The unnaturally modified Fc region of human IgG specifically binds to the unnaturally modified Fc receptor.

Abstract

The present invention provides polypeptides comprising modified Fc regions of IgG, modified Fcγ receptors that specifically bind to the polypeptides, and methods for treating or preventing diseases or disorders in patients using immunotherapy.

Description

【Technical Field】 【0001】 Related Applications This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 067,629, filed on August 19, 2020. The entire content of the said application is incorporated herein by reference. 【Background Art】 【0002】 Background of the Invention In recent years, research and development of cancer immunotherapy that eliminates cancer cells by utilizing the immune system have advanced (Nature Reviews Drug Discovery (2019) 18, pp. 899-900 (Non-Patent Document 1)). In particular, chimeric antigen receptor T cells (CAR-T) expressing a chimeric antigen receptor (CAR) in which an antigen recognition site and an activation signal transduction site are linked have been reported to have a remarkable therapeutic effect after single administration (The New England Journal of Medicine (2017) 377, pp. 2545-2554 (Non-Patent Document 2)), and new cancer therapies are expected in various fields. 【0003】 Several problems have arisen in the development of conventional CAR-T therapies targeting various cancer types (Nature Reviews Clinical Oncology (2020) 17, pp. 147-167 (Non-Patent Literature 3)). First, there are safety concerns, as serious side effects based on immune responses, such as cytokine release syndrome, have been reported in immunotherapy using CAR-T. With conventional CAR-T, it is difficult to selectively regulate the activity of CAR-expressing cells after they have been transferred to the patient, making it difficult to address the occurrence of side effects. Cancer tissue has mechanisms for acquiring treatment resistance, and problems such as the emergence of cancer cells that have lost cancer-related antigens during the treatment period (Cancer Discovery (2018) 8 (10), pp. 1219-1226 (Non-Patent Literature 4)) and cell populations with diverse characteristics (heterogeneity) also occur. Current treatments using conventional CAR-T that target a single antigen cannot overcome these problems. 【0004】 To address these issues, combination therapies using cancer target molecules and immune effector cells such as T cells and natural killer (NK) cells are being studied. Antibody molecules with tag molecules such as FITC and CAR-T cells that recognize these tag molecules have been developed (WO 2012 / 082841, WO 2016 / 030414, and WO 2017 / 091546 (Patent Documents 1-3)). By combining various cancer target molecules with tag-recognizing CAR-T cells, this technology can confer cytotoxic activity against multiple cancer antigens and cancer cells to a single type of tag-recognizing CAR-T cell. However, there is a risk that conferring tag molecules that do not originally exist in vivo to antibodies will increase the immunogenicity of the antibody molecules. 【0005】 On the other hand, technologies utilizing effector cells and antibody molecules themselves without tagged molecules are also being developed. Antibodies can bind to Fc receptors via the Fc region and transmit signals to effector cells. Cells created for use with antibodies that have cancer antigen recognition function include NK cells expressing IgG Fc fragment receptor IIIa (FcγRIIIA, known as CD16A) as the Fc receptor (JCI Insight. (2019) 4 (20): e130688 (Non-Patent Literature 5)), as well as T cells (CD16A CAR-T) or NK cells (CD16A CAR-NK) expressing CAR fused with CD16A and the signaling site (British Journal of Cancer (2019) 120 (1), pp. 79-87 (Non-Patent Literature 6), Oncotarget. (2017) 8 (23), pp. 37128-37139 (Non-Patent Literature 7)). One type of NK and T cell expressing CD16A can be combined with various cancer-targeting antibodies, making these cells a promising therapeutic tool that can acquire cytotoxic activity against cells expressing various antigens. Because the antibody molecules themselves are used, it is believed that this method will yield a therapeutic approach that is less immunogenic and safer than methods using tagged antibodies. 【0006】 However, in vivo, large amounts of endogenous immunoglobulins are present in the serum, and these also bind to CD16A. The presence of soluble Fc receptors in serum has also been confirmed (Journal of Clinical Investigation (1990) 86, pp. 416-423 (Non-Patent Literature 8)), and these bind to therapeutic antibodies. In other words, if CD16A on effector cells is occupied by immunoglobulins in the body, or if the administered antibody is occupied by soluble Fc receptors, the administered antibody will not be able to transmit the activation signal to the effector cells, and a decrease in drug efficacy is expected. Furthermore, if NK cells, CD16A CAR-T cells, or CD16A CAR-NK cells expressing CD16A are administered to a patient together with antibody molecules that recognize the patient's own tissue, such as autoantibodies, these cells may be activated against the patient's own tissue, potentially causing tissue damage. CD16A mutants that bind to afucosylated antibodies but not to unafucosylated endogenous immunoglobulins are known (WO 2017 / 161333 (Patent Document 4)), but these afucosylated antibodies transmit signals not only to CD16A mutants but also to endogenous CD16A. The combinations of CD16A mutants that do not bind to endogenous immunoglobulins and Fc mutants that do not bind to endogenous CD16A, and the combinations of mutants that specifically bind to each other, are currently unknown. [Prior art documents] [Patent Documents] 【0007】 [Patent Document 1] WO 2012 / 082841 [Patent Document 2] WO 2016 / 030414 [Patent Document 3] WO 2017 / 091546 [Patent Document 4] WO 2017 / 161333 【0008】 [Non-Patent Document 1] Nature Reviews Drug Discovery (2019) 18, pp. 899-900 [Non-Patent Document 2] The New England Journal of Medicine (2017) 377, pp. 2545-2554 [Non-Patent Document 3] Nature Reviews Clinical Oncology (2020) 17, pp. 147-167 [Non-Patent Document 4] Cancer Discovery (2018) 8 (10), pp. 1219-1226 [Non-Patent Document 5] JCI Insight. (2019) 4 (20): e130688 [Non-Patent Document 6] British Journal of Cancer (2019) 120 (1), pp. 79-87 [Non-Patent Document 7] Oncotarget. (2017) 8 (23), pp. 37128-37139 [Non-Patent Document 8] Journal of Clinical Investigation (1990) 86, pp. 416-423 [Overview of the project] 【0009】 This invention relates to a polypeptide containing a modified Fc region that can be used as an immunotherapy, and a modified Fcγ receptor that specifically binds to this polypeptide. The object of this invention is to provide an immunotherapy in which drug efficacy is not reduced by endogenous molecules. 【0010】 The present invention is at least partially based on the discovery of combinations of non-natural Fcγ receptor variants that do not bind to endogenous immunoglobulins and non-natural Fc region variants that do not bind to endogenous Fcγ receptors, the use of these combinations to specifically bind to non-natural Fcγ receptor variants and non-natural Fc region variants as immunotherapies to treat subjects, and methods for producing these combinations. For example, the inventors extracted amino acid sites that affect the binding activity between CD16A and the Fc region of an antibody in silico, prepared variants in which mutations were introduced into the CD16A and the Fc region of the antibody, and evaluated the change in binding activity to the wild type, finding that the binding activity to the wild type was reduced (Examples 1-4). Based on this finding, the inventors identified non-natural Fc region variants that do not show binding activity to wild-type CD16A but maintain high binding activity to non-natural mutated CD16A. The inventors also identified combinations of unnaturally modified Fc regions that do not bind to wild-type CD16A and unnaturally mutated CD16A that do not bind to the wild-type antibody Fc region. Similar results were obtained when using multiple types of antibodies against various antigens, indicating that these characteristics do not change depending on the antigen (Examples 5-8). The inventors further established natural killer (NK) cell lines expressing wild-type CD16A or unnaturally mutated CD16A and confirmed that antibody-dependent cell-mediated cytotoxicity (ADCC) reflects the binding activity confirmed in Example 6 (Examples 10, 11). Furthermore, the inventors confirmed the binding activity characteristics in Example 6 in the presence of an excess amount of IgG1 antibody (Example 9). 【0011】 The present invention provides the following aspects of compositions and methods that are expected to be useful in medicine and industry. 【0012】 In one aspect, the present invention provides a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and contains at least one amino acid mutation compared to the Fc region of wild-type or natural IgG. This polypeptide essentially lacks binding activity to wild-type or natural Fcγ receptors and can bind to non-natural Fcγ receptors containing at least one amino acid mutation compared to wild-type or natural Fcγ receptors. 【0013】 In some embodiments, the wild-type or native Fcγ receptor is wild-type or native CD16A, and the non-native Fcγ receptor containing at least one amino acid mutation is non-native CD16A containing at least one amino acid mutation. 【0014】 In some embodiments, wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78. 【0015】 In some embodiments, CD16A containing at least one amino acid mutation includes at least one mutation selected from (i) a lysine-to-aspartate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a lysine-to-glutamate mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a lysine-to-glutamate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation). 【0016】 In some embodiments, CD16A comprising at least one amino acid mutation comprises one or both of the K131D mutation and the K128E mutation. In other embodiments, CD16A comprising at least one amino acid mutation comprises one or both of the K131E mutation and the K128E mutation. In some embodiments, CD16A comprising at least one amino acid mutation comprises the K131D mutation and further comprises at least one mutation selected from (iv) a mutation from asparagine to glutamine (N38Q mutation) at the position corresponding to position 38 of SEQ ID NO: 78 and (v) a mutation from asparagine to glutamine (N74Q mutation) at the position corresponding to position 74 of SEQ ID NO: 78. In other embodiments, CD16A comprising at least one amino acid mutation comprises the amino acid sequence set forth in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0017】 In some embodiments, the polypeptide comprises a modified Fc region of human Igγ1, the modified Fc region comprising (i) a mutation from glutamic acid to arginine (E269R mutation) at the position corresponding to position 269 as per EU numbering and (ii) at least one mutation selected from (a) a mutation from glutamic acid to arginine (E294R mutation) at the position corresponding to position 294 as per EU numbering and (b) a mutation from glutamic acid to lysine (E294K mutation) at the position corresponding to position 294 as per EU numbering. 【0018】 In some embodiments, the polypeptide is an antibody. In other embodiments, the polypeptide is an antibody that binds to a cancer antigen. 【0019】 In another aspect, the present invention provides a method of treating or preventing a disease or disorder in a patient using immunotherapy. The method includes administering to the patient a polypeptide as described herein and a cell expressing a non-natural Fcγ receptor comprising at least one amino acid mutation compared to a wild-type or native Fcγ receptor, wherein the polypeptide is capable of binding to the non-natural Fcγ receptor comprising at least one amino acid mutation. 【0020】 In some embodiments, the cell is a human immune cell. In other embodiments, the human immune cell is a cell selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. In some embodiments, the cell is derived from a stem cell. In other embodiments, the stem cell is selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postnatal stem cells, multipotent stem cells, and embryonic germ cells. In some embodiments, the stem cell is a pluripotent stem cell. In other embodiments, the pluripotent stem cell is an induced pluripotent stem cell (iPS cell) or an embryonic stem cell (ES cell). 【0021】 In some embodiments, the cell comprises a disruption by genetic engineering in the β-2 microglobulin (B2M) gene. 【0022】 In some embodiments, the cell further comprises a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein comprising at least one portion of a B2M protein covalently bound either directly or via a linker sequence to at least one portion of an HLA-1α chain. In some embodiments, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. 【0023】 In some embodiments, the cells contain genetically engineered disruption in human leukocyte antigen (HLA) class II-related genes. In some embodiments, the HLA class II-related genes are selected from regulator X-related ankyrin-containing protein (RFXANK), regulator 5 (RFX5), regulator X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. 【0024】 In some embodiments, the cells contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. 【0025】 In some embodiments, the method is a method for treating or preventing cancer. 【0026】 In one aspect, the present disclosure provides a pharmaceutical composition comprising a polypeptide as described herein and a pharmaceutically acceptable excipient. 【0027】 In some embodiments, the pharmaceutical composition is intended for use in combination with cells for immunotherapy, wherein the cells express a non-natural Fcγ receptor containing at least one amino acid mutation compared to a wild-type or native Fcγ receptor, and the polypeptide is capable of binding to the non-natural Fcγ receptor containing at least one amino acid mutation. 【0028】 In some embodiments, the cells are human immune cells. In some embodiments, the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. In some embodiments, the cells are derived from stem cells. In some embodiments, the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. In some embodiments, the stem cells are pluripotent stem cells. In some embodiments, the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). 【0029】 In some embodiments, the cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene. 【0030】 In some embodiments, the cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein covalently bonded either directly or via a linker sequence to at least one portion of an HLA-1α chain. In some embodiments, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. 【0031】 In some embodiments, the cells contain genetically engineered disruption in human leukocyte antigen (HLA) class II-related genes. In some embodiments, the HLA class II-related genes are selected from regulator X-related ankyrin-containing protein (RFXANK), regulator 5 (RFX5), regulator X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. 