Compositions and methods for treating cancer using cell therapy

Abstract

Various embodiments of this disclosure provide therapeutic compositions and related methods for cell therapies comprising one or more cells, such as allogeneic cells, including chimeric receptors and improved hematopoietic stem cell transplantation, including methods for enhancing protection from graft-versus-host disease while maintaining an effective immune response, such as a graft-versus-tumor immune response. In preferred embodiments, the cell therapy comprises (a) CD19 and CD22 bispecific chimeric antigen receptor (CAR) T cells, and (b) allogeneic hematopoietic stem cell transplantation (HSCT) comprising one or more populations selected from a population of CD34+ hematopoietic stem progenitor cells (HSPCs), a population of conventional CD3+ T cells (Tcon), a population of CD4+CD25+CD127dim regulatory T cells (Treg), and / or a population of CD45RA-memory T cells (Tmem).

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Patent Application No. 63 / 580,701, filed on 5 September 2023, and U.S. Provisional Patent Application No. 63 / 638,850, filed on 25 April 2024, both of which are incorporated herein by reference in their entirety for all purposes.

[0002] Sequence List This application includes a sequence listing submitted electronically in XML format, the entirety of which is incorporated herein by reference. The XML copy created in ### ##,20## is named ###_###_sequencelisting.XML and has a size of #,###,### bytes. [Background technology]

[0003] background For example, patients with hematological malignancies such as leukemia and lymphoma who do not achieve initial remission or experience refractory relapses are rarely cured with standard chemotherapy. Myeloablative allogeneic hematopoietic stem cell transplantation (alloHCT) improves survival rates in these patients. Myeloablative alloHCT is a treatment in which the patient receives chemotherapy or radiation to remove or destroy the bone tissue causing the malignancy. The patient is then administered hematopoietic cells, including hematopoietic stem progenitor cells (HSPCs) derived from the donor's blood. However, alloHCT has a major drawback in that it often causes graft-versus-host disease (GVHD). GVHD is a disease in which the transplanted donor peripheral blood stem cells perceive the patient's body as exogenous, and the donor cells attack the patient's tissues (e.g., skin, GI tissue, liver tissue, and lung tissue), leading to many complications, many of which are serious and can result in morbidity and death. Recent preclinical studies using mouse models have shown that CD19-positive lymphoma-bearing BALB / c mice, recipients of adoptive transfer of donor-derived 19-28z CAR T cells after B6 allogeneic transplantation, were less likely to die from GVHD compared to control mice. Therefore, there is a need for improved methods and therapies for the administration of chimeric receptor-expressing allogeneic cells in combination with hematopoietic stem cell transplantation to reduce the incidence and severity of GVHD, including reduced morbidity and mortality. [Overview of the Initiative]

[0004] overview In certain embodiments, a method for treating a human subject having or suspected of having a malignant tumor is described herein, comprising administering a cell therapy to the human subject, wherein the cell therapy comprises (a) one or more allogeneic cells containing a chimeric receptor, and (b) an immunomodulatory cell therapy product, wherein one or more of the allogeneic cells and / or one or more of the immunomodulatory cell therapy product are derived from an HLA-matched donor to the human subject. In some embodiments, the one or more cells containing a chimeric receptor comprises an isolated nucleic acid encoding the chimeric receptor.

[0005] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + A method comprising (b) a population of hematopoietic stem progenitor cells (HSPCs) and (b) a pharmaceutical composition comprising one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0006] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy is CD34 + A method comprising a population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing a chimeric receptor, is described herein.

[0007] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD4 + CD25 + CD127 dim A method comprising (b) a population of regulatory T cells (Tregs) and (b) a pharmaceutical composition comprising one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0008] In a particular embodiment, a method for treating a human subject having or suspected of having a hematological malignancy, comprising administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy is CD4 + CD25 + CD127 dim A method comprising a population of regulatory T cells (Tregs), wherein the population of Tregs comprises one or more cells containing a chimeric receptor, is described herein.

[0009] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD45RA- Described herein is a method comprising: a population of memory T cells (Tmem); and (b) a pharmaceutical composition comprising one or more cells comprising a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor.

[0010] In certain embodiments, provided is a method of treating a human subject having or suspected of having a hematological malignancy, the method comprising administering to the human subject a multi-component cell therapy, wherein the multi-component cell therapy comprises CD45RA - Described herein is a method comprising a population of memory T cells (Tmem), wherein the population of Tmem comprises one or more cells comprising a chimeric receptor.

[0011] In certain embodiments, provided is a method of treating a human subject having or suspected of having a hematological malignancy, the method comprising administering to the human subject a multi-component cell therapy, wherein the multi-component cell therapy comprises: (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC); and (b) CD4 + CD25 + CD127 dim Described herein is a method comprising: a population of regulatory T cells (Treg); and (c) a pharmaceutical composition comprising one or more cells comprising a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor.

[0012] In certain embodiments, provided is a method of treating a human subject having or suspected of having a hematological malignancy, the method comprising administering to the human subject a multi-component cell therapy, wherein the multi-component cell therapy comprises: (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC); and (b) CD4 + CD25 + CD127 dim Described herein is a method comprising a population of regulatory T cells (Treg), wherein the population of HSPC, the population of Treg, or the population of HSPC and the population of Treg comprises one or more cells comprising a chimeric receptor.

[0013] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD45RA - A method comprising (c) a population of memory T cells (Tmem) and a pharmaceutical composition comprising (c) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0014] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD45RA - A method is described herein that includes a population of memory T cells (Tmem), wherein the population of HSPCs, the population of Tmem, or the population of HSPCs and the population of Tmem comprises one or more cells containing a chimeric receptor.

[0015] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and (b) normal CD3 + A method comprising (c) a population of T cells (Tcon) and a pharmaceutical composition comprising (c) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor is described herein.

[0016] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and (b) normal CD3 +Methods comprising a population of T cells (Tcon), wherein the population of HSPCs, the population of Tcon, or the population of HSPCs and Tcon comprises one or more cells containing a chimeric receptor, are described herein.

[0017] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim (c) A population of regulatory T cells (Tregs) and a normal CD3 + A method comprising (d) a population of T cells (Tcon) and a pharmaceutical composition comprising one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0018] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim (c) A population of regulatory T cells (Tregs) and a normal CD3 + A method is described herein that includes a population of T cells (Tcon), wherein the population of HSPCs, a population of Tregs, and / or the population of Tcon includes one or more cells containing a chimeric receptor.

[0019] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim A population of regulatory T cells (Tregs) and (c)CD45RA -A population of memory T cells (Tmem) and (d) normal CD3 + A method comprising (e) a population of T cells (Tcon) and a pharmaceutical composition comprising one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0020] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim A population of regulatory T cells (Tregs) and (c)CD45RA - A population of memory T cells (Tmem) and (c) normal CD3 + A method is described herein that includes a population of T cells (Tcon), wherein the population of HSPCs, a population of Tregs, a population of Tmems, and / or a population of Tcon cells comprises one or more cells containing a chimeric receptor.

[0021] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim (c) A population of regulatory T cells (Tregs) and a normal CD3 + A method comprising a population of T cells (Tcon), wherein the population of Tcon comprises one or more cells containing a chimeric receptor, is described herein.

[0022] In certain embodiments, a method for treating a human subject having or suspected of having a hematological malignancy comprises administering a multicomponent cell therapy to the human subject, wherein the multicomponent cell therapy comprises (a) CD34 + A population of hematopoietic stem progenitor cells (HSPCs), CD4 + CD25 +CD127 dim A population of regulatory T cells (Tregs), typical CD3 + A method comprising (b) a population of T cells (Tcon) or any combination thereof, and (b) a population of tumor-infiltrating lymphocytes (TILs) is described herein.

[0023] In certain embodiments, methods for treating human subjects having or suspected to have a hematological disorder are described herein, the method comprising (a) administering a single pre-transplant conditioning regimen and (b) administering two or more cell therapies.

[0024] In certain embodiments, multicomponent cell therapy products are described herein, comprising (a) one or more cells containing a chimeric receptor, or a pharmaceutical composition comprising an isolated nucleic acid encoding a chimeric receptor, and (b) an immunomodulatory cell therapy product. In certain embodiments, one or more cells comprising the chimeric receptor or the pharmaceutical composition, and / or one or more cells of the immunomodulatory cell therapy product are allogeneic cells derived from an HLA-matched donor to a human subject receiving the multicomponent cell therapy product.

[0025] In certain embodiments, a multicomponent cell therapy product is described herein, comprising (a) one or more allogeneic cells containing a chimeric receptor, and (b) an immunomodulatory cell therapy product, wherein one or more allogeneic cells and / or one or more cells of the immunomodulatory cell therapy product are derived from an HLA-matched donor to a human subject receiving the multicomponent cell therapy product.

[0026] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + A multicomponent cell therapy product comprising (b) a population of hematopoietic stem progenitor cells (HSPCs) and (b) one or more cells containing a chimeric receptor, or a pharmaceutical composition comprising an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0027] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 +This specification describes a multicomponent cell therapy product comprising a population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing a chimeric receptor.

[0028] In a particular embodiment, the product for multicomponent cell therapy is (a) CD4 + CD25 + CD127 dim A multicomponent cell therapy product comprising (b) a population of regulatory T cells (Tregs) and (b) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0029] In a particular embodiment, CD4 + CD25 + CD127 dim This specification describes a multicomponent cell therapy product containing a population of regulatory T cells (Tregs), wherein the Treg population contains one or more cells containing a chimeric receptor.

[0030] In a particular embodiment, the product for multicomponent cell therapy is (a) CD45RA - Multicomponent cell therapy products comprising (b) a population of memory T cells (Tmem) and (b) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, are described herein.

[0031] In a specific embodiment, CD45RA - This specification describes a multicomponent cell therapy product containing a population of memory T cells (Tmem), wherein the Tmem population contains one or more cells containing a chimeric receptor.

[0032] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dimA multicomponent cell therapy product comprising (c) a population of regulatory T cells (Tregs) and a pharmaceutical composition comprising (c) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0033] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD4 + CD25 + CD127 dim This specification describes multicomponent cell therapy products comprising a population of regulatory T cells (Tregs), wherein the population of HSPCs, the population of Tregs, or the population of HSPCs and the population of Tregs comprises one or more cells containing a chimeric receptor.

[0034] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD45RA - A multicomponent cell therapy product comprising (c) a population of memory T cells (Tmem) and a pharmaceutical composition comprising (c) one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor is described herein.

[0035] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and CD45RA - This specification describes a multicomponent cell therapy product comprising a population of memory T cells (Tmem), wherein the population of HSPCs, the population of Tmem, or the population of HSPCs and Tmem comprises one or more cells containing a chimeric receptor.

[0036] In a particular embodiment, the product for multicomponent cell therapy is (a) CD34 + (b) A population of hematopoietic stem progenitor cells (HSPCs) and (b) normal CD3 +Described herein is a multi-component cell therapy product comprising a population of T cells (Tcon) and (c) a pharmaceutical composition comprising one or more cells comprising a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor.

[0037] In certain embodiments, provided herein is a multi-component cell therapy product, wherein (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC), and (b) conventional CD3 + a population of T cells (Tcon), wherein the population of HSPC, the population of Tcon, or the population of HSPC and the population of Tcon comprises one or more cells comprising a chimeric receptor, is described herein.

[0038] In certain embodiments, provided herein is a multi-component cell therapy product, wherein (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC), and (b) CD4 + CD25 + CD127 dim a population of regulatory T cells (Treg), (c) conventional CD3 + a population of T cells (Tcon), and (d) a pharmaceutical composition comprising one or more cells comprising a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, is described herein.

[0039] In certain embodiments, provided herein is a multi-component cell therapy product, wherein (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC), and (b) CD4 + CD25 + CD127 dim a population of regulatory T cells (Treg), and (c) conventional CD3 + a population of T cells (Tcon), wherein the population of HSPC, the population of Treg, and / or the population of Tcon comprises one or more cells comprising a chimeric receptor, is described herein.

[0040] In certain embodiments, provided herein is a multi-component cell therapy product, wherein (a) CD34 + a population of hematopoietic stem and progenitor cells (HSPC), and (b) CD4 +CD25 + CD127 dim a population of regulatory T cells (Treg), and (c) CD45RA - a population of memory T cells (Tmem), and (d) conventional CD3 + a population of T cells (Tcon), and (e) a pharmaceutical composition comprising one or more cells comprising a chimeric receptor, or an isolated nucleic acid encoding a chimeric receptor, described herein is a product for multicomponent cell therapy.

[0041] In certain embodiments, there is provided herein a product for multicomponent cell therapy, comprising (a) CD34 + a population of hematopoietic stem progenitor cells (HSPC), and (b) CD4 + CD25 + CD127 dim a population of regulatory T cells (Treg), and (c) CD45RA - a population of memory T cells (Tmem), and (d) conventional CD3 + a population of T cells (Tcon), wherein the population of HSPC, the population of Treg, the population of Tmem, and / or the population of Tcon comprises one or more cells comprising a chimeric receptor, described herein is a product for multicomponent cell therapy.

[0042] In certain embodiments, there is provided herein a product for multicomponent cell therapy, comprising (a) CD34 + a population of hematopoietic stem progenitor cells (HSPC), and (b) CD4 + CD25 + CD127 dim a population of regulatory T cells (Treg), and (c) conventional CD3 + a population of T cells (Tcon), wherein the population of Tcon comprises one or more cells comprising a chimeric receptor, described herein is a product for multicomponent cell therapy.

[0043] In certain embodiments, described herein is a kit for use in preparing a cell therapy product, the kit comprising (a) an anti-human CD34 affinity reagent, (b) an anti-human CD25 affinity reagent, (c) a vector comprising a nucleic acid encoding a chimeric receptor, and (d) a transfection reagent.

[0044] Built-in by reference All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent that each individual publication, patent, or patent application is specifically and individually incorporated by reference. [Brief explanation of the drawing]

[0045] The novelty of this disclosure is described in detail in the appended claims. A deeper understanding of the features and advantages of this disclosure will be obtained by referring to the following detailed description illustrating explanatory embodiments in which the principles of this disclosure are utilized, and to the appended drawings.

[0046] [Figure 1] This diagram illustrates the pre-transplant conditioning regimen and administration timeline for CAR T production and immunomodulatory cell therapy products prior to CAR T injection. [Figure 2] The results of a flow cytometry assay evaluating CD19 CAR expression are shown. [Figure 3] The results of a CAR T-cell cytotoxicity assay, which evaluates the percentage of T-cell killing, are shown. [Figure 4] Figure 4A shows a histogram of the CellTrace proliferation assay, which evaluates CD3 T cell proliferation. Figure 4B shows the quantitative results of the CellTrace proliferation assay, which evaluates CD3 T cell proliferation. [Figure 5] Figure 5A shows a histogram of the CellTrace proliferation assay for evaluating CAR T cell proliferation. Figure 5B shows the quantitative results of the CellTrace proliferation assay for evaluating CAR T cell proliferation. [Figure 6] The baseline characteristics and clinical outcomes of patients treated with immunomodulatory cell therapy products are illustrated. [Figure 7]Figure 7A shows the amplification and persistence of donor CD4+CD19 / CD22 CAR T cells in patients treated with immunomodulatory cell therapy products. Figure 7B shows the amplification and persistence of donor CD8+CD19 / CD22 CAR T cells in patients treated with immunomodulatory cell therapy products. Figure 7C shows the amplification and persistence of total donor CD19 / CD22 CAR T cells in patients treated with immunomodulatory cell therapy products. [Figure 8] The qPCR results showing amplification and persistence of donor CD19 / CD22 CAR T cells in patients treated with immunomodulatory cell therapy products are illustrated. [Figure 9] Figure 9A shows the proportion of whole blood donor chimeras in recipients of donor CD19 / CD22 CAR T cells. Figure 9B shows the proportion of CD3+ cell donor chimeras in recipients of donor CD19 / CD22 CAR T cells. [Figure 10] The baseline characteristics and clinical outcomes of patients treated with immunomodulatory cell therapy products are illustrated. [Modes for carrying out the invention]

[0047] Detailed explanation Introduction This specification describes a multicomponent cell therapy product comprising (a) one or more cells (e.g., allogeneic cells) containing a chimeric receptor, and (b) an immunomodulatory cell therapy product, wherein one or more allogeneic cells and / or one or more cells of the immunomodulatory cell therapy product are derived from an HLA-matched donor to a human subject that receives the multicomponent cell therapy product, and a method of using the multicomponent cell therapy product for treating a human subject having or suspected of having a malignant tumor. In some embodiments, the one or more cells containing the chimeric receptor comprises a pharmaceutical composition comprising an isolated nucleic acid encoding the chimeric receptor.

[0048] This specification describes a multicomponent cell therapy product comprising (a) one or more cells (e.g., allogeneic cells) containing a binder, and (b) an immunomodulatory cell therapy product, wherein one or more allogeneic cells and / or one or more cells of the immunomodulatory cell therapy product are derived from an HLA-matched donor to a human subject that receives the multicomponent cell therapy product, and a method of using the multicomponent cell therapy product for treating a human subject having or suspected of having a malignant tumor. In some embodiments, one or more cells containing a chimeric receptor comprises a pharmaceutical composition comprising an isolated nucleic acid encoding the chimeric receptor.

[0049] The immunomodulatory cell therapy product may include regulatory T cell therapy, for example, any of the regulatory T cell therapies described herein. The immunomodulatory cell therapy product may also include allogeneic hematopoietic stem cell transplantation (HSCT), for example, any of the allo-HSCTs described herein.

[0050] Binding agents, chimeric receptors, and cells containing them As provided herein, the cells of the Disclosure may comprise one or more binders that provide means for binding to one or more epitopes of a target of interest, such as target CD19, CD20, CD22, and / or BCMA. In certain embodiments, the means for binding to the target of interest (e.g., CD19, CD20, CD22, and / or BCMA) comprises an antibody or its antigen-binding fragment or equivalent. In certain embodiments, the means for binding to the target of interest (e.g., CD19, CD20, CD22, and / or BCMA) is a binder comprising a T cell receptor (TCR) or a chimeric antigen receptor (CAR). In certain embodiments, the means for binding to the target of interest (e.g., CD19, CD20, CD22, and / or BCMA) is a binder comprising one or more CARs of the Disclosure having antigen specificity for CD19, CD20, CD22, and / or BCMA. In certain embodiments, the means for binding to a target of interest (e.g., CD19, CD20, CD22, and / or BCMA) is a binder comprising one or more of the CDR1, CDR2, and CDR3 regions of a heavy chain variable region, a light chain variable region, or an antigen-binding domain provided in this disclosure, for example, an antigen-binding domain having an antigen specific to CD19, CD20, CD22, and / or BCMA.

[0051] As provided herein, the cells of this disclosure may comprise one or more chimeric receptors. The chimeric receptors of this disclosure may comprise, without limitation, T cell receptors (TCRs) or chimeric antigen receptors (CARs).

[0052] In some embodiments, the cells of this disclosure containing a chimeric receptor may include a chimeric antigen receptor (CAR), which is an artificially constructed hybrid protein or polypeptide containing the antigen-binding domain of one or more antibodies (e.g., single-strand variable fragments (scFv)) linked to a T cell signaling domain. A key feature of CARs is their ability to redirect T cell specificity and reactivity to selected targets in an MHC-independent manner by utilizing the antigen-binding properties of monoclonal antibodies. MHC-independent antigen recognition gives T cells expressing CARs the ability to recognize antigens independently of antigen treatment and thus bypass the major mechanisms of tumor escape. Furthermore, when expressed in T cells, CARs advantageously do not dimerize with the endogenous T cell receptor (TCR) alpha and beta chains.

[0053] As used herein, the terms “antigen(ic) specificity” and “induces antigen-specific response” mean that the CAR specifically binds to an antigen and is immunologically recognized such that the binding of the CAR to the antigen induces an immune response.

[0054] As used herein, the terms “dual specificity” and “dual specific” mean that the same CAR can specifically bind to and immunologically recognize two different antigens such that the binding of the CAR to at least one of two antigens triggers an immune response.

[0055] In some embodiments, the CARs of the Disclosure may have antigen specificity for CD19. In some embodiments, the CARs of the Disclosure may have antigen specificity for CD20. In some embodiments, the CARs of the Disclosure may have antigen specificity for CD22. In some embodiments, the CARs of the Disclosure may have antigen specificity for both CD20 and CD19. In some embodiments, the CARs of the Disclosure may have antigen specificity for both CD22 and CD19. In some embodiments, the CARs of the Disclosure may have antigen specificity for both CD20 and CD22. In some embodiments, the CARs of the Disclosure may have antigen specificity for CD19, CD20, and CD22.

[0056] CD22 is a lineage-specific B-cell antigen belonging to the immunoglobulin (Ig) superfamily. CD22 is expressed in 60-70% of B-cell lymphomas and leukemias (e.g., B-cell chronic lymphocytic leukemia, hairy cell leukemia, acute lymphoblastic leukemia (ALL), and Burkitt lymphoma) and is absent from the cell surface in the early stages of B-cell development or in stem cells. Vaickus et al., Crit. Rev. Oncol. / Hematol., 11:267-297 (1991); Bang et al., Clin. Cancer Res., 11:1545-50 (2005). CD19 (also known as B lymphocyte antigen CD19, B4, and CVID3) is a cell surface molecule expressed only by hematopoietic B lymphocytes and follicular dendritic cells. It is the earliest expressed B-lineage-specific antigen and is present on most precursor B cells and most non-T-cell acute lymphoblastic leukemia cells and B-cell chronic lymphoblastic leukemia cells (Tedder and Isaacs, J.Immun., 143:712-717 (1989)). In other embodiments, the CARs of this disclosure may have antigen specificity for BMCA.

[0057] The bispecific CARs of the present invention may offer several advantages. For example, the bispecific CARs of the present invention may advantageously offer greater efficacy compared to CARs that have antigen specificity to only one of CD19 and CD22 (but not both). The bispecific CARs of the present invention may also advantageously reduce or prevent cancer cell escape caused by the loss of expression of either CD19 or CD22 by cancer cells. For example, the bispecific CARs of the present invention are thought to reduce or prevent recurrence observed in cancer patients who have been treated with a CAR that has antigen specificity to CD19 only, and whose cancer has lost CD19 expression. The bispecific CARs of the present invention may also increase the patient population that can be successfully treated. For example, patients who may not respond to CAR therapy targeting only CD19 may respond to CAR therapy targeting CD22, and patients who may not respond to CAR therapy targeting only CD22 may respond to CAR therapy targeting CD19.

[0058] Although not bound by any particular theory or mechanism, the CAR of the present invention is thought to provide one or more of the following by inducing an antigen-specific response to CD22 and CD19: targeting and destruction of CD22-expressing cancer cells, targeting and destruction of CD19-expressing cancer cells, reduction or elimination of cancer cells, promotion of immune cell infiltration into tumor sites, and enhancement / extension of the anti-cancer response.

[0059] Embodiments of the present disclosure provide a CAR comprising the anti-CD22 antigen-binding domain (m971) of the m971 antibody. The antigen-binding domain of m971 specifically binds to CD22. In this regard, preferred embodiments of the present disclosure provide a CAR comprising an anti-CD22 antigen-binding domain comprising, consisting of, or essentially consisting of a single-stranded variable fragment (scFv) of the antigen-binding domain of m971. The HA22 immunotoxin and the m971 antibody bind to different CD22 epitopes.

[0060] The anti-CD22 antigen-binding domain may include a light chain variable region and / or a heavy chain variable region. In embodiments of this disclosure, the heavy chain variable region includes a CDR1 region, a CDR2 region, and a CDR3 region. In this regard, the anti-CD22 antigen-binding domain may include one or more of the following: a heavy chain CDR1 region containing the amino acid sequence of SEQ ID NO: 1, a heavy chain CDR2 region containing the amino acid sequence of SEQ ID NO: 2, and a heavy chain CDR3 region containing the amino acid sequence of SEQ ID NO: 3. Preferably, the heavy chain of the anti-CD22 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 1 to 3.

[0061] In embodiments of this disclosure, the light chain variable region of the anti-CD22 antigen-binding domain may include a light chain CDR1 region, a light chain CDR2 region, and a light chain CDR3 region. In this regard, the anti-CD22 antigen-binding domain may include one or more of the light chain CDR1 region containing the amino acid sequence of SEQ ID NO: 4, the light chain CDR2 region containing the amino acid sequence of SEQ ID NO: 5, and the light chain CDR3 region containing the amino acid sequence of SEQ ID NO: 6. Preferably, the light chain of the anti-CD22 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 4 to 6. In particularly preferred embodiments, the anti-CD22 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 1 to 6.

[0062] The heavy chain variable region of the anti-CD22 antigen-binding domain includes, consists of, or is essentially composed of, the amino acid sequence of SEQ ID NO: 13. The light chain variable region of the anti-CD22 antigen-binding domain includes, consists of, or is essentially composed of, the amino acid sequence of SEQ ID NO: 14. Therefore, in embodiments of the present disclosure, the anti-CD22 antigen-binding domain includes a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 13 and / or a light chain variable region containing the amino acid sequence of SEQ ID NO: 14. Preferably, the anti-CD22 antigen-binding domain includes both the amino acid sequences of SEQ ID NO: 13 and 14.

[0063] Embodiments of the present disclosure provide a CAR comprising the anti-CD19 antigen-binding domain ("FMC63") of an FMC63 antibody. The antigen-binding domain of FMC63 specifically binds to CD19. In this regard, preferred embodiments of the present disclosure provide a CAR comprising an anti-CD19 antigen-binding domain comprising, consisting of, or essentially consisting of, a single-stranded variable fragment (scFv) of the antigen-binding domain of FMC63.

[0064] The anti-CD19 antigen-binding domain may include a light chain variable region and / or a heavy chain variable region. In embodiments of this disclosure, the heavy chain variable region of the anti-CD19 antigen-binding domain includes a CDR1 region, a CDR2 region, and a CDR3 region. In this regard, the anti-CD19 antigen-binding domain may include one or more of the heavy chain CDR1 region containing the amino acid sequence of SEQ ID NO: 7, the heavy chain CDR2 region containing the amino acid sequence of SEQ ID NO: 8, and the heavy chain CDR3 region containing the amino acid sequence of SEQ ID NO: 9. Preferably, the heavy chain of the anti-CD19 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 7 to 9.

[0065] In embodiments of this disclosure, the light chain variable region of the anti-CD19 antigen-binding domain may include a light chain CDR1 region, a light chain CDR2 region, and a light chain CDR3 region. In this regard, the anti-CD19 antigen-binding domain may include one or more of the light chain CDR1 region containing the amino acid sequence of SEQ ID NO: 10, the light chain CDR2 region containing the amino acid sequence of SEQ ID NO: 11, and the light chain CDR3 region containing the amino acid sequence of SEQ ID NO: 12. Preferably, the light chain of the anti-CD19 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 10 to 12. In a particularly preferred embodiment, the anti-CD19 antigen-binding domain contains all the amino acid sequences of SEQ ID NOs from 7 to 12.

[0066] The heavy chain variable region of the anti-CD19 antigen-binding domain includes, consists of, or is essentially composed of, the amino acid sequence of SEQ ID NO: 15. The light chain variable region of the anti-CD19 antigen-binding domain includes, consists of, or is essentially composed of, the amino acid sequence of SEQ ID NO: 16. Therefore, in embodiments of the present disclosure, the anti-CD19 antigen-binding domain includes a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 15 and / or a light chain variable region containing the amino acid sequence of SEQ ID NO: 16. Preferably, the anti-CD19 antigen-binding domain includes the amino acid sequences of both SEQ ID NO: 15 and 16.

[0067] Some embodiments of this disclosure provide a CAR comprising the anti-CD20 antigen-binding domain of a Leu16 antibody ("Leu16"). Some embodiments of this disclosure provide a CAR comprising the anti-CD20 antigen-binding domain of a 1F5 antibody ("1F5"). In some embodiments, the CAR of this disclosure is C-CAR066.

[0068] A bispecific CAR has an anti-CD22 antigen-binding domain and an anti-CD19 antigen-binding domain. In one embodiment of this disclosure, the CAR includes all six CDR regions of both the anti-CD22 antigen-binding domain and the anti-CD19 antigen-binding domain. In this regard, the CAR may include all of SEQ ID NOs: 1-12. In another embodiment of this disclosure, the CAR includes the light chain variable region and the heavy chain variable region of both the anti-CD22 antigen-binding domain and the anti-CD19 antigen-binding domain. In this regard, the CAR may include all of SEQ ID NOs: 13-16.

[0069] In some embodiments, the sequences of the light and heavy chain CDR regions and the light and heavy chain variable regions are selected from Table 1A. In some embodiments, the CDRs can be annotated and / or defined according to various CDR annotation schemes, such as the IMGT definition (see Lefranc, (1999) The Immunologist, 7, 132-136), the Kabat numbering scheme (see Kabat et al., 1992, Sequences of Proteins of Immunological Interest, DIANE Publishing: 2719), the Chothia numbering scheme (see Chothia and Lesk, J Mol Biol, 1987, 196: 901-917), or a combination of Kabat and Chothia. Other CDR annotation schemes include, but are not limited to, the AbM definition (see Whitelegg & Rees, Protein Eng. 2000, 13:819-824, Whitelegg & Rees, Methods Mol Biol. 2004, 248:51-91) and / or the Contact definition (see MacCallum et al., J.Mol.Biol. 1996, 262:732-745). While the CDR sequences provided herein may conform to one CDR annotation scheme (e.g., IMGT, Kabat, and / or Chothia), those skilled in the art can recognize and determine CDR sequences based on other CDR annotation schemes through the analysis and annotation of light chain and heavy chain variable region sequences described herein.

[0070] (Table 1A) TIFF2026532606000002.tif111152

[0071] The anti-CD22 antigen-binding domain and the anti-CD19 antigen-binding domain may each contain any antigen-binding portion of an anti-CD22 or anti-CD19 antibody. The antigen-binding portion can be any portion having at least one antigen-binding site, such as Fab, F(ab')2, dsFv, scFv, bispecific antibodies, and trispecific antibodies. Preferably, the antigen-binding portion is a single-stranded variable region fragment (scFv) antibody fragment. scFv is a shortened Fab fragment containing a variable (V) domain of an antibody heavy chain linked to the V domain of a light chain antibody via a synthetic peptide linker, which can be produced using techniques of conventional recombinant DNA technology. Similarly, disulfide-stabilized variable region fragments (dsFv) can be prepared by recombinant DNA technology.

[0072] Some embodiments of this disclosure provide a CAR comprising the anti-CD22 antigen-binding domain of a YK-CD22 antibody ("YK-CD22"). Some embodiments of this disclosure provide a CAR comprising the anti-CD22 antigen-binding domain of an m971 antibody ("m971"). Some embodiments of this disclosure provide a CAR comprising the anti-CD22 antigen-binding domain of an FH80 antibody ("FH80").

[0073] In one embodiment of the present disclosure, the light chain variable region and the heavy chain variable region of the anti-CD22 antigen-binding domain are linked to each other by a linker. The linker may comprise any preferred amino acid sequence. In one embodiment of the present disclosure, the linker is a Gly / Ser linker having an amino acid length of about 1 to about 100, about 3 to about 20, about 5 to about 30, about 5 to about 18, or about 3 to about 8 amino acids, and consisting of glycine and / or serine residues in the sequence. Thus, the Gly / Ser linker may consist of glycine and / or serine residues. In some embodiments, the Gly / Ser linker is of the formula: (Xaal) nThe peptide is such that each amino acid residue Xaal is independently selected from glycine and serine, and n is an integer between 3 and 8. Preferably, the Gly / Ser linker contains the amino acid sequence of SEQ ID NO: 19 (GGGGS). In another embodiment of the present disclosure, the linker contains the amino acid sequence of SEQ ID NO: 17 (GSTSGSGKPGSGEGSTKG, also known as the "218 linker").

[0074] In one embodiment of the present disclosure, the light chain variable region and the heavy chain variable region of the anti-CD19 antigen-binding domain are linked to each other by a linker. The linker may be any of the linkers described herein in relation to other embodiments of the present disclosure. In one embodiment of the present disclosure, the light chain variable region and the heavy chain variable region of the anti-CD19 antigen-binding domain are linked to each other by a linker comprising the amino acid sequence of SEQ ID NO: 17.

[0075] In one embodiment, the anti-CD22 antigen-binding domain includes a light chain variable region, a heavy chain variable region, and a linker. In this regard, embodiments of the anti-CD22 antigen-binding domain, including the light chain variable region, the heavy chain variable region, and the 218 linker, include, consist of, or essentially consist of, all of SEQ ID NOs: 1-6 and 17, all of SEQ ID NOs: 13, 14, and 17, or SEQ ID NO: 20 (QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSSGSTSGSGKPGSGEGSTKGDIQMTQSPSSLSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK). Embodiments of the anti-CD22 antigen-binding domain, including the light chain variable region, the heavy chain variable region, and the Gly / Ser linker, include, consist of, or essentially consist of, all of SEQ ID NOs: 1-6 and 19, all of SEQ ID NOs: 13, 14, and 19, or SEQ ID NO: 21 (QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK).

[0076] In one embodiment, the anti-CD19 antigen-binding domain includes a light chain variable region, a heavy chain variable region, and a linker. In this regard, embodiments of the anti-CD19 antigen-binding domain including the light chain variable region, the heavy chain variable region, and the 218 linker include, consist of, or essentially consist of, all of SEQ ID NOs: 7-12 and 17, all of SEQ ID NOs: 15-17, or SEQ ID NO: 22 (DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGSGKPGSGEGSTKGEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS). Embodiments of the anti-CD19 antigen-binding domain, including the light chain variable region, the heavy chain variable region, and the Gly / Ser linker, include, consist of, or essentially consist of, all of SEQ ID NOs: 7-12 and 19, all of SEQ ID NOs: 15, 16, and 19, or SEQ ID NO: 41 (DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS).

[0077] In embodiments of this disclosure, the anti-CD19 antigen-binding domain and the anti-CD22 antigen-binding domain are linked to each other by a linker. The linker may comprise any preferred amino acid sequence. The linker may comprise, for example, any of the linkers described herein in relation to other embodiments of this disclosure. Preferably, the linker linking the anti-CD19 antigen-binding domain and the anti-CD22 antigen-binding domain to each other is of the formula [GGGGS(SEQ ID NO: 19)]m The molecule contains a Gly / Ser linker, which is a peptide, where m is an integer from 1 to 10, 2 to 8, or 3 to 5. Preferably, m is 5. Preferably, the linker that links the anti-CD19 antigen-binding domain and the anti-CD22 antigen-binding domain together contains the amino acid sequence of SEQ ID NO: 18 (GGGGSGGGGSGGGGSGGGGSGGGGS). In this regard, the anti-CD19 antigen-binding domain and the anti-CD22 antigen-binding domain linked to each other by the linker contains the amino acid sequence of SEQ ID NO: 24 (including the m971 heavy and light chain linked by the Gly / Ser linker, and the FMC63 heavy and light chain linked by the 218 linker, with the SEQ ID NO: 18 linker positioned between the anti-CD22 and anti-CD19 antigen-binding domains). The anti-CD19 antigen-binding domain may be located adjacent to the amino terminus of the anti-CD22 antigen-binding domain (with a linker located between them), but in embodiments of this disclosure, the anti-CD22 antigen-binding domain is located adjacent to the amino terminus of the anti-CD19 antigen-binding domain (with a linker located between them).

