis uveitis syndrome (TINU), Ulcerative colitis (UC), Undifferentiated connective tissue disease (UCTD), Uveitis | anterior / intermediate / posterior, Vasculitis, VEXAS Syndrome, Vitiligo, Vogt-Koyanagi- Harada syndrome (VKH), and combinations thereof.

8. The method of claim 7, wherein autoimmune disorder is Systemic lupus erythematosus (SLE).

9. The method of any one of the preceding claims, wherein the patient has lupus nephritis.

10. The method of any of the forgoing claims, wherein the CD38 targeting antibody is daratumumab or isatuximab.

11. The method of any of the forgoing claims, wherein the patient is subjected to lymphodepleting chemotherapy prior to treatment.

12. The method of claim 11, wherein the lymphodepleting chemotherapy is non- myeloablative chemotherapy.

13. The method of any of the forgoing claims, wherein the NK cells are not genetically modified.

14. The method of any of the forgoing claims, wherein at least 70% of the NK cells are CD56+ and CD16+.

15. The method of any of the forgoing claims, wherein at least 85% of the NK cells are CD56+ and CD3-.

16. The method of any of the forgoing claims, wherein 1% or less of the NK cells are CD3+, 1% or less of the NK cells are CD19+ and 1% or less of the NK cells are CD14+.

17. The method of any of the forgoing claims, wherein each administration of NK cells is administration of 1 x 109to 5 x 109NK cells.

18. The method of any of the forgoing claims, wherein each administration of NK cells is administration of 1 x 109to 5 x 109NK cells.

19. The method of any of the forgoing claims wherein the patient receives a dose of the CD38 targeted antibody before the first dose of NK cells.

20. The method of any of the forgoing claims, wherein the expanded natural killer cells are expanded umbilical cord blood natural killer cells.

21. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% CD 16+ cells.

22. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKG2D+ cells.

23. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp46+ cells.

24. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% NKp30+ cells.

25. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%, or 100% DNAM-1+ cells.

26. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises at least 60%, e.g., at least 70%, at least 80%, at least 90% at least 95%, at least 99%. or 100% NKp44+ cells.

27. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises less than 20%, e.g., 10% or less, 5% or less, 1% or less, 0.5% or less, or 0% CD3+ cells.

28. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises less than 20% or less, e.g., 10% or less. 5% or less. 1% or less. 0.5% or less, or 0% CD 14+ cells.

29. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises less than 20% or less, e.g., 10% or less, 5% or less, 1% or less, 0.5% or less, or 0% CD 19+ cells.

30. The method of any of the forgoing claims, wherein the population of expanded natural killer cells comprises 50% or less CD38+ cells, e.g., 20% or less, 10% or less, 5% or less, 1 % or less, 0.5% or less, or 0% CD38+ cells.

31. The method of any of the forgoing claims, wherein the natural killer cells do not comprise a CD 16 transgene.

32. The method of any of the forgoing claims, wherein the natural killer cells do not express an exogenous CD 16 protein.

33. The method of any of the forgoing claims, wherein the expanded natural killer cells are not genetically engineered.

34. The method of any of the forgoing claims, wherein the expanded natural killer cells are derived from the same umbilical cord blood donor.

35. The method of any of the forgoing claims, wherein the population of NK cells comprises at least 100 million expanded natural killer cells, e.g., 200 million. 250 million. 300 million, 400 million, 500 million, 600 million, 700 million, 750 million, 800 million, 900 million, 1 billion, 2 billion, 3 billion, 4 billion, 5 billion, 6 billion, 7 billion, 8 billion, 9 billion, 10 billion, 15 billion, 20 billion, 25 billion, 50 billion. 75 billion, 80 billion, 9- billion, 100 billion, 200 billion, 250 billion, 300 billion. 400 billion, 500 billion, 600 billion, 700 billion, 800 billion, 900 billion, 1 trillion, 2 trillion, 3 trillion, 4 trillion, 5 trillion, 6 trillion, 7 trillion, 8 trillion, 9 trillion, or 10 trillion expanded natural killer cells.

36. The method of any of the forgoing claims, wherein the population of NK cells is produced by a method comprising:(a) obtaining seed cells comprising natural killer cells from umbilical cord blood;(b) depleting the seed cells of CD3+ cells;(c) expanding the natural killer cells by culturing the depleted seed cells with a first plurality of Hut78 cells engineered to express a membrane bound IL-21, a mutated TNFa, and a 4-1 BBL gene to produce expanded natural killer cells, thereby producing the population of expanded natural killer cells.

37. The method of any of the forgoing claims, wherein the population of NK cells is produced by a method comprising:(a) obtaining seed cells comprising natural killer cells from umbilical cord blood;(b) depleting the seed cells of CD3+ cells;(c) expanding the natural killer cells by culturing the depleted seed cells with a first plurality of Hut78 cells engineered to express a membrane bound IL-21, a mutated TNFa,and a 4-1 BBL gene to produce a master cell bank population of expanded natural killer cells; and(d) expanding the master cell bank population of expanded natural killer cells by culturing with a second plurality of Hut78 cells engineered to express a membrane bound IL- 21, a mutated TNFa, and a 4-1BBL gene to produce expanded natural killer cells; thereby producing the population of expanded natural killer cells.

38. The method of claim 36 or claim 37. wherein the population of NK cells is produced by a method further comprising, after step (c),(i) freezing the master cell bank population of expanded natural killer cells in a plurality7of containers; and(ii) thawing a container comprising an aliquot of the master cell bank population of expanded natural killer cells, wherein expanding the master cell bank population of expanded natural killer cells in step (d) comprises expanding the aliquot of the master cell bank population of expanded natural killer cells.

39. The method of any one of claims 36 to 38. wherein the umbilical cord blood is from a donor with the KIR-B haplotype and homozygous for the CD 16 158V polymorphism.

40. The method of any one of claims 36-39, wherein the population of NK cells is produced by a method comprising expanding the natural killer cells from umbilical cord blood at least 10.000 fold, e.g., 15,000 fold, 20,000 fold, 25,000 fold. 30,000 fold, 35,000 fold, 40,000 fold, 45,000 fold. 50.000 fold, 55,000 fold. 60.000 fold, 65,000 fold, or 70,000 fold.

41. The method of any one of claims 36-40, wherein the population of expanded natural killer cells is not enriched or sorted after expansion.

42. The method of any one of claims 36-41, wherein the percentage of NK cells expressing CD 16 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

43. The method of any one of claims 36-42, wherein the percentage of NK cells expressing NKG2D in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

44. The method of any one of claims 36-43, wherein the percentage of NK cells expressing NKp30 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

45. The method of any one of claims 36-44, wherein the percentage of NK cells expressing NKp44 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

46. The method of any one of claims 36-45, wherein the percentage of NK cells expressing NKp46 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

47. The method of any one of claims 36-46, wherein the percentage of NK cells expressing DNAM-1 in the population of expanded natural killer cells is the same or higher than the percentage of natural killer cells in the seed cells from umbilical cord blood.

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