【0032】 In some embodiments, the cells contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. 【0033】 In another aspect, the present invention provides a kit for treating or preventing a patient's disease or disorder using immunotherapy. The kit comprises (i) a polypeptide as described herein and (ii) cells expressing a non-natural Fcγ receptor having at least one amino acid mutation compared to a wild-type or native Fcγ receptor, wherein the polypeptide can bind to the non-natural Fcγ receptor having at least one amino acid mutation. 【0034】 In some embodiments, the cells are human immune cells. In some embodiments, the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. In some embodiments, the cells are derived from stem cells. In some embodiments, the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. In some embodiments, the stem cells are pluripotent stem cells. In some embodiments, the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). 【0035】 In some embodiments, the cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene. 【0036】 In some embodiments, the cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein covalently bonded either directly or via a linker sequence to at least one portion of an HLA-1α chain. In some embodiments, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. 【0037】 In some embodiments, the cells contain genetically engineered disruption in human leukocyte antigen (HLA) class II-related genes. In some embodiments, the HLA class II-related genes are selected from regulator X-related ankyrin-containing protein (RFXANK), regulator 5 (RFX5), regulator X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. 【0038】 In some embodiments, the cells contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. 【0039】 In one aspect, the present invention provides cells expressing a non-natural CD16A that contains at least one amino acid mutation compared to wild-type or natural CD16A, wherein the at least one amino acid mutation is selected from (i) a lysine-to-aspartate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a lysine-to-glutamate mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a lysine-to-glutamate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation), and the non-natural CD16A contains an amino acid sequence having 90% or more amino acid sequence identity with respect to SEQ ID NO: 78. 【0040】 In some embodiments, CD16A containing at least one amino acid mutation includes one or both of the K131D mutation and the K128E mutation. 【0041】 In some embodiments, CD16A containing at least one amino acid mutation includes one or both of the K131E and K128E mutations. 【0042】 In some embodiments, CD16A containing at least one amino acid mutation contains the K131D mutation and further contains at least one mutation selected from (iv) an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (v) an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0043】 In some embodiments, CD16A containing at least one amino acid mutation includes the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0044】 In some embodiments, the cells are human immune cells. In some embodiments, the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. In some embodiments, the cells are derived from stem cells. In some embodiments, the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. In some embodiments, the stem cells are pluripotent stem cells. In some embodiments, the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). 【0045】 In some embodiments, the cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene. 【0046】 In some embodiments, the cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein covalently bonded either directly or via a linker sequence to at least one portion of an HLA-1α chain. In some embodiments, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. 【0047】 In some embodiments, the cells contain genetically engineered disruption in human leukocyte antigen (HLA) class II-related genes. In some embodiments, the HLA class II-related genes are selected from regulator X-related ankyrin-containing protein (RFXANK), regulator 5 (RFX5), regulator X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. 【0048】 In some embodiments, the cells contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. 【0049】 In one aspect, the present invention provides a pharmaceutical composition comprising cells as described herein and pharmaceutically acceptable excipients. 【0050】 In some embodiments, the pharmaceutical composition is intended for use in combination with a polypeptide comprising a modified Fc region of IgG for immunotherapy, wherein the modified Fc region is non-natural and comprises at least one amino acid mutation compared to the Fc region of wild-type or natural IgG, and the polypeptide essentially lacks binding activity to wild-type or natural CD16A and can bind to non-natural CD16A comprising at least one amino acid mutation expressed by cells. 【0051】 In some embodiments, the polypeptide comprises a modified Fc region of human Igγ1, the modified Fc region comprising (i) a glutamate-to-arginine mutation (E269R mutation) at the position corresponding to EU index number 269, and (ii) at least one mutation selected from (a) a glutamate-to-arginine mutation (E294R mutation) at the position corresponding to EU index number 294, and (b) a glutamate-to-lysine mutation (E294K mutation) at the position corresponding to EU index number 294. 【0052】 In some embodiments, the polypeptide is an antibody. In some embodiments, the polypeptide is an antibody that binds to a cancer antigen. 【0053】 In some embodiments, the pharmaceutical composition is for the treatment of cancer. 【0054】 In one aspect, the present invention provides a method for preparing polypeptides comprising a modified Fc region of IgG. The method comprises the steps of: (1) providing a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and contains at least one amino acid mutation compared to wild-type or natural IgG; (2) measuring the binding activity of the polypeptide obtained in (1) to a wild-type or natural Fcγ receptor; (3) measuring the binding activity of the polypeptide obtained in (1) to a non-natural Fcγ receptor containing at least one amino acid mutation compared to a wild-type or natural Fcγ receptor; and (4) selecting from the polypeptides obtained in (1) a polypeptide that essentially does not have binding activity to a wild-type or natural Fcγ receptor and binds to a non-natural Fcγ receptor containing at least one amino acid mutation. 【0055】 In some embodiments, the wild-type or native Fcγ receptor is wild-type or native CD16A, and the non-native Fcγ receptor containing at least one amino acid mutation is non-native CD16A containing at least one amino acid mutation. 【0056】 In some embodiments, wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78. 【0057】 In some embodiments, a non-natural CD16A containing at least one amino acid mutation includes at least one mutation selected from (i) a lysine-to-aspartate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a lysine-to-glutamate mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a lysine-to-glutamate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation). 【0058】 In some embodiments, CD16A containing at least one amino acid mutation includes one or both of the K131D mutation and the K128E mutation. 【0059】 In some embodiments, CD16A containing at least one amino acid mutation includes one or both of the K131E and K128E mutations. 【0060】 In some embodiments, CD16A containing at least one amino acid mutation contains the K131D mutation and further contains at least one mutation selected from (iv) an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (v) an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0061】 In some embodiments, a non-natural polypeptide containing a modified Fc region of IgG is an antibody. In some embodiments, the antibody is an antibody that binds to a cancer antigen. 【0062】 In some embodiments, a polypeptide comprising a modified Fc region of IgG is an antibody, and the method further comprises the steps of contacting the antibody with immune cells expressing a non-natural Fcγ receptor comprising at least one amino acid mutation and cells expressing an antigen to which the antibody binds, and measuring antibody-dependent cell-mediated cytotoxicity (ADCC) activity. 【0063】 In another aspect, the present invention provides a method for preparing non-natural Fcγ receptors. This method comprises the steps of: (1) providing a non-natural Fcγ receptor having at least one amino acid mutation compared to a wild-type or natural Fcγ receptor; (2) providing a polypeptide comprising the Fc region of wild-type or natural IgG and a polypeptide comprising the Fc region of IgG having at least one amino acid mutation compared to wild-type or natural IgG; (3) measuring the binding activity of the non-natural Fcγ receptor obtained in (1) to the polypeptide comprising the Fc region of wild-type or natural IgG; (4) measuring the binding activity of the non-natural Fcγ receptor obtained in (1) to a polypeptide comprising the Fc region of IgG having at least one amino acid mutation; and (5) selecting a non-natural Fcγ receptor from the non-natural Fcγ receptor obtained in (1) that essentially does not have binding activity to a polypeptide comprising the Fc region of wild-type or natural IgG and binds to a polypeptide comprising the Fc region of IgG having at least one amino acid mutation. 【0064】 In some embodiments, the Fcγ receptor is CD16A. 【0065】 In some embodiments, the wild-type or native Fcγ receptor is CD16A, which contains the amino acid sequence shown in SEQ ID NO: 78. 【0066】 In some embodiments, the Fc region of IgG, comprising at least one amino acid mutation, is the Fc region of human Igγ1 comprising at least one amino acid mutation compared to wild-type or natural human Igγ1, and comprises (a) a glutamate-to-arginine mutation at the position corresponding to position 269 according to EU index numbering (E269R mutation), and (b) at least one mutation selected from (i) a glutamate-to-arginine mutation at the position corresponding to position 294 according to EU index numbering (E294R mutation) and (ii) a glutamate-to-lysine mutation at the position corresponding to position 294 according to EU index numbering (E294K mutation). 【0067】 In some embodiments, a polypeptide comprising the Fc region of IgG containing at least one amino acid mutation is an antibody. In some embodiments, the antibody is an antibody that binds to a cancer antigen. 【0068】 In some embodiments, a polypeptide comprising the Fc region of IgG containing at least one amino acid mutation is an antibody, and the method further comprises the step of contacting the antibody comprising the Fc region of IgG containing at least one amino acid mutation obtained in (2) with immune cells expressing an Fcγ receptor containing at least one amino acid mutation obtained in (1) and cells expressing an antigen to which the antibody binds, and measuring antibody-dependent cell-mediated cytotoxicity (ADCC) activity. 【0069】 In another aspect, the present invention provides a method for preparing a binding pair comprising (a) a polypeptide comprising a modified Fc region of IgG and (b) a non-natural modified Fcγ receptor. The method comprises the following steps: (1) providing a polypeptide comprising the Fc region of wild-type or natural IgG and a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and comprises at least one amino acid mutation compared to the Fc region of wild-type or natural IgG; (2) providing a wild-type or natural Fcγ receptor and a non-natural modified Fcγ receptor, wherein the modified Fcγ receptor comprises at least one amino acid mutation compared to the wild-type or natural Fcγ receptor; (3) measuring the binding activity of each Fcγ receptor obtained in (2) to each polypeptide obtained in (1); and (4) selecting (a) a polypeptide comprising a modified Fc region that binds to the modified Fcγ receptor and essentially has no binding activity to the wild-type or natural Fcγ receptor, and (b) a modified Fcγ receptor that binds to the polypeptide comprising the modified Fc region and does not bind to the Fc region of wild-type or natural IgG. 【0070】 In some embodiments, the Fcγ receptor is CD16A. 【0071】 In some embodiments, wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78. 【0072】 In some embodiments, the polypeptide containing a modified IgG Fc is an antibody. In some embodiments, the antibody is an antibody that binds to a cancer antigen. 【0073】 In some embodiments, the polypeptide comprising a modified Fc region of IgG selected in (4) is an antibody, and the method further comprises the step of contacting the antibody with immune cells expressing a modified Fcγ receptor selected in (4) and cells expressing an antigen to which the antibody binds, and measuring antibody-dependent cell-mediated cytotoxicity (ADCC) activity. 