[0078] The heavy and light chains of the anti-CD19 and anti-CD22 antigen-binding domains may be positioned in any preferred orientation with any of the linkers described herein, positioned between the heavy and light chains. In one embodiment of the present disclosure, the CAR includes (i) an FMC63 light chain positioned adjacent to the amino terminus of an m971 heavy chain, with a linker positioned between them; (ii) an m971 light chain positioned adjacent to the amino terminus of an m971 light chain, with a linker positioned between them; and (iii) an m971 light chain positioned adjacent to the amino terminus of an FMC63 heavy chain, with a linker positioned between them (e.g., Sequence ID No. 23, and Sequence IDs No. 29 and 39, described in more detail below). In another embodiment of the present disclosure, the CAR includes (i) an m971 heavy chain located adjacent to the amino terminus of an m971 light chain, with a linker positioned between them; (ii) an m971 light chain located adjacent to the amino terminus of an FMC63 light chain, with a linker positioned between them; and (iii) an FMC63 light chain located adjacent to the amino terminus of an FMC63 heavy chain, with a linker positioned between them (for example, Sequence ID No. 24, and Sequence ID Nos. 30 and 40, which are described in more detail below).

[0079] In one embodiment, the antigen-binding domain includes a leader sequence. In embodiments of this disclosure, the leader sequence may be located at the amino terminus of the light chain variable region of the anti-CD19 antigen-binding domain (e.g., at the amino terminus of the FMC63 light chain). In another embodiment of this disclosure, the leader sequence is located at the amino terminus of the light chain variable region of the anti-CD22 antigen-binding domain (e.g., at the amino terminus of the m971 heavy chain). The leader sequence may include any preferred leader sequence. In one embodiment, the leader sequence includes, consists of, or essentially consists of the amino acid sequence of SEQ ID NO: 25 (MLLLVTSLLLCELPHPAFLLIP). In one embodiment of this disclosure, the leader sequence may promote the expression of CAR on the cell surface, but the presence of the leader sequence in the expressed CAR is not required for the CAR to function. In one embodiment of this disclosure, the leader sequence may be cleaved from the CAR upon expression of the CAR on the cell surface. Therefore, in embodiments of this disclosure, the CAR lacks a leader sequence.

[0080] In one embodiment of the present disclosure, CAR includes a hinge domain. In one embodiment of the present disclosure, the hinge domain is a CDS hinge domain. In a preferred embodiment, the CDS hinge domain is human. Preferably, the CDS hinge domain includes, consists of, or essentially consists of SEQ ID NO: 33 (TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD).

[0081] In one embodiment of the present disclosure, the CAR includes a transmembrane (TM) domain. In one embodiment of the present disclosure, the TM domain is a CDS™ domain. In a preferred embodiment, the CDS™ domain is human. Preferably, the CDS™ domain includes, consists of, or essentially consists of SEQ ID NO: 26 (IYIWAPLAGTCGVLLLSLVITLYC).

[0082] In embodiments of this disclosure, the CAR comprises an intracellular T cell signaling domain. In one embodiment of this disclosure, the intracellular T cell signaling domain comprises a 4-1BB intracellular T cell signaling sequence. 4-1BB, also known as CD137, transmits potent costimulatory signals to T cells, promoting differentiation and enhancing long-term survival of T lymphocytes. Preferably, the 4-1BB intracellular T cell signaling sequence is human. In preferred embodiments, the 4-1BB intracellular T cell signaling sequence comprises, consists of, or essentially consists of the amino acid sequence of SEQ ID NO: 27 (KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL).

[0083] In embodiments of this disclosure, the intracellular T cell signaling domain comprises a CD3 zeta (ζ) intracellular T cell signaling sequence. CD3ζ associates with the TCR to generate a signal and contains an immunoreceptor tyrosine-based activation motif (ITAM). Preferably, the CD3ζ intracellular T cell signaling sequence is human. In preferred embodiments, the CD3ζ intracellular T cell signaling sequence comprises, consists of, or essentially consists of the amino acid sequence of SEQ ID NO: 28 (RVKFSRSADAPAYXQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), where X at position 14 is glutamine or lysine.

[0084] Additional embodiments of this disclosure provide full-length CARs that include, consist of, or essentially consist of any of the amino acid sequences listed in Table 1B.

[0085] (Table 1B) TIFF2026532606000003.tif229150TIFF2026532606000004.tif234150TIFF2026532606000005.tif56150

[0086] In some embodiments, the CAR of this disclosure may be tisagenecleucel, axicapbutagenesilolucel, brexcapbutageneautolucel, or lysocabbutagenemaralucel.

[0087] In some embodiments, the CARs of this disclosure may have antigen specificity for B cell maturation antigens (BCMAs).

[0088] The BCMA-specific CARs of this disclosure may have a BCMA antigen-binding domain. In some embodiments, the BCMA antigen-binding domain is a single-domain antigen-binding domain having a heavy chain variable (VH) region including the CDR region of SEQ ID NOs. 42-44, 54-47, 48-50, or 51-53. In other embodiments of this disclosure, the CAR includes the heavy chain variable region of the BCMA single-domain antigen-binding domain. In this regard, the CAR may include any of SEQ ID NOs. 54-57.

[0089] In some embodiments of the present disclosure, the CAR comprising the BCMA antigen-binding domain comprises a single-stranded variable fragment (scFv) having a heavy-chain variable (VH) region comprising the amino acid sequence of SEQ ID NO: 58 or 60, and a light-chain variable (VL) region comprising the amino acid sequence of any of SEQ ID NOs: 59 and 61-63. In some embodiments, the BCMA CAR of the present disclosure comprises an scFv having any of the amino acid sequences of SEQ ID NOs: 64-67.

[0090] In some embodiments, the CAR of this disclosure includes a BCMA antigen binding domain which is a VHH antibody binding domain. In some embodiments, the VHH BCMA antigen binding domain includes the amino acid sequence of SEQ ID NO: 68 or 69.

[0091] In some embodiments, the CAR of this disclosure includes a BCMA antigen binding comprising the amino acid sequence of SEQ ID NO: 70.

[0092] In some embodiments, the sequence of the BCMA antigen-binding domain is selected from Table 2A. In some embodiments, the CDR can be annotated and / or defined according to various CDR annotation schemes, such as the IMGT definition (see Lefranc, (1999) The Immunologist, 7, 132-136), the Kabat numbering scheme (see Kabat et al., 1992, Sequences of Proteins of Immunological Interest, DIANE Publishing: 2719), the Chothia numbering scheme (see Chothia and Lesk, J Mol Biol, 1987, 196: 901-917), or a combination of Kabat and Chothia. Other CDR annotation schemes include, but are not limited to, the AbM definition (see Whitelegg & Rees, Protein Eng. 2000, 13:819-824, Whitelegg & Rees, Methods Mol Biol. 2004, 248:51-91) and / or the Contact definition (see MacCallum et al., J.Mol.Biol. 1996, 262:732-745). While the CDR sequences provided herein may conform to one CDR annotation scheme (e.g., IMGT, Kabat, and / or Chothia), those skilled in the art can recognize and determine CDR sequences based on other CDR annotation schemes through the analysis and annotation of light chain and heavy chain variable region sequences described herein.

[0093] (Table 2A) TIFF2026532606000006.tif190152TIFF2026532606000007.tif166152

[0094] Additional embodiments of the present disclosure provide full-length anti-BCMA CAR constructs comprising, consisting of, or essentially consisting of any of the amino acid sequences of SEQ ID NOs. 71-92.

[0095] In some embodiments, the sequence of full-length anti-BCMA CAR constructs is selected from Table 2B.

[0096] (Table 2B) TIFF2026532606000008.tif23152TIFF2026532606000009.tif208152TIFF2026532606000010.tif224152TIFF20265326060 00011.tif213152TIFF2026532606000012.tif235152TIFF2026532606000013.tif231152TIFF2026532606000014.tif45152

[0097] In some embodiments, the anti-BCMA CAR constructs of this disclosure may comprise one or more components including any of the amino acid sequences of SEQ ID NOs. 93 to 104.

[0098] In some embodiments, the arrangement of components of the anti-BCMA CAR construct of this disclosure is selected from Table 2C.

[0099] (Table 2C) TIFF2026532606000015.tif81146

[0100] In some embodiments, the CAR of this disclosure may be idekabutagen bicleucel or siltabutagen autoleucel.

[0101] The functional portions of the CARs of the present invention described herein are included in the scope of this disclosure. When used in relation to a CAR, the term “functional portion” means any portion or fragment of a CAR of this disclosure that retains the biological activity of the CAR from which it is a part (parent CAR). A functional portion includes, for example, a portion of a CAR that retains the ability to recognize target cells or to detect, treat, or prevent cancer to a similar, the same, or higher degree as the parent CAR. With respect to the parent CAR, a functional portion may include, for example, about 10%, 25%, 30%, 50%, 68%, 80%, 90%, 95%, or more of the parent CAR.

[0102] The functional moiety may contain additional amino acids at its amino or carboxyl terminus, or at both ends, which are not found in the amino acid sequence of the parent CAR. Preferably, the additional amino acids do not interfere with the biological function of the functional moiety, such as recognizing target cells, detecting cancer, or treating or preventing cancer. More preferably, the additional amino acids enhance the biological activity compared to that of the parent CAR.

[0103] Functional variants of the CARs of the present invention as described herein are included in the scope of this disclosure. As used herein, the term “functional variant” means a CAR, polypeptide, or protein having substantial or significant sequence identity or similarity to the parent CAR, and the functional variant retains the biological activity of the CAR from which it is a variant. Functional variants include, for example, variants of the CAR (parent CAR) described herein that retain the ability to recognize target cells to a similar, the same, or a higher degree than the parent CAR. With respect to the parent CAR, the functional variant can be, for example, identical to the parent CAR in amino acid sequence by at least about 30%, about 50%, about 75%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more.

[0104] A functional variant may include, for example, the amino acid sequence of a parent CAR having at least one conserved amino acid substitution. Alternatively or additionally, a functional variant may include the amino acid sequence of a parent CAR having at least one non-conserved amino acid substitution. In this case, it is preferable that the non-conserved amino acid substitution does not interfere with or inhibit the biological activity of the functional variant. The non-conserved amino acid substitution may enhance the biological activity of the functional variant such that its biological activity is increased compared to that of the parent CAR.

[0105] The amino acid substitutions of CARs in the present invention are preferably conservative amino acid substitutions. Conservative amino acid substitutions are known in the art and include amino acid substitutions in which one amino acid having a particular physical and / or chemical property is replaced with another amino acid having the same or similar chemical or physical property. For example, conservative amino acid substitutions can include substitution of an acidic / negatively polar amino acid with another acidic / negatively polar amino acid (e.g., Asp or Glu), substitution of an amino acid with a nonpolar side chain with another amino acid with a nonpolar side chain (e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Cys, Val, etc.), substitution of a basic / positively polar amino acid with another basic / positively polar amino acid (e.g., Lys, His, Arg, etc.), substitution of an uncharged amino acid with a polar side chain with another uncharged amino acid with a polar side chain (e.g., Asn, Gln, Ser, Thr, Tyr, etc.), substitution of an amino acid with a beta-branched side chain with another amino acid with a beta-branched side chain (e.g., Ile, Thr, and Val), substitution of an amino acid with an aromatic side chain with another amino acid with an aromatic side chain (e.g., His, Phe, Trp, and Tyr), and so on.

[0106] CARs can essentially be derived from specific amino acid sequences described herein, such that other components, for example, other amino acids, do not substantially alter the biological activity of the functional variant.

[0107] The CARs (including functional portions and functional variants) of embodiments of the present disclosure can be of any length, that is, they can contain any number of amino acids, provided that the CAR (or its functional portion or functional variant) retains biological activity, such as the ability to specifically bind to an antigen, detect diseased cells in mammals, or treat or prevent a disease in mammals. For example, a CAR can be about 50 to about 5000 amino acid lengths, for example, 50, 70, 75, 100, 125, 150, 175, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 amino acid lengths.

[0108] The CARs of embodiments of the present disclosure (including the functional parts and functional variants of the present disclosure) may include synthetic amino acids instead of one or more naturally derived amino acids. Such synthetic amino acids are known in the art and include, for example, aminocyclohexanecarboxylic acid, norleucine, α-amino-n-decanoic acid, homoserine, S-acetylaminomethyl-cysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, β-phenylserine, β-hydroxyphenylalanine, phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, and Indian It contains phosphorus-2-carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N'-benzyl-N'-methyllysine, N',N'-dibenzyllysine, 6-hydroxylysine, ornithine, α-aminocyclopentanecarboxylic acid, α-aminocyclohexanecarboxylic acid, α-aminocycloheptanecarboxylic acid, α-(2-amino-2-norbomane)-carboxylic acid, α,γ-diaminobutyric acid, α,β-diaminopropionic acid, homophenylalanine, and α-tert-butylglycine.

[0109] The CARs (including functional portions and functional variants) of embodiments of the present disclosure can be glycosylated, amidated, carboxylic acidified, phosphorylated, esterified, N-acylated, cyclized via disulfide crosslinks, etc., or converted to acid addition salts, and / or optionally dimerized, polymerized, or conjugated.

[0110] The CARs of the embodiments of this disclosure (including their functional parts and functional variants) can be obtained by methods known in the art. The CARs can be produced by any preferred method for producing polypeptides or proteins, including denovation synthesis. Alternatively, the CARs can be recombinantly produced using the nucleic acids described herein, using standard recombination methods. See, for example, Green et al., Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Press, Cold Spring Harbor, NY 2012. Furthermore, some of the CARs of this disclosure (including their functional parts and functional variants) can be isolated and / or purified from sources such as plants, bacteria, insects, mammals, e.g., rats, humans. Methods for isolation and purification are well known in the art. Alternatively, the CARs (including their functional parts and functional variants) described herein can be commercially synthesized by companies such as Synpep (Dublin, CA), Peptide Technologies Corp. (Gaithersburg, MD), and Multiple Peptide Systems (San Diego, CA). Thus, the CARs of the present invention can be synthesized, recombinant, isolated, and / or purified.

[0111] Embodiments of this disclosure further provide nucleic acids comprising nucleotide sequences encoding any of the CARs (including their functional portions and functional variants) described herein. The nucleic acids of this disclosure may comprise nucleotide sequences encoding any of the leader sequences, antigen-binding domains, transmembrane domains, linkers, and / or intracellular T cell signaling domains described herein.

[0112] In one embodiment, the nucleic acid comprises a nucleotide sequence encoding a leader sequence, anti-CD22 and anti-CD19 antigen-binding domains (including linker-bound light chain and heavy chain variable regions), a CD8 hinge domain, a CD8 transmembrane domain, a 4-1BB intracellular T cell signaling domain, and a CD3ζ intracellular T cell signaling domain. For example, the nucleic acid may comprise a nucleotide sequence encoding anti-CD22 and anti-CD19 antigen-binding domains (including linker-bound light chain and heavy chain variable regions), a CD8 hinge domain (SEQ ID NO: 34), a CD8 transmembrane domain (SEQ ID NO: 35), a 4-1BB intracellular T cell signaling domain (SEQ ID NO: 36), and a CD3ζ intracellular T cell signaling domain (SEQ ID NO: 37). In one embodiment of the present disclosure, the nucleic acid comprises a CD8 hinge domain, a CD8 transmembrane domain, a 4-1BB intracellular T cell signaling domain, and a nucleotide sequence encoding a CD3ζ intracellular T cell signaling domain including the nucleotide sequence of SEQ ID NO: 38. In embodiments of this disclosure, the nucleic acid comprises, consists of, or essentially comprises the nucleotide sequence of SEQ ID NO: 31 or 32, which encodes a full-length CAR comprising the amino acid sequence of SEQ ID NO: 29 or 30, respectively. The nucleotide sequences of SEQ ID NOs: 31, 32, 34-38 are shown in Table 3.

[0113] (Table 3) TIFF2026532606000016.tif117143TIFF2026532606000017.tif235143TIFF2026532606000018.tif75143

[0114] As used herein, “nucleic acid” includes “polynucleotide,” “oligonucleotide,” and “nucleic acid molecule,” and generally means a polymer of DNA or RNA, which may be single-stranded or double-stranded, which may be synthesized or obtained (e.g., isolated and / or purified) from natural sources, which may include natural, unnatural, or modified nucleotides, and which may include modified internucleotide bonds, such as phosphoramidate bonds or phosphorothioate bonds, instead of phosphodiesters found between nucleotides in natural, unnatural, or unmodified oligonucleotides. In some embodiments, the nucleic acid does not contain any insertions, deletions, inversions, and / or substitutions. However, in some cases, as discussed herein, it may be preferable for the nucleic acid to contain one or more insertions, deletions, inversions, and / or substitutions. In some embodiments, the nucleic acid may encode an additional amino acid sequence that does not affect the function of CAR, which may or may not be translated when the nucleic acid is expressed by a host cell.

[0115] The nucleic acids in one embodiment of this disclosure may be recombinant. As used herein, the term “recombinant” means (i) a molecule constructed outside a living cell by binding a natural or synthetic nucleic acid segment to a nucleic acid molecule that can replicate in a living cell, or (ii) a molecule resulting from the replication of the one described in (i) above. For the purposes of this specification, replication may be in vitro replication or in vivo replication.

[0116] Recombinant nucleic acids may have sequences that are not of natural origin, or sequences that are created by artificially combining two differently separated segments of a sequence. This artificial combination is often achieved by chemical synthesis, or more generally by artificial manipulation of isolated segments of nucleic acids, such as by genetic engineering techniques, as described above by Green et al. Nucleic acids can be constructed based on chemical synthesis and / or enzymatic ligation reactions using procedures known in the art. See, for example, Green et al., above. For example, nucleic acids can be chemically synthesized using naturally derived nucleotides, or various modified nucleotides (e.g., phosphorothioate derivatives and acridine-substituted nucleotides) designed to increase the biological stability of the molecule or to increase the physical stability of the double helix formed during hybridization. Examples of modified nucleotides that can be used to produce nucleic acids include, but are not limited to, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl)uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylosin, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-substituted adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylosin, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N 6Examples include isopentenyl adenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, quasin, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methyl ester, 3-(3-amino-3-N-2-carboxypropyl)uracil, and 2,6-diaminopurine. Alternatively, one or more of the nucleic acids of this disclosure can be purchased from companies such as Macromolecular Resources (Fort Collins, CO) and Synthegen (Houston, TX).

[0117] Nucleic acids may include any isolated or purified nucleotide sequences that encode either a CAR or a functional part or functional variant thereof. Alternatively, a nucleotide sequence may include a nucleotide sequence that degenerates into any of the sequences, or a combination of degenerate sequences.

[0118] Embodiments of the present disclosure also provide isolated or purified nucleic acids comprising a nucleotide sequence complementary to any of the nucleotide sequences of the nucleic acids described herein, or a nucleotide sequence that hybridizes to any of the nucleotide sequences of the nucleic acids described herein under stringent conditions.

[0119] Nucleotide sequences that hybridize under stringent conditions may hybridize under high-stringency conditions. “High-stringency conditions” means that a nucleotide sequence hybridizes specifically to a target sequence (any nucleotide sequence of the nucleic acids described herein) in a detectably stronger amount than non-specific hybridization. High-stringency conditions include conditions that distinguish polynucleotides with precisely complementary sequences, or polynucleotides containing only a few scattering mismatches, from random sequences that have occurred to have a few small regions (e.g., 3–10 bases) that match the nucleotide sequence. Such small complementary regions are more readily dissociated than full-length complementary chains of 14–17 bases or more, and high-stringency hybridization makes them easily distinguishable. Relatively high-stringency conditions would include low-salt and / or high-temperature conditions, such as those provided by about 0.02–0.1 M NaCl or equivalent at a temperature of about 50–70°C. Such highly stringent conditions tolerate little to no mismatch between the nucleotide sequence and the template or target strand, if any, and are particularly suitable for detecting the expression of any of the CARs of the present invention. Generally, it is understood that the conditions can be made more stringent by adding an increased amount of formamide.

[0120] This disclosure also provides nucleic acids comprising nucleotide sequences that are at least about 70% identical, for example, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% identical to any of the nucleic acids described herein.

[0121] In one embodiment, the nucleic acids of the Disclosure can be incorporated into a recombinant expression vector. In this regard, embodiments of the Disclosure provide recombinant expression vectors comprising any of the nucleic acids of the Disclosure. For the purposes of this Spectrum, the term “recombinant expression vector” means a genetically modified oligonucleotide or polynucleotide construct that enables the expression of mRNA, protein, polypeptide, or peptide by a host cell when the construct comprises a nucleotide sequence encoding mRNA, protein, polypeptide, or peptide and comes into contact with the cell under conditions sufficient to have mRNA, protein, polypeptide, or peptide expressed in the cell. The vectors of the Disclosure are not of natural origin as a whole.

[0122] However, some of the vectors may be of natural origin. The recombinant expression vectors of the present invention may contain any type of nucleotides, including DNA and RNA, which may be single-stranded or double-stranded, may be partially synthesized or obtained from natural sources, and may contain natural, unnatural, or modified nucleotides. The recombinant expression vectors may contain naturally derived or unnatural nucleotide-nucleotide bonds, or both types of bonds. Preferably, unnatural or modified nucleotides or nucleotide-nucleotide bonds do not interfere with the transcription or replication of the vector.

[0123] In one embodiment, the recombinant expression vector of this disclosure can be any suitable recombinant expression vector and can be used to transform or transfect any suitable host cell. Suitable vectors include plasmids and viruses, which are designed for proliferation and amplification, or for expression, or both. Vectors can be selected from the group consisting of the pUC series (Fermentas Life Sciences, Glen Burnie, MD), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Phannacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, CA).

[0124] Bacteriophage vectors such as λGT10, λGT11, λZapII (Stratagene), λEMBL4, and λNM1149 can also be used. Examples of plant expression vectors include pBI101, pBI101.2, pBI101.3, pBI121, and pBIN19 (Clontech). Examples of animal expression vectors include pEUK-C1, pMAM, and pMAMneo (Clontech). Recombinant expression vectors may be viral vectors, such as retroviral or lentiviral vectors, oncolytic viral vectors, adenovirus vectors, adeno-associated virus (AAV) vectors, and virus-like particles (VLPs).

[0125] In one embodiment, the recombinant expression vector of this disclosure can be prepared, for example, using standard recombinant DNA techniques as described above by Green et al. The expression vector construct, which may be circular or linear, can be prepared to contain a functional replication system in prokaryotic or eukaryotic host cells. The replication system may be derived, for example, from ColE1, 2μ plasmid, λ, SV40, bovine papillomavirus, etc.

[0126] Recombinant expression vectors may contain regulatory sequences, such as transcription and translation start and stop codons, which are specific to the type of host cell into which the vector is introduced (e.g., bacteria, fungi, plants, or animals), and whether the vector is DNA or RNA-based. Recombinant expression vectors may also contain restriction sites to facilitate cloning.

[0127] Recombinant expression vectors may contain one or more marker genes that enable the selection of transformed or transfected host cells. Examples of marker genes include biocide resistance, such as resistance to antibiotics and heavy metals, and nutritional support in nutritionally dependent hosts to provide protrophotrophy. Suitable marker genes for the expression vectors of the present invention include, for example, neomycin / G418 resistance genes, hygromycin resistance genes, histidinol resistance genes, tetracycline resistance genes, and ampicillin resistance genes.

[0128] Recombinant expression vectors may include innate or non-natural promoters operably ligated to the CAR-encoding nucleotide sequence (including its functional portion and functional variants), or to nucleotide sequences complementary to or hybridizing with the CAR-encoding nucleotide sequence. The selection of promoters, e.g., strong, weak, inducible, tissue-specific, and development-specific, is within the scope of the art. Similarly, combinations of nucleotide sequences and promoters are also within the scope of the art. Promoters may be non-viral promoters or viral promoters, e.g., cytomegalovirus (CMV) promoters, SV40 promoters, RSV promoters, or promoters found in long-terminal repeats of mouse stem cell viruses.

[0129] The recombinant expression vectors of the present invention can be designed for transient expression, stable expression, or both. Furthermore, recombinant expression vectors can be prepared for constitutive or inducible expression.

[0130] Furthermore, recombinant expression vectors can be constructed to contain suicide genes. As used herein, the term “suicide gene” refers to a gene that kills cells that express a suicide gene. A suicide gene may be a gene that confers sensitivity to a drug, for example, a drug, to cells on which it is expressed, causing the cells to die when they come into contact with or are exposed to the drug. Suicide genes are known in the art and include, for example, the herpes simplex virus (HSV) thymidine kinase (TK) gene, cytosine daminase, purine nucleoside phosphorylase, and nitroreductase.

[0131] Included in the scope of this disclosure are conjugates, such as bioconjugates, comprising any of the CARs of the present invention (including any of their functional portions or variants), nucleic acids, recombinant expression vectors, host cells, or populations of host cells. Conjugates and methods for generally synthesizing conjugates are known in the art.

[0132] Embodiments of this disclosure further provide host cells containing any of the recombinant expression vectors described herein. As used herein, the term “host cell” refers to any type of cell that may contain the recombinant expression vectors of the present invention. The host cell may be a eukaryotic cell, e.g., a plant, animal, fungus, or algae, or a prokaryotic cell, e.g., a bacterium or protist. The host cell may be a cultured cell or a primary cell, i.e., it may be isolated directly from an organism, e.g., a human. The host cell may be an adherent cell or a suspension cell, i.e., a cell that grows in a suspension state. Suitable host cells are known in the art and include, for example, DH5α E. coli cells, Chinese hamster ovary cells, monkey VERO cells, COS cells, HEK293 cells, and the like. With regard to the purpose of amplifying or replicating recombinant expression vectors, the host cell may be a prokaryotic cell, e.g., a DH5α cell. With regard to the purpose of generating recombinant CARs, the host cell may be a mammalian cell. The host cell may be a human cell. The host cell can be of any cell type, may originate from any type of tissue, may be at any developmental stage, and may be a peripheral blood lymphocyte (PBL) or a peripheral blood mononuclear cell (PBMC). In some embodiments, the host cell may be a T cell. For the purposes of this specification, the T cell may be any T cell, such as a cultured T cell, e.g., primary T cell, or a cultured T cell line, e.g., T cells from Jurkat, SupTl, etc., or T cells obtained from a mammal. When obtained from a mammal, the T cell may be obtained from a number of sources, including, but not limited to, blood, bone marrow, lymph nodes, thymus, or other tissues or fluids. The T cell may also be enriched or purified. The T cell may be a human T cell. The T cell may be a T cell isolated from a human. The T cell may be any type of T cell, but not limited to, CD4 + / CDS + Double-positive T cells, CD4+ helper T cells, e.g., Th1 and Th2 cells, CDS +It can be any developmental stage, including T cells (e.g., cytotoxic T cells), tumor-infiltrating cells, memory T cells, naive T cells, etc. T cells are CDS + T cells or CD4 + T cells @.

[0133] In other embodiments, the host cell may be a natural killer (NK) cell. In other embodiments, the host cell may be a cytotoxic T lymphocyte (CTL). In other embodiments, the host cell may be a regulatory T cell. In other embodiments, the host cell may be a natural killer T cell (NKT).

[0134] Embodiments of this disclosure also provide a cell population comprising at least one host cell as described herein. The cell population can be a heterogeneous population comprising at least one other cell, e.g., a host cell not comprising any of the recombinant expression vectors (e.g., a T cell), or a cell other than a T cell, e.g., a B cell, macrophage, neutrophil, erythrocyte, hepatocyte, endothelial cell, epithelial cell, muscle cell, brain cell, etc., in addition to a host cell comprising any of the recombinant expression vectors described herein. Alternatively, the cell population can be a substantially homogeneous population, the population mainly comprising host cells comprising (e.g., essentially consisting of) a recombinant expression vector. The population can also be a clonal cell population in which all cells in the population are clones of a single host cell comprising a recombinant expression vector, such that all cells in the population comprise a recombinant expression vector. In one embodiment of this disclosure, the cell population is a clonal population comprising host cells comprising a recombinant expression vector as described herein.

[0135] CARs (including their functional parts and variants), nucleic acids, recombinant expression vectors, and host cells (including their populations), all of which are collectively referred to hereafter as “CAR materials,” can be isolated and / or purified. As used herein, the term “isolated” means removed from its natural environment. The terms “purified” or “isolated” are intended as relative terms, not requiring absolute purity or isolation. Thus, for example, a purified (or isolated) host cell preparation is one in which the host cells are purer than those in the natural environment within the body. Such host cells can be produced, for example, by standard purification techniques. In some embodiments, the host cell preparation is purified such that the host cells represent at least about 50% of the total cell content of the preparation, for example, at least about 70%. For example, the purity can be at least about 50%, about 60%, about 70%, or more than about 80%, or about 100%.

[0136] CAR materials can be formulated into compositions such as pharmaceutical compositions. In this regard, one embodiment of the present disclosure provides a pharmaceutical composition comprising any of the CAR materials described herein and a pharmaceutically acceptable carrier. A pharmaceutical composition of the present invention containing any of the CAR materials may include two or more CAR materials of the present invention, for example, CAR and nucleic acid, or two or more different CARs. Alternatively, the pharmaceutical composition may include a CAR material of the present invention in combination with other pharmaceutically active agents or drugs, such as chemotherapeutic agents such as asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, and vincristine. In a preferred embodiment, the pharmaceutical composition includes host cells or populations thereof of the present invention.

[0137] With respect to pharmaceutical compositions, pharmaceutically acceptable carriers can be any of those conventionally used and are limited only by chemophysical considerations such as lack of solubility and reactivity with the activator, and by the route of administration. The pharmaceutically acceptable carriers described herein, such as vehicles, adjuvants, excipients, and diluents, are well known to those skilled in the art and are readily available to the public. It is preferable that pharmaceutically acceptable carriers are carriers that do not have harmful side effects or toxicity under the conditions of use.

[0138] The choice of carrier will be determined in part by the CAR material of a particular invention and by the particular method used to administer the CAR material of the present invention. Therefore, a variety of suitable formulations of the pharmaceutical compositions of this disclosure exist. Methods for preparing administerable (e.g., parenterally administerable) compositions are known or obvious to those skilled in the art and are described in detail, for example, Remington: The Science and Practice of Pharmacy, Pharmaceutical Press; 22nd ed. (2012).

[0139] CAR material can be administered in any preferred manner. Preferably, CAR material is administered by injection (e.g., subcutaneous, intravenous, intratumoral, intraarterial, intramuscular, intradermal, intraperitoneal, or subarachnoid). Preferably, CAR material is administered intravenously.

[0140] Suitable pharmaceutically acceptable carriers for the CAR material of the present invention for injection may include any isotonic carrier, such as physiological saline (about 0.90% w / v NaCl in water, about 300 mOsm / L NaCl in water, or about 9.0 g NaCl per liter of water), NORMOSOL R electrolyte solution (Abbott, Chicago, IL), PLASMA-LYTE A (Baxter, Deerfield, IL), about 5% dextrose in water, or Ringer's lactate solution. In one embodiment, the pharmaceutically acceptable carrier is supplemented with human serum albumin.

[0141] "Effective dose" or "therapeutic dose" refers to a dose appropriate for preventing or treating cancer in an individual. The dose effective for therapeutic or prophylactic use will depend, for example, on the stage and severity of the disease or disorder being treated, the patient's age, weight, and general health, as well as the judgment of the prescribing physician. The size of the dose will also be determined by the selected activity, method of administration, timing and frequency of administration, the presence, nature, and extent of any adverse side effects that may accompany the administration of a particular activity, and the desired physiological effect. It will be understood by those skilled in the art that various diseases or disorders may require long-term treatment with multiple doses, and that CAR material may be used at each dose or at various stages of administration. As an example, and not intended to limit this disclosure, if the CAR material of the present invention is host cells, an exemplary dose of host cells would be at least 1 million cells (1 × 10⁶). 6 It may be (cells / dose). In some embodiments, the subject of this disclosure (e.g., human subject) is about 1.0 × 10 per kilogram of the subject's body weight. 5 ~Approx. 5.0×10 6 individual CAR + Cells, or approximately 4.0 × 10 5 ~1.0×10 9 Total CAR + CAR in the range of cells + The cells are administered in doses.

[0142] For the purposes of this disclosure, the amount or dose of the CAR material administered should be sufficient to produce a therapeutic or prophylactic response in the subject or animal over a reasonable time frame. For example, the dose of the CAR material of the present invention should be sufficient to bind to an antigen or to detect, treat, or prevent cancer within about two hours or more from the time of administration, for example, within about 12 to about 24 hours or more. In certain embodiments, the period may be longer. The dose will be determined by the efficacy of the CAR material of the particular invention, the condition of the animal (e.g., human), and the body weight of the animal being treated (e.g., human).

[0143] For the purposes of this disclosure, for example, an assay may be used to determine an initial dose to be administered to a mammal, which involves comparing the extent to which target cells are lysed and / or the extent to which IFN-γ is secreted by T cells expressing the CAR of the present invention among a series of mammals, each being administered different doses of T cells, upon administration of a given dose of such T cells to the mammal. The extent to which target cells are lysed and / or the extent to which IFN-γ is secreted upon administration of a particular dose can be measured by methods known in the art.

[0144] When a CAR material is administered with one or more additional therapeutic agents, these additional agents can be administered co-administered to mammals. “Co-administration” means administering the additional therapeutic agents and the CAR material in sufficiently close proximity in time so that the CAR material can enhance the effects of the additional therapeutic agents, or vice versa. In this regard, the CAR material may be administered first, followed by the additional therapeutic agents, or vice versa. Alternatively, the CAR material and the additional therapeutic agents may be administered simultaneously. An exemplary therapeutic agent that can be administered co-administered with a CAR material is IL-2, which is thought to enhance the therapeutic effect of the CAR material. As described herein, the administration of allogeneic cells containing chimeric receptors, which may include the CAR materials described herein, may be administered together (e.g., as part of a treatment regime or multicomponent cell therapy product) with immunomodulatory cell therapy products (e.g., any of the regulatory T cell therapies and / or allogeneic HSCTs described herein), including but not limited to (a) allogeneic cells containing chimeric receptors and (b) immunomodulatory cell therapy products, including concurrent, sequential, or combined administration of the immunomodulatory cell therapy products. In exemplary, non-limiting examples, any of the Tregs and HSPCs described herein may be administered on day 0 of a treatment regimen, and then cells containing chimeric receptors (e.g., any of the CAR materials described herein) may be administered on day 2. In further examples, cells containing chimeric receptors may be administered together with any of the Tcons described herein on the same day (e.g., day 2), such as a separate administration of cells containing chimeric receptors after administration of Tcon.

[0145] CAR materials are intended to be used in methods for treating or preventing diseases in mammals. While not bound by any particular theory or mechanism, CAR materials possess biological activity, such as the ability to recognize an antigen, such as BCMA, or one or both of CD19 and CD22, so that when expressed by a cell, the CAR can mediate an immune response against cells expressing a bispecific antigen, such as BCMA, or one or both of CD19 and CD22. In this regard, one embodiment of the present disclosure provides a method for treating or preventing cancer in a mammal, comprising administering to a mammal in an amount effective for treating or preventing cancer in the mammal, any of the CAR, nucleic acids, recombinant expression vectors, host cells, cell populations, and / or pharmaceutical compositions of the present disclosure.

[0146] One embodiment of the present disclosure further includes lymphocyte depletion of a mammal before administration of CAR material. Examples of lymphocyte depletion include, but are not limited to, non-myelosuppressive lymphocyte depletion chemotherapy, myelosuppressive lymphocyte depletion chemotherapy, and total body irradiation. Pre-transplant conditioning regimens including myelosuppressive pre-transplant conditioning are further described herein.

[0147] With regard to the object of the method of the present invention in which host cells or a population of cells are administered, the cells may be homogeneous or self-cells of the mammal. Preferably, the cells are self-cells of the mammal.

[0148] The mammals referred to herein may be any mammal. As used herein, the term “mammal” refers to any mammal, including, but is not limited to, rodents such as mice and hamsters, and serratus mammals such as rabbits. Mammals may be Carnivores, including Felidae (cats) and Canidae (dogs). Mammals may be Artiodactyla, including Bovidae (cats) and Suidae (pigs), or Perissodactyla, including Equidae (horse). Mammals may be Primates, Platyrrhines, or Phanerophia (monkeys), or Homorhini (humans and apes). Preferably, the mammal is human.

[0149] With respect to the method of the present invention, cancer can be any cancer, including acute lymphoblastic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bladder cancer (e.g., bladder cancer), bone cancer, brain cancer (e.g., medulloblastoma), breast cancer, cancer of the anus, anal canal, or anorectum, eye cancer, intrahepatic cholangiocarcinoma, joint cancer, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, oral cancer, vulvar cancer, chronic lymphocytic leukemia (CLL), chronic myeloid cancer, colon cancer, esophageal cancer, cervical cancer, fibrosarcoma, gastrointestinal carcinoid tumor, head and neck cancer (e.g., head and neck squamous cell carcinoma), Hodgkin lymphoma, hypopharyngeal cancer, kidney cancer, pharyngeal cancer, leukemia, humoral tumor, liver cancer. This includes any of the following: visceral cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, malignant mesothelioma, mast cell tumor, melanoma, multiple myeloma, nasopharyngeal cancer, non-Hodgkin lymphoma, B-cell chronic lymphocytic leukemia, precursor B-cell acute lymphoblastic leukemia (B-ALL), precursor B-cell acute lymphoblastic leukemia (BCP-ALL), B-cell lymphoma, hairy cell leukemia, acute lymphoblastic leukemia (ALL), and Burkitt lymphoma, ovarian cancer, pancreatic cancer, cancers of the peritoneum, omentum, and mesentery, pharyngeal cancer, prostate cancer, rectal cancer, kidney cancer, skin cancer, small intestine cancer, soft tissue cancer, solid tumors, gastric cancer, testicular cancer, thyroid cancer, and ureteral cancer. Preferably, the cancer is a hematological malignancy (e.g., leukemia or lymphoma including, but not limited to, Hodgkin lymphoma, non-Hodgkin lymphoma, CLL, acute lymphoblastic carcinoma, acute myeloid leukemia, B-cell chronic lymphocytic leukemia, hairy cell leukemia, acute lymphoblastic leukemia (ALL) (also known as acute lymphoblastic leukemia), B-ALL, BCP-ALL, B-cell lymphoma, Burkitt lymphoma, and myeloma, but not limited to multiple myeloma (MM)). In some embodiments, the cancer may be characterized by BCMA expression, or by one or both of CD22 and CD19, or it may be a hematological malignancy characterized by BCMA or by one or both of CD19 and CD22.

[0150] As used herein, the terms “treatment” and “prevention,” and words derived therefrom, do not necessarily mean 100% or complete treatment or prevention. Rather, there are varying degrees of treatment or prevention that a person skilled in the art would recognize as having potential benefits or therapeutic effects. In this regard, the methods of the present invention can provide any amount or level of treatment or prevention of cancer in mammals. Furthermore, the treatment or prevention provided by the methods of the present invention may include the treatment or prevention of one or more conditions or symptoms of the disease being treated or prevented, for example, cancer. Also, for the purposes of this specification, “prevention” may include delaying the onset of the disease, or its symptoms or conditions.

[0151] Another embodiment of this disclosure provides the use of the CAR, nucleic acid, recombinant expression vector, host cell, cell population, or pharmaceutical composition of the present invention for the treatment or prevention of cancer in mammals.

[0152] Methods for testing CARs for their ability to recognize target cells and antigen specificity are known in the art. For example, Clay et al., J.Immunol., 163:507-513 (1999) teach a method for measuring the release of cytokines (e.g., interferon-γ, granulocyte / monocyte colony-stimulating factor (GM-CSF), tumor necrosis factor α (TNF-α), or interleukin-2 (IL-2)). In addition, CAR function can be evaluated by measuring cytotoxicity, as described in Zhao et al., J.Immunol., 174:4415-4423 (2005).

[0153] Allogeneic hematopoietic stem cell transplantation Various embodiments of this disclosure provide compositions, multicomponent medical procedures, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods relating to improved allogeneic hematopoietic stem cell transplantation (alloHSCT), including, for example, the transplantation of separate cell populations enriched with hematopoietic stem progenitor cells (HSPCs), regulatory T cells (Tregs), and conventional T cells (Tcones) for use in combination with allogeneic cells containing chimeric receptors, e.g., TCRs or CAR-T cells ("CAR materials" as described herein).

[0154] Standard alloHCT typically involves transplanting pluripotent hematopoietic stem progenitor cells (HSPCs), usually derived from donor bone marrow, peripheral blood, or umbilical cord blood, into the recipient. The recipient may undergo myeloablative conditioning, which kills hematopoietic cells, including tumor cells and host immune cells. The transplanted HSPCs then reorganize the hematopoietic compartment. HSCT is useful as a cancer treatment, possessing the ability to induce antitumor activity in donor T cells and is referred to as graft-versus-tumor (GVT). In patients with hematological malignancies refractory to chemotherapy, HSCT has been associated with improved survival.

[0155] Remarkably, recipients of the improved alloHSCT described herein, while receiving monotherapy with graft-versus-host disease (GVHD) prophylactic agents, such as tacrolimus, had significantly better clinical outcomes than existing standard alloHSCT regimens and standard treatments. These recipients experienced improved clinical outcomes, including, for example, increased overall survival, increased relapse-free survival, increased GVHD and relapse-free survival (GRFS), faster and / or complete engraftment of various hematopoietic components (e.g., neutrophils, platelets, T cells, B cells), improved donor chimerism (e.g., T cell chimerism), reduced relapses, reduced primary graft failure, reduced secondary graft failure, reduced procedure-related mortality, reduced acute and / or chronic GVHD, and shorter time to discharge after monotherapy with GVHD prophylactic agents such as tacrolimus.

[0156] While alloHSCT is associated with improved survival in patients with hematological malignancies refractory to chemotherapy, some subjects treated with existing alloHSCT regimens exhibit cancer recurrence, and numerous complications may limit the efficacy of alloHSCT. The efficacy of standard alloHSCT may be limited by, for example, primary or secondary graft failure, limited or delayed engraftment of various hematopoietic components (e.g., neutrophils, platelets, T cells, or B cells), and limited donor chimerism (e.g., T cell chimerism). Furthermore, standard alloHSCT may have ethical or toxic implications associated with the procedure. However, donor T cells may also attack non-tumor host cells, leading to graft-versus-host disease (GVHD). GVHD is a major cause of post-HCT complications and can be fatal. Management of GVHD may require immunosuppressive therapy or cytotoxic mediation, which can result in toxicity, increased susceptibility to infection, and / or blunting of anti-tumor immunity. Early morbidity and death associated with acute graft-versus-host disease (aGVHD; occurring within the first 100 days after transplantation), as well as prolonged morbidity associated with chronic GVHD (cGVHD), are major limiting factors in the success of HCT. GVHD is a risk for both HLA-matched and HLA-mismatched transplants. GVHD can occur even when the donor and recipient are HLA-matched, because the immune system can still recognize other differences between donor tissues.

[0157] Both GVT and GVHD are primarily mediated by conventional T cells (Tcon) that initiate an immune response upon recognition of congenital antigens by T cell receptors. Depleting T cells from hematopoietic stem cell transplant (HCT) grafts can reduce GVHD, but may also reduce GVT and increase the likelihood of cancer recurrence. In addition to Tcon, Tregs are a further subset of T cells that negatively modulate inflammation and promote immune tolerance. Tregs can prevent or reduce GVHD by negatively modulating inflammation, including inflammation induced by them when donor Tcon recognize recipient antigens, for example.

[0158] This specification provides compositions and methods for improved alloHSCT, which target CD45 containing at least HSPC. + The first population of cells, CD45, including at least Treg cells. + A second population of cells, and CD45 containing at least Tcon + This includes administering a specific cell population, which includes a third population of cells. In some embodiments, CD45 + The second population of cells is also called the Treg-enriched cell population. While I don't want to be constrained by theory, CD45 + Administering a second population of cells reduced the incidence and / or severity of GVHD, while CD45 containing Tcon was also reduced. + GVT is enhanced by administering a third population of cells. Therefore, embodiments of this disclosure provide compositions, multicomponent medical treatments, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods for enhancing GVT while minimizing GVHD by administering at least both populations of T cells. Accordingly, the compositions, multicomponent medical treatments, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods disclosed herein can prevent or reduce graft-versus-host disease (GVHD) in subjects while maintaining the graft-versus-tumor response (GVT) effect of alloHSCT administered to subjects with cancer (e.g., hematological malignancies). In some embodiments, two or more cell populations are administered at different time points, for example, CD45 containing at least HSPC. + The first population of cells, and the Treg-enriched cell population, contain at least CD45 containing Tcon. + It may be administered before the third population of cells.

[0159] cell population Embodiments of this disclosure provide multicomponent medical treatments or multicomponent cell therapy products administered to human subjects in need. In some embodiments, the multicomponent treatment includes (a) hematopoietic stem progenitor cells (HSPCs) and CD45 cells including granule cells. + A solution containing a first population of cells, CD45+ (b) a solution containing a cell population enriched with regulatory T cells (Tregs), and (c) CD45 + A solution containing a third population of cells, CD45 + The third population of cells is at least about 20% CD3 + (d) a solution comprising (d) a solution comprising (a) a normal T cell (Tcon), at least about 10% monocytes, and at least about 10% granule cells, and (a) a solution comprising one or more doses of a graft-versus-host disease (GVHD) prophylactic agent, such as tacrolimus. In various embodiments, the HSPC is CD34 + In some embodiments, the multicomponent cell therapy product is (a) approximately 1.0 × 10¹⁶ units per kilogram of body weight of a human subject receiving the product. 5 ~Approx. 1.0×10 8 individual CD34 + Hematopoietic stem progenitor cells (HSPC), approximately 1.5×10 7 ~Approx. 1.5×10 10 HSPC, or approximately 5.0 × 10 5 ~Approx. 5.0×10 8 Isolated CD45 containing doses of individual HSPCs + A first single-dose transfer bag containing a first population of cells, CD45 + The first population of cells is formulated with excipients at a neutral pH into a first single-dose transfer bag, and (b) approximately 1.0 × 10⁶ units per kilogram of body weight of a human subject receiving the product. 5 ~Approx. 2.0×10 7 Individual fresh CD4 + CD25 + CD127 dim Regulatory T cells (Treg), approximately 1.5 x 10 7 ~Approx. 3.0×10 9 Individual fresh Tregs, or approximately 5.0 x 10 5 ~Approx. 1.0×10 8 Isolated CD45 containing a dose of fresh Treg + A second single-dose transfer bag containing a second population of cells, isolated CD45 +The second population of cells is formulated with excipients at a neutral pH in a second single-dose transfer bag, and (c) approximately 1.0 × 10⁶ units per kilogram of body weight in human subjects receiving the product. 5 ~Approx. 4.0×10 7 Individual standard CD3 + T cells (Tcon), approximately 1.5×10 7 ~Approx. 6.0×10 9 Tcon, or approximately 5.0 × 10 5 ~Approx. 2.0×10 8 Isolated CD45 containing doses of Tcon + A third single-dose transfer bag containing a third population of cells, isolated CD45 + A third population of cells comprises a third single-dose transfer bag formulated with excipients at a neutral pH, the excipients comprising one or more cryoprotective agents.

[0160] In some embodiments, CD45 + The first group of cells is CD34 + Contains hematopoietic stem progenitor cells (HSPCs). When used herein, CD45 + The first population of cells, and CD34 + Populations of hematopoietic stem progenitor cells (HSPCs) can be used interchangeably. In other embodiments, CD45 + The first population of cells contains at least one dose of HSPC. In some embodiments, CD45 contains HSPC or at least one dose of HSPC. + The first population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects. 5 ~Approx. 5.0×10 8 The amount of HSPC per kilogram of body weight in humans is approximately 1.0 × 10⁻¹⁴. 5 ~Approx. 1.0×10 8 Approximately 5.0 × 10¹⁴ HSPCs per kilogram of human body weight. 5 ~Approx. 2.0×10 7 Includes HSPCs. In some embodiments, CD45 includes HSPCs or at least one dose of HSPCs. + The first population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects.5 For HSPCs of one or more individuals, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 2.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 3.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 4.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 4.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 5.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 5.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 6.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 6.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 7.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 7.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 8.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 8.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 9.0 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of 1 or more individuals, approximately 9.5 × 10¹⁶ per kilogram of body weight in humans. 5 For HSPCs of one or more individuals, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 1.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 1.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 6For HSPCs of one or more individuals, approximately 2.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 2.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 2.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 3.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 3.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 3.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 4.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 4.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 4.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 4.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 5.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 5.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 5.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 5.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 6.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 6.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 6.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 6.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 7.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 7.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 7.5 × 10¹⁶ per kilogram of body weight in humans. 6For HSPCs of one or more individuals, approximately 7.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 8.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 8.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 8.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 8.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 9.0 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 9.25 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 9.5 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of 1 or more individuals, approximately 9.75 × 10¹⁶ per kilogram of body weight in humans. 6 For HSPCs of one or more individuals, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 1.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 1.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 2.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 2.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 2.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 3.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 3.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 3.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 4.0 × 10¹⁶ per kilogram of body weight in humans. 7For HSPCs of one or more individuals, approximately 4.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 4.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 4.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 5.0 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 5.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 5.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 5.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 6.0 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 6.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 6.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 6.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 7.0 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 7.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 7.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 7.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 8.0 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 8.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 8.5 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 8.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 9.0 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 9.25 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of 1 or more individuals, approximately 9.5 × 10¹⁶ per kilogram of body weight in humans. 7For HSPCs of 1 or more individuals, approximately 9.75 × 10¹⁶ per kilogram of body weight in humans. 7 For HSPCs of one or more individuals, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 1.25 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 1.75 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 2.25 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 2.5 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 2.75 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 3.25 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 3.5 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 3.75 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of 1 or more individuals, approximately 4.0 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 4.25 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 4.5 × 10¹⁶ per kilogram of body weight in humans. 8 For HSPCs of one or more individuals, approximately 4.75 × 10¹⁶ per kilogram of body weight in humans. 8 One or more HSPCs, or approximately 5.0 × 10 per kilogram of human body weight 8 It includes one or more HSPCs. In some embodiments, the weight of the human subject is the actual weight of the human subject. In other embodiments, the weight of the human subject is the ideal actual weight of the human subject.

[0161] In some embodiments, CD45 +The first population of cells includes at least about 0.5% granule cells, at least about 1% granule cells, up to about 5% granule cells, up to about 3% granule cells, up to about 3% monocytes, up to about 2% monocytes, up to about 0.5% lymphocytes, up to about 2% lymphocytes, at least about 15% granule cells, at least about 20% granule cells, up to about 35% granule cells, up to about 30% granule cells, up to about 25% granule cells, at least about 15% monocytes, at least about 20% monocytes, up to about 35% monocytes, up to about 30% monocytes, up to about 25% monocytes, at least about 0.5% NK cells, and / or at least about 2% NK cells. In various embodiments, CD45 + The first population of cells, the Treg-enriched cell population, and CD45 + A third population of cells is obtained from one donor. In some embodiments, CD45 + The first population of cells, the Treg-enriched cell population, and / or CD45 + The third population of cells is homogeneous with respect to the human subject. In several embodiments, CD45 + The first population of cells, the Treg-enriched cell population, and / or CD45 + A third population of cells is obtained from an HLA-matched donor for a human subject. In various embodiments, CD45 + The first population of cells, the Treg-enriched cell population, and / or CD45 + A third population of cells is obtained from HLA-mismatched donors for human subjects. In some embodiments, CD45 + The first population of cells, the Treg-enriched cell population, and / or CD45 + The third population of cells is obtained from half-matched donors for the human subjects.

[0162] In some embodiments, Treg is CD4 + CD25 + CD127 dim or CD4 + FOXP3 + In some embodiments, Treg is CD4 + CD25 + CD127dim And FOXP3 + Also, in some cases, Treg-enriched cell populations are CD45 + Includes cells, for example, CD45 + Over 90% of the cells are Tregs. In some embodiments, CD45 + The second group of cells is CD4 + CD25 + CD127 dim Contains regulatory T cells (Treg). When used herein, CD45 + A second population of cells, a population of cells enriched with Treg cells, and CD4 + CD25 + CD127 dim A population of regulatory T cells (Tregs) can be used interchangeably. In other embodiments, CD45 + The second population of cells contains one or more doses of Treg. In various embodiments, a Treg-enriched cell population or CD45 containing at least one dose of Treg is used. + The second population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects. 5 ~Approx. 1.0×10 8 Each Treg cell has approximately 1.0 × 10¹⁶ units of body weight per kilogram in humans. 5 ~Approx. 2.0×10 7 Each Treg cell has approximately 1.0 × 10¹⁶ units of body weight per kilogram in humans. 5 ~Approx. 1.0×10 7 Each Treg, or approximately 5.0 × 10⁶ per kilogram of body weight in a human subject. 5 ~Approx. 4.0×10 6 Contains Treg cells. In some embodiments, a Treg-enriched cell population or CD45 containing at least one dose of Treg cells. + The second population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects. 5 One or more Tregs, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 5 One or more Tregs, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 2.5 × 10⁶ per kilogram of body weight in humans. 5One or more Tregs, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 3.5 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 4.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 4.5 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 5.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 5.5 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 6.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 6.5 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 7.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 7.5 × 10⁶ per kilogram of body weight in humans. 5 One or more Tregs, approximately 8.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 8.5 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 9.0 × 10⁶ per kilogram of body weight in humans. 5 More than 10 Tregs, approximately 9.5 × 10⁶ per kilogram of body weight in humans. 5 One or more Tregs, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 1.25 × 10¹⁶ units per kilogram of body weight. 6 One or more Tregs, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 1.75 × 10¹⁶ Treg cells per kilogram of body weight. 6 One or more Tregs, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 2.25 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 2.5 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 2.75 × 10¹⁶ units per kilogram of body weight. 6One or more Tregs, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 3.25 × 10¹⁶ units per kilogram of body weight. 6 More than 10 Tregs, approximately 3.5 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 3.75 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 4.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more individuals, approximately 4.25 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 4.5 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 4.75 × 10¹⁶ units per kilogram of body weight. 6 More than 10 Tregs, approximately 5.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 5.25 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 5.5 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 5.75 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 6.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 6.25 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 6.5 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 6.75 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 7.0 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, approximately 7.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 7.5 × 10⁶ per kilogram of body weight in humans. 6 For more than one Treg cell, the human population yields approximately 7.75 × 10¹⁶ units per kilogram of body weight. 6 One or more Tregs, approximately 8.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 8.25 × 10¹⁶ per kilogram of body weight in humans. 6More than 10 Tregs, approximately 8.5 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more individuals, approximately 8.75 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 9.0 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 9.25 × 10¹⁶ per kilogram of body weight in humans. 6 More than 10 Tregs, approximately 9.5 × 10⁶ per kilogram of body weight in humans. 6 For Tregs of 1 or more, approximately 9.75 × 10¹⁶ per kilogram of body weight in humans. 6 One or more Tregs, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 1.25 × 10¹⁶ units per kilogram of body weight. 7 One or more Tregs, approximately 1.5 × 10¹⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 1.75 × 10¹⁶ Treg cells per kilogram of body weight. 7 One or more Tregs, approximately 2.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 2.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 2.5 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 2.75 × 10¹⁶ units per kilogram of body weight. 7 One or more Tregs, approximately 3.0 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 3.25 × 10¹⁶ units per kilogram of body weight. 7 More than 10 Tregs, approximately 3.5 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 3.75 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 4.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more individuals, approximately 4.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 4.5 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 4.75 × 10¹⁶ units per kilogram of body weight. 7More than 10 Tregs, approximately 5.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 5.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 5.5 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 5.75 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 6.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 6.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 6.5 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 6.75 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 7.0 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, approximately 7.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 7.5 × 10⁶ per kilogram of body weight in humans. 7 For more than one Treg cell, the human population yields approximately 7.75 × 10¹⁶ units per kilogram of body weight. 7 One or more Tregs, approximately 8.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 8.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 8.5 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more individuals, approximately 8.75 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 9.0 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 9.25 × 10¹⁶ per kilogram of body weight in humans. 7 More than 10 Tregs, approximately 9.5 × 10⁶ per kilogram of body weight in humans. 7 For Tregs of 1 or more, approximately 9.75 × 10¹⁶ per kilogram of body weight in humans. 7 One or more Tregs, approximately 1.0 × 10⁶ per kilogram of body weight in humans. 8It contains one or more Tregs. In some embodiments, the weight of the human subject is the actual weight of the human subject. In other embodiments, the weight of the human subject is the ideal actual weight of the human subject.

[0163] In some embodiments, CD45 + The third group of cells is normal CD3 + Includes T cells (Tcon). In other embodiments, CD45 + The third population contains at least one dose of Tcon. When used herein, CD45 + The third population of cells and normal CD3 + Populations of T cells (Tcon) can be used interchangeably. In some embodiments, CD45 contains Tcon, or contains at least one dose of Tcon. + The third population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects. 5 ~Approx. 1.0×10 8 Each Tcon is approximately 1.0 × 10⁻¹⁶ per kilogram of body weight in humans. 5 ~Approx. 4.0×10 7 Each Tcon is approximately 1.0 × 10⁻¹⁶ per kilogram of body weight in humans. 5 ~Approx. 1.0×10 7 Each Tcon, or approximately 5.0 × 10⁻¹⁴ units per kilogram of body weight in a human subject. 5 ~Approx. 5×10 6 Contains Tcon. In some embodiments, CD45 contains Tcon, or contains at least one dose of Tcon. + The third population of cells is approximately 1.0 × 10⁶ per kilogram of body weight in human subjects. 5 Tcon of more than 1.5 × 10¹⁶ per kilogram of human body weight 5 Tcon of more than 10 units, approximately 2.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 2.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 3.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 3.5 × 10 units per kilogram of body weight in humans. 5Tcon of more than 10 units, approximately 4.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 4.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 5.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 5.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 6.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 6.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 7.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 7.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 8.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 8.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 9.0 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 10 units, approximately 9.5 × 10 units per kilogram of body weight in humans. 5 Tcon of more than 1.0 × 10¹ per kilogram of human body weight 6 For Tcon of any size, approximately 1.25 × 10¹⁶ per kilogram of body weight in humans. 6 Tcon of more than 1.5 × 10¹⁶ per kilogram of human body weight 6 For Tcon of any size, approximately 1.75 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 2.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 2.25 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 2.5 × 10 units per kilogram of body weight in humans. 6 For more than one Tcon, approximately 2.75 × 10¹⁶ units per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 3.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 3.25 × 10¹⁶ per kilogram of human body weight. 6Tcon of more than 10 units, approximately 3.5 × 10 units per kilogram of body weight in humans. 6 For more than one Tcon, approximately 3.75 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 4.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 4.25 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 4.5 × 10 units per kilogram of body weight in humans. 6 Tcon of more than 10 units, approximately 4.75 × 10⁻¹⁴ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 5.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 5.25 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 5.5 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 5.75 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 6.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 6.25 × 10¹⁶ units per kilogram of body weight in humans. 6 Tcon of more than 10 units, approximately 6.5 × 10 units per kilogram of body weight in humans. 6 For more than one Tcon, approximately 6.75 × 10¹⁶ units per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 7.0 × 10 units per kilogram of body weight in humans. 6 For more than one Tcon, approximately 7.25 × 10¹⁶ units per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 7.5 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 7.75 × 10¹⁶ units per kilogram of body weight in humans. 6 Tcon of more than 10 units, approximately 8.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 8.25 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 8.5 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 8.75 × 10¹⁶ per kilogram of human body weight. 6Tcon of more than 10 units, approximately 9.0 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 9.25 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 10 units, approximately 9.5 × 10 units per kilogram of body weight in humans. 6 For Tcon of any size, approximately 9.75 × 10¹⁶ per kilogram of human body weight. 6 Tcon of more than 1.0 × 10¹ per kilogram of human body weight 7 For Tcon of any size, approximately 1.25 × 10¹⁶ per kilogram of body weight in humans. 7 Tcon of more than 1.5 × 10¹⁶ per kilogram of human body weight 7 For Tcon of any size, approximately 1.75 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 2.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 2.25 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 2.5 × 10 units per kilogram of body weight in humans. 7 For more than one Tcon, approximately 2.75 × 10¹⁶ units per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 3.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 3.25 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 3.5 × 10 units per kilogram of body weight in humans. 7 For more than one Tcon, approximately 3.75 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 4.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 4.25 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 4.5 × 10 units per kilogram of body weight in humans. 7 Tcon of more than 10 units, approximately 4.75 × 10⁻¹⁴ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 5.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 5.25 × 10¹⁶ per kilogram of human body weight. 7Tcon of more than 10 units, approximately 5.5 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 5.75 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 6.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 6.25 × 10¹⁶ units per kilogram of body weight in humans. 7 Tcon of more than 10 units, approximately 6.5 × 10 units per kilogram of body weight in humans. 7 For more than one Tcon, approximately 6.75 × 10¹⁶ units per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 7.0 × 10 units per kilogram of body weight in humans. 7 For more than one Tcon, approximately 7.25 × 10¹⁶ units per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 7.5 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 7.75 × 10¹⁶ units per kilogram of body weight in humans. 7 Tcon of more than 10 units, approximately 8.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 8.25 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 8.5 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 8.75 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 9.0 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 9.25 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 10 units, approximately 9.5 × 10 units per kilogram of body weight in humans. 7 For Tcon of any size, approximately 9.75 × 10¹⁶ per kilogram of human body weight. 7 Tcon of more than 1.0 × 10¹ per kilogram of human body weight 8 It contains one or more Tcon units. In some embodiments, the weight of the human subject is the actual weight of the human subject. In other embodiments, the weight of the human subject is the ideal actual weight of the human subject.

[0164] In some embodiments, CD45+ The third population of cells contains at least about 0.1% CD34 + Cells, or approximately 0.2% to 20% of CD34 + The mixture contains cells and / or at least about 0.1% Treg. In various embodiments, CD45 + The third group of cells is memory T cells (Tmem), such as CD3 + CD45RA-CD45RO + This includes a population of Tmem. In some embodiments, the population of Tmem is approximately 3 × 10¹⁶ per kilogram of the actual or ideal body weight of the human subject. 5 It contains more than 10 Tmem. In some embodiments, the population of Tmem is about 3 x 10 per kilogram of the actual or ideal body weight of the human subject. 5 ~approximately 1x10 9 Includes several Tmem. In various embodiments, CD45 + The third group of cells is invariant natural killer T cells (iNKTs), such as CD3 + Vα24Jα18 + This includes a population of iNKTs. In some embodiments, the iNKT population has approximately 5 × 10⁶ times the actual or ideal body weight of a human subject per kilogram. 2 It contains more than 10 iNKTs. In some embodiments, the population of iNKTs is about 5 × 10 per kilogram of the actual or ideal body weight of the human subject. 2 ~Approx. 1×10 7 Includes iNKT. In some embodiments, CD45 + A third population of cells is co-cultured with donor cancer antigen / peptides and / or antigen-presenting cells.

[0165] In some embodiments, the cell population of this disclosure is approximately 1.0 × 10⁶ per kilogram of body weight of a human subject. 5 ~Approx. 5.0×10 8 Each Tmem is approximately 1.0 × 10¹⁶ per kilogram of body weight in humans. 5 ~Approx. 1.0×10 8 Each Tmem, or approximately 5.0 × 10⁻¹⁴ per kilogram of human body weight. 5 ~Approx. 2.0×10 7Contains Tmem. In some embodiments, Tmem is present in the composition, cell therapy product, and / or about 1.0 × 10¹⁶ per kilogram of body weight of a human subject. 5 More than 10¹Tmem units, approximately 1.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 2.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 2.5 × 10⁻¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 3.0 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 3.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 4.0 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 4.5 × 10⁻¹ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 5.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 5.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 6.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 6.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 7.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 5 More than 10¹ Tmem units, approximately 7.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 8.0 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 8.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 9.0 × 10⁻¹⁴ units per kilogram of body weight in humans. 5 More than 10¹ Tmem units, approximately 9.5 × 10¹⁶ units per kilogram of body weight in humans. 5 More than 10 Tmem units, approximately 1.0 × 10⁻¹⁶ per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 1.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹Tmem units, approximately 1.5 × 10¹⁶ units per kilogram of body weight in humans. 6More than 10¹ Tmem units, approximately 1.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 2.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 2.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 2.5 × 10⁻¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 2.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 3.0 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 3.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 3.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 3.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 4.0 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 4.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 4.5 × 10⁻¹ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 4.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 5.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 5.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 5.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 5.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 6.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 6.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 6.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 6.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 7.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 6More than 10¹ Tmem units, approximately 7.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 7.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 7.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 8.0 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 8.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 8.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 8.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 9.0 × 10⁻¹⁴ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 9.25 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 9.5 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10¹ Tmem units, approximately 9.75 × 10¹⁶ units per kilogram of body weight in humans. 6 More than 10 Tmem units, approximately 1.0 × 10⁻¹⁶ per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 1.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹Tmem units, approximately 1.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 1.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 2.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 2.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 2.5 × 10⁻¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 2.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 3.0 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 3.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 3.5 × 10¹⁶ units per kilogram of body weight in humans. 7More than 10¹ Tmem units, approximately 3.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 4.0 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 4.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 4.5 × 10⁻¹ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 4.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 5.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 5.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 5.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 5.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 6.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 6.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 6.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 6.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 7.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 7.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 7.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 7.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 8.0 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 8.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 8.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 8.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 9.0 × 10⁻¹⁴ units per kilogram of body weight in humans. 7More than 10 Tmem units, approximately 9.25 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 9.5 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10¹ Tmem units, approximately 9.75 × 10¹⁶ units per kilogram of body weight in humans. 7 More than 10 Tmem units, approximately 1.0 × 10⁻¹⁶ per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 1.25 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10¹Tmem units, approximately 1.5 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 1.75 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 2.0 × 10⁻¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 2.25 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 2.5 × 10⁻¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 2.75 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 3.0 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 3.25 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 3.5 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 3.75 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 4.0 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 4.25 × 10¹⁶ units per kilogram of body weight in humans. 8 More than 10 Tmem units, approximately 4.5 × 10⁻¹ units per kilogram of body weight in humans. 8 More than 10¹ Tmem units, approximately 4.75 × 10¹⁶ units per kilogram of body weight in humans. 8 One or more Tmem units, or approximately 5.0 × 10 units per kilogram of body weight in humans. 8The subject is administered an amount or dose containing one or more Tmem particles. In some embodiments, the body weight of the human subject is the actual body weight of the human subject. In other embodiments, the body weight of the human subject is the ideal actual body weight of the human subject.

[0166] In some embodiments, the cell population of the present disclosure, for example, CD45 + The first group of cells, CD45 + A second population of cells, and / or CD45 + A third population of cells may include one or more cells containing one or more chimeric receptors. In some embodiments, the one or more cells may be T cells, NK cells, CTLs, Tregs, and / or NKT cells. In some embodiments, the one or more chimeric receptors include one or more extracellular antigen-binding domains that bind to one or more cancer-related antigens. Any suitable cancer-related antigen known in the art may be targeted. In some embodiments, the chimeric receptor is a chimeric antigen receptor or a T cell receptor.

[0167] This specification provides compositions, multicomponent medical procedures, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods for improved hematopoietic stem cell transplantation (HCT), such as allogeneic hematopoietic stem cell transplantation (alloHSCT). The compositions, multicomponent medical procedures, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods disclosed herein may comprise one or more cell populations that can be administered in combination with GVHD prophylactic agents to achieve positive clinical outcomes. The cell populations may comprise one or more cell types, such as hematopoietic stem progenitor cells (HSPCs), conventional T cells (Tcon), regulatory T cells (Treg), invariant natural killer T cells (iNKT), memory T cells (Tmem), and combinations thereof.

[0168] This disclosure provides parameters relating to cell populations and methods for administering cell populations that may contribute to favorable clinical outcomes in HCT recipients. While we do not wish to be bound by theory, parameters that may contribute to favorable clinical outcomes in HCT recipients include, for example, the co-administration of GVHD prophylactic agents (e.g., tacrolimus) as described herein, the population to be administered, the order and timing of administration of different populations, the purity standards of the populations, methods for obtaining the populations, methods for handling or selecting the populations, the dosage of the population to be administered, methods for obtaining the populations, and combinations thereof.

[0169] In various embodiments, the administration involves CD45 in human subjects. + The first population of cells, the Treg-enriched cell population (e.g., CD45 + (Second population of cells), and CD45 + This involves injecting a third population of cells.