【0074】 It is anticipated that the present invention can provide immunotherapy using a combination of a mutagenerated Fcγ receptor and a mutagenerated Fc region that exhibits a specific binding pattern, and that drug efficacy is not reduced by endogenous molecules. [Invention 1001] A polypeptide comprising a modified Fc region of IgG, The modified Fc region is non-natural and contains at least one amino acid mutation compared to the Fc region of wild-type or natural IgG, and The polypeptide essentially lacks binding activity to wild-type or native Fcγ receptors, and can bind to non-native Fcγ receptors that have at least one amino acid mutation compared to wild-type or native Fcγ receptors. Polypeptide. [Invention 1002] The polypeptide of the present invention 1001, wherein the wild-type or natural Fcγ receptor is wild-type or natural CD16A, and the non-natural Fcγ receptor containing at least one amino acid mutation is non-natural CD16A containing at least one amino acid mutation. [Invention 1003] The polypeptide of the present invention 1002, wherein wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78. [Invention 1004] CD16A containing at least one amino acid mutation, (i) A mutation from lysine to aspartic acid at position 131 of SEQ ID NO: 78 (K131D mutation), (ii) A mutation from lysine to glutamate at position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) A mutation from lysine to glutamate at position 131 of SEQ ID NO: 78 (K131E mutation) Polypeptides of the present invention 1002 or 1003 comprising at least one more selected mutation. [Invention 1005] The polypeptide of the present invention 1004, wherein CD16A containing at least one amino acid mutation contains one or both of the K131D mutation and the K128E mutation. [Invention 1006] The polypeptide of the present invention 1004, wherein CD16A containing at least one amino acid mutation contains one or both of the K131E mutation and the K128E mutation. [Invention 1007] CD16A containing at least one amino acid mutation contains the K131D mutation, and (iv) A mutation from asparagine to glutamine at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (v) SEQ ID NO: Mutation from asparagine to glutamine at position 74 of 78 (N74Q mutation) Polypeptide of the present invention 1004, further comprising at least one more selected mutation. [Invention 1008] The polypeptide of the present invention 1004, wherein CD16A containing at least one amino acid mutation comprises the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. [Invention 1009] The polypeptide comprises a modified Fc region of human Igγ1, The modified Fc region is (i) A mutation from glutamate to arginine at the position corresponding to position 269 according to the EU index number assignment (E269R mutation), (ii) At least one mutation selected from (a) a glutamate-to-arginine mutation at the position corresponding to position 294 according to EU index number assignment (E294R mutation) and (b) a glutamate-to-lysine mutation at the position corresponding to position 294 according to EU index number assignment (E294K mutation) A polypeptide comprising any of the inventions 1001 to 1008, including the above. [Invention 1010] An antibody, which is a polypeptide according to any of the invention 1001 to 1009. [Invention 1011] A polypeptide of the present invention 1010, which is an antibody that binds to a cancer antigen. [Invention 1012] A method of treating or preventing a patient's disease or disorder using immunotherapy, The present invention includes the step of administering to a patient a polypeptide according to any of the invention 1001 to 1011 and cells expressing a non-natural Fcγ receptor containing at least one amino acid mutation compared to the wild-type or natural Fcγ receptor, A method by which the polypeptide can bind to a non-native Fcγ receptor comprising at least one amino acid mutation. [Invention 1013] The method of the present invention 1012, wherein the cells are human immune cells. [Invention 1014] The method of the present invention 1013, wherein the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. [Invention 1015] The method according to any of items 1012 to 1014 of the present invention, wherein the cells are derived from stem cells. [Invention 1016] The method of the present invention 1015, wherein the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. [Invention 1017] The method of the present invention 1016, wherein the stem cells are pluripotent stem cells. [Invention 1018] The method of the present invention 1017, wherein the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). [Invention 1019] The method according to any of items 1012 to 1018 of the present invention, wherein the cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene. [Invention 1020] The aforementioned cells, A single-chain fusion human leukocyte antigen (HLA) class I protein comprising at least one portion of a B2M protein covalently bound to at least one portion of an HLA-1α chain, either directly or via a linker sequence. The method of the present invention 1019 further comprises a polynucleotide capable of encoding a polynucleotide. [Invention 1021] The method of the present invention 1020, wherein the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. [Invention 1022] The method according to any one of items 1012 to 1021 of the present invention, wherein the cells contain genetically modified disruption of human leukocyte antigen (HLA) class II related genes. [Invention 1023] The method of the present invention 1022, wherein the HLA class II related gene is selected from regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. [Invention 1024] The aforementioned cells, A single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. A method of the present invention, including any of the methods described in items 1012 to 1023. [Invention 1025] A method for treating or preventing cancer, according to any of the present invention 1012 to 1024. [Invention 1026] A pharmaceutical composition comprising any polypeptide according to invention 1001 to 1011 and a pharmaceutically acceptable excipient. [Invention 1027] A pharmaceutically acceptable composition of the present invention 1026 for use in combination with cells for immunotherapy, wherein cells express a non-natural Fcγ receptor containing at least one amino acid mutation compared to a wild-type or native Fcγ receptor, and the polypeptide can bind to the non-natural Fcγ receptor containing at least one amino acid mutation. [Invention 1028] The pharmaceutical composition of the present invention 1027, wherein the cells are human immune cells. [Invention 1029] A pharmaceutical composition of the present invention 1028, wherein the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. [Invention 1030] A pharmaceutical composition according to any one of the present invention 1027 to 1029, wherein the aforementioned cells are derived from stem cells. [Invention 1031] A pharmaceutical composition of the present invention 1030, wherein the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. [Invention 1032] A pharmaceutical composition according to the present invention 1031, wherein the stem cells are pluripotent stem cells. [Invention 1033] The pharmaceutical composition of the present invention 1032, wherein the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). [Invention 1034] The aforementioned cells contain genetically modified disruption of the β-2 microglobulin (B2M) gene, and are part of any of the pharmaceutical compositions according to invention 1027 to 1033. [Invention 1035] The aforementioned cells, A single-chain fusion human leukocyte antigen (HLA) class I protein comprising at least one portion of a B2M protein covalently bound to at least one portion of an HLA-1α chain, either directly or via a linker sequence. A pharmaceutical composition of the present invention 1034, further comprising a polynucleotide capable of encoding a nucleotide. [Invention 1036] A pharmaceutical composition according to Invention 1035, wherein the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. [Invention 1037] The aforementioned cells contain genetically modified disruption of human leukocyte antigen (HLA) class II related genes, and are part of any of the pharmaceutical compositions according to invention 1027 to 1036. [Invention 1038] The pharmaceutical composition of Invention 1037, wherein the HLA class II-related gene is selected from regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. [Invention 1039] The aforementioned cells, A single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. A pharmaceutical composition according to any of the invention 1027 to 1038, comprising [the specified element]. [Invention 1040] A kit for treating or preventing a patient's disease or disability using immunotherapy, (i) A polypeptide according to any of the present invention 1001 to 1011, and (ii) a cell expressing a non-natural Fcγ receptor having at least one amino acid mutation compared to the wild-type or natural Fcγ receptor, A kit comprising a polypeptide capable of binding to a non-native Fcγ receptor containing at least one amino acid mutation. [Invention 1041] The kit of the present invention 1040, wherein the cells are human immune cells. [Invention 1042] The kit of the present invention 1041, wherein the human immune cells are selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. [Invention 1043] A kit according to any of invention 1040 to 1042, wherein the aforementioned cells are derived from stem cells. [Invention 1044] A kit according to the present invention 1043, wherein the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. [Invention 1045] A kit according to the present invention 1044, wherein the stem cells are pluripotent stem cells. [Invention 1046] A kit according to Invention 1045, wherein the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). [Invention 1047] The aforementioned cells contain genetically modified disruption of the β-2 microglobulin (B2M) gene, according to any of the kits 1040 to 1046 of the present invention. [Invention 1048] The aforementioned cells, A single-chain fusion human leukocyte antigen (HLA) class I protein comprising at least one portion of a B2M protein covalently bound to at least one portion of an HLA-1α chain, either directly or via a linker sequence. The kit of the present invention 1047 further comprises a polynucleotide capable of encoding. [Invention 1049] A kit according to Invention 1048, wherein the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. [Invention 1050] The aforementioned cells contain genetically modified disruption of human leukocyte antigen (HLA) class II related genes, according to any one of the kits 1040 to 1049 of the present invention. [Invention 1051] The kit of Invention 1050, wherein the HLA class II-related gene is selected from regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. [Invention 1052] The aforementioned cells, A single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. A kit according to any of inventions 1040 to 1051, including the above. [Invention 1053] Cells expressing a non-natural CD16A that contains at least one amino acid mutation compared to wild-type or natural CD16A, The at least one amino acid mutation is selected from (i) a lysine-to-aspartic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a lysine-to-glutamic acid mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a lysine-to-glutamic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation), and The non-natural CD16A contains an amino acid sequence that has more than 90% amino acid sequence identity with SEQ ID NO: 78. cell. [Invention 1054] The cells of the present invention 1053, wherein CD16A containing at least one amino acid mutation contains one or both of the K131D mutation and the K128E mutation. [Invention 1055] The cells of the present invention 1053, wherein CD16A containing at least one amino acid mutation contains one or both of the K131E mutation and the K128E mutation. [Invention 1056] CD16A containing at least one amino acid mutation contains the K131D mutation, and (iv) A mutation from asparagine to glutamine at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (v) SEQ ID NO: Mutation from asparagine to glutamine at position 74 of 78 (N74Q mutation) Cells of the present invention 1053, further comprising at least one more selected mutation. [Invention 1057] A cell according to the present invention 1053, wherein CD16A containing at least one amino acid mutation comprises the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. [Invention 1058] Human immune cells, which are any of the cells described in items 1053 to 1057 of this invention. [Invention 1059] The cells of the present invention 1058, wherein the human immune cells are cells selected from T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. [Invention 1060] Cells derived from stem cells, according to any of the invention items 1053 to 1059. [Invention 1061] The cells of the present invention 1060, wherein the stem cells are selected from pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells. [Invention 1062] The cells of the present invention 1061, wherein the stem cells are pluripotent stem cells. [Invention 1063] The cells of the present invention 1062, wherein the pluripotent stem cells are induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). [Invention 1064] A cell according to any of the invention 1053 to 1063, which contains a genetically modified disruption in the β-2 microglobulin (B2M) gene. [Invention 1065] A single-chain fusion human leukocyte antigen (HLA) class I protein comprising at least one portion of a B2M protein covalently bound to at least one portion of an HLA-1α chain, either directly or via a linker sequence. A cell of the present invention 1064 further comprising a polynucleotide capable of encoding. [Invention 1066] Cells of the present invention 1065, wherein the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. [Invention 1067] Cells according to any of the invention 1053 to 1066, which contain genetically modified disruption in human leukocyte antigen (HLA) class II related genes. [Invention 1068] Cells of the present invention 1067, wherein the HLA class II-related gene is selected from regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB. [Invention 1069] A single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. A cell comprising any of the present invention 1053 to 1068, including the cell comprising [Invention 1070] A pharmaceutical composition comprising any of the cells described in invention 1053 to 1069 and a pharmaceutically acceptable excipient. [Invention 1071] The modified Fc region is non-natural and contains at least one amino acid mutation compared to the Fc region of wild-type or natural IgG, and The polypeptide does not inherently possess binding activity to wild-type or native CD16A, and can bind to non-native CD16A containing at least one amino acid mutation expressed by the cell. A pharmaceutical composition of the present invention 1070 for use in combination with a polypeptide containing a modified Fc region of IgG for immunotherapy. [Invention 1072] The polypeptide comprises a modified Fc region of human Igγ1, The modified Fc region is (i) A mutation from glutamate to arginine at the position corresponding to position 269 according to the EU index number assignment (E269R mutation), (ii) At least one mutation selected from (a) a glutamate-to-arginine mutation at the position corresponding to position 294 according to EU index number assignment (E294R mutation) and (b) a glutamate-to-lysine mutation at the position corresponding to position 294 according to EU index number assignment (E294K mutation) A pharmaceutical composition of the present invention 1071, comprising [the specified element]. [Invention 1073] A pharmaceutical composition according to invention 1071 or 1072, wherein the polypeptide is an antibody. [Invention 1074] The pharmaceutical composition of the present invention 1073, wherein the polypeptide is an antibody that binds to a cancer antigen. [Invention 1075] A pharmaceutical composition according to any of invention 1071 to 1074 for the treatment of cancer. [Invention 1076] A method for preparing a polypeptide containing a modified Fc region of IgG, (1) A step of providing a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and comprises at least one amino acid mutation compared to wild-type or natural IgG; (2) A step of measuring the binding activity of the polypeptide obtained in (1) to the wild-type or native Fcγ receptor; (3) A step of measuring the binding activity of the polypeptide obtained in (1) to a non-natural Fcγ receptor that contains at least one amino acid mutation compared to the wild-type or natural Fcγ receptor; and (4) Selecting from the polypeptides obtained in (1) a polypeptide that binds to a non-natural Fcγ receptor, which does not inherently possess binding activity to the wild-type or natural Fcγ receptor and contains at least one amino acid mutation. Methods that include... [Invention 1077] The method of the present invention 1076, wherein the wild-type or native Fcγ receptor is wild-type or native CD16A, and