[0170] In some embodiments, a Treg-enriched cell population, or CD45 + The second population of cells (disclosed herein) is CD45 + It is administered approximately 5 minutes to approximately 5 hours after administration of the first population of cells (disclosed herein). In some embodiments, a Treg-enriched cell population or CD45 + The second group of cells is CD45 +Approximately 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 17 minutes, 18 minutes, 19 minutes, 20 minutes, 21 minutes, 22 minutes, 23 minutes, 24 minutes, 25 minutes, 26 minutes, 27 minutes, 28 minutes, 29 minutes, 30 minutes, 31 minutes, and 32 minutes after administration of the first population of cells. Approximately 33 minutes later, approximately 34 minutes later, approximately 35 minutes later, approximately 36 minutes later, approximately 37 minutes later, approximately 38 minutes later, approximately 39 minutes later, approximately 40 minutes later, approximately 41 minutes later, approximately 42 minutes later, approximately 43 minutes later, approximately 44 minutes later, approximately 45 minutes later, approximately 46 minutes later, approximately 47 minutes later, approximately 48 minutes later, approximately 49 minutes later, approximately 50 minutes later, approximately 51 minutes later, approximately 52 minutes later, approximately 53 minutes later, approximately 54 minutes later, approximately 55 minutes later, approximately 56 minutes later, approximately 57 minutes later, approximately 58 minutes later, approximately 59 minutes later, approximately 1 hour later, approximately 1.10 After a certain time, approximately 1.20 hours later, approximately 1.30 hours later, approximately 1.40 hours later, approximately 1.50 hours later, approximately 1.60 hours later, approximately 1.70 hours later, approximately 1.80 hours later, approximately 1.90 hours later, approximately 2 hours later, approximately 2.10 hours later, approximately 2.20 hours later, approximately 2.30 hours later, approximately 2.40 hours later, approximately 2.50 hours later, approximately 2.60 hours later, approximately 2.70 hours later, approximately 2.80 hours later, approximately 2.90 hours later, approximately 3 hours later, approximately 3.10 hours later Subsequently, it is administered at approximately 3.20 hours, 3.30 hours, 3.40 hours, 3.50 hours, 3.60 hours, 3.70 hours, 3.80 hours, 3.90 hours, 4 hours, 4.10 hours, 4.20 hours, 4.30 hours, 4.40 hours, 4.50 hours, 4.60 hours, 4.70 hours, 4.80 hours, 4.90 hours, or approximately 5 hours later.

[0171] In some embodiments, CD45 + A third population of cells (disclosed herein) is CD45 + The first population of cells (disclosed herein) is administered at least about 12 hours later, and CD45 + The third group of cells is CD45 + The first population of cells was administered approximately 24 to 96 hours later, and CD45 + The third group of cells is CD45 + The first population of cells was administered approximately 36 to 60 hours later, and CD45 +A third population of cells was administered at least about 12 hours after the Treg-enriched cell population (disclosed herein), and CD45 + A third population of cells was administered approximately 24–96 hours after the Treg-enriched cell population, and / or CD45 + The third population of cells is a Treg-enriched cell population or CD45 + The second population of cells is administered approximately 36 to 60 hours later.

[0172] In some embodiments, CD45 + A third population of cells (disclosed herein) is isolated CD45 + It is administered approximately 12 to 120 hours after administration of the first population of cells (disclosed herein). In some embodiments, CD45 + The third group of cells is CD45 +Approximately 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, and 38 hours after administration of the first population of cells. Then, approximately 39 hours later, approximately 40 hours later, approximately 41 hours later, approximately 42 hours later, approximately 43 hours later, approximately 44 hours later, approximately 45 hours later, approximately 46 hours later, approximately 47 hours later, approximately 48 hours later, approximately 49 hours later, approximately 50 hours later, approximately 51 hours later, approximately 52 hours later, approximately 53 hours later, approximately 54 hours later, approximately 55 hours later, approximately 56 hours later, approximately 57 hours later, approximately 58 hours later, approximately 59 hours later, approximately 60 hours later, approximately 61 hours later, approximately 62 hours later, approximately 63 hours later, approximately 64 hours later, approximately 65 hours later, approximately 66 hours later, approximately 67 Hours later, approximately 68 hours later, approximately 69 hours later, approximately 70 hours later, approximately 71 hours later, approximately 72 hours later, approximately 73 hours later, approximately 74 hours later, approximately 75 hours later, approximately 76 hours later, approximately 77 hours later, approximately 78 hours later, approximately 79 hours later, approximately 80 hours later, approximately 81 hours later, approximately 82 hours later, approximately 83 hours later, approximately 84 hours later, approximately 85 hours later, approximately 86 hours later, approximately 87 hours later, approximately 88 hours later, approximately 89 hours later, approximately 90 hours later, approximately 91 hours later, approximately 92 hours later, approximately 93 hours later, approximately 94 hours later, approximately 95 hours later, approximately It is administered at approximately 96 hours, 97 hours, 98 hours, 99 hours, 100 hours, 101 hours, 102 hours, 103 hours, 104 hours, 105 hours, 106 hours, 107 hours, 108 hours, 109 hours, 110 hours, 111 hours, 112 hours, 113 hours, 114 hours, 115 hours, 116 hours, 117 hours, 118 hours, 119 hours, and 120 hours.

[0173] In some embodiments, CD45 + The third population of cells (disclosed herein) is a Treg-enriched cell population or CD45 + It is administered approximately 12 to 120 hours after administration of the second population of cells (disclosed herein). In some embodiments, CD45 +The third population of cells is a Treg-enriched cell population or CD45 + Approximately 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 31 hours, 32 hours, 33 hours, 34 hours, 35 hours, 36 hours, 37 hours, and 38 hours after administration of the second population of cells. Then, approximately 39 hours later, approximately 40 hours later, approximately 41 hours later, approximately 42 hours later, approximately 43 hours later, approximately 44 hours later, approximately 45 hours later, approximately 46 hours later, approximately 47 hours later, approximately 48 hours later, approximately 49 hours later, approximately 50 hours later, approximately 51 hours later, approximately 52 hours later, approximately 53 hours later, approximately 54 hours later, approximately 55 hours later, approximately 56 hours later, approximately 57 hours later, approximately 58 hours later, approximately 59 hours later, approximately 60 hours later, approximately 61 hours later, approximately 62 hours later, approximately 63 hours later, approximately 64 hours later, approximately 65 hours later, approximately 66 hours later, approximately 67 Hours later, approximately 68 hours later, approximately 69 hours later, approximately 70 hours later, approximately 71 hours later, approximately 72 hours later, approximately 73 hours later, approximately 74 hours later, approximately 75 hours later, approximately 76 hours later, approximately 77 hours later, approximately 78 hours later, approximately 79 hours later, approximately 80 hours later, approximately 81 hours later, approximately 82 hours later, approximately 83 hours later, approximately 84 hours later, approximately 85 hours later, approximately 86 hours later, approximately 87 hours later, approximately 88 hours later, approximately 89 hours later, approximately 90 hours later, approximately 91 hours later, approximately 92 hours later, approximately 93 hours later, approximately 94 hours later, approximately 95 hours later, approximately It is administered at approximately 96 hours, 97 hours, 98 hours, 99 hours, 100 hours, 101 hours, 102 hours, 103 hours, 104 hours, 105 hours, 106 hours, 107 hours, 108 hours, 109 hours, 110 hours, 111 hours, 112 hours, 113 hours, 114 hours, 115 hours, 116 hours, 117 hours, 118 hours, 119 hours, and 120 hours.

[0174] HSPCs may possess broad regenerative capacity and the ability to differentiate into specialized cell types, such as the ability to reconstruct all hematopoietic cell lineages. HSPCs may accept asynchronous replication, resulting in two daughter cells with different phenotypes. HSPC cells may exist in a quiescent mitotic state. HSPCs may originate from bone marrow, peripheral blood, and / or umbilical cord blood.

[0175] Subsets of immune cells, such as normal T cells (Tcon), regulatory T cells (Treg), invariant natural killer T cells (iNKT), and memory T cells (Tmem), may contribute to the characteristics of GVHD after HCT, for example, to the GVT immune response, immune rearrangement, susceptibility to infection, and patient survival.

[0176] GVHD is primarily mediated by donor T cells, which can recognize recipient antigens and trigger an inflammatory response. The likelihood of acute and / or chronic GVHD can be reduced by T cell depletion (TCD) from the cell population intended for transplantation into the target. T cells can be depleted using methods including, but not limited to, physical adsorption of T cells to protein ligands such as lectins, immunodepletion with T cell-specific antibodies, and immunoaffinity techniques (e.g., use of T cell or lymphocyte-specific antibodies in immunoadsorption columns, magnetically activated cell sorting (MACS), or fluorescence-activated cell sorting (FACS)). The application of TCD techniques to donor grafts can, for example, reduce the number of T cells by 1 / 10 to 1 / 10. 5 This can lead to a reduction in T cell depletion to 1 / 2 and a decrease in the incidence of GVHD. However, TCD may also lead to an increased rate of cancer recurrence because T cells are deficient and the immune response to graft-versus-tumor (GVT) may be impaired. Furthermore, TCD may lead to impaired immune recovery and increased susceptibility to infection.

[0177] Both GVT and GVHD are largely mediated by conventional T cells (Tcon), and when T cell receptors recognize congenital antigens (tumor antigens in the case of GVT, and non-tumor recipient antigens in the case of GVHD), conventional T cells initiate an immune response. Tcon can, for example, cause GVT, GVHD, or a combination thereof. In some embodiments, administering Tcon after Treg administration may enhance GVT immunity and / or reduce susceptibility to infection.

[0178] Tcon, in a broad sense, refers to all CD3 + This can refer to T cells, cells expressing CD3 and CD4, or cells expressing CD3 and CD8, cells expressing moderate to high levels of CD127, cells expressing CD3 and moderate to high levels of CD127, cells expressing CD3, cells expressing moderate to high levels of CD127, and cells expressing CD4 or CD8. In some embodiments, Tcon does not express Vα24Jα18TCR. Tcon and regulatory T cells ("Treg") can be non-mutually exclusive cell populations. In some embodiments, Tcon and Treg are mutually exclusive cell populations.

[0179] Regulatory T cells ("Tregs") are a specialized subpopulation of T cells that negatively modulate (e.g., suppress) the activation of the immune system, thereby promoting immune tolerance. While we do not wish to be bound by theory, the Treg-enriched cell populations of this disclosure contribute to positive clinical outcomes, for example, by reducing the incidence and / or severity of GVHD in transplant recipient subjects and / or by improving immune rearrangement in transplant recipients. CD45, including at least HSPCs. +By administering a Treg-enriched cell population along with a cell population, for example, the retention of graft-versus-tumor response (GVT) may be promoted, and the incidence and / or severity of GVHD may be promoted. While we do not wish to be bound by theory, for example, compared to alternative hematopoietic stem cell transplantation (HCT) methods, i.e., methods separate from the compositions, multicomponent medical procedures, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods disclosed herein, administering a Treg-enriched cell population may prevent GVHD, and at least CD45 containing Tcon. + The GVT effect can be enhanced by administering a third population of cells. In some embodiments, compared to alternative HCT methods, i.e., methods separate from the compositions, multicomponent medical treatments, multicomponent cell therapy products, cell populations, solutions, formulations, kits, and / or methods disclosed herein, administering a Treg-enriched cell population reduces the risk of developing GVHD and contains at least Tcon and CD45 + Administering a third population of cells enhances the GVT effect.

[0180] When used herein, alternative compositions lack one or more cell populations and / or prophylactic agents disclosed herein. For example, an alternative composition lacks one or more of the following: a cell population containing HSPCs, a cell population containing Tregs, a cell population containing Tcones, and a prophylactic agent. In some embodiments, the alternative composition or treatment regimen includes additional cell populations or agents, such as additional or different GVHD prophylactic agents, compared to the composition or treatment regimen of the Disclosure.

[0181] Several subsets of Treg exist, for example, TCRαβ + CD4 +Regulatory T cells include natural regulatory T cells (nTregs) and induced regulatory T cells (iTregs). nTregs may be T cells produced in the thymus and delivered to the periphery as a long-lived lineage of autoantigen-specific lymphocytes. iTregs may be recruited from circulating lymphocytes and acquire regulatory properties under specific stimuli in the periphery. Both nTregs and iTregs are CD4 + CD25 + And both of them are dose-dependent in CD4 + CD25-T cell proliferation can be inhibited. In some embodiments, Tregs are anergistic and do not proliferate in response to TCR stimulation. In addition to being positive for CD4 and CD25, Tregs may also be positive for the intracellular marker foxp3 transcription factor. Tregs can be identified or selected based on various marker expression profiles. Non-limiting examples of marker expression profiles that may be used to select Tregs include (1) CD4 + CD25 + CD127 dim (2) CD4 + FOXP3 + (3) CD3 + CD4 + CD25 + (4) CD3 + CD4 + CD25 + CD127 dim (5) CD3 + CD4 + CD25 + CD127 dim FOXP3 + (6) CD3 + FOXP3 + (7) CD3 + CD4 + FOXP3 + (8) CD3 + CD4 + CD25 + FOXP3 + (9) CD3 + CD25 + FOXP3 + (10) CD3 + CD25+ CD127 dim (11) CD4 + CD25 + (12) CD4 + CD25 + CD127 dim FOXP3 + (13) FOXP3 + (14)CD4 + FOXP3 + (15) CD4 + CD25 + FOXP3 + (16)CD25 + FOXP3 + , and (17)CD25 + CD127 dim These are some examples.

[0182] Selection based on specific expression profiles, based on extracellular markers, does not require cell permeability, for example, CD4 + CD25 + CD127 dim This can be achieved through choices based on that.

[0183] Cell populations containing Tregs can, for example, reduce the incidence of transplant rejection, reduce the incidence and / or severity of GVHD, promote hematopoietic rearrangement, promote immune rearrangement, promote mixed chimerism, or achieve a combination of these.

[0184] The cell populations of this disclosure may include invariant natural killer T cells (iNKTs). iNKTs are a subclass of CD1d-limited natural killer T (NKT) cells that express highly conserved αβ-T cell receptors, including those of the Vα24Jα18TCRα chain in humans (referred to herein as "Vα24Jα18"). +iNKT cells are referred to as "CD3". iNKT cells can be identified by binding to CD1d polymers such as those carrying α-galactosylceramide (GalCer), PBS-57, PBS-44, or other natural or synthetic glycolipids. Another identification method is an antibody or a combination of antibodies that specifically recognize the Vα24Jα18 region. An example is the monoclonal antibody clone 6B11, which specifically binds to a Vα24 antibody, a Jα18 antibody, or a specific region of the Vα24Jα18TCR and can be used to identify iNKT cells. iNKT cells are CD3 + Vα24Jα18 + It can be done this way.

[0185] In some embodiments, iNKT can promote engraftment, promote GVT, reduce the incidence and / or severity of GVHD, reduce susceptibility to cancer recurrence, reduce susceptibility to infection, or a combination thereof. In some embodiments, iNKT promotes Treg activity. In some embodiments, iNKT promotes HSPC activity.

[0186] The cell populations of this disclosure may include memory T cells (Tmem). Tmem may refer to antigen-experienced T cells expressing, for example, the phenotypic marker CD45RO, TCRα, TCRβ, CD3, CD4, CD95, and IL-2Rβ, or the phenotypic marker CD45RO, TCRα, TCRβ, CD3, CD8, CD95, and IL-2Rβ. Tmem can confer immunity and persist in an inactive state for a long period of time. Tmem can rapidly acquire effector function when reloaded with the antigen. The population of Tmem may include any combination of the subclasses central memory T cells and effector memory T cells. In some embodiments, Tmem may express CD3 + CD45RA - CD45RO +In various ways, Tmem administered to HCT recipients can achieve, for example, promoting GVT, reducing GVHD, decreasing susceptibility to cancer recurrence, decreasing susceptibility to infection, or a combination of these effects.

[0187] Cell acquisition, processing, and preparation Certain aspects of this disclosure relate to methods for preparing compositions, multicomponent medical treatments, multicomponent cell therapy products, cell populations, solutions, formulations, and / or kits.

[0188] In some embodiments, at least one mobilized peripheral blood donor donation is collected from the donor, or up to two mobilized peripheral blood donor donations are collected from the donor. In some embodiments, the mobilized peripheral blood donor donation is an HSPC mobilized peripheral blood apheresis donor donation. In some embodiments, prior to peripheral blood donation, the donor may be vaccinated with tumor antigens and / or pathogens to enhance graft-versus-infection.

[0189] In several embodiments, at least one of the mobilized peripheral blood donor providers is CD34 + Cells and Tregs are processed and sorted to enrich them. In some embodiments, peripheral blood donor samples are further processed and sorted, or CD3 + Cells (e.g., Tcon) are alternatively processed and sorted to enrich them. In some embodiments, at least one of the mobilized peripheral blood donor offerings is CD34 + Cells are processed and sorted to enrich them with Treg and / or Tcon. CD34 + The processing and sorting of cells, Tregs, and Tcones may be carried out in any order. For example, at least one of the recruited peripheral blood samples may be CD34 first. + The cells are processed and sorted to enrich them with Tcon, and then Treg. Alternatively, at least one of the mobilized peripheral blood donor offerings is first CD34 +Cells may be processed and sorted to enrich cells, then Tregs, and then Tcones. Alternatively, at least one of the mobilized peripheral blood donor offerings may be processed and sorted to enrich Tregs first, then CD34s. + Cells and Tcon enrichment may be processed and sorted. Alternatively, at least one of the mobilized peripheral blood donor offerings may be enriched first with Treg, then Tcon, and then CD34. + Cells may be processed and sorted to enrich them. Alternatively, at least one of the mobilized peripheral blood donor offerings may be first enriched with Tcon, and then with CD34. + Cells and Tregs may be processed and sorted to enrich them. Alternatively, at least one of the mobilized peripheral blood donor offerings may be processed and sorted to enrich Tcones first, then Tregs, and then CD34. + Cells may be processed and sorted to enrich them. In some embodiments, at least one of the recruited peripheral blood donor offerings is enriched with Treg first, then CD34, without enriching Tcon. + Cells may be processed and sorted to enrich them. In other embodiments, at least one of the mobilized peripheral blood donor offerings is initially CD34 without enriching Tcon. + Cells can then be treated and sorted to enrich them with Treg cells.

[0190] In some embodiments, the time required to process and sort one or more mobilized peripheral blood donor offerings is less than approximately 40 hours, less than approximately 35 hours, less than approximately 30 hours, less than approximately 25 hours, less than approximately 20 hours, and The sorting time is less than approximately 15 hours, and / or the processing and sorting time for one or more mobilized peripheral blood donor donations is up to approximately 35 hours, the processing and sorting time for one or more mobilized peripheral blood donor donations is up to approximately 30 hours, the processing and sorting time for one or more mobilized peripheral blood donor donations is up to approximately 25 hours, the processing and sorting time for one or more mobilized peripheral blood donor donations is up to approximately 20 hours, or the processing and sorting time for one or more mobilized peripheral blood donor donations is up to approximately 15 hours.

[0191] In various embodiments, one or more of the mobilized peripheral blood donor offerings are processed and sorted using one or more immunoisolated particles (ISPs). For example, an ISP may comprise an affinity reagent such as immunomagnetic separation particles, where the particles may each be antibodies conjugated to iron-containing particles, and / or an affinity reagent such as immunofluorescence separation particles, where the particles may each be antibodies conjugated to fluorophores or other fluorescent particles.

[0192] In some embodiments, the cell population of the Disclosure is obtained from whole blood. The cell population of the Disclosure can be obtained from mobilized peripheral blood apheresis products, such as GCSF, GM-CSF, Mozovir® (Prelixafor), and combinations thereof, mobilized by administering these to a donor. The cell population of the Disclosure can be obtained from at least one apheresis product, two apheresis products, three apheresis products, four apheresis products, five apheresis products, six apheresis products, or more products. In some embodiments, the cell population of the Disclosure is obtained from one apheresis product. In some embodiments, the cell population of the Disclosure is obtained from two apheresis products. In some embodiments, the cell population of the Disclosure is obtained from apheresis products from one donor and apheresis products from at least a second donor.

[0193] In some embodiments, the cell populations of the Disclosure are obtained from bone marrow samples. In some embodiments, the cell populations of the Disclosure can be obtained from bone marrow samples, such as mobilized bone marrow samples, which are mobilized by administering GCSF, GM-CSF, mozovir (plelixafor), and combinations thereof to a donor.

[0194] In some embodiments, the cell population of this disclosure is obtained from umbilical cord blood.

[0195] The cell populations of this disclosure can be purified by selection from a cell population, for example, peripheral blood or peripheral blood apheresis products. The method for selecting the cell population may include a method involving positive or negative selection of the cell population of interest. The method for selecting the cell population may include affinity reagents, including but not limited to antibodies, full-length antibodies, antibody fragments, naturally occurring antibodies, synthetic antibodies, genetically engineered antibodies, full-length aphibodies, aphibody fragments, full-length affilins, affilin fragments, full-length antikalins, antikalin fragments, full-length avimers, avimer fragments, full-length DARPin, DARPin fragments, full-length finomers, finomer fragments, full-length kunitz domain peptides, kunitz domain peptide fragments, full-length monobodies, monobody fragments, peptides, or polyamino acids. In some embodiments, the affinity reagent is directly bound to the detection reagent and / or purification reagent. In some examples, the detection reagent and the purification reagent are the same. In some examples, the detection reagent and the purification reagent are different. For example, detection reagents and / or purification reagents are fluorescent, magnetic, or otherwise. In some cases, detection reagents and / or purification reagents are magnetic particles for column purification. For example, magnetic column purification can be performed using columns, antibodies, buffers, preparation materials, and reagents from a Miltenyi system (CliniMAC).

[0196] In various embodiments, at least one of the cell populations has a plurality of immunoisolated particles (ISPs) bound to the receptors of the cells of the cell population. In some examples, the plurality of ISPs are immunomagnetic isolation particles. In some embodiments, the plurality of ISPs contain antibodies conjugated to iron-containing particles. In some examples, at least a portion of the plurality of ISPs are CD34 of the HSPC cell population. +By binding to the receptor and selectively, the average number of ISPs per HSPC in an HSPC cell population is less than approximately 6,000, the average number of ISPs per HSPC in an HSPC cell population is equal to or less than approximately 3,000, and / or the average number of ISPs per HSPC in an HSPC cell population is approximately 1,700 to approximately 3,000. In some cases, at least a portion of multiple ISPs are bound to the CD25 cells of a Treg cell population. + By binding to receptors and selectively determining the average number of ISPs per T-reg cell in a Treg population, the average number of ISPs per T-reg cell in a Treg population is equal to or less than approximately 1700, or the average number of ISPs per T-reg cell in a Treg population is approximately 1400 to approximately 1700. In some cases, at least a portion of multiple ISPs are attached to the CD3 of cells in a heterogeneous cell population. + When bound to receptors and selectively observed, the average number of ISPs per cell in a heterogeneous T cell population is less than approximately 1,000.

[0197] Affinity reagents include immunoaffinity reagents that utilize the binding specificity of antibodies or their fragments or derivatives, and can positively or negatively select a cell population of interest. Methods for selecting cell populations may include affinity agents and columns, such as magnetically activated cell sorting (MACS) using specific antibodies and microbeads. Methods for selecting cell populations may include fluorescence-activated cell sorting (FACS), where cell populations are sorted based on staining profiles using one or more fluorescently bound antibodies. Methods for selecting cell populations may include physical adsorption, such as the physical adsorption of T cells to protein ligands like lectins.

[0198] HSPCs can be obtained by collection from bone marrow or peripheral blood. Bone marrow can be aspirated from the posterior or anterior iliac crest while the donor is under local or general anesthesia. HSPCs can also be obtained by collection from peripheral blood, for example, by peripheral blood apheresis. The number of recovered stem cells can be increased by treating the donor with a mobilizing agent, i.e., a drug that mobilizes stem cells from bone marrow to peripheral blood. Non-limiting examples of mobilizers include granule cell colony-stimulating factor (G-CSF), granule cell macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF), SDF-1 antagonists, CXCR4 antagonists (e.g., POL6326, BKT-140, TG-0054, NOX-A12), mozovir (plelixafor), CXCR2 ligands (e.g., GROβ), sphingosine-1-phosphatase (S1P) agonists (e.g., SEW2871), VCAM / VLA-4 inhibitors (e.g., BIO5192), proteosome inhibitors (e.g., bortezomib), parathyroid hormone, hypoxia-inducible factor (HIF) stabilizers (e.g., FG-4497), and combinations thereof. Techniques for recruiting stem cells into peripheral blood may include, for example, administering a recruiting agent to a donor at a rate of 10-40 μg / kg / day. The recruiting agent may be administered to the donor in, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 doses. The apheresis product may be isolated from the donor approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 26, 28, or 30 hours after administration of the recruiting agent.

[0199] CD45 of this disclosure + The cell population may include HSPCs. HSPCs can be selected based on CD34 expression. For example, the HSPCs of this disclosure can be selected using an anti-CD34 antibody as part of a magnetically activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS) system.

[0200] CD45 +The number of HSPCs in a cell population can be determined, for example, by flow cytometry using CD34 + This can be determined by quantifying the cells. In some embodiments, dose calculations are adjusted based on a measure of cell viability, such as viability determined by flow cytometry using propidium iodide or 7-AAD, or by trypan blue exclusion.

[0201] The cell population disclosed herein is Treg (e.g., CD45 + A second population of cells can be enriched. Tregs can be selected based on the expression of markers including CD3, CD4, CD25, CD127, FOXP3, and combinations thereof.

[0202] Treg cells can be selected using magnetically activated cell sorting (MACS). Treg cells can be selected using fluorescence-activated cell sorting (FACS). Treg cells can be selected using multiple procedures, such as multiple MACS selections, multiple FACS selections, or a combination of MACS and FACS selections. For example, the first selection may be based on CD25 expression, for example, by using MACS to select cells with CD25 expression from a hematopoietic cell sample. + The cells may be isolated. A second option is CD25 + This may also be carried out by contacting cells with antibodies specific to CD4 and CD127, where FACS is performed on CD4 + CD127 dim It is used to isolate certain cells.

[0203] Treg cells can be isolated from whole blood. Treg cells can be isolated from peripheral blood apheresis products. Treg cells can be isolated from cell populations that are pre-enriched and / or depleted of one or more other cell types, e.g., CD34 + Cells can be isolated from a depleted cell population. In some embodiments, Treg is CD34 + It is isolated from the MACS-selected flow-through fraction.

[0204] The number of Tregs in a cell population can be determined, for example, by flow cytometry, and Tregs are, for example, CD4 + CD25 + CD127 dim or CD4 + FOXP3 + It can be identified as such. Dose calculations can be adjusted based on a measure of cell viability, such as flow cytometry using propidium iodide or 7-AAD, or viability determined by trypan blue dye exclusion.

[0205] In some embodiments, CD45 + The third population of cells may include a population of Tcon. CD45 containing at least Tcon. + A third population of cells can draw its supply from peripheral blood. CD45 + A third population of cells can draw its supply from peripheral blood apheresis products.

[0206] In some embodiments, the selection step is not performed, and CD45 including at least Tcon is not performed. + The cell population is supplied directly from peripheral blood or aliquots of apheresis products. In some embodiments, the cell population can be enriched with Tcon by sorting based on the expression of various markers, for example, MACS, FACS, or a combination thereof. In some embodiments, CD45 + The third group of cells is CD3 + Cells can be enriched by sorting. In some embodiments, CD45 + The third group of cells is CD4 + and CD8 + Cells can be enriched by selection. In some embodiments, CD45 + A third population of cells can be enriched by negative selection, and non-Tcon cells are eliminated by depleting, for example, cells expressing CD34, CD19, CD25, or combinations thereof, using MACS.

[0207] CD45 + The number of Tcon present in the third population of cells can be determined, for example, by flow cytometry using CD3 + This can be quantified by quantifying the cells. CD3 in aliquots + The number of cells can be determined, and the volume containing an appropriate dose of CD3 cells to be administered to the recipient can be determined. The dose calculation can be adjusted based on a measure of cell viability, such as flow cytometry using propidium iodide or 7-AAD, or viability determined by trypan blue dye exclusion.

[0208] The apheresis product of this disclosure consists of two parts, namely CD45 including at least Tcon. + One portion used to obtain a third population of cells, as well as CD45 containing at least HSPC + The cell population and the Treg-enriched cell population can be separated into two parts for isolation and purification. In an alternative embodiment, CD34 + Cells are isolated and purified from apheresis products to create a CD34-negative cell fraction, from which Treg cells are then isolated to assist in obtaining a Treg-enriched cell population.

[0209] The cell populations of this disclosure may include a population of iNKT cells. The iNKT population may be supplied from peripheral blood. The iNKT population may be supplied from peripheral blood apheresis products.

[0210] Cell populations can be enriched with iNKTs by selecting them based on the expression of various markers, for example, MACS, FACS, or a combination thereof. The iNKT population can be enriched with, for example, CD3 + Vα24Jα18 + Cells can be enriched through selection.

[0211] The number of iNKTs present in a population can be determined, for example, by flow cytometry using CD3 +Vα24Jα18 + This can be quantified by quantifying cells. CD3 in aliquots + Vα24Jα18 + The number of cells can be determined, and the appropriate volume containing iNKT to be administered to the recipient can be determined. In some embodiments, the dose calculation is adjusted based on a measure of cell viability, such as viability determined by flow cytometry using propidium iodide or 7-AAD, or by trypan blue exclusion.

[0212] The cell populations of this disclosure may include populations of Tmem. Populations of Tmem may be supplied from peripheral blood. Populations of Tmem may be supplied from peripheral blood apheresis products.

[0213] Cell populations can be enriched with Tmem by sorting them based on the expression of various markers, for example, MACS, FACS, or a combination thereof. The Tmem population can be enriched with, for example, CD3 + CD45RA-CD45RO + Cells can be enriched through selection.

[0214] The number of Tmem present in a population can be determined, for example, by flow cytometry using CD3 + CD45RA-CD45RO + This can be quantified by quantifying the cells. CD3 in aliquots + CD45RA-CD45RO + The number of cells can be determined, and the appropriate volume containing Tmem to be administered to the recipient can be determined. The dose calculation can be adjusted based on a measure of cell viability, such as flow cytometry using propidium iodide or 7-AAD, or viability determined by trypan blue dye exclusion.

[0215] The cell populations described herein can be administered fresh after isolation, or after cryopreservation and subsequent thawing.

[0216] Fresh cells isolated from a donor ("fresh cells") can be administered to the recipient. Fresh cells can be stored in a buffer, such as CliniMACs PBS-EDTA buffer containing 0.5% human serum albumin, or in Plasma-Lyte-A at pH 7.4 supplemented with 2% human serum albumin. Fresh cells can be stored at low temperatures (e.g., 2-8°C) without cryopreservation or freezing.

[0217] After obtaining a population of fresh cells from a donor, the fresh cells should be allowed to stand for at least approximately 1 hour, at least approximately 2 hours, at least approximately 3 hours, at least approximately 4 hours, at least approximately 5 hours, at least approximately 6 hours, at least approximately 7 hours, at least approximately 8 hours, at least approximately 9 hours, at least approximately 10 hours, at least approximately 11 hours, at least approximately 12 hours, at least approximately 13 hours, at least approximately 14 hours, at least approximately 15 hours, at least approximately 16 hours, at least approximately 17 hours, at least approximately 18 hours, at least approximately 19 hours, at least approximately 20 hours, at least approximately 21 hours, at least approximately 22 hours, at least approximately 23 hours, at least approximately 24 hours, at least approximately 25 hours, at least approximately 26 hours, at least approximately 27 hours, at least approximately 28 hours, and at least It can be stored for approximately 29 hours, at least approximately 30 hours, at least approximately 31 hours, at least approximately 32 hours, at least approximately 33 hours, at least approximately 34 hours, at least approximately 35 hours, at least approximately 36 hours, at least approximately 37 hours, at least approximately 38 hours, at least approximately 39 hours, at least approximately 40 hours, at least approximately 44 hours, at least approximately 48 hours, at least approximately 50 hours, at least approximately 55 hours, at least approximately 60 hours, at least approximately 61 hours, at least approximately 62 hours, at least approximately 65 hours, at least approximately 70 hours, at least approximately 72 hours, at least approximately 80 hours, at least approximately 90 hours, at least approximately 96 hours, at least approximately 120 hours, at least approximately 150 hours, at least approximately 200 hours, at least approximately 300 hours, or longer.

[0218] After obtaining a fresh cell population from a donor, the fresh cells can be stored for up to approximately 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 40 hours, 48 ​​hours, 60 hours, 70 hours, 72 hours, 80 hours, 90 hours, 96 hours, 120 hours, 150 hours, 200 hours, or 300 hours before administration to the subject.

[0219] In some embodiments, after processing, one or more cell populations of the present disclosure, for example, CD45 + The first group of cells, CD45 + A second population of cells, and / or CD45 + A third population of cells may be formulated with one or more excipients and / or cryoprotectants. In some embodiments, the one or more excipients include, for example, a buffer such as a transport buffer or an infusion buffer. In some embodiments, the cell population is formulated at a neutral pH. In some embodiments, the HSPC of the Disclosure is formulated with one or more excipients at a neutral pH. In some embodiments, the Treg of the Disclosure is formulated with one or more excipients at a neutral pH. In some embodiments, the Tcon of the Disclosure is formulated with one or more excipients at a neutral pH. In some embodiments, the HSPC and Treg of the Disclosure are formulated together with one or more excipients at a neutral pH. In some embodiments, the HSPC and Tcon of the Disclosure are formulated together with one or more excipients at a neutral pH. In some embodiments, the Treg and Tcon of the Disclosure are formulated together with one or more excipients at a neutral pH. In some embodiments, the HSPC, Treg, and Tcon of this disclosure are formulated together with one or more excipients at a neutral pH. In some embodiments, the one or more excipients include one or more transport buffers.

[0220] In some embodiments, the neutral pH is in the range of about 6.8 to about 7.6. In some embodiments, the neutral pH is about 6.8, about 6.85, about 6.9, about 6.95, about 7, about 7.05, about 7.1, about 7.15, about 7.2, about 7.25, about 7.3, about 7.35, about 7.4, about 7.45, about 7.5, about 7.55, or about 7.6.

[0221] donor In some embodiments, one or more cell populations of the Disclosure, for example, CD45 of the Disclosure + The first population of cells, CD45 of this disclosure + The second population of cells, CD45 of this disclosure + A third population of cells, and / or one or more cells containing chimeric receptors, are derived from one human blood donor. In some embodiments, each cell population is provided as a separate cell population and is derived from one human blood donor.

[0222] One or more cells comprising a cell population and / or chimeric receptor may include, for example, cells derived from one or more donors, each HLA-typed, to determine the degree of HLA matching with the recipient of the cell population. In some embodiments, the donor is unrelated to the recipient. In some embodiments, the donor is related to the recipient, for example, the donor is a parent, child, sibling, grandparent, grandchild, aunt, uncle, or cousin. In some embodiments, the donor is a first-degree relative of the recipient. In some embodiments, the donor is a second-degree relative of the recipient. In some embodiments, related or unrelated donors are HLA-matched to the recipient. In some embodiments, related or unrelated donors are HLA-mismatched to the recipient. In some embodiments, related or unrelated donors are HLA half-matched to the recipient. In some embodiments, related or unrelated donors are at least 16 years of age. In some embodiments, the related or unrelated donor is at least 18 years of age.

[0223] Human leukocyte antigens (HLA), also broadly known as major histocompatibility complex (MHC) antigens, are protein molecules expressed on the cell surface that can confer antigen specificity to a cell. HLA / MHC antigens are target molecules that can be recognized by T cells and natural killer (NK) cells as either originating from the same hematopoietic stem cell source as immune effector cells ("self") or from a different hematopoietic cell source ("non-self"). HLA class I antigens (A, B, and C in humans) can be expressed by the majority of cells, while HLA class II antigens (DR, DP, and DQ in humans) can be expressed mainly by professional antigen-presenting cells. Both HLA classes may be involved in GVHD.

[0224] HLA antigens are encoded by highly polymorphic genes, and various alleles exist for each HLA class I and II gene. The gene products of alleles may differ in one or more amino acids in the α and / or β domains. An individual's HLA haplotype can be determined using a panel of specific antibodies or nucleic acid reagents, for example, using leukocytes expressing class I and class II molecules. HLA alleles can be described at various levels. Most nomenclature begins with the name of the HLA and locus, followed by several (even) digits that identify the allele. The first two digits may identify a group of alleles. The third and fourth digits, if present, may identify synonymous alleles. The fifth and sixth digits, if present, may indicate any synonymous variation within the gene's coding frame. The seventh and eighth digits, if present, may identify variations outside the coding region. Letters such as L, N, Q, or S follow the allele name to identify the expression level or other known non-genomic data related thereto. Therefore, a fully described allele, excluding HLA prefixes and locus notations, can be up to nine words long.

[0225] A set of HLA alleles inherited from one parent forms a haplotype. HLA haploidentical can refer to a donor-recipient pair in which one chromosome matches at least in HLA-A, HLA-B, and HLA-DR. Haploidentical pairs may or may not match in other alleles, such as other HLA genes on other chromosomes, or additional histocompatibility loci on either chromosome. Such donors can often be found within a family; for example, both parents may be haploidentical to their child, and siblings may also be haploidentical.

[0226] The cell population may be derived from related or unrelated donors who have been HLA-typed with any number of HLA alleles. The donor and subject may be HLA-matched, for example, matching in all typed HLA alleles. The donor and subject may be HLA-mismatched, for example, at least one HLA antigen may be mismatched between the donor and recipient.

[0227] In some embodiments, related or unrelated donors and subjects can be classified by HLA type using six alleles, such as HLA-A, HLA-B, and HLA-DR alleles. Donors and subjects can match in, for example, 3 / 6, 4 / 6, 5 / 6, or 6 / 6 alleles. In some embodiments, donors and subjects match in at least 5 / 6 alleles. In some embodiments, donors and subjects match in 6 / 6 alleles.

[0228] In some embodiments, related or unrelated donors and subjects can be HLA-typed using eight alleles, such as HLA-A, HLA-B, HLA-C, and HLA-DR alleles (e.g., HLA-DRB1 allele). Donors and subjects can match in, for example, 4 / 8, 5 / 8, 6 / 8, 7 / 8, or 8 / 8 alleles. In some embodiments, donors and subjects match in at least 6 / 8 alleles. In some embodiments, donors and subjects match in at least 7 / 8 alleles. In some embodiments, donors and subjects match in 8 / 8 alleles.

[0229] In some embodiments, related or unrelated donors and subjects can be classified by HLA type using 10 alleles, such as HLA-A, HLA-B, HLA-C, and HLA-DR alleles (e.g., HLA-DRB1 allele). Donors and subjects can match in, for example, 5 / 10, 6 / 10, 7 / 10, 8 / 10, 9 / 10, or 10 / 10 alleles. In some embodiments, donors and subjects match in at least 7 / 10 alleles. In some embodiments, donors and subjects match in at least 8 / 10 alleles. In some embodiments, donors and subjects match in at least 9 / 10 alleles. In some embodiments, donors and subjects match in 10 / 10 alleles.

[0230] In some embodiments, related or unrelated donors and subjects can be classified by HLA type using 12 alleles, such as HLA-A, HLA-B, HLA-C, HLA-DR allele (e.g., HLA-DRB1 allele), and HLA-DP allele (e.g., HLA-DPB1 allele). Donors and subjects can match in, for example, 6 / 12, 7 / 12, 8 / 12, 9 / 12, 10 / 12, 11 / 12, or 12 / 12 alleles. In some embodiments, donors and subjects match in at least 9 / 12 alleles. In some embodiments, donors and subjects match in at least 10 / 12 alleles. In some embodiments, donors and subjects match in at least 11 / 12 alleles. In some embodiments, donors and subjects match in 12 / 12 alleles.

[0231] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched unrelated donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched unrelated donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched unrelated donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched unrelated donors with 12 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.Cell populations can be generated from matched unrelated donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched unrelated donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0232] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched related donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched related donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched related donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched related donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched related donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched related donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched related donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched related donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched related donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and all are typed using DNA-based, highly analytical methods.Cell populations can be generated from matched related donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched related donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using high-resolution DNA-based methods.

[0233] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched parent donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched parent donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched parent donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched parent donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched parent donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched parent donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched parent donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched parent donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched parent donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based, high-resolution methods. Cell populations can be generated from matched parent donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based, high-resolution methods.Cell populations can be generated from matching parental donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and all are typed using DNA-based, highly analytical methods.

[0234] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched child donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors that are 11 / 12 matched in terms of HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based, highly analytical methods.Cell populations can be generated from matched child donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0235] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched sibling donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched sibling donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched sibling donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched sibling donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched sibling donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched sibling donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched sibling donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched child donors that are 11 / 12 matched in terms of HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based, highly analytical methods.Cell populations can be generated from matched child donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched child donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0236] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched grandparent donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandparent donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandparent donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandparent donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandparent donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched grandparent donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched grandparent donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched grandparent donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matching grandparent donors that are 11 / 12 matched in terms of HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and all are typed using DNA-based, highly analytical methods.Cell populations can be generated from matched grandparent donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandparent donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0237] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched grandchild donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandchild donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandchild donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandchild donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandchild donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched grandchild donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched grandchild donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched grandchild donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched grandchild donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched grandchild donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.Cell populations can be generated from matched grandchild donors with 9 / 12 matching in terms of HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and all are typed using DNA-based, highly analytical methods.

[0238] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched aunt donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched aunt donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched aunt donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched aunt donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched aunt donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched aunt donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched aunt donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched aunt donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched aunt donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and all are typed using DNA-based, highly analytical methods.Cell populations can be generated from matched aunt donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched aunt donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0239] Cell populations and / or one or more cells containing chimeric receptors can be generated from a matched uncle donor with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using a DNA-based high-resolution method. Cell populations can be generated from a matched uncle donor with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using a DNA-based high-resolution method. Cell populations can be generated from a matched uncle donor with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using a DNA-based high-resolution method. Cell populations can be generated from a matched uncle donor with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a DNA-based high-resolution method. Cell populations can be generated from matched uncle donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched uncle donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched uncle donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched uncle donors with a 12 / 12 match for HLA-A, -B, -C, -DQB1, -DRB1, and -DRB1, and they are all typed using high-resolution DNA-based methods. Cell populations can be generated from matched uncle donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based, highly analytical methods.Cell populations can be generated from matched uncle donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched uncle donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0240] Cell populations and / or one or more cells containing chimeric receptors can be generated from matched cousin donors with 8 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with 7 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with 6 / 8 matching for HLA-A, -B, -C, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with 10 / 10 matching for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with a 9 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched cousin donors with an 8 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched cousin donors with a 7 / 10 match for HLA-A, -B, -C, -DQB1, and -DRB1, and they are all typed using a high-resolution DNA-based method. Cell populations can be generated from matched cousin donors with 12 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with 11 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.Cell populations can be generated from matched cousin donors with 10 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods. Cell populations can be generated from matched cousin donors with 9 / 12 matching for HLA-A, -B, -C, -DQB1, -DRB1, and DPB1, and they are all typed using DNA-based high-resolution methods.

[0241] In some embodiments, HLA mismatch occurs because a related or unrelated donor of the same species is homozygous with respect to the HLA allele, while the recipient is heterozygous with respect to the HLA allele. In some embodiments, HLA mismatch occurs because a related or unrelated donor of the same species is heterozygous with respect to the HLA allele, while the recipient is homozygous with respect to the HLA allele. In some embodiments, HLA mismatch occurs because both the related or unrelated donor and the recipient are heterozygous with respect to the HLA allele.

[0242] In various embodiments, CD45 + The first population of cells, the Treg-enriched cell population (e.g., CD45 + (Second group of cells), CD45 + A third population of cells, and / or one or more cells containing chimeric receptors, are homogeneous with respect to human subjects.

[0243] In some embodiments, CD45 + The first population of cells, the Treg-enriched cell population (e.g., CD45 + (Second group of cells), CD45 + A third population of cells, and / or one or more cells containing chimeric receptors, are obtained from a donor that is HLA-matched to a human subject.

[0244] In some embodiments, CD45 + The first population of cells, the Treg-enriched cell population (e.g., CD45 +(Second group of cells), CD45 + A third population of cells, and / or one or more cells containing chimeric receptors, are obtained from a donor that is HLA-mismatched to a human subject.

[0245] In various embodiments, CD45 + The first population of cells, the Treg-enriched cell population (e.g., CD45 + (Second group of cells), CD45 + A third population of cells, and / or one or more cells containing chimeric receptors, are obtained from a donor that is half-matched to the human subject.

[0246] A cell population containing chimeric receptors and / or one or more cells may be derived from an allogeneic donor. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is a first-degree relative of the subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is a second-degree relative of the subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is not related to the subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is HLA-matched to the recipient subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is HLA-mismatched to the recipient subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is half-matched to the recipient subject. A cell population containing chimeric receptors and / or one or more cells may be generated from a donor who is related to the recipient subject by blood, such as a parent, child, sibling, grandparent, grandchild, uncle, aunt, or cousin. A cell population containing chimeric receptors and / or one or more cells can be generated from a donor who is at least 16 years old. The cell population can be generated from a donor who is at least 18 years old.

[0247] Cell populations containing chimeric receptors and / or one or more cells may be generated from donors that meet the eligibility criteria for viable leukocyte-rich cell or tissue donors as defined in 21 CFR §1271 2018 and related FDA Guidance for Industry. For example, cell populations may be generated from donors that meet one or more of the eligibility criteria outlined below. Eligibility Determination for Donors of Human Cells,Tissues,and Cellular and Tissue-Based Products,2007;Use of Donor Screening Tests to Test Donors of Human Cells,Tissues and Cellular and Tissue-Based Products for Infection with Treponema pallidum(Syphilis),2015;Use of Nucleic Acid Tests to Reduce the Risk of Transmission of Hepatitis B Virus from Donors of Human Cells,Tissues,and Cellular and Tissue-Based Products,2016;Use of Nucleic Acid Tests to Reduce the Risk of Transmission of West Nile Virus from Living Donors of Human Cells,Tissues,and Cellular and Tissue-Based Products(HCT / Ps),2016;and Donor Screening Recommendations to Reduce the Risk of Transmission of Zika Virus by Human Cells, Tissues, and Cellular and Tissue-Based Products, 2018).Cell populations can be generated from donors who meet any of the donation criteria, as identified by the standard NMDP guidelines (NMDP donors).

[0248] Cell populations containing chimeric receptors and / or one or more cells can be generated from donors who show no evidence of active infection. Cell populations can be generated from donors who are not seropositive for HIV-1 or -2, or HTLV-1 or 2. Cell populations containing chimeric receptors and / or one or more cells can be generated from donors who are not positive for anti-hepatitis C (HCV) antibodies or HCV NAT. Cell populations containing chimeric receptors and / or one or more cells can be generated from donors who are negative for chronic HBV infection. Cell populations containing chimeric receptors and / or one or more cells can be generated from donors who are not likely to have Zika virus infection as defined by any of the following: (i) a medical diagnosis of Zika virus infection within the past six months; (ii) residence in or travel to an area with active Zika virus transmission within the past six months; (iii) unprotected sexual intercourse with a person known to have either risk factor (i) or (ii) within the past six months. A population of cells containing chimeric receptors and / or one or more cells can be generated from a donor that does not exhibit signs or symptoms consistent with active Zika virus infection.

[0249] One or more cell populations and / or one or more cells containing the chimeric receptors of this disclosure can be obtained from one donor, for example, from mobilized peripheral blood apheresis from one donor. HSPC, Treg, Tcon, iNKT, Tmem, or any combination thereof can be obtained from one donor.

[0250] One or more cell populations and / or one or more cells containing the chimeric receptor of this disclosure can be obtained from one donor, and one or more additional cell populations and / or one or more cells containing the chimeric receptor of this disclosure can be obtained from a second donor. One cell population and / or one or more cells containing the chimeric receptor of this disclosure can be obtained from one donor, and a second cell population and / or one or more cells containing the chimeric receptor of this disclosure can be obtained from multiple donors. The populations and / or one or more cells of this disclosure containing the chimeric receptor can be obtained from multiple donors, for example, from mobilized peripheral blood apheresis from multiple donors. HSPCs can be obtained from multiple donors. Tregs can be obtained from multiple donors. Tcones can be obtained from multiple donors. One or more cells containing the chimeric receptor can be obtained from multiple donors. iNKTs can be obtained from multiple donors. Tmems can be obtained from multiple donors.

[0251] Dose of cell population The dose of a cell population containing a chimeric receptor and / or one or more cells administered to a subject (e.g., a human subject) may be based on the subject's body weight. In some cases, the subject's body weight may be used to determine the dose of one or more cell populations administered to the subject. In some cases, the cell dose may be based on the subject's actual weight, e.g., ideal body weight, instead of actual body weight. Ideal body weight may be a preferred dose calculation method to avoid erroneous cell doses due to excess body fat and / or muscle mass. The subject's ideal body weight may be calculated using the subject's height and sex. Other methods may be used to calculate the subject's ideal body weight, e.g., other methods for determining the subject's body fat percentage. One or more cell populations of this disclosure, e.g., CD45 + The first group of cells, CD45 + A second population of cells, and / or CD45 + The dose for the third population of cells may be based on the subject's adjusted body weight (ABW) if the subject's actual body weight exceeds 120% of the subject's ideal body weight (IBW).

[0252] In some embodiments, the human subject has a weight in the range of approximately 4 kilograms (kg) to approximately 200 kilograms (kg). In some embodiments, the human subject has a weight in the range of approximately 4 kilograms, approximately 5 kilograms, approximately 6 kilograms, approximately 7 kilograms, approximately 8 kilograms, approximately 9 kilograms, approximately 10 kilograms, approximately 11 kilograms, approximately 12 kilograms, approximately 13 kilograms, approximately 14 kilograms, approximately 15 kilograms, approximately 16 kilograms, approximately 17 kilograms, approximately 18 kilograms, approximately 19 kilograms, approximately 20 kilograms, approximately 21 kilograms, approximately 22 kilograms, approximately 23 kilograms, approximately 24 kilograms, approximately 25 kilograms, approximately 26 kilograms, approximately 27 kilograms, approximately 28 kilograms, approximately 29 kilograms, approximately 30 kilograms, approximately 31 kilograms, approximately 32 kilograms, approximately 33 kilograms, approximately 34 kilograms, approximately 35 kilograms, approximately 36 kilograms, approximately 37 kilograms, approximately 38 kilograms, approximately 39 kilograms, approximately 40 kilograms, approximately 41 kilograms, approximately 42 kilograms, approximately 43 kilograms, approximately 44 kilograms, approximately 45 kilograms, approximately 46 kilograms, approximately 47 kilograms, approximately 48 kilograms, approximately 49 kilograms, approximately 50 kilograms, approximately 51 kilograms, approximately 52 kilograms, approximately 53 kilograms, approximately 54 kilograms, approximately 55 kilograms, approximately 56 kilograms, approximately 57 kilograms, approximately 58 kilograms, approximately 59 kilograms, approximately 60 kilograms, approximately 61 kilograms, approximately 62 kilograms, approximately 63 kilograms, approximately 64 kilograms, approximately 65 kilograms, approximately 66 kilograms, approximately 67 kilograms, approximately 68 kilograms, approximately 69 kilograms, approximately 70 kilograms, approximately 71 kilograms, approximately 72 kilograms, approximately 73 kilograms, approximately 74 kilograms, approximately 75 kilograms, approximately 76 kilograms, approximately 77 kilograms, approximately 78 kilograms, approximately 79 kilograms, approximately 80 kilograms, approximately 81 kilograms Grams, approximately 82 kilograms, approximately 83 kilograms, approximately 84 kilograms, approximately 85 kilograms, approximately 86 kilograms, approximately 87 kilograms, approximately 88 kilograms, approximately 89 kilograms, approximately 90 kilograms, approximately 91 kilograms, approximately 92 kilograms, approximately 93 kilograms, approximately 94 kilograms, approximately 95 kilograms, approximately 96 kilograms, approximately 97 kilograms, approximately 98 kilograms, approximately 99 kilograms, approximately 100 kilograms, approximately 105 kilograms, approximately 110 kilograms, approximately 115 kilograms, approximately 120 kilograms, approximately 125 kilograms, approximately 130 kilograms,They have a weight of approximately 135 kilograms, 140 kilograms, 145 kilograms, 150 kilograms, 155 kilograms, 160 kilograms, 165 kilograms, 170 kilograms, 175 kilograms, 180 kilograms, 185 kilograms, 190 kilograms, 195 kilograms, or 200 kilograms.

[0253] HSPC CD45 containing at least HSPC or at least one dose of HSPC + The first population of cells, or CD34 + The population of hematopoietic stem progenitor cells (HSPCs) is at least approximately 1 × 10¹⁶ per kilogram (kg) of actual or ideal body weight of the recipient subject (e.g., human subject). 4 each, at least about 2 × 10 4 Each, at least approximately 3 × 10 4 Each, at least approximately 4 × 10 4 Each, at least approximately 5 x 10 4 Each, at least approximately 6 x 10 4 Each, at least approximately 7 x 10 4 Each, at least approximately 8 x 10 4 Each, at least approximately 9 x 10 4 each, at least about 1 × 10 5 each, at least about 2 × 10 5 Each, at least approximately 3 × 10 5 Each, at least approximately 4 × 10 5 Each, at least approximately 5 x 10 5 Each, at least approximately 6 x 10 5 Each, at least approximately 7 x 10 5 Each, at least approximately 8 x 10 5 Each, at least approximately 9 x 10 5 each, at least about 1 × 10 6 Each, at least about 1.1 × 10 6 Each, at least approximately 1.2 × 10 6 Each, at least approximately 1.3 × 10 6 Each, at least approximately 1.4 × 10 6 Each, at least approximately 1.5 × 10 6 Each, at least approximately 1.6 × 10 6 Each, at least approximately 1.7 × 106 Each, at least approximately 1.8 × 10 6 Each, at least approximately 1.9 × 10 6 each, at least about 2 × 10 6 Each, at least approximately 2.1 × 10 6 Each, at least approximately 2.2 × 10 6 Each, at least approximately 2.3 × 10 6 Each, at least approximately 2.4 × 10 6 Each, at least approximately 2.5 × 10 6 Each, at least approximately 2.6 × 10 6 Each, at least approximately 2.7 × 10 6 Each, at least approximately 2.8 × 10 6 Each, at least approximately 2.9 × 10 6 Each, at least approximately 3 × 10 6 Each, at least approximately 3.1 × 10 6 Each, at least approximately 3.2 × 10 6 Each, at least approximately 3.3 × 10 6 Each, at least approximately 3.4 × 10 6 Each, at least approximately 3.5 × 10 6 Each, at least approximately 3.6 × 10 6 Each, at least approximately 3.7 × 10 6 Each, at least approximately 3.8 × 10 6 Each, at least approximately 3.9 × 10 6 Each, at least approximately 4 × 10 6 Each, at least approximately 4.1 × 10 6 Each, at least approximately 4.2 × 10 6 Each, at least approximately 4.3 × 10 6 Each, at least approximately 4.4 × 10 6 Each, at least approximately 4.5 × 10 6 Each, at least approximately 4.6 × 10 6 Each, at least approximately 4.7 × 10 6 Each, at least approximately 4.8 × 10 6 Each, at least approximately 4.9 × 10 6 Each, at least approximately 5 x 10 6 Each, at least approximately 5.1 × 10 6 Each, at least approximately 5.2 × 10 6 Each, at least approximately 5.3 × 10 6 Each, at least approximately 5.4 × 10 6cells, at least about 5.5×10 6 cells, at least about 5.6×10 6 cells, at least about 5.7×10 6 cells, at least about 5.8×10 6 cells, at least about 5.9×10 6 cells, at least about 6×10 6 cells, at least about 6.1×10 6 cells, at least about 6.2×10 6 cells, at least about 6.3×10 6 cells, at least about 6.4×10 6 cells, at least about 6.5×10 6 cells, at least about 6.6×10 6 cells, at least about 6.7×10 6 cells, at least about 6.8×10 6 cells, at least about 6.9×10 6 cells, at least about 7×10 6 cells, at least about 7.1×10 6 cells, at least about 7.2×10 6 cells, at least about 7.3×10 6 cells, at least about 7.4×10 6 cells, at least about 7.5×10 6 cells, at least about 7.6×10 6 cells, at least about 7.7×10 6 cells, at least about 7.8×10 6 cells, at least about 7.9×10 6 cells, at least about 8×10 6 cells, at least about 8.1×10 6 cells, at least about 8.2×10 6 cells, at least about 8.3×10 6 cells, at least about 8.4×10 6 cells, at least about 8.5×10 6 cells, at least about 8.6×10 6 cells, at least about 8.7×10 6 cells, at least about 8.8×10 6 cells, at least about 8.9×10 6 cells, at least about 9×10 6 cells, at least about 9.1×10 6cells, at least about 9.2×10 6 cells, at least about 9.3×10 6 cells, at least about 9.4×10 6 cells, at least about 9.5×10 6 cells, at least about 9.6×10 6 cells, at least about 9.7×10 6 cells, at least about 9.8×10 6 cells, at least about 9.9×10 6 cells, at least about 1×10 7 cells, at least about 1.5×10 7 cells, at least about 2×10 7 cells, at least about 2.5×10 7 cells, at least about 3×10 7 cells, at least about 3.5×10 7 cells, at least about 4×10 7 cells, at least about 4.5×10 7 cells, at least about 5×10 7 cells, at least about 5.5×10 7 cells, at least about 6×10 7 cells, at least about 6.5×10 7 cells, at least about 7×10 7 cells, at least about 7.5×10 7 cells, at least about 8×10 7 cells, at least about 8.5×10 7 cells, at least about 9×10 7 cells, at least about 9.5×10 7 cells, at least about 1×10 8 cells, at least about 1.5×10 8 cells, at least about 2×10 8 cells, at least about 2.5×10 8 cells, at least about 3×10 8 cells, at least about 3.5×10 8 cells, at least about 4×10 7 cells, at least about 4.5×10 8 cells, at least about 5×10 8 cells, at least about 5.5×10 8 cells, at least about 6×10 8 cells, at least about 6.5×10 8Each, at least approximately 7 x 10 8 Each, at least approximately 7.5 × 10 8 Each, at least approximately 8 x 10 8 Each, at least approximately 8.5 × 10 8 Each, at least approximately 9 x 10 8 Each, at least approximately 9.5 × 10 8 each, at least about 1 × 10 9 A single cell or more CD45 + The first population of cells, and / or HSPC or HSPC (e.g., CD34) + It can include the dose of cells.

[0254] CD45 + The first population of cells or CD34 + The population of hematopoietic stem progenitor cells (HSPCs) is up to approximately 1 × 10¹⁶ per kg of the recipient's actual or ideal body weight. 4 Each, up to approximately 2 x 10 4 Each, up to approximately 3 x 10 4 Each, up to approximately 4 x 10 4 Each, up to approximately 5 x 10 4 Each, up to approximately 6 x 10 4 Each, up to approximately 7 x 10 4 Each, up to approximately 8 x 10 4 Each, up to approximately 9 x 10 4 Each, up to approximately 1 x 10 5 Each, up to approximately 2 x 10 5 Each, up to approximately 3 x 10 5 Each, up to approximately 4 x 10 5 Each, up to approximately 5 x 10 5 Each, up to approximately 6 x 10 5 Each, up to approximately 7 x 10 5 Each, up to approximately 8 x 10 5 Each, up to approximately 9 x 10 5 Each, up to approximately 1 x 10 6 Each, up to approximately 1.1 x 10 6 Each, up to approximately 1.2 x 10 6 Each, up to approximately 1.3 x 10 6 Each, up to approximately 1.4 x 10 6 Each, up to approximately 1.5 x 10 6 Each, up to approximately 1.6 x 10 6 Each, up to approximately 1.7 x 106 Each, up to approximately 1.8 x 10 6 Each, up to approximately 1.9 x 10 6 Each, up to approximately 2 x 10 6 Each, up to approximately 2.1 x 10 6 Each, up to approximately 2.2 x 10 6 Each, up to approximately 2.3 x 10 6 Each, up to approximately 2.4 x 10 6 Each, up to approximately 2.5 x 10 6 Each, up to approximately 2.6 x 10 6 Each, up to approximately 2.7 x 10 6 Each, up to approximately 2.8 x 10 6 Each, up to approximately 2.9 x 10 6 Each, up to approximately 3 x 10 6 Each, up to approximately 3.1 x 10 6 Each, up to approximately 3.2 x 10 6 Each, up to approximately 3.3 x 10 6 Each, up to approximately 3.4 x 10 6 Each, up to approximately 3.5 x 10 6 Each, up to approximately 3.6 x 10 6 Each, up to approximately 3.7 x 10 6 Each, up to approximately 3.8 x 10 6 Each, up to approximately 3.9 x 10 6 Each, up to approximately 4 x 10 6 Each, up to approximately 4.1 x 10 6 Each, up to approximately 4.2 x 10 6 Each, up to approximately 4.3 x 10 6 Each, up to approximately 4.4 x 10 6 Each, up to approximately 4.5 x 10 6 Each, up to approximately 4.6 x 10 6 Each, up to approximately 4.7 x 10 6 Each, up to approximately 4.8 x 10 6 Each, up to approximately 4.9 x 10 6 Each, up to approximately 5 x 10 6 Each, up to approximately 5.1 x 10 6 Each, up to approximately 5.2 x 10 6 Each, up to approximately 5.3 x 10 6 Each, up to approximately 5.4 x 10 6 Each, up to approximately 5.5 x 10 6 Each, up to approximately 5.6 x 10 6 Each, up to approximately 5.7 x 10 6Each, up to approximately 5.8 x 10 6 Each, up to approximately 5.9 x 10 6 Each, up to approximately 6 x 10 6 Each, up to approximately 6.1 x 10 6 Each, up to approximately 6.2 x 10 6 Each, up to approximately 6.3 x 10 6 Each, up to approximately 6.4 x 10 6 Each, up to approximately 6.5 x 10 6 Each, up to approximately 6.6 x 10 6 Each, up to approximately 6.7 x 10 6 Each, up to approximately 6.8 x 10 6 Each, up to approximately 6.9 x 10 6 Each, up to approximately 7 x 10 6 Each, up to approximately 7.1 x 10 6 Each, up to approximately 7.2 x 10 6 Each, up to approximately 7.3 x 10 6 Each, up to approximately 7.4 x 10 6 Each, up to approximately 7.5 x 10 6 Each, up to approximately 7.6 x 10 6 Each, up to approximately 7.7 x 10 6 Each, up to approximately 7.8 x 10 6 Each, up to approximately 7.9 x 10 6 Each, up to approximately 8 x 10 6 Each, up to approximately 8.1 x 10 6 Each, up to approximately 8.2 x 10 6 Each, up to approximately 8.3 x 10 6 Each, up to approximately 8.4 x 10 6 Each piece is approximately 8.5 x 10 6 Each, up to approximately 8.6 x 10 6 Each, up to approximately 8.7 x 10 6 Each, up to approximately 8.8 x 10 6 Each, up to approximately 8.9 x 10 6 Each, up to approximately 9 x 10 6 Each, up to approximately 9.1 x 10 6 Each, up to approximately 9.2 x 10 6 Each, up to approximately 9.3 x 10 6 Each, up to approximately 9.4 x 10 6 Each, up to approximately 9.5 x 10 6 Each, up to approximately 9.6 x 10 6 Each, up to approximately 9.7 x 10 6Each, up to approximately 9.8 x 10 6 Each, up to approximately 9.9 x 10 6 Each, up to approximately 1 x 10 7 Each, up to approximately 1.5 x 10 7 Each, up to approximately 2 x 10 7 Each, up to approximately 2.5 x 10 7 Each, up to approximately 3 x 10 7 Each, up to approximately 3.5 x 10 7 Each, up to approximately 4 x 10 7 Each, up to approximately 4.5 x 10 7 Each, up to approximately 5 x 10 7 Each, up to approximately 5.5 x 10 7 Each, up to approximately 6 x 10 7 Each, up to approximately 6.5 x 10 7 Each, up to approximately 7 x 10 7 Each, up to approximately 7.5 x 10 7 Each, up to approximately 8 x 10 7 Each piece is approximately 8.5 x 10 7 Each, up to approximately 9 x 10 7 Each, up to approximately 9.5 x 10 7 Each, up to approximately 1 x 10 8 Each, up to approximately 1.5 x 10 8 Each, up to approximately 2 x 10 8 Each, up to approximately 2.5 x 10 8 Each, up to approximately 3 x 10 8 Each, up to approximately 3.5 x 10 8 Each, up to approximately 4 x 10 7 Each, up to approximately 4.5 x 10 8 Each, up to approximately 5 x 10 8 Each, up to approximately 5.5 x 10 8 Each, up to approximately 6 x 10 8 Each, up to approximately 6.5 x 10 8 Each, up to approximately 7 x 10 8 Each, up to approximately 7.5 x 10 8 Each, up to approximately 8 x 10 8 Each piece is approximately 8.5 x 10 8 Each, up to approximately 9 x 10 8 Each, up to approximately 9.5 x 10 8 individual, or at most about 1 x 10 9 CD45 of individual cells + The first population of cells, and / or HSPC or HSPC (e.g., CD34)+ It can include the dose of cells.

[0255] For example, CD45 + The first population of cells or CD34 + The population of hematopoietic stem progenitor cells (HSPCs) is 1 × 10¹⁶ per kg of the recipient's actual or ideal body weight. 4 ~1 × 10 9 pieces, 1×10 5 ~1 × 10 8 pieces, 1×10 5 ~2×10 7 pieces, 5×10 5 ~2×10 7 pieces, 5×10 5 ~1.5×10 7 pieces, 5×10 5 ~1 × 10 7 pieces, 5×10 5 ~9×10 6 pieces, 5×10 5 ~8×10 6 pieces, 5×10 5 ~7×10 6 pieces, 5×10 5 ~6×10 6 pieces, 5×10 5 ~5×10 6 pieces, 5×10 5 ~4×10 6 pieces, 5×10 5 ~3×10 6 pieces, 5×10 5 ~2×10 6 pieces, 5×10 5 ~1 × 10 6 pieces, 1×10 6 ~1.5×10 7 pieces, 1×10 6 ~1 × 10 7 pieces, 1×10 6 ~9×10 6 pieces, 1×10 6 ~8×10 6 pieces, 1×10 6 ~7×10 6 pieces, 1×10 6 ~6×10 6 pieces, 1×10 6 ~5×10 6 pieces, 1×10 6 ~4×106 1×10 6 ~3×10 6 1×10 6 ~2×10 6 1.5 × 10 6 ~1.5×10 7 1.5 × 10 6 ~1×10 7 1.5 × 10 6 ~9×10 6 1.5 × 10 6 ~8×10 6 1.5 × 10 6 ~7×10 6 1.5 × 10 6 ~6×10 6 1.5 × 10 6 ~5×10 6 1.5 × 10 6 ~4×10 6 1.5 × 10 6 ~3×10 6 1.5 × 10 6 ~2×10 6 1, 2×10 6 ~1.5×10 7 1, 2×10 6 ~1×10 7 1, 2×10 6 ~9×10 6 1, 2×10 6 ~8×10 6 1, 2×10 6 ~7×10 6 1, 2×10 6 ~6×10 6 1, 2×10 6 ~5×10 6 1, 2×10 6 ~4×10 6 1, 2×10 6 ~3×10 6 1, 2.5 × 10 6 ~1.5×10 7 1, 2.5 × 10 6 ~1×10 7 1, 2.5 × 10 6 ~9×10 6 1, 2.5 × 10 6 ~8×10 6 1, 2.5 × 10 6 ~7×106 pieces, 2.5×10 6 ~6×10 6 pieces, 2.5×10 6 ~5×10 6 pieces, 2.5×10 6 ~4×10 6 1, or 2.5 × 10 6 ~3×10 6 CD45 of individual cells + A first population of cells, a population of HSPCs, and / or HSPCs or HSPCs (e.g., CD34) + It can include the dose of cells.