the non-native Fcγ receptor containing at least one amino acid mutation is non-native CD16A containing at least one amino acid mutation. [Invention 1078] The method of the present invention 1077, wherein the wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78. [Invention 1079] Non-natural CD16A containing at least one amino acid mutation, (i) A mutation from lysine to aspartic acid at position 131 of SEQ ID NO: 78 (K131D mutation), (ii) A mutation from lysine to glutamate at position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) A mutation from lysine to glutamate at position 131 of SEQ ID NO: 78 (K131E mutation) The method of the present invention 1077 or 1078, comprising at least one more selected mutation. [Invention 1080] The method of the present invention 1079, wherein CD16A containing at least one amino acid mutation contains one or both of the K131D mutation and the K128E mutation. [Invention 1081] The method of the present invention 1079, wherein CD16A containing at least one amino acid mutation contains one or both of the K131E mutation and the K128E mutation. [Invention 1082] CD16A containing at least one amino acid mutation contains the K131D mutation, and (iv) A mutation from asparagine to glutamine at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (v) SEQ ID NO: Mutation from asparagine to glutamine at position 74 of 78 (N74Q mutation) The method of the present invention 1079, further comprising at least one more selected mutation. [Invention 1083] A method according to any one of the present invention 1076 to 1082, wherein the antibody is a non-natural polypeptide containing a modified Fc region of IgG. [Invention 1084] The method of the present invention 1083, wherein the antibody is an antibody that binds to a cancer antigen. [Invention 1085] A polypeptide containing a modified Fc region of IgG is an antibody. The method described above is The antibody is brought into contact with immune cells expressing a non-native Fcγ receptor containing at least one amino acid mutation and cells expressing an antigen to which the antibody binds, and antibody-dependent cell-mediated cytotoxicity (ADCC) activity is measured. Further including, The method of the present invention 1083 or 1084. [Invention 1086] A method for preparing a non-natural Fcγ receptor, (1) A step of providing a non-natural Fcγ receptor that contains at least one amino acid mutation compared to the wild-type or natural Fcγ receptor; (2) A step of providing a polypeptide comprising the Fc region of wild-type or natural IgG, and a polypeptide comprising the Fc region of IgG comprising at least one amino acid mutation compared to wild-type or natural IgG; (3) A step of measuring the binding activity of the non-natural Fcγ receptor obtained in (1) to polypeptides containing the Fc region of wild-type or natural IgG; (4) A step of measuring the binding activity of the non-native Fcγ receptor obtained in (1) to a polypeptide containing the Fc region of IgG containing at least one amino acid mutation; and (5) Selecting from the unnatural Fcγ receptors obtained in (1) a non-natural Fcγ receptor that does not inherently possess binding activity to a polypeptide containing the Fc region of wild-type or natural IgG and that binds to a polypeptide containing the Fc region of IgG containing at least one amino acid mutation. Methods that include... [Invention 1087] The method of the present invention 1086, wherein the Fcγ receptor is CD16A. [Invention 1088] The method of the present invention 1087, wherein the wild-type or native Fcγ receptor is CD16A containing the amino acid sequence shown in SEQ ID NO: 78. [Invention 1089] The Fc region of IgG containing at least one amino acid mutation is the Fc region of human Igγ1 containing at least one amino acid mutation compared to wild-type or natural human Igγ1, and (a) A mutation from glutamate to arginine at the position corresponding to position 269 according to the EU index number assignment (E269R mutation), (b) At least one mutation selected from (i) a glutamate-to-arginine mutation at the position corresponding to position 294 according to EU index number assignment (E294R mutation) and (ii) a glutamate-to-lysine mutation at the position corresponding to position 294 according to EU index number assignment (E294K mutation) Any method of the present invention 1086 to 1088, including the above. [Invention 1090] A method according to any one of the present invention 1086 to 1089, wherein the antibody is a polypeptide comprising the Fc region of IgG containing at least one amino acid mutation. [Invention 1091] The method of the present invention 1090, wherein the antibody is an antibody that binds to a cancer antigen. [Invention 1092] A polypeptide containing the Fc region of IgG with at least one amino acid mutation is an antibody. The method described above is (2) The antibody containing the Fc region of IgG containing at least one amino acid mutation obtained in (2) is brought into contact with immune cells expressing the Fcγ receptor containing at least one amino acid mutation obtained in (1) and cells expressing the antigen to which the antibody binds, and antibody-dependent cell-mediated cytotoxicity (ADCC) activity is measured. Further including, The method of the present invention 1090 or 1091. [Invention 1093] A method for preparing a binding pair comprising (a) a polypeptide containing a modified Fc region of IgG and (b) a non-natural modified Fcγ receptor, (1) A step of providing a polypeptide comprising the Fc region of wild-type or natural IgG and a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and comprises at least one amino acid mutation compared to the Fc region of wild-type or natural IgG; (2) A step of providing a wild-type or native Fcγ receptor and a non-native modified Fcγ receptor, wherein the modified Fcγ receptor comprises at least one amino acid mutation compared to the wild-type or native Fcγ receptor; (3) A step of measuring the binding activity of each Fcγ receptor obtained in (2) to each polypeptide obtained in (1); and (4) The step of selecting (a) a polypeptide comprising a modified Fc region that binds to a modified Fcγ receptor and essentially does not have binding activity to wild-type or native Fcγ receptors, and (b) a modified Fcγ receptor that binds to a polypeptide comprising a modified Fc region and does not bind to the Fc region of wild-type or native IgG. Methods that include... [Invention 1094] The method of invention 1093, wherein the Fcγ receptor is CD16A. [Invention 1095] The method of the present invention 1093 or 1094, wherein the wild-type or natural CD16A comprises the amino acid sequence shown in SEQ ID NO: 78. [Invention 1096] A method according to any one of the present invention 1093 to 1095, wherein the polypeptide containing a modified Fc region of IgG is an antibody. [Invention 1097] The method of the present invention 1096, wherein the antibody is an antibody that binds to a cancer antigen. [Invention 1098] The polypeptide containing the modified Fc region of IgG selected in (4) is the antibody. The method described above is The antibody is brought into contact with immune cells expressing the modified Fcγ receptor selected in (4) and cells expressing the antigen to which the antibody binds, and antibody-dependent cell-mediated cytotoxicity (ADCC) activity is measured. Further including, Any method of the present invention 1093 to 1097. [Brief explanation of the drawing] 【0075】 [Figure 1] This graph shows the binding activity of wild-type Fc-type anti-HER2 antibodies or mutant Fc-type anti-HER2 antibodies against CD16V or CD16V mutants. The vertical axis represents the difference between the absorbance at 450 nm and the absorbance at the reference wavelength of 570 nm, and the horizontal axis represents the concentration (ng / mL) of CD16V or CD16V mutant. [Figure 2] This graph shows the binding activity of wild-type Fc-type anti-HER2 antibodies or mutant Fc-type anti-HER2 antibodies against CD16V or CD16V mutants. The vertical axis represents the difference between the absorbance at 450 nm and the absorbance at the reference wavelength of 650 nm, and the horizontal axis represents the concentration (ng / mL) of CD16V or CD16V mutants. [Figure 3] This graph shows the binding activity of wild-type Fc-type anti-EGFR antibodies or mutant Fc-type anti-EGFR antibodies against CD16V or CD16V mutants. The vertical axis represents the difference between the absorbance at 450 nm and the absorbance at the reference wavelength of 650 nm, and the horizontal axis represents the concentration (ng / mL) of CD16V or CD16V mutant. [Figure 4]This graph shows the binding activity of wild-type Fc-type anti-EpCAM antibodies or mutant Fc-type anti-EpCAM antibodies against CD16V or CD16V mutants. The vertical axis represents the difference between the absorbance at 450 nm and the absorbance at the reference wavelength of 650 nm, and the horizontal axis represents the concentration (ng / mL) of CD16V or CD16V mutant. [Figure 5] This graph shows the binding activity of wild-type Fc-type anti-HER2 antibodies or mutant Fc-type anti-HER2 antibodies against CD16V or CD16V mutants under conditions of competition with anti-KLH antibodies. The vertical axis represents the difference between absorbance at 450 nm and absorbance at the reference wavelength of 650 nm, and the horizontal axis represents the concentration (ng / mL) of CD16V or CD16V mutant. [Figure 6] This graph shows the expression levels of CD16V or the CD16V mutant in KHYG-1 cells expressing CD16V or KHYG-1 cells expressing the CD16V mutant, as analyzed by flow cytometry. The vertical axis represents the number of cells, and the horizontal axis represents the fluorescence intensity (CD16V expression level). The numbers in the figure indicate the percentage of cells expressing CD16V or the CD16V mutant. [Figure 7] This graph shows the ADCC activity of KHYG-1 cells against HER2-positive SK-BR-3 cells in the presence of an anti-HER2 antibody. The vertical axis represents cytotoxic activity (%), and the horizontal axis represents antibody concentration (ng / mL). [Figure 8] This graph shows the ADCC activity of KHYG-1 cells against HER2-positive SK-BR-3 cells, induced by an anti-HER2 antibody, in the presence of human serum. The vertical axis represents cytotoxic activity (%). [Figure 9] This graph shows the cytotoxic activity of CD16V CAR-T against HER2-positive SK-BR-3 cells in the presence of an anti-HER2 antibody. The vertical axis represents cytotoxic activity (%), and the horizontal axis represents antibody concentration (ng / mL). [Modes for carrying out the invention] 【0076】 Detailed description of the invention The following is a detailed description of the present invention. It should be noted that the present invention is not limited to this description. In this specification, scientific and technical terms used in connection with the present invention have the meanings generally understood by those skilled in the art, unless otherwise specified. 【0077】 I definition As used herein, the term "antibody" means immunoglobulin, a biomolecule comprising two heavy chains (H chains) and two light chains (L chains) stabilized by disulfide bonds. The heavy chain consists of a heavy chain variable region (VH), a heavy chain constant region (CH1, CH2, CH3), and a hinge region located between CH1 and CH2, while the light chain consists of a light chain variable region (VL) and a light chain constant region (CL). Of these, the variable region fragment (Fv), composed of VH and VL, is directly involved in antigen binding and is the region that gives diversity to antibodies. Of the variable regions, the region that directly contacts the antigen is particularly volatile and is known as the complementarity-determining region (CDR). The regions other than the CDR, which are relatively less volatile, are known as the framework region (FR). The light chain variable region and the heavy chain variable region each have three CDRs, which are known as heavy chain CDR1 to CDR3 and light chain CDR1 to CDR3, respectively, in order from the N-terminus. 【0078】 As used herein, the term "IgG" refers to one of the five classes of immunoglobulins (IgG, IgM, IgA, IgD, and IgE). IgG has subclasses IgG1, IgG2, IgG3, and IgG4, and their corresponding heavy chains are called Igγ1, Igγ2, Igγ3, and Igγ4. 【0079】 As used herein, the term “Fc region” means the region consisting of the hinge region, CH2, and CH3 in the heavy chain of an antibody, or the region consisting of CH2 and CH3 in the heavy chain of an antibody. The Fc region may contain genetic polymorphisms. (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., 1991, NIH Publication No. 91-3242). 【0080】 As used herein, the term “antigen-binding fragment” means a fragment of an antibody that can bind to an antigen. Specific examples of antigen-binding fragments include Fab, consisting of a VL region, a VH region, a CL region, and a CH1 region; F(ab')2, in which two Fabs are linked by a disulfide bond in a hinge region; Fv, consisting of VL and VH; scFv, a single-chain antibody in which VL and VH are linked by an artificial polypeptide linker; and bispecific antibodies such as diabodies, single-chain diabodies (scDb), tandem scFv, and leucine zippers. 【0081】 As used herein, the term “human antibody” means an antibody having a human immunoglobulin amino acid sequence. In this specification, “humanized antibody” means an antibody in which some, most, or all of the amino acid residues other than the CDR are replaced by amino acid residues derived from a human immunoglobulin molecule. There are no particular limitations on the method of humanization, and humanized antibodies can be prepared, for example, by reference to U.S. Patent No. 5,225,539 or U.S. Patent No. 6,180,370. The entirety of each of the aforementioned patents is incorporated herein by reference. 【0082】 In addition to conventional full-length antibodies, antibodies include various forms such as single-arm antibodies constructed by combining full-length antibodies, antigen-binding fragments, and / or Fc regions (Proceedings of the National Academy of Sciences, (2013) 110 (32), pp. E2987-2996), and bispecific antibodies (Nature Reviews Drug Discovery, (2019) 18, pp. 585-608). 【0083】 The amino acid residue numbers of antibodies used herein are specified using EU index numbering (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed., 1991, NIH Publication No. 91-3242), and can be determined according to this numbering scheme. 【0084】 As used herein, the term "Fcγ receptor (FcγR)" refers to a protein belonging to the immunoglobulin superfamily that is expressed in many immune cells. FcγRs are receptor proteins for the Fc region of IgG and have binding activity to the Fc region of IgG. FcγRs include FcγRI(CD64), FcγRIIA(CD32A), FcγRIIB(CD32B), FcγRIIIA(CD16A), and FcγRIIIB(CD16B). FcγRI(CD64), expressed in macrophages and dendritic cells, is known to bind strongly to the Fc region of IgG. FcγRIIA(CD32A), FcγRIIB(CD32B), and FcγRIIC(CD32C), expressed on monocytes and neutrophils, as well as FcγRIIIA(CD16A) and FcγRIIIB(CD16B), expressed in macrophages and NK cells, are known to bind weakly to the Fc region of IgG. CD16A is known to be involved in the initiation of ADCC (described later) by binding to the Fc region of IgG. 