[0256] In some embodiments, HSPC, CD34 + CD45 containing a population of hematopoietic stem progenitor cells (HSPCs) or containing at least one dose of HSPCs + The first group of cells is, for example, CD45 + In the first population of cells, approximately 1.0 × 10⁻⁶ 5 ~Approx. 5.0×10 10 pieces, approximately 5.0×10 5 ~Approx. 1.5×10 10 pieces, approximately 5.0×10 5 ~Approx. 5.0×10 8 1.5 x 10 7 ~Approx. 1.5×10 10 Contains HSPCs. In some embodiments, CD45 contains HSPCs, or contains at least one dose of HSPCs. + The first population of cells is approximately 1.0 × 10⁶ 5 One or more HSPCs, approximately 2.0 x 10 5 More than one HSPC, approximately 3.0 x 10 5 More than one HSPC, approximately 4.0 x 10 5 More than one HSPC, approximately 5.0 x 10 5 More than one HSPC, approximately 6.0 x 10 5 More than one HSPC, approximately 7.0 x 10 5 More than one HSPC, approximately 8.0 x 10 5 More than one HSPC, approximately 9.0 x 10 5 More than one HSPC, approximately 1.0 x 10 6 More than one HSPC, approximately 1.0 x 10 6More than HSPCs, approximately 1.1 x 10 6 More than HSPCs, approximately 1.2 x 10 6 More than one HSPC, approximately 1.3 x 10 6 More than one HSPC, approximately 1.4 x 10 6 More than one HSPC, approximately 1.5 x 10 6 More than one HSPC, approximately 1.6 x 10 6 More than one HSPC, approximately 1.7 × 10 6 More than HSPC, approximately 1.8 x 10 6 More than one HSPC, approximately 1.9 × 10 6 One or more HSPCs, approximately 2.0 x 10 6 More than HSPC, approximately 2.1 x 10 6 More than one HSPC, approximately 2.2 x 10 6 More than HSPCs, approximately 2.3 x 10 6 More than HSPCs, approximately 2.4 x 10 6 More than one HSPC, approximately 2.5 x 10 6 More than one HSPC, approximately 2.6 x 10 6 More than one HSPC, approximately 2.7 x 10 6 More than one HSPC, approximately 2.8 x 10 6 More than one HSPC, approximately 2.9 x 10 6 More than one HSPC, approximately 3.0 x 10 6 More than one HSPC, approximately 3.1 x 10 6 More than one HSPC, approximately 3.2 x 10 6 More than one HSPC, approximately 3.3 x 10 6 More than one HSPC, approximately 3.4 x 10 6 More than one HSPC, approximately 3.5 x 10 6 More than one HSPC, approximately 3.6 x 10 6 More than one HSPC, approximately 3.7 x 10 6 More than one HSPC, approximately 3.8 x 10 6 More than one HSPC, approximately 3.9 x 10 6 More than one HSPC, approximately 4.0 x 10 6 More than one HSPC, approximately 4.1 x 10 6 More than one HSPC, approximately 4.2 x 10 6 More than one HSPC, approximately 4.3 x 10 6 More than one HSPC, approximately 4.4 x 10 6More than one HSPC, approximately 4.5 x 10 6 More than one HSPC, approximately 4.6 x 10 6 More than one HSPC, approximately 4.7 x 10 6 More than one HSPC, approximately 4.8 x 10 6 More than one HSPC, approximately 4.9 x 10 6 More than one HSPC, approximately 5.0 x 10 6 More than HSPCs, approximately 5.1 x 10 6 More than one HSPC, approximately 5.2 x 10 6 More than one HSPC, approximately 5.3 x 10 6 More than one HSPC, approximately 5.4 x 10 6 More than one HSPC, approximately 5.5 x 10 6 More than one HSPC, approximately 5.6 x 10 6 More than one HSPC, approximately 5.7 x 10 6 More than one HSPC, approximately 5.8 x 10 6 More than one HSPC, approximately 5.9 x 10 6 More than one HSPC, approximately 6.0 x 10 6 More than HSPCs, approximately 6.1 x 10 6 More than one HSPC, approximately 6.2 x 10 6 More than one HSPC, approximately 6.3 x 10 6 More than one HSPC, approximately 6.4 x 10 6 More than one HSPC, approximately 6.5 x 10 6 More than one HSPC, approximately 6.6 x 10 6 More than one HSPC, approximately 6.7 x 10 6 More than one HSPC, approximately 6.8 x 10 6 More than one HSPC, approximately 6.9 x 10 6 More than one HSPC, approximately 7.0 x 10 6 More than one HSPC, approximately 7.1 x 10 6 More than one HSPC, approximately 7.2 x 10 6 More than one HSPC, approximately 7.3 x 10 6 More than one HSPC, approximately 7.4 x 10 6 More than one HSPC, approximately 7.5 x 10 6 More than one HSPC, approximately 7.6 x 10 6 More than one HSPC, approximately 7.7 x 10 6 More than one HSPC, approximately 7.8 x 10 6More than one HSPC, approximately 7.9 x 10 6 More than one HSPC, approximately 8.0 x 10 6 More than one HSPC, approximately 8.1 x 10 6 More than one HSPC, approximately 8.2 x 10 6 More than one HSPC, approximately 8.3 x 10 6 More than one HSPC, approximately 8.4 x 10 6 More than one HSPC, approximately 8.5 x 10 6 More than one HSPC, approximately 8.6 x 10 6 More than one HSPC, approximately 8.7 x 10 6 More than one HSPC, approximately 8.8 x 10 6 More than one HSPC, approximately 8.9 x 10 6 More than one HSPC, approximately 9.0 x 10 6 More than one HSPC, approximately 9.1 x 10 6 More than one HSPC, approximately 9.2 x 10 6 More than one HSPC, approximately 9.3 x 10 6 More than one HSPC, approximately 9.4 x 10 6 More than one HSPC, approximately 9.5 x 10 6 More than one HSPC, approximately 9.6 x 10 6 More than one HSPC, approximately 9.7 x 10 6 More than one HSPC, approximately 9.8 x 10 6 More than one HSPC, approximately 9.9 x 10 6 More than one HSPC, approximately 1.0 x 10 7 More than HSPCs, approximately 1.1 x 10 7 More than HSPCs, approximately 1.2 x 10 7 More than one HSPC, approximately 1.3 x 10 7 More than one HSPC, approximately 1.4 x 10 7 More than one HSPC, approximately 1.5 x 10 7 More than one HSPC, approximately 1.6 x 10 7 More than one HSPC, approximately 1.7 × 10 7 More than HSPC, approximately 1.8 x 10 7 More than one HSPC, approximately 1.9 × 10 7 One or more HSPCs, approximately 2.0 x 10 7 More than HSPC, approximately 2.1 x 10 7 More than one HSPC, approximately 2.2 x 10 7More than HSPCs, approximately 2.3 x 10 7 More than HSPCs, approximately 2.4 x 10 7 More than one HSPC, approximately 2.5 x 10 7 More than one HSPC, approximately 2.6 x 10 7 More than one HSPC, approximately 2.7 x 10 7 More than one HSPC, approximately 2.8 x 10 7 More than one HSPC, approximately 2.9 x 10 7 More than one HSPC, approximately 3.0 x 10 7 More than one HSPC, approximately 3.1 x 10 7 More than one HSPC, approximately 3.2 x 10 7 More than one HSPC, approximately 3.3 x 10 7 More than one HSPC, approximately 3.4 x 10 7 More than one HSPC, approximately 3.5 x 10 7 More than one HSPC, approximately 3.6 x 10 7 More than one HSPC, approximately 3.7 x 10 7 More than one HSPC, approximately 3.8 x 10 7 More than one HSPC, approximately 3.9 x 10 7 More than one HSPC, approximately 4.0 x 10 7 More than one HSPC, approximately 4.1 x 10 7 More than one HSPC, approximately 4.2 x 10 7 More than one HSPC, approximately 4.3 x 10 7 More than one HSPC, approximately 4.4 x 10 7 More than one HSPC, approximately 4.5 x 10 7 More than one HSPC, approximately 4.6 x 10 7 More than one HSPC, approximately 4.7 x 10 7 More than one HSPC, approximately 4.8 x 10 7 More than one HSPC, approximately 4.9 x 10 7 More than one HSPC, approximately 5.0 x 10 7 More than HSPCs, approximately 5.1 x 10 7 More than one HSPC, approximately 5.2 x 10 7 More than one HSPC, approximately 5.3 x 10 7 More than one HSPC, approximately 5.4 x 10 7 More than one HSPC, approximately 5.5 x 10 7 More than one HSPC, approximately 5.6 x 10 7More than one HSPC, approximately 5.7 x 10 7 More than one HSPC, approximately 5.8 x 10 7 More than one HSPC, approximately 5.9 x 10 7 More than one HSPC, approximately 6.0 x 10 7 More than HSPCs, approximately 6.1 x 10 7 More than one HSPC, approximately 6.2 x 10 7 More than one HSPC, approximately 6.3 x 10 7 More than one HSPC, approximately 6.4 x 10 7 More than one HSPC, approximately 6.5 x 10 7 More than one HSPC, approximately 6.6 x 10 7 More than one HSPC, approximately 6.7 x 10 7 More than one HSPC, approximately 6.8 x 10 7 More than one HSPC, approximately 6.9 x 10 7 More than one HSPC, approximately 7.0 x 10 7 More than one HSPC, approximately 7.1 x 10 7 More than one HSPC, approximately 7.2 x 10 7 More than one HSPC, approximately 7.3 x 10 7 More than one HSPC, approximately 7.4 x 10 7 More than one HSPC, approximately 7.5 x 10 7 More than one HSPC, approximately 7.6 x 10 7 More than one HSPC, approximately 7.7 x 10 7 More than one HSPC, approximately 7.8 x 10 7 More than one HSPC, approximately 7.9 x 10 7 More than one HSPC, approximately 8.0 x 10 7 More than one HSPC, approximately 8.1 x 10 7 More than one HSPC, approximately 8.2 x 10 7 More than one HSPC, approximately 8.3 x 10 7 More than one HSPC, approximately 8.4 x 10 7 More than one HSPC, approximately 8.5 x 10 7 More than one HSPC, approximately 8.6 x 10 7 More than one HSPC, approximately 8.7 x 10 7 More than one HSPC, approximately 8.8 x 10 7 More than one HSPC, approximately 8.9 x 10 7 More than one HSPC, approximately 9.0 x 10 7More than one HSPC, approximately 9.1 x 10 7 More than one HSPC, approximately 9.2 x 10 7 More than one HSPC, approximately 9.3 x 10 7 More than one HSPC, approximately 9.4 x 10 7 More than one HSPC, approximately 9.5 x 10 7 More than one HSPC, approximately 9.6 x 10 7 More than one HSPC, approximately 9.7 x 10 7 More than one HSPC, approximately 9.8 x 10 7 More than one HSPC, approximately 9.9 x 10 7 More than one HSPC, approximately 1.0 x 10 8 More than HSPCs, approximately 1.1 x 10 8 More than HSPCs, approximately 1.2 x 10 8 More than one HSPC, approximately 1.3 x 10 8 More than one HSPC, approximately 1.4 x 10 8 More than one HSPC, approximately 1.5 x 10 8 More than one HSPC, approximately 1.6 x 10 8 More than one HSPC, approximately 1.7 × 10 8 More than HSPC, approximately 1.8 x 10 8 More than one HSPC, approximately 1.9 × 10 8 One or more HSPCs, approximately 2.0 x 10 8 More than HSPC, approximately 2.1 x 10 8 More than one HSPC, approximately 2.2 x 10 8 More than HSPCs, approximately 2.3 x 10 8 More than HSPCs, approximately 2.4 x 10 8 More than one HSPC, approximately 2.5 x 10 8 More than one HSPC, approximately 2.6 x 10 8 More than one HSPC, approximately 2.7 x 10 8 More than one HSPC, approximately 2.8 x 10 8 More than one HSPC, approximately 2.9 x 10 8 More than one HSPC, approximately 3.0 x 10 8 More than one HSPC, approximately 3.1 x 10 8 More than one HSPC, approximately 3.2 x 10 8 More than one HSPC, approximately 3.3 x 10 8 More than one HSPC, approximately 3.4 × 10 8More than one HSPC, approximately 3.5 x 10 8 More than one HSPC, approximately 3.6 x 10 8 More than one HSPC, approximately 3.7 x 10 8 More than one HSPC, approximately 3.8 x 10 8 More than one HSPC, approximately 3.9 x 10 8 More than one HSPC, approximately 4.0 x 10 8 More than one HSPC, approximately 4.1 x 10 8 More than one HSPC, approximately 4.2 x 10 8 More than one HSPC, approximately 4.3 x 10 8 More than one HSPC, approximately 4.4 x 10 8 More than one HSPC, approximately 4.5 x 10 8 More than one HSPC, approximately 4.6 x 10 8 More than one HSPC, approximately 4.7 x 10 8 More than one HSPC, approximately 4.8 x 10 8 More than one HSPC, approximately 4.9 x 10 8 More than one HSPC, approximately 5.0 x 10 8 More than HSPCs, approximately 5.1 x 10 8 More than one HSPC, approximately 5.2 x 10 8 More than one HSPC, approximately 5.3 x 10 8 More than one HSPC, approximately 5.4 x 10 8 More than one HSPC, approximately 5.5 x 10 8 More than one HSPC, approximately 5.6 x 10 8 More than one HSPC, approximately 5.7 x 10 8 More than one HSPC, approximately 5.8 x 10 8 More than one HSPC, approximately 5.9 x 10 8 More than one HSPC, approximately 6.0 x 10 8 More than HSPCs, approximately 6.1 x 10 8 More than one HSPC, approximately 6.2 x 10 8 More than one HSPC, approximately 6.3 x 10 8 More than one HSPC, approximately 6.4 x 10 8 More than one HSPC, approximately 6.5 x 10 8 More than one HSPC, approximately 6.6 x 10 8 More than one HSPC, approximately 6.7 x 10 8 More than one HSPC, approximately 6.8 x 10 8More than one HSPC, approximately 6.9 x 10 8 More than one HSPC, approximately 7.0 x 10 8 More than one HSPC, approximately 7.1 x 10 8 More than one HSPC, approximately 7.2 x 10 8 More than one HSPC, approximately 7.3 x 10 8 More than one HSPC, approximately 7.4 x 10 8 More than one HSPC, approximately 7.5 x 10 8 More than one HSPC, approximately 7.6 x 10 8 More than one HSPC, approximately 7.7 x 10 8 More than one HSPC, approximately 7.8 x 10 8 More than one HSPC, approximately 7.9 x 10 8 More than one HSPC, approximately 8.0 x 10 8 More than one HSPC, approximately 8.1 x 10 8 More than one HSPC, approximately 8.2 x 10 8 More than one HSPC, approximately 8.3 x 10 8 More than one HSPC, approximately 8.4 x 10 8 More than one HSPC, approximately 8.5 x 10 8 More than one HSPC, approximately 8.6 x 10 8 More than one HSPC, approximately 8.7 x 10 8 More than one HSPC, approximately 8.8 x 10 8 More than one HSPC, approximately 8.9 x 10 8 More than one HSPC, approximately 9.0 x 10 8 More than one HSPC, approximately 9.1 x 10 8 More than one HSPC, approximately 9.2 x 10 8 More than one HSPC, approximately 9.3 x 10 8 More than one HSPC, approximately 9.4 x 10 8 More than one HSPC, approximately 9.5 x 10 8 More than one HSPC, approximately 9.6 x 10 8 More than one HSPC, approximately 9.7 x 10 8 More than one HSPC, approximately 9.8 x 10 8 More than one HSPC, approximately 9.9 x 10 8 More than one HSPC, approximately 1.0 x 10 9 More than HSPCs, approximately 1.1 x 10 9 More than HSPCs, approximately 1.2 x 10 9More than one HSPC, approximately 1.3 x 10 9 More than one HSPC, approximately 1.4 x 10 9 More than one HSPC, approximately 1.5 x 10 9 More than one HSPC, approximately 1.6 x 10 9 More than one HSPC, approximately 1.7 × 10 9 More than HSPC, approximately 1.8 x 10 9 More than one HSPC, approximately 1.9 × 10 9 One or more HSPCs, approximately 2.0 x 10 9 More than HSPC, approximately 2.1 x 10 9 More than one HSPC, approximately 2.2 x 10 9 More than HSPCs, approximately 2.3 x 10 9 More than HSPCs, approximately 2.4 x 10 9 More than one HSPC, approximately 2.5 x 10 9 More than one HSPC, approximately 2.6 x 10 9 More than one HSPC, approximately 2.7 x 10 9 More than one HSPC, approximately 2.8 x 10 9 More than one HSPC, approximately 2.9 x 10 9 More than one HSPC, approximately 3.0 x 10 9 More than one HSPC, approximately 3.1 x 10 9 More than one HSPC, approximately 3.2 x 10 9 More than one HSPC, approximately 3.3 x 10 9 More than one HSPC, approximately 3.4 x 10 9 More than one HSPC, approximately 3.5 x 10 9 More than one HSPC, approximately 3.6 x 10 9 More than one HSPC, approximately 3.7 x 10 9 More than one HSPC, approximately 3.8 x 10 9 More than one HSPC, approximately 3.9 x 10 9 More than one HSPC, approximately 4.0 x 10 9 More than one HSPC, approximately 4.1 x 10 9 More than one HSPC, approximately 4.2 x 10 9 More than one HSPC, approximately 4.3 x 10 9 More than one HSPC, approximately 4.4 x 10 9 More than one HSPC, approximately 4.5 x 10 9 More than one HSPC, approximately 4.6 x 10 9More than one HSPC, approximately 4.7 x 10 9 More than one HSPC, approximately 4.8 x 10 9 More than one HSPC, approximately 4.9 x 10 9 More than one HSPC, approximately 5.0 x 10 9 More than HSPCs, approximately 5.1 x 10 9 More than one HSPC, approximately 5.2 x 10 9 More than one HSPC, approximately 5.3 x 10 9 More than one HSPC, approximately 5.4 x 10 9 More than one HSPC, approximately 5.5 x 10 9 More than one HSPC, approximately 5.6 x 10 9 More than one HSPC, approximately 5.7 x 10 9 More than one HSPC, approximately 5.8 x 10 9 More than one HSPC, approximately 5.9 x 10 9 More than one HSPC, approximately 6.0 x 10 9 More than HSPCs, approximately 6.1 x 10 9 More than one HSPC, approximately 6.2 x 10 9 More than one HSPC, approximately 6.3 x 10 9 More than one HSPC, approximately 6.4 x 10 9 More than one HSPC, approximately 6.5 x 10 9 More than one HSPC, approximately 6.6 x 10 9 More than one HSPC, approximately 6.7 x 10 9 More than one HSPC, approximately 6.8 x 10 9 More than one HSPC, approximately 6.9 x 10 9 More than one HSPC, approximately 7.0 x 10 9 More than one HSPC, approximately 7.1 x 10 9 More than one HSPC, approximately 7.2 x 10 9 More than one HSPC, approximately 7.3 x 10 9 More than one HSPC, approximately 7.4 x 10 9 More than one HSPC, approximately 7.5 x 10 9 More than one HSPC, approximately 7.6 x 10 9 More than one HSPC, approximately 7.7 x 10 9 More than one HSPC, approximately 7.8 x 10 9 More than one HSPC, approximately 7.9 x 10 9 More than one HSPC, approximately 8.0 x 10 9More than one HSPC, approximately 8.1 x 10 9 More than one HSPC, approximately 8.2 x 10 9 More than one HSPC, approximately 8.3 x 10 9 More than one HSPC, approximately 8.4 x 10 9 More than one HSPC, approximately 8.5 x 10 9 More than one HSPC, approximately 8.6 x 10 9 More than one HSPC, approximately 8.7 x 10 9 More than one HSPC, approximately 8.8 x 10 9 More than one HSPC, approximately 8.9 x 10 9 More than one HSPC, approximately 9.0 x 10 9 More than one HSPC, approximately 9.1 x 10 9 More than one HSPC, approximately 9.2 x 10 9 More than one HSPC, approximately 9.3 x 10 9 More than one HSPC, approximately 9.4 x 10 9 More than one HSPC, approximately 9.5 x 10 9 More than one HSPC, approximately 9.6 x 10 9 More than one HSPC, approximately 9.7 x 10 9 More than one HSPC, approximately 9.8 x 10 9 More than one HSPC, approximately 9.9 x 10 9 More than one HSPC, approximately 1.0 x 10 10 More than HSPCs, approximately 1.1 x 10 10 More than HSPCs, approximately 1.2 x 10 10 More than one HSPC, approximately 1.3 x 10 10 More than one HSPC, approximately 1.4 x 10 10 More than one HSPC, approximately 1.5 x 10 10 More than one HSPC, approximately 1.6 x 10 10 More than one HSPC, approximately 1.7 × 10 10 More than HSPC, approximately 1.8 x 10 10 More than one HSPC, approximately 1.9 × 10 10 One or more HSPCs, approximately 2.0 x 10 10 More than HSPC, approximately 2.1 x 10 10 More than one HSPC, approximately 2.2 x 10 10 More than HSPCs, approximately 2.3 x 10 10 More than HSPCs, approximately 2.4 x 10 10More than one HSPC, approximately 2.5 x 10 10 More than one HSPC, approximately 2.6 x 10 10 More than one HSPC, approximately 2.7 x 10 10 More than one HSPC, approximately 2.8 x 10 10 More than one HSPC, approximately 2.9 x 10 10 More than one HSPC, approximately 3.0 x 10 10 More than one HSPC, approximately 3.1 x 10 10 More than one HSPC, approximately 3.2 x 10 10 More than one HSPC, approximately 3.3 x 10 10 More than one HSPC, approximately 3.4 x 10 10 More than one HSPC, approximately 3.5 x 10 10 More than one HSPC, approximately 3.6 x 10 10 More than one HSPC, approximately 3.7 x 10 10 More than one HSPC, approximately 3.8 x 10 10 More than one HSPC, approximately 3.9 x 10 10 More than one HSPC, approximately 4.0 x 10 10 More than one HSPC, approximately 4.1 x 10 10 More than one HSPC, approximately 4.2 x 10 10 More than one HSPC, approximately 4.3 x 10 10 More than one HSPC, approximately 4.4 x 10 10 More than one HSPC, approximately 4.5 x 10 10 More than one HSPC, approximately 4.6 x 10 10 More than one HSPC, approximately 4.7 x 10 10 More than one HSPC, approximately 4.8 x 110 10 More than one HSPC, approximately 4.9 x 10 10 One or more HSPCs, or approximately 5.0 × 10 10 Includes one or more HSPCs.

[0257] T's Reg A cell population enriched with Tregs, or a cell population containing at least one dose of Tregs, for example, CD45 containing at least one dose of Tregs. + A second population of cells, or CD4 + CD25 + CD127 dim The population of regulatory T cells (Tregs) is at least approximately 1 × 10⁶ 4each, at least about 1 × 10 5 each, at least about 2 × 10 4 Each, at least approximately 3 × 10 4 Each, at least approximately 4 x 10 4 Each, at least approximately 5 x 10 4 Each, at least approximately 6 x 10 4 Each, at least approximately 7 x 10 4 Each, at least approximately 8 x 10 4 , at least about 9 x 10 4 each, at least about 1 × 10 5 each, at least about 2 × 10 5 Each, at least approximately 3 × 10 5 Each, at least approximately 4 x 10 5 Each, at least approximately 5 x 10 5 Each, at least approximately 6 x 10 5 Each, at least approximately 7 x 10 5 Each, at least approximately 8 x 10 5 Each, at least approximately 9 x 10 5 each, at least about 1 × 10 6 Each, at least about 1.1 × 10 6 Each, at least approximately 1.2 × 10 6 Each, at least approximately 1.3 × 10 6 Each, at least approximately 1.4 × 10 6 Each, at least approximately 1.5 × 10 6 Each, at least approximately 1.6 × 10 6 Each, at least approximately 1.7 × 10 6 Each, at least approximately 1.8 × 10 6 Each, at least approximately 1.9 × 10 6 each, at least about 2 × 10 6 Each, at least approximately 2.1 × 10 6 Each, at least approximately 2.2 × 10 6 Each, at least approximately 2.3 × 10 6 Each, at least approximately 2.4 × 10 6 Each, at least approximately 2.5 × 10 6 Each, at least approximately 2.6 × 10 6 Each, at least approximately 2.7 × 10 6 Each, at least approximately 2.8 × 10 6 Each, at least approximately 2.9 × 10 6 Each, at least approximately 3 × 106 Each, at least approximately 3.1 × 10 6 Each, at least approximately 3.2 × 10 6 Each, at least approximately 3.3 × 10 6 Each, at least approximately 3.4 × 10 6 Each, at least approximately 3.5 × 10 6 Each, at least approximately 3.6 × 10 6 Each, at least approximately 3.7 × 10 6 Each, at least approximately 3.8 × 10 6 Each, at least approximately 3.9 × 10 6 Each, at least approximately 4 x 10 6 Each, at least approximately 4.1 × 10 6 Each, at least approximately 4.2 × 10 6 Each, at least approximately 4.3 × 10 6 Each, at least approximately 4.4 × 10 6 Each, at least approximately 4.5 × 10 6 Each, at least approximately 4.6 × 10 6 Each, at least approximately 4.7 × 10 6 Each, at least approximately 4.8 × 10 6 Each, at least approximately 4.9 × 10 6 Each, at least approximately 5 x 10 6 Each, at least approximately 5.1 × 10 6 Each, at least approximately 5.2 × 10 6 Each, at least approximately 5.3 × 10 6 Each, at least approximately 5.4 × 10 6 Each, at least approximately 5.5 × 10 6 Each, at least approximately 5.6 × 10 6 Each, at least approximately 5.7 × 10 6 Each, at least approximately 5.8 × 10 6 Each, at least approximately 5.9 × 10 6 Each, at least approximately 6 x 10 6 Each, at least approximately 6.1 × 10 6 Each, at least approximately 6.2 × 10 6 Each, at least approximately 6.3 × 10 6 Each, at least approximately 6.4 × 10 6 Each, at least approximately 6.5 × 10 6 Each, at least approximately 6.6 × 10 6 Each, at least approximately 6.7 × 106 Each, at least approximately 6.8 × 10 6 Each, at least approximately 6.9 × 10 6 , pieces, at least about 7 × 10 6 Each, at least approximately 7.1 × 10 6 Each, at least approximately 7.2 × 10 6 Each, at least approximately 7.3 × 10 6 Each, at least approximately 7.4 × 10 6 Each, at least approximately 7.5 × 10 6 Each, at least approximately 7.6 × 10 6 Each, at least approximately 7.7 × 10 6 Each, at least approximately 7.8 × 10 6 Each, at least approximately 7.9 × 10 6 Each, at least approximately 8 x 10 6 Each, at least approximately 8.1 × 10 6 Each, at least approximately 8.2 × 10 6 Each, at least approximately 8.3 × 10 6 Each, at least approximately 8.4 × 10 6 Each, at least approximately 8.5 × 10 6 Each, at least approximately 8.6 × 10 6 Each, at least approximately 8.7 × 10 6 Each, at least approximately 8.8 × 10 6 Each, at least approximately 8.9 × 10 6 Each, at least approximately 9 x 10 6 Each, at least approximately 9.1 × 10 6 Each, at least approximately 9.2 × 10 6 Each, at least approximately 9.3 × 10 6 Each, at least approximately 9.4 × 10 6 Each, at least approximately 9.5 × 10 6 Each, at least approximately 9.6 × 10 6 Each, at least approximately 9.7 × 10 6 Each, at least approximately 9.8 × 10 6 Each, at least approximately 9.9 × 10 6 each, at least about 1 × 10 7 Each, at least approximately 1.5 × 10 7 each, at least about 2 × 10 7 Each, at least approximately 2.5 × 10 7 Each, at least approximately 3 × 10 7Each, at least approximately 3.5 × 10 7 Each, at least approximately 4 x 10 7 Each, at least approximately 4.5 × 10 7 Each, at least approximately 5 x 10 7 Each, at least approximately 5.5 × 10 7 Each, at least approximately 6 x 10 7 Each, at least approximately 6.5 × 10 7 Each, at least approximately 7 x 10 7 Each, at least approximately 7.5 × 10 7 Each, at least approximately 8 x 10 7 Each, at least approximately 8.5 × 10 7 Each, at least approximately 9 x 10 7 Each, at least approximately 9.5 × 10 7 each, at least about 1 × 10 8 Each, at least approximately 1.5 × 10 8 each, at least about 2 × 10 8 Each, at least approximately 2.5 × 10 8 Each, at least approximately 3 × 10 8 Each, at least approximately 3.5 × 10 8 Each, at least approximately 4 x 10 7 Each, at least approximately 4.5 × 10 8 Each, at least approximately 5 x 10 8 Each, at least approximately 5.5 × 10 8 Each, at least approximately 6 x 10 8 Each, at least approximately 6.5 × 10 8 Each, at least approximately 7 x 10 8 Each, at least approximately 7.5 × 10 8 Each, at least approximately 8 x 10 8 Each, at least approximately 8.5 × 10 8 Each, at least approximately 9 x 10 8 Each, at least approximately 9.5 × 10 8 each, at least about 1 × 10 9 A population of cells enriched with Treg cells (e.g., CD45) + (a second population of cells) and / or may contain Treg or Treg doses (e.g., Treg is CD4 + CD25 + CD127 dim CD3 + CD4+ CD25 + CD3 + CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD127 dim FOXP3 + CD3 + FOXP3 + CD3 + CD4 + FOXP3 + CD3 + CD4 + CD25 + FOXP3 + CD3 + CD25 + FOXP3 + CD3 + CD25 + CD127 dim CD4 + CD25 + CD4 + CD25 + CD127 dim FOXP3 + FOXP3 + CD4 + FOXP3 + CD4 + CD25 + FOXP3 + CD25 + FOXP3 + , or CD25 + CD127 dim (That is.)

[0258] A cell population enriched with Tregs, or a cell population containing at least one dose of Tregs, for example, CD45 containing at least one dose of Tregs. + A second population of cells, or CD4 + CD25 + CD127 dim The regulatory T cell (Treg) population is up to approximately 1 × 10¹⁶ per kg of the recipient's actual or ideal body weight. 4 Each, up to approximately 2 x 10 4Each, up to approximately 3 x 10 4 Each, up to approximately 4 x 10 4 Each, up to approximately 5 x 10 4 Each, up to approximately 6 x 10 4 Each, up to approximately 7 x 10 4 Each, up to approximately 8 x 10 4 Each, up to approximately 9 x 10 4 Each, up to approximately 1 x 10 5 Each, up to approximately 2 x 10 5 Each, up to approximately 3 x 10 5 Each, up to approximately 4 x 10 5 Each, up to approximately 5 x 10 5 Each, up to approximately 6 x 10 5 Each, up to approximately 7 x 10 5 Each, up to approximately 8 x 10 5 Each, up to approximately 9 x 10 5 Each, up to approximately 1 x 10 6 Each, up to approximately 1.1 x 10 6 Each, up to approximately 1.2 x 10 6 Each, up to approximately 1.3 x 10 6 Each, up to approximately 1.4 x 10 6 Each, up to approximately 1.5 x 10 6 Each, up to approximately 1.6 x 10 6 Each, up to approximately 1.7 x 10 6 Each, up to approximately 1.8 x 10 6 Each, up to approximately 1.9 x 10 6 Each, up to approximately 2 x 10 6 Each, up to approximately 2.1 x 10 6 Each, up to approximately 2.2 x 10 6 Each, up to approximately 2.3 x 10 6 Each, up to approximately 2.4 x 10 6 Each, up to approximately 2.5 x 10 6 Each, up to approximately 2.6 x 10 6 Each, up to approximately 2.7 x 10 6 Each, up to approximately 2.8 x 10 6 Each, up to approximately 2.9 x 10 6 Each, up to approximately 3 x 10 6 Each, up to approximately 3.1 x 10 6 Each, up to approximately 3.2 x 10 6 Each, up to approximately 3.3 x 10 6 Each, up to approximately 3.4 x 10 6 Each, up to approximately 3.5 x 106 Each, up to approximately 3.6 x 10 6 Each, up to approximately 3.7 x 10 6 Each, up to approximately 3.8 x 10 6 Each, up to approximately 3.9 x 10 6 Each, up to approximately 4 x 10 6 Each, up to approximately 4.1 x 10 6 Each, up to approximately 4.2 x 10 6 Each, up to approximately 4.3 x 10 6 Each, up to approximately 4.4 x 10 6 Each, up to approximately 4.5 x 10 6 Each, up to approximately 4.6 x 10 6 Each, up to approximately 4.7 x 10 6 Each, up to approximately 4.8 x 10 6 Each, up to approximately 4.9 x 10 6 Each, up to approximately 5 x 10 6 Each, up to approximately 5.1 x 10 6 Each, up to approximately 5.2 x 10 6 Each, up to approximately 5.3 x 10 6 Each, up to approximately 5.4 x 10 6 Each, up to approximately 5.5 x 10 6 Each, up to approximately 5.6 x 10 6 Each, up to approximately 5.7 x 10 6 Each, up to approximately 5.8 x 10 6 Each, up to approximately 5.9 x 10 6 Each, up to approximately 6 x 10 6 Each, up to approximately 6.1 x 10 6 Each, up to approximately 6.2 x 10 6 Each, up to approximately 6.3 x 10 6 Each, up to approximately 6.4 x 10 6 Each, up to approximately 6.5 x 10 6 Each, up to approximately 6.6 x 10 6 Each, up to approximately 6.7 x 10 6 Each, up to approximately 6.8 x 10 6 Each, up to approximately 6.9 x 10 6 , pieces, maximum approximately 7 x 10 6 Each, up to approximately 7.1 x 10 6 Each, up to approximately 7.2 x 10 6 Each, up to approximately 7.3 x 10 6 Each, up to approximately 7.4 x 10 6 Each, up to approximately 7.5 x 106 Each, up to approximately 7.6 x 10 6 Each, up to approximately 7.7 x 10 6 Each, up to approximately 7.8 x 10 6 Each, up to approximately 7.9 x 10 6 Each, up to approximately 8 x 10 6 Each, up to approximately 8.1 x 10 6 Each, up to approximately 8.2 x 10 6 Each, up to approximately 8.3 x 10 6 Each, up to approximately 8.4 x 10 6 Each piece is approximately 8.5 x 10 6 Each, up to approximately 8.6 x 10 6 Each, up to approximately 8.7 x 10 6 Each, up to approximately 8.8 x 10 6 Each, up to approximately 8.9 x 10 6 Each, up to approximately 9 x 10 6 Each, up to approximately 9.1 x 10 6 Each, up to approximately 9.2 x 10 6 Each, up to approximately 9.3 x 10 6 Each, up to approximately 9.4 x 10 6 Each, up to approximately 9.5 x 10 6 Each, up to approximately 9.6 x 10 6 Each, up to approximately 9.7 x 10 6 Each, up to approximately 9.8 x 10 6 Each, up to approximately 9.9 x 10 6 Each, up to approximately 1 x 10 7 Each, up to approximately 1.5 x 10 7 Each, up to approximately 2 x 10 7 Each, up to approximately 2.5 x 10 7 Each, up to approximately 3 x 10 7 Each, up to approximately 3.5 x 10 7 Each, up to approximately 4 x 10 7 Each, up to approximately 4.5 x 10 7 Each, up to approximately 5 x 10 7 Each, up to approximately 5.5 x 10 7 Each, up to approximately 6 x 10 7 Each, up to approximately 6.5 x 10 7 Each, up to approximately 7 x 10 7 Each, up to approximately 7.5 x 10 7 Each, up to approximately 8 x 10 7 Each piece is approximately 8.5 x 10 7 Each, up to approximately 9 x 107 Each, up to approximately 9.5 x 10 7 Each, up to approximately 1 x 10 8 Each, up to approximately 1.5 x 10 8 Each, up to approximately 2 x 10 8 Each, up to approximately 2.5 x 10 8 Each, up to approximately 3 x 10 8 Each, up to approximately 3.5 x 10 8 Each, up to approximately 4 x 10 7 Each, up to approximately 4.5 x 10 8 Each, up to approximately 5 x 10 8 Each, up to approximately 5.5 x 10 8 Each, up to approximately 6 x 10 8 Each, up to approximately 6.5 x 10 8 Each, up to approximately 7 x 10 8 Each, up to approximately 7.5 x 10 8 Each, up to approximately 8 x 10 8 Each piece is approximately 8.5 x 10 8 Each, up to approximately 9 x 10 8 Each, up to approximately 9.5 x 10 8 , or at most about 1 x 10 9 A population of cells enriched with Treg cells (e.g., CD45) + It may include a second population of cells or a population of Tregs and / or Tregs or Treg doses (for example, Tregs may be CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD3 + CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD127 dim FOXP3 + CD3 + FOXP3 + CD3 + CD4 + FOXP3 + CD3 + CD4 + CD25 + FOXP3 +CD3 + CD25 + FOXP3 + CD3 + CD25 + CD127 dim CD4 + CD25 + CD4 + CD25 + CD127 dim FOXP3 + FOXP3 + CD4 + FOXP3 + CD4 + CD25 + FOXP3 + CD25 + FOXP3 + , or CD25 + CD127 dim (That is.)

[0259] For example, a cell population enriched with Treg, or a cell population containing at least one dose of Treg, for example, CD45 containing at least one dose of Treg. + A second population of cells, or CD4 + CD25 + CD127 dim The regulatory T cell (Treg) population is 1 × 10¹⁶ per kg of the recipient's actual or ideal body weight. 4 ~1 × 10 9 pieces, 1×10 5 ~1 × 10 8 pieces, 1×10 5 ~2×10 7 pieces, 5×10 5 ~2×10 7 pieces, 5×10 5 ~1.5×10 7 pieces, 5×10 5 ~1 × 10 7 pieces, 5×10 5 ~9×10 6 pieces, 5×10 5 ~8×10 6 pieces, 5×10 5 ~7×10 6 pieces, 5×10 5 ~6×10 6 pieces, 5×105 ~5×10 6 5×10 5 ~4×10 6 5×10 5 ~3×10 6 5×10 5 ~2×10 6 5×10 5 ~1×10 6 1×10 6 ~1.5×10 7 1×10 6 ~1×10 7 1×10 6 ~9×10 6 1×10 6 ~8×10 6 1×10 6 ~7×10 6 1×10 6 ~6×10 6 1×10 6 ~5×10 6 1×10 6 ~4×10 6 1×10 6 ~3×10 6 1×10 6 ~2×10 6 1.5 × 10 6 ~1.5×10 7 1.5 × 10 6 ~1×10 7 1.5 × 10 6 ~9×10 6 1.5 × 10 6 ~8×10 6 1.5 × 10 6 ~7×10 6 1.5 × 10 6 ~6×10 6 1.5 × 10 6 ~5×10 6 1.5 × 10 6 ~4×10 6 1.5 × 10 6 ~3×10 6 1.5 × 10 6 ~2×10 6 1, 2×10 6 ~1.5×10 7 1, 2×10 6~1 × 10 7 pieces, 2×10 6 ~9×10 6 pieces, 2×10 6 ~8×10 6 pieces, 2×10 6 ~7×10 6 pieces, 2×10 6 ~6×10 6 pieces, 2×10 6 ~5×10 6 pieces, 2×10 6 ~4×10 6 pieces, 2×10 6 ~3×10 6 pieces, 2.5×10 6 ~1.5×10 7 pieces, 2.5×10 6 ~1 × 10 7 pieces, 2.5×10 6 ~9×10 6 pieces, 2.5×10 6 ~8×10 6 pieces, 2.5×10 6 ~7×10 6 pieces, 2.5×10 6 ~6×10 6 pieces, 2.5×10 6 ~5×10 6 pieces, 2.5×10 6 ~4×10 6 1, or 2.5 × 10 6 ~3×10 6 A population of cells enriched with Treg cells (e.g., CD45) + It may include a second population of cells or a population of Tregs and / or Tregs or Treg doses (for example, Tregs may be CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD3 + CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD127 dim FOXP3 + CD3 +FOXP3 + CD3 + CD4 + FOXP3 + CD3 + CD4 + CD25 + FOXP3 + CD3 + CD25 + FOXP3 + CD3 + CD25 + CD127 dim CD4 + CD25 + CD4 + CD25 + CD127 dim FOXP3 + FOXP3 + CD4 + FOXP3 + CD4 + CD25 + FOXP3 + CD25 + FOXP3 + , or CD25 + CD127 dim (That is.)

[0260] In some embodiments, a Treg-enriched cell population, or a cell population containing at least one dose of Treg, for example, CD45 containing at least one dose of Treg. + A second population of cells, or CD4 + CD25 + CD127 dim The regulatory T cell (Treg) population is approximately 1.0 × 10⁶ in the Treg-enriched cell population. 5 ~Approx. 5.0×10 9 pieces, approximately 5.0×10 5 ~Approx. 3.0×10 9 pieces, approximately 1.5×10 7 ~Approx. 3.0×10 9 1 piece, or approximately 5.0 x 10 5 ~Approx. 1.0×10 8The Treg comprises, for example, fresh isolated Tregs. In some embodiments, a Treg-enriched cell population or a cell population containing at least one dose of Treg, for example, a second population of CD45+ cells containing at least one dose of Treg, is approximately 1.0 × 10⁶ 5 One or more Tregs, approximately 2.0 × 10⁻¹⁶ 5 One or more Tregs, approximately 3.0 × 10⁻¹⁶ 5 One or more Tregs, approximately 4.0 × 10⁻⁶ 5 One or more Tregs, approximately 5.0 × 10⁻¹⁶ 5 One or more Treg cells, approximately 6.0 × 10⁻⁶ 5 One or more Tregs, approximately 7.0 × 10⁻¹⁶ 5 One or more Treg cells, approximately 8.0 × 10⁻⁶ 5 One or more Tregs, approximately 9.0 × 10⁻¹⁶ 5 One or more Treg cells, approximately 1.0 × 10⁻⁶ 6 One or more Treg cells, approximately 1.0 × 10⁻⁶ 6 One or more Treg cells, approximately 1.1 × 10⁻⁶ 6 One or more Treg cells, approximately 1.2 × 10⁻⁶ 6 One or more Treg cells, approximately 1.3 × 10⁻⁶ 6 More than 10 Treg cells, approximately 1.4 × 10⁻⁶ 6 One or more Treg cells, approximately 1.5 × 10⁻⁶ 6 One or more Treg cells, approximately 1.6 × 10⁻⁶ 6 More than 10 Treg cells, approximately 1.7 × 10⁻⁶ 6 One or more Treg cells, approximately 1.8 × 10⁻⁶ 6 One or more Treg cells, approximately 1.9 × 10⁻⁶ 6 One or more Tregs, or approximately 2.0 × 10⁻⁶ 6 One or more Treg cells, approximately 2.1 × 10⁻⁶ 6 One or more Tregs, approximately 2.2 × 10⁻¹⁶ 6 One or more Tregs, approximately 2.3 × 10⁻¹⁶ 6 One or more Tregs, approximately 2.4 × 10⁻¹⁶ 6 One or more Treg cells, approximately 2.5 × 10⁻⁶ 6 One or more Treg cells, approximately 2.6 × 10⁻⁶ 6 One or more Treg cells, approximately 2.7 × 10⁻⁶ 6 One or more Treg cells, approximately 2.8 × 10⁻⁶ 6 One or more Treg cells, approximately 2.9 × 10⁻⁶ 6 One or more Tregs, approximately 3.0 × 10⁻¹⁶ 6One or more Tregs, approximately 3.1 × 10⁻¹⁶ 6 One or more Tregs, approximately 3.2 × 10⁻⁶ 6 One or more Tregs, approximately 3.3 × 10⁻⁶ 6 More than one Treg, approximately 3.4 × 10⁻⁶ 6 One or more Treg cells, approximately 3.5 × 10⁻⁶ 6 One or more Treg cells, approximately 3.6 × 10⁻⁶ 6 One or more Tregs, approximately 3.7 × 10⁻⁶ 6 One or more Treg cells, approximately 3.8 × 10⁻⁶ 6 One or more Treg cells, approximately 3.9 × 10⁻⁶ 6 One or more Tregs, approximately 4.0 × 10⁻⁶ 6 One or more Tregs, approximately 4.1 × 10⁻⁶ 6 More than 10 Tregs, approximately 4.2 × 10 6 More than one Treg, approximately 4.3 × 10⁻⁶ 6 More than one Treg, approximately 4.4 × 10⁻⁶ 6 One or more Tregs, approximately 4.5 × 10⁻⁶ 6 One or more Tregs, approximately 4.6 × 10⁻⁶ 6 One or more Tregs, approximately 4.7 × 10⁻⁶ 6 One or more Tregs, approximately 4.8 × 10⁻⁶ 6 More than one Treg, approximately 4.9 × 10⁻⁶ 6 One or more Tregs, approximately 5.0 × 10⁻¹⁶ 6 One or more Tregs, approximately 5.1 × 10⁻¹⁶ 6 More than one Treg, approximately 5.2 × 10⁻⁶ 6 One or more Tregs, approximately 5.3 × 10⁻⁶ 6 More than one Treg, approximately 5.4 × 10⁻⁶ 6 One or more Tregs, approximately 5.5 × 10 6 One or more Tregs, approximately 5.6 × 10⁻⁶ 6 One or more Tregs, approximately 5.7 × 10⁻⁶ 6 One or more Tregs, approximately 5.8 × 10⁻¹⁶ 6 One or more Treg cells, approximately 5.9 × 10⁻⁶ 6 One or more Treg cells, approximately 6.0 × 10⁻⁶ 6 More than 10 Tregs, approximately 6.1 × 10⁻⁶ 6 More than 10 Tregs, approximately 6.2 × 10⁻⁶ 6 More than one Treg, approximately 6.3 × 10⁻⁶ 6 More than one Treg, approximately 6.4 × 10⁻⁶ 6One or more Tregs, approximately 6.5 × 10⁻⁶ 6 More than one Treg, approximately 6.6 × 10⁻⁶ 6 One or more Tregs, approximately 6.7 × 10⁻⁶ 6 One or more Tregs, approximately 6.8 × 10⁻⁶ 6 More than one Treg cell, approximately 6.9 × 10⁻⁶ 6 One or more Tregs, approximately 7.0 × 10⁻¹⁶ 6 One or more Tregs, approximately 7.1 × 10⁻⁶ 6 One or more Tregs, approximately 7.2 × 10⁻⁶ 6 One or more Treg cells, approximately 7.3 × 10⁻⁶ 6 One or more Tregs, approximately 7.4 × 10⁻⁶ 6 One or more Treg cells, approximately 7.5 × 10⁻⁶ 6 More than one Treg, approximately 7.6 × 10⁻¹⁶ 6 One or more Tregs, approximately 7.7 × 10⁻⁶ 6 One or more Tregs, approximately 7.8 × 10⁻⁶ 6 One or more Treg cells, approximately 7.9 × 10⁻⁶ 6 One or more Treg cells, approximately 8.0 × 10⁻⁶ 6 One or more Tregs, approximately 8.1 × 10⁻¹⁶ 6 One or more Tregs, approximately 8.2 × 10⁻⁶ 6 One or more Tregs, approximately 8.3 × 10⁻⁶ 6 One or more Tregs, approximately 8.4 × 10⁻⁶ 6 One or more Treg cells, approximately 8.5 × 10⁻⁶ 6 One or more Treg cells, approximately 8.6 × 10⁻⁶ 6 More than one Treg, approximately 8.7 × 10⁻⁶ 6 One or more Tregs, approximately 8.8 × 10⁻⁶ 6 One or more Treg cells, approximately 8.9 × 10⁻⁶ 6 One or more Tregs, approximately 9.0 × 10⁻¹⁶ 6 One or more Tregs, approximately 9.1 × 10⁻⁶ 6 One or more Tregs, approximately 9.2 × 10⁻⁶ 6 One or more Treg cells, approximately 9.3 × 10⁻⁶ 6 One or more Tregs, approximately 9.4 × 10⁻⁶ 6 One or more Treg cells, approximately 9.5 × 10⁻⁶ 6 One or more Treg cells, approximately 9.6 × 10⁻⁶ 6 One or more Treg cells, approximately 9.7 × 10⁻⁶ 6 One or more Treg cells, approximately 9.8 × 10⁻⁶ 6One or more Treg cells, approximately 9.9 × 10⁻⁶ 6 One or more Treg cells, approximately 1.0 × 10⁻⁶ 7 One or more Treg cells, approximately 1.1 × 10⁻⁶ 7 One or more Treg cells, approximately 1.2 × 10⁻⁶ 7 One or more Treg cells, approximately 1.3 × 10⁻⁶ 7 More than 10 Treg cells, approximately 1.4 × 10⁻⁶ 7 One or more Treg cells, approximately 1.5 × 10⁻⁶ 7 One or more Treg cells, approximately 1.6 × 10⁻⁶ 7 More than 10 Treg cells, approximately 1.7 × 10⁻⁶ 7 One or more Treg cells, approximately 1.8 × 10⁻⁶ 7 One or more Treg cells, approximately 1.9 × 10⁻⁶ 7 One or more Tregs, or approximately 2.0 × 10⁻⁶ 7 One or more Treg cells, approximately 2.1 × 10⁻⁶ 7 One or more Tregs, approximately 2.2 × 10⁻¹⁶ 7 One or more Tregs, approximately 2.3 × 10⁻¹⁶ 7 One or more Tregs, approximately 2.4 × 10⁻¹⁶ 7 One or more Treg cells, approximately 2.5 × 10⁻⁶ 7 One or more Treg cells, approximately 2.6 × 10⁻⁶ 7 One or more Treg cells, approximately 2.7 × 10⁻⁶ 7 One or more Treg cells, approximately 2.8 × 10⁻⁶ 7 One or more Treg cells, approximately 2.9 × 10⁻⁶ 7 One or more Tregs, approximately 3.0 × 10⁻¹⁶ 7 One or more Tregs, approximately 3.1 × 10⁻¹⁶ 7 One or more Tregs, approximately 3.2 × 10⁻⁶ 7 One or more Tregs, approximately 3.3 × 10⁻⁶ 7 More than one Treg, approximately 3.4 × 10⁻⁶ 7 One or more Treg cells, approximately 3.5 × 10⁻⁶ 7 One or more Treg cells, approximately 3.6 × 10⁻⁶ 7 One or more Tregs, approximately 3.7 × 10⁻⁶ 7 One or more Treg cells, approximately 3.8 × 10⁻⁶ 7 One or more Treg cells, approximately 3.9 × 10⁻⁶ 7 One or more Tregs, approximately 4.0 × 10⁻⁶ 7 One or more Tregs, approximately 4.1 × 10⁻⁶ 7 More than 10 Tregs, approximately 4.2 × 10 7More than one Treg, approximately 4.3 × 10⁻⁶ 7 More than one Treg, approximately 4.4 × 10⁻⁶ 7 One or more Tregs, approximately 4.5 × 10⁻⁶ 7 One or more Tregs, approximately 4.6 × 10⁻⁶ 7 One or more Tregs, approximately 4.7 × 10⁻⁶ 7 One or more Tregs, approximately 4.8 × 10⁻⁶ 7 More than one Treg, approximately 4.9 × 10⁻⁶ 7 One or more Tregs, approximately 5.0 × 10⁻¹⁶ 7 One or more Tregs, approximately 5.1 × 10⁻¹⁶ 7 One or more Tregs, approximately 5.2 × 10⁻⁶ 7 One or more Tregs, approximately 5.3 × 10⁻⁶ 7 More than one Treg, approximately 5.4 × 10⁻⁶ 7 One or more Tregs, approximately 5.5 × 10 7 One or more Tregs, approximately 5.6 × 10⁻⁶ 7 One or more Tregs, approximately 5.7 × 10⁻⁶ 7 One or more Tregs, approximately 5.8 × 10⁻¹⁶ 7 One or more Treg cells, approximately 5.9 × 10⁻⁶ 7 One or more Treg cells, approximately 6.0 × 10⁻⁶ 7 More than 10 Tregs, approximately 6.1 × 10⁻⁶ 7 More than 10 Tregs, approximately 6.2 × 10⁻⁶ 7 More than one Treg, approximately 6.3 × 10⁻⁶ 7 More than one Treg, approximately 6.4 × 10⁻⁶ 7 One or more Tregs, approximately 6.5 × 10⁻⁶ 7 More than one Treg, approximately 6.6 × 10⁻⁶ 7 One or more Tregs, approximately 6.7 × 10⁻⁶ 7 One or more Tregs, approximately 6.8 × 10⁻⁶ 7 More than one Treg cell, approximately 6.9 × 10⁻⁶ 7 One or more Tregs, approximately 7.0 × 10⁻¹⁶ 7 One or more Tregs, approximately 7.1 × 10⁻⁶ 7 One or more Tregs, approximately 7.2 × 10⁻⁶ 7 One or more Treg cells, approximately 7.3 × 10⁻⁶ 7 One or more Tregs, approximately 7.4 × 10⁻⁶ 7 One or more Treg cells, approximately 7.5 × 10⁻⁶ 7 More than one Treg, approximately 7.6 × 10⁻¹⁶ 7One or more Tregs, approximately 7.7 × 10⁻⁶ 7 One or more Tregs, approximately 7.8 × 10⁻⁶ 7 One or more Treg cells, approximately 7.9 × 10⁻⁶ 7 One or more Treg cells, approximately 8.0 × 10⁻⁶ 7 One or more Tregs, approximately 8.1 × 10⁻¹⁶ 7 One or more Tregs, approximately 8.2 × 10⁻⁶ 7 One or more Tregs, approximately 8.3 × 10⁻⁶ 7 One or more Tregs, approximately 8.4 × 10⁻⁶ 7 One or more Treg cells, approximately 8.5 × 10⁻⁶ 7 One or more Treg cells, approximately 8.6 × 10⁻⁶ 7 More than one Treg, approximately 8.7 × 10⁻⁶ 7 One or more Tregs, approximately 8.8 × 10⁻⁶ 7 One or more Treg cells, approximately 8.9 × 10⁻⁶ 7 One or more Tregs, approximately 9.0 × 10⁻¹⁶ 7 One or more Tregs, approximately 9.1 × 10⁻⁶ 7 One or more Tregs, approximately 9.2 × 10⁻⁶ 7 One or more Treg cells, approximately 9.3 × 10⁻⁶ 7 One or more Tregs, approximately 9.4 × 10⁻⁶ 7 One or more Treg cells, approximately 9.5 × 10⁻⁶ 7 One or more Treg cells, approximately 9.6 × 10⁻⁶ 7 One or more Treg cells, approximately 9.7 × 10⁻⁶ 7 One or more Treg cells, approximately 9.8 × 10⁻⁶ 7 One or more Treg cells, approximately 9.9 × 10⁻⁶ 7 One or more Treg cells, approximately 1.0 × 10⁻⁶ 8 One or more Treg cells, approximately 1.1 × 10⁻⁶ 8 One or more Treg cells, approximately 1.2 × 10⁻⁶ 8 One or more Treg cells, approximately 1.3 × 10⁻⁶ 8 More than 10 Treg cells, approximately 1.4 × 10⁻⁶ 8 One or more Treg cells, approximately 1.5 × 10⁻⁶ 8 One or more Treg cells, approximately 1.6 × 10⁻⁶ 8 More than 10 Treg cells, approximately 1.7 × 10⁻⁶ 8 One or more Treg cells, approximately 1.8 × 10⁻⁶ 8 One or more Treg cells, approximately 1.9 × 10⁻⁶ 8 One or more Tregs, or approximately 2.0 × 10⁻⁶ 8One or more Treg cells, approximately 2.1 × 10⁻⁶ 8 One or more Tregs, approximately 2.2 × 10⁻¹⁶ 8 One or more Tregs, approximately 2.3 × 10⁻¹⁶ 8 One or more Tregs, approximately 2.4 × 10⁻¹⁶ 8 One or more Treg cells, approximately 2.5 × 10⁻⁶ 8 One or more Treg cells, approximately 2.6 × 10⁻⁶ 8 One or more Treg cells, approximately 2.7 × 10⁻⁶ 8 One or more Treg cells, approximately 2.8 × 10⁻⁶ 8 One or more Treg cells, approximately 2.9 × 10⁻⁶ 8 One or more Tregs, approximately 3. 0 x 10 8 One or more Tregs, approximately 3.1 × 10⁻¹⁶ 8 One or more Tregs, approximately 3.2 × 10⁻⁶ 8 One or more Tregs, approximately 3.3 × 10⁻⁶ 8 More than one Treg, approximately 3.4 × 10⁻⁶ 8 One or more Treg cells, approximately 3.5 × 10⁻⁶ 8 One or more Treg cells, approximately 3.6 × 10⁻⁶ 8 One or more Tregs, approximately 3.7 × 10⁻⁶ 8 One or more Treg cells, approximately 3.8 × 10⁻⁶ 8 One or more Treg cells, approximately 3.9 × 10⁻⁶ 8 One or more Tregs, approximately 4.0 × 10⁻⁶ 8 One or more Tregs, approximately 4.1 × 10⁻⁶ 8 More than 10 Tregs, approximately 4.2 × 10 8 More than one Treg, approximately 4.3 × 10⁻⁶ 8 More than one Treg, approximately 4.4 × 10⁻⁶ 8 One or more Tregs, approximately 4.5 × 10⁻⁶ 8 One or more Tregs, approximately 4.6 × 10⁻⁶ 8 One or more Tregs, approximately 4.7 × 10⁻⁶ 8 One or more Tregs, approximately 4.8 × 10⁻⁶ 8 More than one Treg, approximately 4.9 × 10⁻⁶ 8 One or more Tregs, approximately 5.0 × 10⁻¹⁶ 8 One or more Tregs, approximately 5.1 × 10⁻¹⁶ 8 One or more Tregs, approximately 5.2 × 10⁻⁶ 8 One or more Tregs, approximately 5.3 × 10⁻⁶ 8 More than one Treg, approximately 5.4 × 10⁻⁶ 8One or more Tregs, approximately 5.5 × 10 8 One or more Tregs, approximately 5.6 × 10⁻⁶ 8 One or more Tregs, approximately 5.7 × 10⁻⁶ 8 One or more Tregs, approximately 5.8 × 10⁻¹⁶ 8 One or more Treg cells, approximately 5.9 × 10⁻⁶ 8 One or more Treg cells, approximately 6.0 × 10⁻⁶ 8 More than 10 Tregs, approximately 6.1 × 10⁻⁶ 8 More than 10 Tregs, approximately 6.2 × 10⁻⁶ 8 More than one Treg, approximately 6.3 × 10⁻⁶ 8 More than one Treg, approximately 6.4 × 10⁻⁶ 8 One or more Tregs, approximately 6.5 × 10⁻⁶ 8 More than one Treg, approximately 6.6 × 10⁻⁶ 8 One or more Tregs, approximately 6.7 × 10⁻⁶ 8 One or more Tregs, approximately 6.8 × 10⁻⁶ 8 More than one Treg cell, approximately 6.9 × 10⁻⁶ 8 One or more Tregs, approximately 7.0 × 10⁻¹⁶ 8 One or more Tregs, approximately 7.1 × 10⁻⁶ 8 One or more Tregs, approximately 7.2 × 10⁻⁶ 8 One or more Treg cells, approximately 7.3 × 10⁻⁶ 8 One or more Tregs, approximately 7.4 × 10⁻⁶ 8 One or more Treg cells, approximately 7.5 × 10⁻⁶ 8 More than one Treg, approximately 7.6 × 10⁻¹⁶ 8 One or more Tregs, approximately 7.7 × 10⁻⁶ 8 One or more Tregs, approximately 7.8 × 10⁻⁶ 8 One or more Treg cells, approximately 7.9 × 10⁻⁶ 8 One or more Treg cells, approximately 8.0 × 10⁻⁶ 8 One or more Tregs, approximately 8.1 × 10⁻¹⁶ 8 One or more Tregs, approximately 8.2 × 10⁻⁶ 8 One or more Tregs, approximately 8.3 × 10⁻⁶ 8 One or more Tregs, approximately 8.4 × 10⁻⁶ 8 One or more Treg cells, approximately 8.5 × 10⁻⁶ 8 One or more Treg cells, approximately 8.6 × 10⁻⁶ 8 More than one Treg, approximately 8.7 × 10⁻⁶ 8 One or more Tregs, approximately 8.8 × 10⁻⁶ 8One or more Treg cells, approximately 8.9 × 10⁻⁶ 8 One or more Tregs, approximately 9.0 × 10⁻¹⁶ 8 One or more Tregs, approximately 9.1 × 10⁻⁶ 8 One or more Tregs, approximately 9.2 × 10⁻⁶ 8 One or more Treg cells, approximately 9.3 × 10⁻⁶ 8 One or more Tregs, approximately 9.4 × 10⁻⁶ 8 One or more Treg cells, approximately 9.5 × 10⁻⁶ 8 One or more Treg cells, approximately 9.6 × 10⁻⁶ 8 One or more Treg cells, approximately 9.7 × 10⁻⁶ 8 One or more Treg cells, approximately 9.8 × 10⁻⁶ 8 One or more Treg cells, approximately 9.9 × 10⁻⁶ 8 One or more Treg cells, approximately 1.0 × 10⁻⁶ 9 One or more Treg cells, approximately 1.1 × 10⁻⁶ 9 One or more Treg cells, approximately 1.2 × 10⁻⁶ 9 One or more Treg cells, approximately 1.3 × 10⁻⁶ 9 More than 10 Treg cells, approximately 1.4 × 10⁻⁶ 9 One or more Treg cells, approximately 1.5 × 10⁻⁶ 9 One or more Treg cells, approximately 1.6 × 10⁻⁶ 9 More than 10 Treg cells, approximately 1.7 × 10⁻⁶ 9 One or more Treg cells, approximately 1.8 × 10⁻⁶ 9 One or more Treg cells, approximately 1.9 × 10⁻⁶ 9 One or more Tregs, or approximately 2.0 × 10⁻⁶ 9 One or more Treg cells, approximately 2.1 × 10⁻⁶ 9 One or more Tregs, approximately 2.2 × 10⁻¹⁶ 9 One or more Tregs, approximately 2.3 × 10⁻¹⁶ 9 One or more Tregs, approximately 2.4 × 10⁻¹⁶ 9 One or more Treg cells, approximately 2.5 × 10⁻⁶ 9 One or more Treg cells, approximately 2.6 × 10⁻⁶ 9 One or more Treg cells, approximately 2.7 × 10⁻⁶ 9 One or more Treg cells, approximately 2.8 × 10⁻⁶ 9 One or more Treg cells, approximately 2.9 × 10⁻⁶ 9 One or more Tregs, approximately 3.0 × 10⁻¹⁶ 9 One or more Tregs, approximately 3.1 × 10⁻¹⁶ 9 One or more Tregs, approximately 3.2 × 10⁻⁶ 9One or more Tregs, approximately 3.3 × 10⁻⁶ 9 More than one Treg, approximately 3.4 × 10⁻⁶ 9 One or more Treg cells, approximately 3.5 × 10⁻⁶ 9 One or more Treg cells, approximately 3.6 × 10⁻⁶ 9 One or more Tregs, approximately 3.7 × 10⁻⁶ 9 One or more Treg cells, approximately 3.8 × 10⁻⁶ 9 One or more Treg cells, approximately 3.9 × 10⁻⁶ 9 One or more Tregs, approximately 4.0 × 10⁻⁶ 9 One or more Tregs, approximately 4.1 × 10⁻⁶ 9 More than 10 Tregs, approximately 4.2 × 10 9 More than one Treg, approximately 4.3 × 10⁻⁶ 9 More than one Treg, approximately 4.4 × 10⁻⁶ 9 One or more Tregs, approximately 4.5 × 10⁻⁶ 9 One or more Tregs, approximately 4.6 × 10⁻⁶ 9 One or more Tregs, approximately 4.7 × 10⁻⁶ 9 One or more Tregs, approximately 4.8 × 10⁻⁶ 9 More than one Treg, approximately 4.9 × 10⁻⁶ 9 One or more Tregs, or approximately 5.0 × 10⁻⁶ 9 It contains one or more Tregs. In some embodiments, the Treg is CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD3 + CD4 + CD25 + CD127 dim CD3 + CD4 + CD25 + CD127 dim FOXP3 + CD3 + FOXP3 + CD3 + CD4 + FOXP3 + CD3 + CD4 + CD25 + FOXP3 + CD3 + CD25 + FOXP3+ CD3 + CD25 + CD127 dim CD4 + CD25 + CD4 + CD25 + CD127 dim FOXP3 + FOXP3 + CD4 + FOXP3 + CD4 + CD25 + FOXP3 + CD25 + FOXP3 + , or CD25 + CD127 dim That is the case.

[0261] Tcon CD45 containing at least Tcon or at least one dose of Tcon + A third population of cells, or typical CD3 + The population of T cells (Tcon) is at least approximately 1 × 10¹⁶ per kilogram of the recipient's actual or ideal body weight. 4 each, at least about 2 × 10 4 Each, at least approximately 3 × 10 4 Each, at least approximately 4 × 10 4 Each, at least approximately 5 x 10 4 Each, at least approximately 6 x 10 4 Each, at least approximately 7 x 10 4 Each, at least approximately 8 x 10 4 Each, at least approximately 9 x 10 4 each, at least about 1 × 10 5 each, at least about 2 × 10 5 Each, at least approximately 3 × 10 5 Each, at least approximately 4 × 10 5 Each, at least approximately 5 x 10 5 Each, at least approximately 6 x 10 5 Each, at least approximate...

Claims

1. Products for multicomponent cell therapy, (a) A pharmaceutical composition comprising one or more cells containing a chimeric receptor, or an isolated nucleic acid encoding the chimeric receptor, (b) Products for immunomodulatory cell therapy, The multicomponent cell therapy product wherein, optionally, one or more cells containing the chimeric receptor or the pharmaceutical composition and / or one or more cells of the immunomodulatory cell therapy product are allogeneic cells derived from an HLA-matched donor for a human subject receiving the multicomponent cell therapy product.

2. a) Whether the HLA-matched donor is an allogeneic HLA-matched donor for the human subject, The HLA-matched donor is an HLA-mismatched donor of the same species as the human subject, and / or b) One or more of the allogeneic cells are not genetically modified to reduce the risk of graft-versus-host disease (GVHD) and / or to prevent allogeneic rejection of the human subject, and / or c) The immunomodulatory cell therapy product includes regulatory T cell therapy, or The immunomodulatory cell therapy product comprises allogeneic hematopoietic stem cell transplantation (HSCT), Selectively, the same type of HSCT is CD34 ＋ A population of hematopoietic stem progenitor cells (HSPCs), or the said allogeneic HSCTs, CD4 ＋ CD25 ＋ CD127 ｄｉｍ A population of regulatory T cells (Treg), or the homogeneous HSCT comprises HSPC and Treg, and / or Selectively, the same type of HSCT is CD45RA － The homogeneous HSCT includes a population of memory T cells (Tmem), or the homogeneous HSCT includes a population of HSPCs and a population of Tmem, or the homogeneous HSCT includes a population of Tregs and a population of Tmem, or the homogeneous HSCT includes a population of HSPCs, a population of Tregs, and a population of Tmem, and / or Optionally, the same type of HSCT is used for naive, conventional CD3 ＋ CD25 － CD45RA ＋ T cells are depleted, and / or Optionally, the allogeneic HSCT is conventional CD3 ＋ comprises a population of T cells (Tcon), or said allogeneic HSCT comprises a population of HSPC and a population of Tcon, or said allogeneic HSCT comprises a population of Treg and a population of Tcon, or said allogeneic HSCT comprises a population of Tmem and a population of Tcon, or said allogeneic HSCT comprises a population of HSPC, a population of Treg, and a population of Tcon, or said allogeneic HSCT comprises a population of HSPC, a population of Treg, a population of Tmem, and a population of Tcon, and / or Selectively, the allogeneic HSCT is a T-cell depleted allogeneic HSCT, and / or d) The immunomodulatory cell therapy product is CD34 ＋ A population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, Selectively, the immunomodulatory cell therapy product is CD4 ＋ CD25 ＋ CD127 ｄｉｍ The present invention further comprises a population of regulatory T cells (Treg), wherein the population of Tregs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. and / or the immunomodulatory cell therapy product is CD45RA － The present invention further comprises a population of memory T cells (Tmem), wherein the population of Tmem comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, and / or the immunomodulatory cell therapy product is a conventional CD3 ＋ The present invention further comprises a population of T cells (Tcon), wherein the population of Tcon comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, and / or e) The immunomodulatory cell therapy product is CD4 ＋ CD25 ＋ CD127 ｄｉｍ A population of regulatory T cells (Tregs), wherein the population of Tregs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, Selectively, the immunomodulatory cell therapy product is CD34 ＋ The present invention further comprises a population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. And / or the immunomodulatory cell therapy product is CD45RA － The present invention further comprises a population of memory T cells (Tmem), wherein the population of Tmem comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, And / or the immunomodulatory cell therapy product is a conventional CD3 ＋ The present invention further comprises a population of T cells (Tcon), wherein the population of Tcon comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, and / or f) The immunomodulatory cell therapy product is CD45RA － A population of memory T cells (Tmem), wherein the population of Tmem comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, Selectively, the immunomodulatory cell therapy product is CD34 ＋ The present invention further comprises a population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. And / or the immunomodulatory cell therapy product is CD4 ＋ CD25 ＋ CD127 ｄｉｍ The present invention further comprises a population of regulatory T cells (Treg), wherein the population of Tregs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. And / or the immunomodulatory cell therapy product is a conventional CD3 ＋ The present invention further comprises a population of T cells (Tcon), wherein the population of Tcon comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, and / or g) The immunomodulatory cell therapy product is a conventional CD3 ＋ A population of T cells (Tcon), wherein the population of Tcon comprises one or more cells containing a chimeric receptor, Selectively, the immunomodulatory cell therapy product is CD34 ＋ The present invention further comprises a population of hematopoietic stem progenitor cells (HSPCs), wherein the population of HSPCs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. And / or the immunomodulatory cell therapy product is CD4 ＋ CD25 ＋ CD127 ｄｉｍ The present invention further comprises a population of regulatory T cells (Treg), wherein the population of Tregs comprises one or more cells containing the chimeric receptor or the pharmaceutical composition. And / or the immunomodulatory cell therapy product is CD45RA － The present invention further comprises a population of memory T cells (Tmem), wherein the population of Tmem comprises one or more cells containing the chimeric receptor or the pharmaceutical composition, and / or h) The multicomponent cell therapy product according to claim 1, wherein the immunomodulatory cell therapy product further comprises a population of tumor-infiltrating lymphocytes (TILs), and optionally the population of Tcon comprises the population of TILs.

3. The pharmaceutical composition comprises a vector containing the nucleic acid, optionally the vector being a viral vector, optionally selected from the group consisting of lentiviral vectors, retroviral vectors, oncolytic viral vectors, adenovirus vectors, adeno-associated virus (AAV) vectors, and virus-like particles (VLPs), or The pharmaceutical composition comprises nanoparticles containing the nucleic acid, wherein the nanoparticles are optionally lipid nanoparticles (LNPs). The multicomponent cell therapy product according to claim 1 or 2, wherein the pharmaceutical composition optionally further comprises a pharmaceutically acceptable carrier, a pharmaceutically acceptable excipient, or a combination thereof.