【0085】 As used herein, the term “antibody-dependent cell-mediated cytotoxicity (ADCC)” refers to one of the effector actions that can be attributed to the Fc region of an antibody. ADCC is the action by which immune cells destroy target cells through the binding of antibodies to antigens on target cells, as well as to immune cells such as macrophages and NK cells. The binding of antibodies to immune cells occurs via the Fc region of the antibody and the Fcγ receptor on the immune cell. ADCC is induced by the binding of the Fc region to the Fcγ receptor IIIA (CD16A). 【0086】 As used herein, the term “combined administration” means the simultaneous or individual administration of various types of active pharmaceutical ingredients to the same subject for therapeutic purposes. During combined administration, the various types of active pharmaceutical ingredients may be contained in the same composition or separately in different compositions. 【0087】 As used herein, the term "pharmaceutical composition" means a single composition containing one or more pharmaceutically active ingredients. "Combination drug" means a combination of pharmaceutical compositions in which different active pharmaceutically active ingredients are contained separately in different compositions. 【0088】 As used herein, the term “natural” means that which exists in nature without any artificial or anthropogenic modification. “Natural” and “wild-type” may be used synonymously. In this specification, “unnatural” means an artificial or anthropogenic product that does not originally exist in nature, for example, a product of the present invention that has at least one amino acid mutation compared to its wild-type or natural product. In the context of the unnatural polypeptides and Fcγ receptors of the present invention, “unnatural” may also mean that it does not essentially bind to its wild-type or natural counterpart, but does bind to an unnatural counterpart that has at least one amino acid mutation compared to its wild-type or natural counterpart. 【0089】 II. The Invention Polypeptide The present invention provides a polypeptide comprising an IgG Fc region, wherein the Fc region comprises at least one amino acid mutation, the polypeptide essentially lacks binding activity to a wild-type Fcγ receptor, and binds to an Fcγ receptor comprising at least one amino acid mutation. 【0090】 This polypeptide contains the Fc region of IgG. In some embodiments, the Fc region of IgG is the Fc region of human IgG and may contain genetic polymorphisms. In some embodiments, the Fc region of IgG is the Fc region of human Igγ1. Those skilled in the art can readily obtain the amino acid sequence of the Fc region from public databases such as UniProt. The polypeptide of the present invention may be any form of polypeptide, as long as it contains the Fc region of IgG. For example, the polypeptide of the present invention may be an antibody, and an Fc fusion protein of a biomolecule or fragment thereof with an Fc region. 【0091】 In some embodiments, polypeptides are antibodies. In some embodiments, polypeptides are human antibodies and humanized antibodies. In the present invention, “antibody” includes ordinary IgG antibodies as well as antibodies of various forms, such as one-arm antibodies and bispecific antibodies, insofar as they have an Fc region. 【0092】 In some embodiments, the Fc region is a modified Fc region of IgG, and the Fc region is non-natural and contains at least one amino acid mutation compared to the Fc region of wild-type or natural IgG. In the present invention, “amino acid mutation” in the Fc region means an amino acid substitution, deletion, or insertion at a predetermined amino acid position in the Fc region. In the present invention, the polypeptide essentially lacks binding activity to the wild-type Fcγ receptor and binds to an Fcγ receptor containing at least one amino acid mutation. The amino acid mutation in the Fc region of the polypeptide of the present invention may be any amino acid mutation, insofar as the polypeptide of the present invention exhibits these binding properties. In one embodiment of the present invention, the amino acid mutation in the Fc region is an amino acid mutation at an amino acid residue within the Fc region that is involved in the binding of the Fc region to the wild-type Fcγ receptor. In one embodiment of the present invention, the Fc region is the Fc region of human Igγ1, and the amino acid mutation in the Fc region is an amino acid mutation at at least one amino acid position selected from the amino acids at positions 233, 234, 235, 236, 237, 239, 265, 269, 294, 297, 299, 328, and 329 within the constant region of human Igγ1 as determined by EU index numbering (Nature (2000) 406, pp. 267-273, Proceedings of the National Academy of Sciences of the United States of America (2015) 112, pp. 833-838). In one embodiment of the present invention, the Fc region is the Fc region of human Igγ1, and the amino acid mutation in the Fc region is an amino acid mutation at at least one or more amino acid positions selected from the amino acids at positions 269 and 294 within the constant region of human Igγ1 as determined by EU index numbering.In one aspect of the present invention, the human Igγ1 constant region before introducing the mutation includes the amino acid sequence shown in SEQ ID NO: 24. In one aspect of the present invention, the human Igγ1 Fc region before introducing the mutation includes residues 1 to 330 of the amino acid sequence shown in SEQ ID NO: 24. 【0093】 In the polypeptide of the present invention, "amino acid mutation" in the target Fcγ receptor means substitution, deletion, or insertion of an amino acid at a predetermined amino acid position in the corresponding wild-type Fcγ receptor. The wild-type Fcγ receptor means a natural Fcγ receptor that may include genetic polymorphisms. 【0094】 The Fcγ receptor containing at least one amino acid mutation includes an amino acid sequence containing at least one amino acid mutation as compared to the amino acid sequence of the wild-type Fcγ receptor. In one aspect of the present invention, the Fcγ receptor containing at least one amino acid mutation includes an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity to the amino acid sequence of the wild-type Fcγ receptor. In one aspect of the present invention, the Fcγ receptor containing at least one amino acid mutation includes 5, 4, 3, 2, or 1 amino acid mutation as compared to the amino acid sequence of the wild-type Fcγ receptor. 【0095】 As used herein, the term "identity" means the value of identity obtained by EMBOSS Needle (Nucleic Acids Res. (2015) 43, pp. W580-W584) using the parameters provided in the initial settings. These parameters are as follows. Gap opening penalty = 10 Gap extension penalty = 0.5 Matrix = EBLOSUM62 【0096】 The binding properties of the polypeptide of the present invention to the Fcγ receptor can be confirmed using any known binding activity measurement method. For example, binding activity can be measured using enzyme-linked immunosorbent assay (ELISA). In one example, the binding properties of the polypeptide of the present invention to the Fcγ receptor can be confirmed by performing the following steps: Prepare wild-type Fcγ receptors and Fcγ receptors containing at least one amino acid mutation as Fcγ receptors targeted by the polypeptide of the present invention. Prepare a polypeptide (e.g., an antibody) containing an Fc region containing at least one amino acid mutation. Immobilize the protein targeted by the polypeptide (e.g., an antigen protein) on an ELISA plate, add the polypeptide, and react with it. After reacting with the polypeptide, add each Fcγ receptor and react with it. After these reactions, react with a secondary antibody, for example, an anti-Fcγ receptor antibody labeled with an enzyme such as horseradish peroxidase (HRP). After these reactions, a washing procedure is performed, and the binding of the secondary antibody is confirmed by measuring the activity using a reagent that detects this activity (for example, TMB reagent (DAKO, catalog S1599) in the case of HRP labeling) so that it is possible to evaluate whether the antibody containing an Fc region with at least one amino acid mutation binds to the wild-type Fcγ receptor and the Fcγ receptor containing one or more amino acid mutations. The specific evaluation method used may be the method described in Example 4 below. 【0097】 As used herein, the expression “essentially lacking binding activity” to the wild-type Fcγ receptor means that the binding activity of the polypeptide of the present invention to the wild-type Fcγ receptor is not significantly higher than the binding activity of the polypeptide of the present invention to the Fcγ receptor containing at least one amino acid mutation to which the polypeptide of the present invention binds. In one embodiment of the present invention, when the “essentially lacking binding activity” state to the wild-type Fcγ receptor is measured by an ELISA method, this is less than 10%, less than 5%, less than 3%, less than 2%, less than 1%, less than 0.5%, less than 0.1%, or less than 0.05% of the binding activity to the Fcγ receptor containing at least one amino acid mutation used as a control. 【0098】 The Fcγ receptor targeted by the polypeptide of the present invention can be selected by those skilled in the art based on the intended use of the polypeptide and other factors. In one embodiment of the present invention, the Fcγ receptor may be FcγRI(CD64), FcγRIIA(CD32A), FcγRIIB(CD32B), FcγRIIIA(CD16A), or FcγRIIIB(CD16B). In one embodiment of the present invention, the Fcγ receptor targeted by the polypeptide of the present invention is CD16. In one embodiment of the present invention, the Fcγ receptor is CD16A. Wild-type CD16A includes, but is not limited to, two genetic polymorphisms, namely CD16A V158 and CD16A F158. In one embodiment of the present invention, wild-type CD16A is CD16A V158. 【0099】 In one embodiment of the polypeptide of the present invention, the target Fcγ receptor is CD16A, and wild-type CD16A comprises the amino acid sequence shown in SEQ ID NO: 78. 【0100】 In one embodiment of the polypeptide of the present invention, the target Fcγ receptor is CD16A comprising at least one amino acid mutation, wherein the CD16A comprising at least one amino acid mutation comprises at least one mutation selected from a lysine-to-aspartic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), a lysine-to-glutamic acid mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and a lysine-to-glutamic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation). In one embodiment of the present invention, the CD16A comprising at least one amino acid mutation comprises one or both of the K131D mutation and the K128E mutation. In one embodiment of the present invention, the CD16A comprising at least one amino acid mutation comprises one or both of the K131E mutation and the K128E mutation. In one embodiment of the present invention, CD16A comprising at least one amino acid mutation comprises the K131D mutation and at least one mutation selected from an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0101】 As used herein, the term "corresponding position" to a position in SEQ ID NO: 78 means that when the amino acid sequence of CD16A is aligned with the amino acid sequence of SEQ ID NO: 78 using a sequence alignment program such as BLAST, that amino acid position in CD16A is aligned to the same position in SEQ ID NO: 78. 【0102】 In one embodiment of the present invention, CD16A containing at least one amino acid mutation comprises the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0103】 In one embodiment of the present invention, the polypeptide of the present invention essentially has no binding activity to CD16A containing the amino acid sequence of SEQ ID NO: 78, and binds to CD16A containing the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0104】 In one embodiment of the present invention, the polypeptide comprises the Fc region of human Igγ1, wherein the Fc region contains an amino acid mutation at at least one or more amino acid positions selected from the amino acids at positions 269 and 294 within the constant region of human Igγ1 as assigned EU index numbers. 【0105】 In one embodiment of the present invention, the polypeptide comprises the Fc region of human Igγ1, the Fc region comprising a glutamate-to-arginine mutation (E269R mutation) at the position corresponding to EU index number 269, and at least one mutation selected from a glutamate-to-arginine mutation (E294R mutation) at the position corresponding to EU index number 294, and a glutamate-to-lysine mutation (E294K mutation) at the position corresponding to EU index number 294. 【0106】 The amino acid sequence information for the polypeptide of the present invention and the Fcγ receptor targeted by the polypeptide can be obtained from public databases such as UniProt, and they can be easily prepared based on this amino acid sequence information using any method known to those skilled in the art or using the methods described herein. 【0107】 When the polypeptide of the present invention is an antibody, an antibody against any antigen can be used. When the polypeptide of the present invention is an antibody, the antibody can be obtained by immunization with a target antigen according to any antibody preparation method known to those skilled in the art, or by introducing amino acid mutations into the Fc region of a known antibody. 【0108】 In one embodiment of the present invention, the polypeptide is an antibody that binds to a cancer antigen. Known antibodies that can be used to prepare the polypeptide of the present invention include anti-CD19 antibody (tafacitamab, Drug Bank accession number: DB15044), anti-HER2 antibody, anti-EpCAM antibody, or anti-EGFR antibody. 【0109】 In one embodiment of the present invention, the polypeptide is an anti-CD19 antibody, anti-HER2 antibody, anti-EpCAM antibody, or anti-EGFR antibody comprising the Fc region of human Igγ1, wherein the Fc region comprises at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the human Igγ constant region as assigned by EU index numbering (E269R mutation), a glutamate-to-arginine mutation at position 294 in the human Igγ constant region as assigned by EU index numbering (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the human Igγ constant region as assigned by EU index numbering (E294K mutation). 【0110】 III. Cells expressing CD16A containing amino acid mutations The present invention also provides CD16A-expressing cells that can be used in combination with the polypeptide of the present invention. 【0111】 In one aspect, the present invention provides cells expressing a non-natural CD16A that contains at least one amino acid mutation compared to wild-type or natural CD16A, wherein the at least one amino acid mutation is selected from (i) a lysine-to-aspartate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a lysine-to-glutamate mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a lysine-to-glutamate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation), and the non-natural CD16A contains an amino acid sequence having 90% or more amino acid sequence identity with respect to SEQ ID NO: 78. 【0112】 In one embodiment of the present invention, the CD16A expressed by the CD16A-expressing cells of the present invention comprises one or both of the K131D mutation and the K128E mutation. In one embodiment of the present invention, the CD16A expressed by the CD16A-expressing cells of the present invention comprises one or both of the K131E mutation and the K128E mutation. In one embodiment of the present invention, the CD16A expressed by the CD16A-expressing cells of the present invention comprises the K131D mutation and at least one mutation selected from an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0113】 In one embodiment of the present invention, the CD16A expressed by the CD16A-expressing cells of the present invention comprises an amino acid sequence having at least 95%, 96%, 97%, 98%, or 99% amino acid sequence identity with respect to the amino acid sequence of SEQ ID NO: 78. 【0114】 In one aspect of the present invention, the CD16A expressed by the CD16A-expressing cells of the present invention comprises the amino acid sequence set forth in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0115】 The cells used as the CD16A-expressing cells of the present invention can be any cells as long as they can regulate (e.g., induce or suppress) an immune response when used together with the polypeptide of the present invention. The cells may be immune cells of the innate immune system or the acquired immune system, and examples include NK cells, NKT cells, macrophages, microglia, osteoclasts, granulocytes (including neutrophils, eosinophils, and basophils), monocytes, dendritic cells, T cells, and B cells. In one aspect of the present invention, the cells are cells of human origin. In one aspect of the present invention, the cells are human immune cells. In one aspect of the present invention, the cells are human NK cells or human T cells. 【0116】 In one aspect of the present invention, the cells used as the CD16A-expressing cells of the present invention include, but are not limited to, stem cells such as pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, or embryonic germ cells, or cells derived from such stem cells, such as the aforementioned immune cells. In one aspect of the present invention, the stem cells are pluripotent stem cells. The pluripotent stem cells may be induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells), and may be of human origin. The iPS cells or ES cells can be prepared by those skilled in the art using any known method. The method used to differentiate stem cells such as iPS cells or ES cells into the CD16A-expressing cells of the present invention can be any method known to those skilled in the art. In one aspect of the present invention, the CD16A-expressing cells of the present invention are human immune cells derived from pluripotent stem cells. In one aspect of the present invention, the CD16A-expressing cells are human NK cells or human T cells derived from pluripotent stem cells. 【0117】 In one embodiment of the present invention, the stem cells may be universal donor cells in which the stem cells have been gene-edited to avoid allogeneic response and lysis by NK cells. Universal donor cells and cells derived from said universal donor cells may contain genetically engineered disruption in the β-2 microglobulin (B2M) gene to eliminate the expression of an HLA class I molecule, for example, as described in WO 2012 / 145384, which is incorporated herein by reference in its entirety. Universal donor cells and cells derived from said universal donor cells may further contain polynucleotides capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, comprising at least one portion of the B2M protein, which is covalently bonded either directly or via a linker sequence to at least one portion of the HLA-1α chain. In one embodiment, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. See also WO 2012 / 145384. Universal donor cells and cells derived from such universal donor cells may additionally contain genetically modified disruption in HLA class II-related genes, for example, by knocking out one or more transcription factors required for the expression of HLA class II genes, such as regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, as described in WO 2013 / 158292, which is also incorporated herein by reference in its entirety. The cells may further contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. See also WO 2013 / 158292. In one embodiment of the present invention, universal donor cells and cells derived from said universal donor cells express CD16A containing at least one amino acid mutation.In one embodiment of the present invention, the universal donor cells are iPS cells or ES cells, and the CD16A-expressing cells are cells derived from such universal donor cells, such as the aforementioned immune cells. In one embodiment of the present invention, the CD16A-expressing cells are human immune cells derived from universal donor cells. In one embodiment of the present invention, the CD16A-expressing cells are human NK cells or human T cells derived from universal donor cells. 【0118】 In one embodiment of the present invention, CD16A-expressing cells can be isolated and / or purified. In one embodiment of the present invention, CD16A-expressing cells are immortalized cells or established cell lines, and those skilled in the art can prepare these immortalized cells or established cell lines using any known method. In one embodiment of the present invention, the cells used as CD16A-expressing cells of the present invention are immortalized cells or established cell lines of human origin. In one embodiment of the present invention, the cells used as CD16A-expressing cells of the present invention are patient-derived cells. 【0119】 In one embodiment of the present invention, the CD16A-expressing cells of the present invention are cells that exogenously express CD16A. The CD16A-expressing cells of the present invention can be prepared by introducing a gene encoding CD16A containing a desired amino acid mutation into cells. For example, the gene can be synthesized using the phosphoramidite method based on its nucleotide sequence, or it can be prepared by combining DNA fragments obtained from a cDNA library using polymerase chain reaction (PCR). The target gene can be introduced into cells using an expression vector containing the gene in the form of cDNA. The gene can also be introduced into cells using polynucleotides in the form of mRNA. Alternatively, the target gene may be introduced directly into cells using methods such as electroporation or lipofection. After introducing the target gene into cells, these cells may be cultured and grown. 【0120】 The expression vector used to prepare CD16A-expressing cells according to the present invention is not particularly limited as long as it can express a target protein, such as CD16A containing amino acid mutations, in the target cells. Examples of expression vectors that can be used include plasmid vectors (e.g., Thermo Fisher Scientific's pcDNA series, Promega's pALTER®-MAX vector, and Takara's pHEK293 Ultra expression vector) or viral vectors (e.g., lentivirus, adenovirus, retrovirus, or adeno-associated virus). For the preparation of viral vectors, the pLVSIN-CMV / EF1α vector (Takara Bio) or pLenti vector (Thermo Fisher Scientific) used for lentivirus preparation can be used. 【0121】 The expression vector may include a start codon and a stop codon. In this case, the expression vector may include an enhancer sequence, an untranslated region, a splicing junction, a polyadenylation site, or a repeatable unit. The expression vector may also include a gene that can act as a marker to confirm the expression of a target gene (e.g., a drug resistance gene, a gene encoding a reporter enzyme, or a gene encoding a fluorescent protein). 【0122】 To obtain or maintain the CD16A-expressing cells of the present invention, culture can be carried out by known methods. Examples of usable basic media include MEM medium (Science (1955) 122, pp. 501-504), DMEM medium (Virology (1959) 8, pp. 396-397), RPMI1640 medium (The Journal of the American Medical Association (1967) 199, pp. 519-524), 199 medium (Proceedings of the Society for Experimental Biology and Medicine (1950) 73, pp. 1-8), FreeStyle® 293 expression medium (Thermo Fisher Scientific, catalog 12338026), CD293 medium (Thermo Fisher Scientific, catalog 11913019), or Expi293® expression medium (Thermo Fisher Scientific, catalog A1435101). The culture medium may also contain serum (e.g., fetal bovine serum: FBS), serum substitutes (e.g., knockout serum substitutes: KSR), fatty acids or lipids, amino acids, vitamins, growth factors, cytokines, antioxidants, 2-mercaptoethanol, pyruvate, buffers, inorganic salts, and antibiotics. In one embodiment of the present invention, the culture medium is serum-free or synthetic. Culture conditions (such as culture time, temperature, pH of the medium, and CO2 concentration) can be appropriately selected by those skilled in the art. The pH of the medium is preferably about 6 to 8. There are no particular restrictions on the culture temperature, but cultures can be used at about 30 to 40°C, preferably about 37°C. The CO2 concentration may be about 1 to 10%, and preferably about 5%. There are no particular restrictions on the culture time, but it may be about 15 to 336 hours. The culture may be aerated or stirred as needed. If an inducible promoter induced by a drug such as tetracycline or doxycycline is used, the process may include the step of culturing cells in a medium containing the drug, and then inducing the expression of a gene functionally linked to the inducible promoter, such as a cancer antigen.This stage can be carried out according to gene induction methods using a general gene transfer system. 【0123】 IV Methods for treating and preventing a patient's disease or disability using immunotherapy The present invention also provides a method for treating or preventing a patient's disease or disorder using immunotherapy. This method comprises the step of administering to a patient cells expressing a polypeptide of the present invention and a non-natural Fcγ receptor comprising at least one amino acid mutation, wherein the polypeptide can bind to the non-natural Fcγ receptor comprising at least one amino acid mutation. 【0124】 In this specification, the term “immunotherapy” means a method for preventing or treating autoimmune diseases, cancer, and various bacterial or viral infections by utilizing the function of self / non-self recognizing immune cells to eliminate foreign substances such as exogenous bacteria, viruses, and cancer cells. 【0125】 The cells used in the therapeutic method of the present invention are cells expressing an Fcγ receptor containing at least one amino acid mutation, the Fcγ receptor being able to bind to the polypeptide of the present invention. The cells expressing the Fcγ receptor can be any cell, insofar as they can modulate (e.g., induce or suppress) an immune response when used together with the polypeptide of the present invention. The cells may be immune cells of the innate or adaptive immune system, and examples include NK cells, NKT cells, macrophages, microglia, osteoclasts, granulocytes (including neutrophils, eosinophils, and basophils), monocytes, dendritic cells, T cells, and B cells. In one embodiment of the present invention, the cells are cells of human origin. In one embodiment of the present invention, the cells are human immune cells. In one embodiment of the present invention, the cells are human NK cells or human T cells. 【0126】 In one aspect of the present invention, the cells used in the therapeutic method of the present invention, such as the aforementioned immune cells, may be derived from stem cells, including, but not limited to, pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, or embryonic germ cells. In one aspect of the present invention, the stem cells are engineered to express an Fcγ receptor containing at least one amino acid mutation, the Fcγ receptor capable of binding to the polypeptide of the present invention. In one aspect of the present invention, the stem cells are pluripotent stem cells. The pluripotent stem cells may be induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells), and may be of human origin. iPS cells or ES cells can be prepared using any known method for those skilled in the art. The method for differentiating stem cells, such as iPS cells or ES cells, into cells used in the therapeutic method of the present invention may be any method known to those skilled in the art. In one aspect of the present invention, the cells are human immune cells derived from pluripotent stem cells. In one aspect of the present invention, the cells are human NK cells or human T cells derived from pluripotent stem cells. 【0127】 In one embodiment of the present invention, the cells used in the therapeutic method of the present invention may be derived from universal donor cells in which the stem cells have been gene-edited to avoid allogeneic response and lysis by NK cells. Universal donor cells and cells derived from said universal donor cells may contain genetically engineered disruption in the β-2 microglobulin (B2M) gene to eliminate the expression of an HLA class I molecule, for example, as described in WO 2012 / 145384, which is incorporated herein by reference in its entirety. Universal donor cells and cells derived from said universal donor cells may further contain polynucleotides capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, comprising at least one portion of the B2M protein, which is covalently bonded either directly or via a linker sequence to at least one portion of the HLA-1α chain. In one embodiment, the HLA-1α chain is selected from HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. See also WO 2012 / 145384. Universal donor cells and cells derived from such universal donor cells may additionally contain genetically modified disruption in HLA class II-related genes, for example, by knocking out one or more transcription factors required for the expression of HLA class II genes, such as regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, as described in WO 2013 / 158292, which is also incorporated herein by reference in its entirety. The cells may also further contain a single-chain fusion HLA class II protein or one or more polynucleotides encoding an HLA class II protein. See also WO 2013 / 158292.In another embodiment, universal donor cells and cells derived from said universal donor cells are engineered to express an Fcγ receptor containing at least one amino acid mutation, the Fcγ receptor capable of binding to the polypeptide of the present invention. In one embodiment of the present invention, the universal donor cells are iPS cells or ES cells. In one embodiment of the present invention, the cells used in the therapeutic method of the present invention are human immune cells derived from universal donor cells. In one embodiment of the present invention, the cells used in the therapeutic method of the present invention are human NK cells or human T cells derived from universal donor cells. 