4. a) The Tcon is formulated for administration approximately 12 hours or more after the administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, or The Tcon is formulated for administration approximately 24 to 120 hours after the administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, or The Tcon is formulated for administration approximately 36 to 72 hours after the administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, and / or b) The Treg, the Tmem, or the Treg and the Tmem are formulated for administration approximately 5 minutes to 5 hours after the administration of the HSPC, and / or c) The one or more cells containing the chimeric receptor or the pharmaceutical composition is formulated for administration approximately 12 hours or more after administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, or The one or more cells containing the chimeric receptor or the pharmaceutical composition may be formulated for administration approximately 24 to 120 hours after administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, or The one or more cells containing the chimeric receptor or the pharmaceutical composition may be formulated for administration approximately 36 to 72 hours after administration of the HSPC, the Treg, the Tmem, or any combination of the HSPC, the Treg, and the Tmem, or The multicomponent cell therapy product according to any one of claims 1 to 3, wherein the one or more cells containing the chimeric receptor or the pharmaceutical composition are formulated for administration approximately 5 minutes to approximately 5 hours after administration of Tcon.

5. a) The multicomponent cell therapy product further comprises a pharmaceutical composition containing one or more doses of a graft-versus-host disease (GVHD) prophylactic agent, optionally comprising tacrolimus and / or b) The multicomponent cell therapy product is present in an amount of approximately 1.0 × 10¹⁶ units per kilogram of body weight in a human subject receiving the product. ５ ~Approx. 1.0×10 ８ Each HSPC, or approximately 5.0 x 10 ５ ~Approx. 1.5×10 １０ The dose of HSPC is within the range of individual HSPCs, and / or c) The multicomponent cell therapy product provides approximately 1.0 × 10¹⁶ units per kilogram of body weight to a human subject receiving the product. ５ ~Approx. 2.0×10 ７ Each Treg, or approximately 5.0 × 10 ５ ~Approx. 3.0×10 ９ The dose of Treg is within the range of individual Tregs, and optionally, the Treg is FOXP3 ＋ and / or d) The multicomponent cell therapy product is present in an amount of approximately 1.0 × 10¹⁶ units per kilogram of body weight in a human subject receiving the product. ５ ~Approx. 1.0×10 ８ Tmem, or approximately 5.0 x 10 ５ ~Approx. 1.5×10 １０ A dose of Tmem that is within the range of Tmem, and / or e) The multicomponent cell therapy product provides approximately 1.0 × 10¹⁶ units per kilogram of body weight to a human subject receiving the product. ５ ~Approx. 4.0×10 ７ Tcon, or approximately 5.0 x 10 ５ ~Approx. 6.0×10 ９ A multicomponent cell therapy product according to any one of claims 1 to 4, comprising a dose of Tcon which is within the range of individual Tcon.

6. a) The one or more cells containing the chimeric receptor or the pharmaceutical composition comprises one or more gene modifications that increase immune tolerance with respect to a human subject receiving the product, and / or b) The one or more cells containing the chimeric receptor or the pharmaceutical composition are not modified to increase immune tolerance with respect to a human subject receiving the product, and / or c) The one or more cells containing the chimeric receptor or the pharmaceutical composition are donor-derived from an allogeneic donor to a human subject receiving the product, and / or d) The HSPC, the Treg, the Tmem, the Tcon, or any combination thereof is isolated from a donor, and the donor is optionally homogeneous with respect to a human subject receiving the product, and / or e) The one or more cells or pharmaceutical composition comprising the HSPC, the Treg, the Tmem, the Tcon, or any combination thereof, and the chimeric receptor, are derived from the same homogeneous donor. Selectively, the allogeneic donor is HLA-matched to a human subject receiving the product in one or more alleles selected from the group consisting of HLA-A, HLA-B, HLA-C, and HLA-DRB1. Selectively, the same donor is 8 / 8 HLA matched with respect to the alleles HLA-A, HLA-B, HLA-C, and HLA-DRB1 in relation to the human subject receiving the product. Selectively, the allogeneic donor has at least one HLA mismatch with respect to a human subject receiving the product, and selectively, the at least one HLA mismatch is in an allele selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-DRB1, and any combination thereof. Optionally, the allogeneic donor is an allohapjoint donor to a human subject receiving the product, and optionally, the allohapjoint donor is at least 4 / 8 HLA mismatched, at least 5 / 8 HLA mismatched, or at least 6 / 8 HLA mismatched in an allele selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-DRB1, and any combination thereof, and / or f) The multicomponent cell therapy product according to any one of claims 1 to 5, wherein the one or more cells or pharmaceutical composition comprising the HSPC, the Treg, the Tmem, the Tcon, or any combination thereof, and / or the chimeric receptor, are collected from one or more umbilical cord blood samples derived from the donor, or from one or more bone marrow samples derived from the donor, and optionally, at least one of the two or more samples is a recruited sample, optionally, the HSPC, the Treg, the Tmem, the Tcon, or any combination thereof, is obtainable from the recruited sample, and optionally, at least one recruited sample is recruitable by granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), mozovir, or any combination thereof.

7. The multicomponent cell therapy product further comprises a pre-transplant conditioning regimen, the pre-transplant conditioning regimen being formulated for administration prior to the administration of the cell therapy. The pre-transplant conditioning regimen is optionally formulated for administration approximately 2 to 10 days prior to the administration of the multicomponent cell therapy. Optionally, the pre-transplant conditioning regimen may be a whole-body irradiation-based (TBI-based) regimen or a whole-bone marrow and lymph node irradiation-based (TMLI-based) regimen. Optionally, the TBI-based regimen includes fractionated whole-body irradiation (fTBI), Optionally, the fTBI may include doses of 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more doses. Optionally, the fTBI includes a total dose in the range of approximately 500 to approximately 1600 cGy. Optionally, the TBI-based or TMLI-based regimen further comprises one or more doses of pre-transplant conditioning reagents, and optionally, the pre-transplant conditioning reagent comprises one or more pre-transplant conditioning reagents selected from the group consisting of cyclophosphamide, etoposide, thiotepa, busulfan, melphalan, fludarabine, and any combination thereof. Optionally, the pre-transplant conditioning regimen is a myeloablative pre-transplant conditioning regimen or a reduced-intensity pre-transplant conditioning (RIC) regimen, and optionally, the myeloablative pre-transplant conditioning regimen or RIC regimen comprises one or more pre-transplant conditioning reagents, and optionally, the one or more pre-transplant conditioning reagents are selected from the group consisting of thiotepa, busulfan, melphalan, fludarabine, cyclophosphamide, anti-thymocyte globulin (ATG), and any combination thereof. The multicomponent cell therapy product according to any one of claims 1 to 6, wherein the myeloablative pre-transplant conditioning regimen optionally comprises three or more pre-transplant conditioning reagents, at least one of which comprises thiotepa, and optionally the myeloablative pre-transplant conditioning regimen comprises one or more doses of busulfan, one or more doses of fludarabine, and one or more doses of thiotepa.

8. a) The chimeric receptors include BCMA, PSMA, CD8, CD10, CD19, CD20, CD21, CD22, CD25, CD30, CD33, CD34, CD37, CD44v6, CD45, CD52, CSF1R (CD115), c-Kit (CD117), CD123, CD133, Fms-like tyrosine kinase 3 (FLT-3, CD135), PDGFR-alpha (CD140a), PDGFR-beta (CD140b), CD186, CD195, VEGFR3 (FLT4, CD309), GPRC5D, TMEFF2, ROR1, B 7-H6, B7-H3, HM1.24, SLAMF7, Chondroitin sulfate proteoglycan 4 (CSPG4, melanoma-associated chondroitin sulfate proteoglycan), Epidermal growth factor receptor (EGFR), Her2, Her3, IGFR, IL3R, Fibroblast-activating protein (FAP), CDCP1, Derlin1, Tenascin, frizzled1-10, VEGFR2 (KDR / FLK1), Endoglin, CLEC14, Temp1-8, Tie2, A33, CAMPATH-1 (CDw52), CEA, Carbonic anhydrase IX (MN / CA It comprises one or more extracellular antigen-binding domains that bind to one or more antigens selected from the group consisting of IX), de2-7, EGFRvIII, EpCAM, Ep-CAM, folate-binding protein, G250, HLA-DR, IGFR, IL-2 receptor, IL3R, MCSP (melanoma-associated cell surface chondroitin sulfate proteoglycan), Muc-1, prostate stem cell antigen (PSCA), prostate-specific antigen (PSA), hK2, TAG-72, KI2L4, NKG2E, NKG2D, NKG2F, BTNL3, BTNL8, PD-1, NKG2, tumor cell neoantigens, and any combination thereof, and / or b) The multicomponent cell therapy product according to any one of claims 1 to 7, wherein the chimeric receptor is a chimeric antigen receptor (CAR) or a T cell receptor (TCR).

9. a) The chimeric receptor is a chimeric antigen receptor (CAR), and the CAR further comprises a transmembrane domain and one or more intracellular signaling domains, and / or b) The transmembrane domain is selected from the group consisting of CD8 transmembrane domain, CD28 transmembrane domain, CD3 zeta chain transmembrane domain, CD4 transmembrane domain, 4-IBB transmembrane domain, 0X40 transmembrane domain, ICOS transmembrane domain, CTLA-4 transmembrane domain, PD-1 transmembrane domain, LAG-3 transmembrane domain, 2B4 transmembrane domain, BTLA transmembrane domain, 0X40 transmembrane domain, DAP10 transmembrane domain, DAP12 transmembrane domain, CD16a transmembrane domain, DNAM-1 transmembrane domain, KIR2DSl transmembrane domain, KIR3DSl transmembrane domain, NKp44 transmembrane domain, NKp46 transmembrane domain, FceRlg transmembrane domain, and NKG2D transmembrane domain, and / or c) The one or more intracellular signaling domains are, each, a CD3 zeta chain intracellular signaling domain, a CD97 intracellular signaling domain, a CD1la-CD18 intracellular signaling domain, a CD2 intracellular signaling domain, an ICOS intracellular signaling domain, a CD27 intracellular signaling domain, a CD154 intracellular signaling domain, a CDS intracellular signaling domain, an 0X40 intracellular signaling domain, a 4-1BB intracellular signaling domain, a CD28 intracellular signaling domain, a ZAP40 intracellular signaling domain, a CD30 intracellular signaling domain, a GITR intracellular signaling domain, and an HVEM intracellular signaling domain. A multicomponent cell therapy product according to any one of claims 1 to 8, selected from the group consisting of a signaling domain, a DAP10 intracellular signaling domain, a DAP12 intracellular signaling domain, a MyD88 intracellular signaling domain, a 2B4 intracellular signaling domain, a CD16a intracellular signaling domain, a DNAM-1 intracellular signaling domain, a KIR2DS1 intracellular signaling domain, a KIR3DS1 intracellular signaling domain, an NKp44 intracellular signaling domain, an NKp46 intracellular signaling domain, a FceRlg intracellular signaling domain, an NKG2D intracellular signaling domain, and an EAT-2 intracellular signaling domain.

10. a) The CAR is a bispecific CAR comprising two or more extracellular antigen-binding domains, and optionally the CAR comprises a first antigen-binding domain specific to CD22 and a second antigen-binding domain specific to CD19. Optionally, the first antigen-binding domain includes a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region includes a heavy chain complementarity determination region 1 (CDR-H1) having the amino acid sequence of GDSVSNSANSAA (SEQ ID NO: 1), a heavy chain complementarity determination region 2 (CDR-H2) having the amino acid sequence of TYYRSKWYN (SEQ ID NO: 2), and a heavy chain complementarity determination region 3 (CDR-H3) having the amino acid sequence of AREVTGDLEDAFDI (SEQ ID NO: 3), and the VL region includes a light chain complementarity determination region 1 (CDR-L1) having the amino acid sequence of QTIWSY (SEQ ID NO: 4), a light chain complementarity determination region 2 (CDR-L2) having the amino acid sequence of AAS (SEQ ID NO: 5), and a light chain complementarity determination region 3 (CDR-L3) having the amino acid sequence of QQSYSIPQT (SEQ ID NO: 6). , optionally, the second antigen-binding domain includes a heavy chain variable (VH) region and a light chain variable (VL) region, wherein the VH region includes a heavy chain complementarity determination region 1 (CDR-H1) having the amino acid sequence of GVSLPDYG (SEQ ID NO: 7), a heavy chain complementarity determination region 2 (CDR-H2) having the amino acid sequence of IWGSETT (SEQ ID NO: 8), and a heavy chain complementarity determination region 3 (CDR-H3) having the amino acid sequence of AKHYYYGGSYAMDY (SEQ ID NO: 9), and the VL region includes a light chain complementarity determination region 1 (CDR-L1) having the amino acid sequence of QDISKY (SEQ ID NO: 10), a light chain complementarity determination region 2 (CDR-L2) having the amino acid sequence of HTS (SEQ ID NO: 11), and a light chain complementarity determination region 3 (CDR-L3) having the amino acid sequence of QQGNTLPYT (SEQ ID NO: 12). Optionally, the first antigen-binding domain comprises VH having the amino acid sequence QVQLQQSGPGLVKPSQTLSLTCCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSS (SEQ ID NO: 13), Optionally, the first antigen-binding domain includes a VL having the amino acid sequence DIQMTQSPSSLSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK (SEQ ID NO: 14), Optionally, the second antigen-binding domain comprises VH having the amino acid sequence EVKLQESGPGLVAPSQSLSVTCCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS (SEQ ID NO: 15), Optionally, the second antigen-binding domain includes a VL having the amino acid sequence DIQMTQTTSSLSASLLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID NO: 16), Selectively, the first antigen-binding domain may be QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTTPEDTAVYYCAREVTGDLEDAFDIWG QGTMVTVSSGSGSGKPGGSGEGSTKGDIQMTQSPSSLSASVGGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK (array number) CD22scFv having the amino acid sequence of (Sequence No. 20) or QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVVTVSSSGGGGGSDIQMTQSPSSLSSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK (Sequence No. 21), Selectively, the second antigen-binding domain may be DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGS KPGSGEGSTKGEVKLQESGPPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS(Sequence No. 2) 2) Or DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEEQEDIAATYFCQQGNTLPYTFGGGTKLEITGGGGGSEVKLQESGPGLVAPSQSLS CD19scFv having the amino acid sequence VTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVVSS (SEQ ID NO: 41) and / or b) The CAR is, i. DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRF SGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGGSQVQLQQSGPGLV KPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITIN PDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSSGSTSGSGKPGSG EGSTKGDIQMTQSPSSLSASVGDRVTITCCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPPSRFSGRGSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIKGGGGGSEVKLQESGPGLVAPSQSLSVTCCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSS (Sequence ID 23), ii. QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYA VSVKSRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSSGGGG SDIQMTQSPSSLSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPSRFS GRGSGTDFTLISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGGSG GGGSDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGSGKPGGSGEGSSTKGEVKLQESGPGLVAPSQSLLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSS (Sequence No. 24) iii. DIQMTQTTSSLSSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQ GNTLPYTFGGGTKLEITGGGGGSQVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVK SRITINPDTSKNQFSLQLNSVTPEDTAVYYCAREVTGDLEDAFDIWGQGTMVTVSSGSTSGSGKPGSGEGSTKGDIQMTQSPSSLSASVG DRVTITCRASQTIWSYLNWYQQRPGKAPNLIYAASSLQSGVPSRFSGRGSGTDFTLISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIK GGGGSEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSL QTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSSSGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFACDIYIWAPLA GTCGVLLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPR (Sequence ID 29), or iv. QVQLQQSGPGLVKPSQTLSLTCAISGDSVSSNSAAWNWIRQSPSRGLEWLGRTYYRSKWYNDYAVSVKSRITINPDTSKNQFSLQLNSVTPEDTAVY YCAREVTGDLEDAFDIWGQGTMVTVSSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQTIWSYLNWYQQRPGKAPNLLIYAASSLQSGVPSRFSGR GSGTDFTLTISSLQAEDFATYYCQQSYSIPQTFGQGTKLEIKGGGGSGGGGSGGGGSGGGGSGGGSDIQMTQTTSSLSSASLGDRVTISCRASQDIS KYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGSGKPGSGEGSTKGEV KLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHY YYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKL A multicomponent cell therapy product according to any one of claims 1 to 9, comprising the amino acid sequence of LYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 30).

11. a) The CAR includes an antigen-binding domain specific to B cell maturation antigen (BCMA), and the antigen-binding domain includes a heavy chain variable (VH) region comprising a heavy chain complementarity-determining region 1 (CDR-H1), a complementarity-determining region 2 (CDR-H2), and a complementarity-determining region 3 (CDR-H3), and the VH is i. GFTFTNHA (SEQ ID NO: 42), ISGNGRTT (SEQ ID NO: 43), and AKDGGETLVDS (SEQ ID NO: 44), ii. GFTFSSHA (SEQ ID NO: 45), ISGSGDFT (SEQ ID NO: 46), and AKDEDGGGSLLLGY ​​(SEQ ID NO: 47), iii. GFTFSSYA (SEQ ID NO: 48), ISGSGDYI (SEQ ID NO: 49), and AKEGTGANSSLADY (SEQ ID NO: 50), and iv. Containing amino acid sequences of CDR-H1, CDR-H2, and CDR-H3 selected from the group consisting of GFTFSSHA (SEQ ID NO: 51), ISGSGDYT (SEQ ID NO: 52), and AKDEDGGGSLLGH (SEQ ID NO: 53), The antigen-binding domain is optionally selected. i. QVQLVESGGGGLVQPGGSLRLSCAASGFTFFTNHAMSWVRQAPGKGLELVSSISGNGRTTTYYADSVKGRFTISRDISKNTLDLQMNSLRAEDTAVYYCAKDGGETLVDSRGQGTLVTVSS (Sequence ID 54), ii. QVQLVESGGGGLVQPGGSLRLSCAASGFTFSSSHAMTWVRQAPGKGLEWVAAISGSGDFTTHYADSVKGRFTISRDNSKNTVSLQMNNLRAEDTAVYYCAKDEDGGGSLLLGYRGQGTLVTVSS (Sequence ID 55), iii. EVQLLESGGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISGSGDYIYYADSVKGRFTISRDISKNTLYLQMNSLRAEDTAVYYCAKEGTGANSSLLADYRGQGTLVTVSS (Sequence ID 56), and iv. Contains a VH having an amino acid sequence selected from the group consisting of EVQLLESGGGLIQPGGSLRLSCAASGFTFSSSHAMTWVRQAPGKGLEWVSAISGSGDYTHYADSVKGRFTISRDNSKNTVYLQMNSLRAEDSAVYYCAKDEDGGGSLLGHRGQGTLVTVSS (SEQ ID NO: 57), and / or b) The CAR comprises an antigen-binding domain specific to B cell maturation antigen (BCMA), and the antigen-binding domain comprises a heavy chain variable (VH) region and a light chain variable (VL) region. The aforementioned VH is, i. QIQLVQSGPELKKPGETVKISCKASGYTFRHYSMNWVKQAPGKGLKWMGRINTESGVPIYAADDFKGRFAFSVETSASTAYLVINNLKDEDTASYFCSNDYLYSLLDFWGQGTALTVSS (Sequence ID 58), and ii. Containing an amino acid sequence selected from the group consisting of QIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYYEDTATYFCALDYSYAMDYWGQGTSVTVSS (SEQ ID NO: 60), VL is i. DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIK (Sequence ID 59), ii. DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIK (Sequence ID 61), iii. DIVLTQSPASLAVSLGQRATISCRASESVDNYGFSFMHWYQQKPGQPPKLLLIYRASNLESGIPARFSGSGSRTDFALTINPVETDDVATYYCQQSNKDPRTFGGGTKLEIK (Sequence ID 62), and iv. Containing an amino acid sequence selected from the group consisting of DIVLTQSPASLAVSLGQRATISCRASESVDNYGFSFMHWYQQKPGQPPKLLLIYRASNLESGIPARFSGSGSRTDFALTINPVETDDDVATYYCQQSNKDPRTFGGGTKLEIK (SEQ ID NO: 63), The antigen-binding domain is optionally selected. i. DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKGSTGSGSGKPGGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFRHYSMNWVKQAPGKGLKWMGRINTESGVPIYAADDFKGRFAFSVETSASTAYLVINNLKDEDTTASYFCSNDYLYSLLDFWWGQGTALTVSS (Sequence ID 64), ii. DIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKGSTGSGSGKPGGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRRFAFSLETSASTAYLQINNLKYYEDTATYFCALDYSYAMDYWGQGTSVTVSS (Sequence ID 65), iii. DIVLTQSPASLAVSLGQRATISCRASESVDNYGFSFMHWYQQKPGQPPKLLLIYRASNLESGIIPARFSGSGSRTDFALTINPVETDDVATYYCQQSNKDPRTFGGGTKLEIKGSTGSGSGKPGGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYYEDTATYFCALDYSYAMDYWGQGTSVTVSS (Sequence ID 66), and iv. BCMA having an amino acid sequence selected from the group consisting of DIVLTQSPASLAMSLGKRATISCRASESVSVIGAHLIHWYQQKPGQPPKLLLIYLASNLETGVPARFSGSGSGTDFTLTIDPVEEDDVAIYSCLQSRIFPRTFGGGTKLEIKGSTSGSGKPGGSGEGSSTKGQIQLVQSGPELCKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPAYAYDFRGRFAFSLETSASTAYLQINNLKYYEDTATYFCALDYSYAMDYWGQGTSVTVSS (SEQ ID NO: 67) Includes scFv and / or c) The CAR comprises one or more single-domain binding regions specific to B cell maturation antigen (BCMA), and each single-domain binding region is i. VKLEESGGGGLVQAGRSLRLSCAASEHTFSSHVMGWFRQAPGKERESVAVIGWRDISTSYADSVKGRFTISRDNAKKTLYLQMNSLKPEDTAVYYCAARRIDAADFDSWGQGTQVTVSS (Sequence ID 68), and ii. Containing an amino acid sequence selected from the group consisting of EVQLVESGGGLLVQAGGSLRLSCAASGRRTFTMGWFRQAPGKEREFVAAISLSPTLAYYAESVKGRFTISRDNAKNTVVLQMNSLKPEDTALYYCAADRKSVMSIRPDYWGQGTQVTVSS (Sequence ID 69), and / or d) The CAR comprises an antigen-binding domain specific to B cell maturation antigen (BCMA), wherein the binding domain comprises the amino acid sequence MGSWSEFWVRLGAIRERLDALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEKLRYTAATIRRFLQAYRHN (SEQ ID NO: 70), and / or e) The CAR is, i. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFTNHAMSWVRQAPGKGLELVSSISGNGRTTYYADSVKGRFTISRDIS KNTLDLQMNSLRAEDTAVYYCAKDGGETLVDSRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFA CDIYIWAAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPR (Sequence ID 71), ii. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVAAISGSGDFTHYADSVKGRFTISRDNS KNTVSLQMNNLRAEDTAVYYCAKDEDGGSLLGYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGGLDF ACDIYIWAAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 72), iii. MALPVTALLPLALLLHAARPEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISGSGDYIYYADSVKGRFTISRDIS KNTLYLQMNSLRAEDTAVYYCAKEGTGANSSLADYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLD FACDIYIWAAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPR (Sequence ID 73), iv. MALPVTALLPLALLLHAARPEVQLLESGGGLIQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVSAISGSGDYTHYADSVKGRFTISRDNS KNTVYLQMNSLRAEDSAVYYCAKDEDGGSLLGHRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGGLDF ACDIYIWAAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 74), v. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFTNHAMSWVRQAPGKGLELVSSISGNGRTTYYADSVKGRFTISRDIS KNTLDLQMNSLRAEDTAVYYCAKDGGETLVDSRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFA CDIYIWAPLAGTCGVLLLLSLVITLYCNHRNKRGRKKKLLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPPR (Sequence ID 75), vi. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVAAISGSGDFTHYADSVKGRFTISRDNS KNTVSLQMNNLRAEDTAVYYCAKDEDGGSLLGYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFA CDIYIWAPLAGTCCGVLLLLSLVITLYCNHRNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 76), vii. MALPVTALLPLALLLHAARPEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISGSGDYIYYADSVKGRFTISRDISK NTLYLQMNSLRAEDTAVYYCAKEGTGANSSLADYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGGLDF ACDIYIWAPLAGTCCGVLLLLSLVITLYCNHRNKRGRKKKLLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 77), viii. MALPVTALLPLALLLHAARPEVQLLESGGGLIQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVSAISGSGDYTHYADSVKGRFTISRDNS KNTVYLQMNSLRAEDSAVYYCAKDEDGGSLLGHRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFA CDIYIWAPLAGTCCGVLLLLSLVITLYCNHRNKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 78), ix. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFTNHAMSWVRQAPGKGLELVSSISGNGRTTYYADSVKGRFTISRDI SKNTLDLQMNSLRAEDTAVYYCAKDGGETLVDSRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLD FACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNNCWLTKKKYSSSSVHDPNGEYMFMRAVNTAKKSRLTDVTLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 79), x. MALPVTALLPLALLHAARPQVQLVESGGGLVQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVAAISGSGDFTHYADSVKGRFTISRDN SKNTVSLQMNNLRAEDTAVYYCAKDEDGGSLLGYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLD FACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNNCWLTKKKYSSSSVHDPNGEYMFMRAVNTAKKSRLTDVTLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 80), xi. MALPVTALLPLALLLHAARPEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSSISGSGDYIYYADSVKGRFTISRDIS KNTLYLQMNSLRAEDTAVYYCAKEGTGANSSLADYRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGL DFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNNCWLTKKKYSSSSVHDPNGEYMFMRAVNTAKKSRLTDVTLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 81), xii. MALPVTALLPLALLHAARPEVQLLESGGGLIQPGGSLRLSCAASGFTFSSHAMTWVRQAPGKGLEWVSAISGSGDYTHYADSVKGRFTISRDN SKNTVYLQMNSLRAEDSAVYYCAKDEDGGSLLGHRGQGTLVTVSSFVPVFLPAKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLD FACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNNCWLTKKKYSSSSVHDPNGEYMFMRAVNTAKKSRLTDVTLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPPR (Sequence ID 82), xiii. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQK PGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTK LEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFRHYSMNWVKQAPG KGLKWMGRINTESGVPIYADDFKGRFAFSVETSASTAYLVINNNLKDEDTASYFCSNDYLYSLDF WGQGTALTVSSSFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPPR (Sequence ID 83), xiv. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQK PGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTK LEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFTDYSINWVKRAPG KGLKWMGWINTETREPAYAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMDY WGQGTSVTVSSFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPPR (Sequence ID 84), xv. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIYWYQQK PGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTK LEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFTHYSMNWVKQAPG KGLKWMGRINTETGEPLYADDFKGRFAFSLETSASTAYLVINNNLKNEDTATFFCSNDYLYSCDY WGQGTTTLTVSSSFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 85), xvi. MLLLVTSLLLCELPHPAFLLIPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQ KPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTK LEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFTDYSINWVKRAPG KGLKWMGWINTETREPAYAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMDYW GQGTSVTVSSAAAAAFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 86), xvii. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQK PGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTK LEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFTDYSINWVKRAPG KGLKWMGWINTETREPAYAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMDYW GQGTSVTVSSAAAAAFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRRPEACRPPAAGGAVHTRGLLDFACDIYIWAPLAGTCGVLLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATATKDTYDALHMQALPPR (Sequence ID 87), xviii. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKP GQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLE IKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKKPGETTVKISCKASGYTFTDYSINWVKRAPGKGL KWMGWINTETREPAYAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQG TSVTVSSAAAAFVPVFLPAKPTTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCCGVLLLLSLVITLYCNHRNRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 88), xix. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQ KPGQPPTLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGT KLEIKGSTSGSGKPGSGEGSTKGQIQLVQSGPELKPGETVKISCKASGYTFTDYSINWVKRAP GKGLKWMGWINTETREPAYAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMD YWGQGTSVTVSSAAAAAFVPVFLPAKPTTTTPAPRPPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAAPLAGTCGVLLLLSLVITLYCNHRNRRRDQRLPPDAHKPPGGGSFRTPPIQEEQADAHSTLAKIRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPPR (Sequence ID 89), xx. MALPVTALLPLALLLLHAARPDIVLTQSPPSLAMSLGKRATISCRASESVTILGSHLIHWYQQKPGQPP TLLIQLASNVQTGVPARFSGSGSRTDFTLTIDPVEEDDVAVYYCLQSRTIPRTFGGGTKLEIKGSTSGSG KPGSGEGSTKGQIQLVQSGPELKKPGETVKISCKASGYTFTDYSINWVKRAPGKGLKWMGWINTETREPA YAYDFRGRFAFSLETASTAYLQINNLKYEDTATYFCALDYSYAMDYWGQGTSVTVSSAAAFVPVFLPAK PTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLY CNHRNRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRRDQRLPPPDAHKPPGGGS FRTPIQEEQADAHSTLAKIRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 90), xxi. MALPVTALLPLALLLHAARPQVKLEESGGGLVQAGRSLRLSCAASEHTFSSHVMGWFRQAP GKERESVAVIGWRDISTSYADSVKGRFTISRDNAKKTLYLQMNSLKPEDTAVYYCAARRIDA ADFDSWGQGTQVTVSSGGGGGSEVQLVESGGGLVQAGGSLRLSCAAASGRTFTMGWFRQAPGKE REFVAAISLSPTLAYYAESVKGRFTISRDNAKNTVVLQMNSLKPEDTALYYCAADRKSVMSIR PDYWGQGTQVTVSSTSTTTPAAPRPPPTPAAPTIASQPLSLRPEACRPAAGGAVHTRGLLDFACDIYIWAAPLAGTCGVLLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQQGQNQLYNELNLGRREEYDVLDKRRGRRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 91), and xxii. MAFLWLLSCWALLGTTFGDYKDDDDKGGGGSGGGGSMGSWEFWVRLGAIRERLDALGGSEAELAAFEKEIAAFESELQAYKGKGNPEVEKLRYTAATIR RFLQAYRHNGGGGSGGGGSGTTTPAPRPTPAPTIASQPLSLRPEACRPAAGGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIF A multicomponent cell therapy product according to any one of claims 1 to 9, comprising an amino acid sequence selected from the group consisting of KQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTATKDTYDALHMQALPPR (Sequence ID 92).

12. a) The product provides approximately 1.0 × 10¹⁶ units per kilogram of body weight to a human subject receiving the product. ５ ~Approx. 5.0×10 ６ Individual CAR ＋ Cells, or approximately 4.0 x 10⁻⁶ ５ ~1.0 x 10 ９ Individual CAR ＋ CAR of the cell range ＋ Includes a dose of cells and / or b) The one or more cells containing the chimeric receptor or the pharmaceutical composition is selected from the group consisting of T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), regulatory T cells, and natural killer T (NKT) cells, and / or c) The multicomponent cell therapy product according to any one of claims 1 to 11, wherein the one or more cells containing the chimeric receptor or the pharmaceutical composition are derived from an allogeneic donor and are genetically modified to reduce the risk of graft-versus-host disease (GVHD) and / or prevent allogeneic rejection.

13. A pharmaceutical composition comprising a multicomponent cell therapy product according to any one of claims 1 to 12.

14. A method for treating a human subject having or suspected of having a malignant tumor, comprising administering to the human subject a multicomponent cell therapy product according to any one of claims 1 to 12 or a pharmaceutical composition according to claim 13, The method wherein the hematological malignancy is arbitrarily selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), multiple myeloma, lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, myelodysplastic syndrome, myelofibrosis, and blastic plasmacytoid dendritic cell neoplasm (BPDCN).

15. A multicomponent cell therapy product according to any one of claims 1 to 12 or a pharmaceutical composition according to claim 13, for use in a method of treating a human subject having or suspected of having a malignant tumor, The multicomponent cell therapy product or pharmaceutical composition, wherein the hematological malignancy is optionally selected from the group consisting of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), multiple myeloma, lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, myelodysplastic syndrome, myelofibrosis, and blastic plasmacytoid dendritic cell neoplasm (BPDCN).

16. An engineered cell graft comprising a multicomponent cell therapy product according to any one of claims 1 to 12.

17. A cell therapy kit comprising a multicomponent cell therapy product according to any one of claims 1 to 12 or a pharmaceutical composition according to claim 13, The cell therapy kit further comprises, optionally, a written instruction for using the cell therapy to treat hematological malignancies in human subjects.

18. A kit for use in the preparation of cell therapy products, wherein the kit is (a) Anti-human CD34 affinity reagent, (b) Anti-human CD25 affinity reagent, (c) A vector containing nucleic acid encoding a chimeric receptor, (d) A transfection reagent, comprising Optionally, the kit further includes written instructions for isolating HSPCs from donor blood samples or donor bone marrow samples using the anti-human CD34 affinity reagent. Optionally, the written instructions further include instructions for isolating Treg from the donor blood sample or the donor bone marrow sample using the anti-human CD25 affinity reagent. The kit wherein the written instructions optionally further include instructions for introducing the vector into one or more cells derived from the donor blood sample or the donor bone marrow sample using the transfection reagent.

19. A method for preparing a product for multicomponent cell therapy, the following: a) Obtaining one or more peripheral blood samples or bone marrow samples from donors of the same species for a human subject that will receive the product, b) Collecting a first cell population from one or more peripheral blood samples or bone marrow samples, and transfecting the cell population with a vector containing a nucleic acid encoding a chimeric receptor according to any one of claims 1 to 12, c) Combining the transfected cell population with one or more cryoprotective agents and cryopreserving the transfected cell population, d) Collecting a second cell population from one or more peripheral blood samples or bone marrow samples, combining the second cell population with one or more cryoprotective agents, and cryopreserving the second cell population, wherein the second cell population is of type CD3 ＋ The above-mentioned collection, combination, and cryopreservation of T cells (Tcon), e) In order to obtain a CD34-enriched cell population and a CD34-depleted cell population, a third cell population is selected from one or more peripheral blood samples or bone marrow samples using one or more CD34-specific immunoisolated particles (ISPs), wherein the CD34-enriched cell population is CD34 ＋ The above selection includes hematopoietic stem progenitor cells (HSPCs), f) In order to obtain CD25-enriched cell populations and CD25-depleted cell populations, the CD34-depleted cell population is selected using one or more ISPs specific to CD25, g) Select the CD25-enriched cell population using one or more ISPs specific to CD4 and one or more ISPs specific to CD127, and CD4 ＋ and CD127 ｄｉｍ The selection of a cell population, and the CD4 ＋ and CD127 ｄｉｍ A group of cells, CD4 ＋ CD25 ＋ CD127 ｄｉｍ The process includes selecting and choosing regulatory T cells (Treg), The multicomponent cell therapy product comprises the transfected cell population, the Tcon, the HSPC, and the Treg. Selectively, the first cell population may include the Tcon, or the first cell population may include the HSPC. Optionally, the CD4 ＋ and CD127 ｄｉｍ The method wherein the cell population is the cell population that has been transfected with the vector.

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