【0128】 In one embodiment of the present invention, cells used in the therapeutic method of the present invention can be isolated and / or purified. Here, “isolation” means separation from living tissue, and “purification” means separation of the cells from one or more additional components in the tissue from which the cells originate. In one embodiment of the present invention, the cells used in the therapeutic method of the present invention are immortalized cells or established cell lines, and those skilled in the art can prepare such immortalized cells or established cell lines using any known method. In one embodiment of the present invention, the cells used in the therapeutic method of the present invention are immortalized cells or established cell lines of human origin. In one embodiment of the present invention, the cells used in the therapeutic method of the present invention are cells derived from a patient. 【0129】 The Fcγ receptor expressed in cells used in the therapeutic method of the present invention is the Fcγ receptor targeted by the polypeptide of the present invention and can be selected by those skilled in the art based on the intended use of the polypeptide of the present invention and other factors. In one embodiment of the present invention, the Fcγ receptor may be FcγRI(CD64), FcγRIIA(CD32A), FcγRIIB(CD32B), FcγRIIIA(CD16A), or FcγRIIIB(CD16B). In one embodiment of the present invention, the Fcγ receptor targeted by the polypeptide of the present invention is CD16. In one embodiment of the present invention, the Fcγ receptor is CD16A. CD16A includes, but is not limited to, two genetic polymorphisms, namely CD16A V158 and CD16A F158. In one embodiment of the present invention, CD16A is CD16A V158. 【0130】 In one embodiment of the present invention, the Fcγ receptor expressed by cells used in the therapeutic method of the present invention is CD16A containing at least one amino acid mutation selected from a lysine-to-aspartic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), a lysine-to-glutamic acid mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and a lysine-to-glutamic acid mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation). In one embodiment of the present invention, the Fcγ receptor expressed by cells used in the therapeutic method of the present invention is CD16A containing one or both of the K131D mutation and the K128E mutation. In one embodiment of the present invention, the Fcγ receptor expressed by cells used in the therapeutic method of the present invention is CD16A containing one or both of the K131E mutation and the K128E mutation. In one embodiment of the present invention, the Fcγ receptor expressed by cells used in the therapeutic method of the present invention is CD16A comprising the K131D mutation and at least one mutation selected from the asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and the asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0131】 In one embodiment of the present invention, the Fcγ receptor expressed by cells used in the therapeutic method of the present invention is CD16A comprising the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0132】 In one embodiment of the present invention, the therapeutic method is: (A) a polypeptide comprising the Fc region of human Igγ1, wherein the Fc region comprises at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), a glutamate-to-arginine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation); (B) (i) (a) a lysine-to-aspartate mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (b) a lysine-to-glutamate mutation at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (c) SEQ ID NO: The step of administering to a patient cells selected from: (ii) cells expressing CD16A containing at least one mutation selected from a lysine-to-glutamine mutation at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation); (iii) cells expressing CD16A containing one or both of the K131D mutation and the K128E mutation; and (iv) cells expressing CD16A containing at least one mutation selected from the K131D mutation and an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0133】 In one embodiment of the present invention, a therapeutic method comprises administering to a patient (A) a polypeptide comprising the Fc region of human Igγ1, wherein the Fc region comprises at least one mutation selected from a mutation of glutamate to arginine at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), a mutation of glutamate to arginine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a mutation of glutamate to lysine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation); and (B) cells expressing CD16A comprising the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0134】 In one embodiment of the present invention, the polypeptide used in the therapeutic method is an antibody. In one embodiment of the present invention, the antibody is an antibody that binds to a cancer antigen. In one embodiment of the present invention, the polypeptide used in the therapeutic method is an anti-CD19 antibody, an anti-HER2 antibody, an anti-EpCAM antibody, or an anti-EGFR antibody having an Fc region, wherein the Fc region is the Fc region of human Igγ1 and includes at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), a glutamate-to-arginine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation). 【0135】 Cells expressing an Fcγ receptor containing at least one amino acid mutation used in the therapeutic method of the present invention can be prepared by introducing a gene encoding an Fcγ receptor containing the target amino acid mutation into the cells. The gene encoding an Fcγ receptor containing the amino acid mutation can be prepared using standard molecular biology and / or chemical techniques, in which the nucleotide sequence encoding the amino acid sequence of the target Fcγ receptor is obtained from an NCBI reference sequence ID or GenBank accession number, and this nucleotide sequence is used to design the sequence of the gene encoding an Fcγ receptor containing the target amino acid mutation. The introduction of the gene into cells and the culture of the cells can be carried out using any method known to those skilled in the art and the method described herein. 【0136】 There are no particular limitations on the diseases that can be treated by the therapeutic methods of the present invention, and may include bacterial infections, viral infections, autoimmune diseases, and cancer. In one embodiment of the present invention, the therapeutic method is used to treat or prevent cancer. In one embodiment of the present invention, the therapeutic method is a method of treatment or prevention for a patient using cancer immunotherapy. In this specification, "cancer immunotherapy" means a method for preventing or treating cancer by activating or increasing immune cells that are involved in identifying cancer cells present in the body as foreign substances through the immune system and eliminating them. 【0137】 In the method of the present invention, the polypeptide of the present invention and cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide binds can be administered to a subject in need of immunotherapy using any method known to those skilled in the art. When the polypeptide of the present invention is administered to a subject, it can be administered in the form of a pharmaceutical composition comprising the polypeptide of the present invention and a pharmaceutically acceptable excipient. When cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide binds can be administered to a subject, they can be administered in the form of a pharmaceutical composition comprising the cells and a pharmaceutically acceptable excipient. In the therapeutic method of the present invention, a pharmaceutical composition comprising the polypeptide of the present invention, cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide binds, and a pharmaceutically acceptable excipient can be administered to a subject. Pharmaceutical compositions described herein can be used. 【0138】 The dose and frequency of administration of the polypeptide of the present invention and cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide binds, administered to the subject, can be appropriately adjusted depending on the target disease, the age, weight, and condition of the subject being treated, the dosage form, the type and potency of the polypeptide of the present invention, and the type of cells used in the therapeutic method of the present invention. The dose of the polypeptide of the present invention may be, for example, about 0.001 mg / kg to 100 mg / kg. The dose of cells used in this method may be, for example, 1 × 10⁶ per administration to the subject. 3 cells / kg~1×10 9 It can be cells / kg. 【0139】 The polypeptides and cells used in the methods of the present invention can be administered to a subject by any suitable route of administration, for example, by intravenous injection, intratumor injection, intradermal injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, or intra-arterial injection. 【0140】 In the method of the present invention, the polypeptide of the present invention and cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide binds, which are administered to a subject, can be administered simultaneously, sequentially, or sequentially. In one embodiment of the present invention, administration of the polypeptide of the present invention to a subject is initiated, followed by administration of the cells used in the therapeutic method of the present invention. In one embodiment of the present invention, administration of the cells used in the therapeutic method of the present invention to a subject is initiated, followed by administration of the polypeptide of the present invention. In one embodiment of the present invention, administration of the polypeptide of the present invention to a subject is completed, followed by administration of the cells used in the therapeutic method of the present invention. In one embodiment of the present invention, administration of the cells used in the therapeutic method of the present invention to a subject is completed, followed by administration of the polypeptide of the present invention. 【0141】 V. Pharmaceutical compositions and compound drugs of the present invention The present invention also provides the following pharmaceutical compositions and compound drugs: (1) A pharmaceutical composition comprising the polypeptide of the present invention and a pharmaceutically acceptable excipient; (2) A pharmaceutical composition comprising CD16A-expressing cells of the present invention and a pharmaceutically acceptable excipient; (3) A pharmaceutical composition comprising the polypeptide of the present invention, cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide is bound, and a pharmaceutically acceptable excipient; and (4) A compound drug comprising a pharmaceutical composition comprising the polypeptide of the present invention and a pharmaceutically acceptable excipient, and a pharmaceutical composition comprising cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide is bound, and a pharmaceutically acceptable excipient. 【0142】 The compound drug of the present invention may be in the form of a kit in which each of the constituent pharmaceutical compositions is contained in a single packaging container. 【0143】 The pharmaceutical compositions of the present invention can be prepared by methods common in the art using excipients common in the art, i.e., pharmaceutically acceptable excipients or pharmaceutically acceptable carriers. The dosage forms of these pharmaceutical compositions may be injections or parenteral drugs such as infusions. During formulation, appropriate excipients, carriers, and additives for the dosage form may be used within pharmaceutically acceptable limits. 【0144】 In one embodiment of the pharmaceutical composition of (1), the pharmaceutical composition of (1) is a pharmaceutical composition for use in combination with cells in a method of treating or preventing a patient by immunotherapy, wherein the cells are cells expressing an Fcγ receptor containing at least one amino acid mutation to which a polypeptide binds. In one embodiment of the present invention, the cells are human immune cells. In one embodiment of the present invention, the human immune cells are human T cells or human NK cells. In one embodiment of the present invention, the cells are CD16A-expressing cells containing at least one amino acid mutation. In one embodiment of the present invention, the cells are the CD16A-expressing cells of the present invention. 【0145】 In one embodiment of the present invention, the pharmaceutical composition of (1) may be a pharmaceutical composition comprising a polypeptide comprising the Fc region of human Igγ1, wherein the Fc region comprises at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the constant region of human Igγ1 as assigned by EU index numbers (E269R mutation), a glutamate-to-arginine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbers (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbers (E294K mutation), and the pharmaceutical composition is a pharmaceutical composition for use in combination with cells selected from the following: (i) Cells expressing CD16A containing at least one mutation selected from the following: a lysine-to-aspartate mutation at position 131 of SEQ ID NO: 78 (K131D mutation), a lysine-to-glutamate mutation at position 128 of SEQ ID NO: 78 (K128E mutation), and a lysine-to-glutamate mutation at position 131 of SEQ ID NO: 78 (K131E mutation); (ii) Cells expressing CD16A containing one or both of the K131D and K128E mutations; (iii) Cells expressing CD16A containing one or both of the K131E and K128E mutations; or (iv) Cells expressing CD16A containing the K131D mutation and at least one mutation selected from the following: an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation). 【0146】 In one embodiment of the present invention, the pharmaceutical composition of (1) is a pharmaceutical composition comprising a polypeptide comprising the Fc region of human Igγ1, wherein the Fc region comprises a mutation from glutamate to arginine at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), a mutation from glutamate to arginine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a mutation from glutamate to lysine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation), and the pharmaceutical composition is a pharmaceutical composition for use in combination with cells expressing CD16A comprising the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88. 【0147】 In one embodiment of the present invention, the polypeptide used in the pharmaceutical composition of (1) is an antibody. In one embodiment of the present invention, the antibody is an antibody that binds to a cancer antigen. In one embodiment of the present invention, the polypeptide used in the pharmaceutical composition of (1) is an anti-CD19 antibody, an anti-HER2 antibody, an anti-EpCAM antibody, or an anti-EGFR antibody having an Fc region, wherein the Fc region is the Fc region of human Igγ1 and includes at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), a glutamate-to-arginine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation). 【0148】 In one embodiment of the pharmaceutical composition of (2), the pharmaceutical composition of (2) is a pharmaceutical composition for use in combination with a polypeptide when treating or preventing a patient's disease or disorder by immunotherapy, wherein the peptide is a polypeptide comprising the Fc region of IgG, the Fc region comprising at least one amino acid mutation, and the polypeptide essentially lacks binding activity to wild-type CD16A and binds to CD16A comprising at least one amino acid mutation expressed in cells. In one embodiment of the present invention, the polypeptide comprises the Fc region of human Igγ1, the Fc region comprising a mutation from glutamate to arginine at position 269 in the constant region of human Igγ1 as assigned by EU index numbering (E269R mutation), at least one mutation selected from a mutation from glutamate to arginine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294R mutation), and a mutation from glutamate to lysine at position 294 in the constant region of human Igγ1 as assigned by EU index numbering (E294K mutation). In one embodiment of the present invention, the polypeptide is an antibody. In one embodiment of the present invention, the antibody is an antibody that binds to a cancer antigen. In one embodiment of the present invention, the polypeptide is an anti-CD19 antibody, anti-HER2 antibody, anti-EpCAM antibody, or anti-EGFR antibody having an Fc region, wherein the Fc region is the Fc region of human Igγ1 and includes a mutation from glutamate to arginine at position 269 as determined by EU index numbering (E269R mutation), a mutation from glutamate to arginine at position 294 within the constant region of human Igγ1 as determined by EU index numbering (E294R mutation), and a mutation from glutamate to lysine at position 294 within the constant region of human Igγ1 as determined by EU index numbering (E294K mutation). 【0149】 In one embodiment of the pharmaceutical composition of (3), the pharmaceutical composition of (3) is a pharmaceutical composition for preventing and treating a patient's disease or disorder using immunotherapy. In one embodiment of the combination drug of (4), the combination drug of (4) is a combination drug for preventing and treating a patient's disease or disorder using immunotherapy. 【0150】 In one embodiment of the pharmaceutical composition of (3) and the compound drug of (4), the cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide of the present invention binds are human immune cells. In one embodiment of the present invention, the human immune cells are human T cells or human NK cells. In one embodiment of the present invention, the cells are cells expressing CD16A containing at least one amino acid mutation. In one embodiment of the present invention, the cells are CD16A-expressing cells of the present invention. 【0151】 In one embodiment of the pharmaceutical composition of (3) and the compound drug of (4), if the cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide of the present invention binds are CD16A-expressing cells of the present invention, the polypeptide of the present invention comprises the Fc region of IgG, the Fc region comprising a mutation from glutamate to arginine at position 269 in the human Igγ1 constant region as assigned by EU index numbering (E269R mutation), at least one mutation selected from a mutation from glutamate to arginine at position 294 in the human Igγ1 constant region as assigned by EU index numbering (E294R mutation), and a mutation from glutamate to lysine at position 294 in the human Igγ1 constant region as assigned by EU index numbering (E294K mutation). In one embodiment of the present invention, the polypeptide is an antibody. In one embodiment of the present invention, the antibody is an antibody that binds to a cancer antigen. In one embodiment of the present invention, the polypeptide is an anti-CD19 antibody, anti-HER2 antibody, anti-EpCAM antibody, or anti-EGFR antibody having an Fc region, wherein the Fc region is the Fc region of human Igγ1 and includes at least one mutation selected from a glutamate-to-arginine mutation at position 269 in the human Igγ1 constant region as determined by EU index numbering (E269R mutation), a glutamate-to-arginine mutation at position 294 in the human Igγ1 constant region as determined by EU index numbering (E294R mutation), and a glutamate-to-lysine mutation at position 294 in the human Igγ1 constant region as determined by EU index numbering (E294K mutation). 【0152】 While there are no specific limitations, the pharmaceutical compositions or combinations of the present invention can be used to treat or prevent bacterial infections, viral infections, autoimmune diseases, and cancer. 【0153】 The present invention also provides the following combinations of CD16A-expressing cells and polypeptides described herein. (1) The polypeptide of the present invention for treating or preventing a patient's disease or disorder by immunotherapy, used in combination with cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide of the present invention binds. In one embodiment of the present invention, the polypeptide is used to treat or prevent cancer. (2) CD16A-expressing cells of the present invention for treating or preventing a patient's disease or disorder using immunotherapy, in combination with a polypeptide comprising an Fc region of IgG that is not inherently bound to wild-type CD16A but can bind to CD16A containing at least one amino acid mutation expressed in the CD16A-expressing cells of the present invention. In one embodiment of the present invention, CD16A-expressing cells are used to treat or prevent cancer. (3) Use of the polypeptide of the present invention in the production of a pharmaceutical composition for treating or preventing a patient's disease or disorder by immunotherapy, in combination with cells expressing an Fcγ receptor containing at least one amino acid mutation to which the polypeptide of the present invention binds. In one embodiment of the present invention, the polypeptide is used to treat or prevent cancer. (4) Use of CD16A-expressing cells of the present invention in the production of a pharmaceutical composition for treating or preventing a patient's disorder by immunotherapy, in combination with a polypeptide comprising an Fc region of IgG that does not inherently possess binding activity to wild-type CD16A but can bind to CD16A containing at least one amino acid mutation expressed in the CD16A-expressing cells of the present invention. In one embodiment of the present invention, CD16A-expressing cells are used to treat or prevent bacterial infections, viral infections, autoimmune diseases, and / or cancer. 【0154】 The embodiments relating to the polypeptides and CD16A-expressing cells of the present invention also apply to this description of the pharmaceutical compositions and compound drugs of the present invention. 【0155】 VI. Methods for preparing modified Fc regions of IgG, modified Fcγ receptors, and combinations thereof. In one aspect, the present invention also provides a method for obtaining a polypeptide comprising a modified Fc region of IgG, the method comprising the following steps: (1) A step of providing a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region is non-natural and comprises at least one amino acid mutation compared to wild-type or natural IgG; (2) A step of measuring the binding activity of the polypeptide obtained in (1) to the wild-type or native Fcγ receptor; (3) A step of measuring the binding activity of the polypeptide obtained in (1) to a non-natural Fcγ receptor that contains at least one amino acid mutation compared to the wild-type or natural Fcγ receptor; and (4) A step of selecting from the polypeptides obtained in (1) a polypeptide that does not essentially have binding activity to wild-type or native Fcγ receptors and that binds to a non-native Fcγ receptor containing at least one amino acid mutation. 【0156】 In some embodiments, a polypeptide comprising a modified Fc region of IgG is an antibody. Step (4) may further include contacting the antibody with immune cells expressing an Fcγ receptor and cells expressing an antigen to which the antibody binds, and then measuring ADCC activity. 【0157】 Embodiments relating to the polypeptides and CD16A-expressing cells of the present invention used in the methods of the present invention are similarly applicable to this description of the polypeptides of the present invention. 【0158】 In another aspect, the present invention provides a method for preparing a non-native Fcγ receptor. This method includes the following steps: (1) A step of providing a non-natural Fcγ receptor that contains at least one amino acid mutation compared to the wild-type or natural Fcγ receptor; (2) A step of providing a polypeptide comprising the Fc region of wild-type or natural IgG, and a polypeptide comprising the Fc region of IgG comprising at least one amino acid mutation compared to wild-type or natural IgG; (3) A step of measuring the binding activity of the non-natural Fcγ receptor obtained in (1) to polypeptides containing the Fc region of wild-type or natural IgG; (4) A step of measuring the binding activity of the non-native Fcγ receptor obtained in (1) to a polypeptide containing the Fc region of IgG containing at least one amino acid mutation; and (5) A step of selecting from the unnatural Fcγ receptors obtained in (1) a nonnatural Fcγ receptor that does not essentially have binding activity to a polypeptide containing the Fc region of wild-type or natural IgG, and that binds to a polypeptide containing the Fc region of IgG containing at least one amino acid mutation. 【0159】 Embodiments relating to the polypeptides and CD16A-expressing cells of the present invention used in the methods of the present invention are similarly applicable to this description of the polypeptides of the present invention. 【0160】 In one aspect, the present invention provides a method for preparing a binding pair comprising (a) a polypeptide containing a modified Fc region of IgG and (b) a non-native modified Fcγ receptor. The method comprises the following steps: (1) A step of providing a polypeptide comprising the Fc region of wild-type or natural IgG and a polypeptide comprising a modified Fc region of IgG, wherein the modified Fc region of IgG is non-natural and comprises at least one amino acid mutation compared to the Fc region of wild-type or natural IgG; (2) A step of providing a wild-type or native Fcγ receptor and a non-native modified Fcγ receptor, wherein the modified Fcγ receptor comprises at least one amino acid mutation compared to the wild-type or native Fcγ receptor; (3) A step of measuring the binding activity of each Fcγ receptor obtained in (2) to each polypeptide obtained in (1); and (4) The step of selecting (a) a polypeptide comprising a modified Fc region that binds to a modified Fcγ receptor and which essentially has no binding activity to wild-type or native Fcγ receptors, and (b) a modified Fcγ receptor that binds to a polypeptide comprising a modified Fc region and which does not bind to the Fc region of wild-type or native IgG. 【0161】 In some embodiments, a polypeptide comprising a modified Fc region of IgG is an antibody. Step (4) may further include contacting the antibody with immune cells expressing an Fcγ receptor and cells expressing an antigen to which the antibody binds, and then measuring ADCC activity. 【0162】 Embodiments relating to the polypeptides and CD16A-expressing cells of the present invention used in the methods of the present invention are similarly applicable to this description of the polypeptides of the present invention. 【0163】 In the method described above, each step can be carried out by a person skilled in the art using any method known to a person skilled in the art or by a method described herein. 【0164】 Next, to further understand the present invention, specific examples are provided for reference, but these are provided for illustrative purposes only, and the present invention is not limited to these examples. [Examples] 【0165】 Human CD16A has two genetic polymorphisms: CD16A V158 and CD16A F158. The following examples were carried out using CD16A V158 (hereinafter referred to as "CD16V"), which has higher binding activity to antibody Fc. Experiments using commercially available kits or reagents were carried out according to the accompanying protocol unless the method was specifically described. 【0166】 Example 1: Protein design using computer calculations By analyzing the three-dimensional structure of complex proteins, it has been reported that introducing mutations to charged amino acids, which are considered important for complex formation, affects the binding activity and stability of the complex, and that the effect of introducing mutations to charged amino acids on binding activity can be predicted using computer calculations (Scientific Reports (2019) 9, pp. 4482). Therefore, the inventors analyzed the three-dimensional structure of the CD16V-antibody Fc complex (PDB code 3ay4) using MOE software (Chemical Computing Group) and extracted basic amino acid residues or acidic amino acid residues present at the binding interface between CD16V and antibody Fc. From these amino acid residues, mutants were designed in which basic amino acids on CD16V were replaced with acidic amino acids, or acidic amino acids on antibody Fc were replaced with basic amino acids (Table 1). The names of the CD16V mutants described in the following examples follow the amino acid residue numbers of the CD16A protein registered as 3ay4. This is based on the sequence of the CD16V protein (GenBank accession number AAH17865.1) excluding the amino acid sequence from positions 1 to 18 (SEQ ID NO: 78) ("3ay4" in the table). The mutation corresponding to the mutation shown in "3ay4" in the CD16V protein, which includes the amino acid sequence from positions 1 to 18, is shown in the "AAH17865.1" column of Table 1. 【0167】 The numbers below Fc in the table indicate the amino acid position within the constant region of human Igγ1, according to EU index number assignment. 【0168】 (Table 1) TIFF0007875855000001.tif76160 【0169】 Example 2: Preparation of CD16V protein and CD16V mutant protein In this embodiment, the protein into which the CD16V mutation has been introduced will be denoted as "CD16V_introduced_mutation" and will be collectively referred to as the CD16V mutant below. 【0170】 To obtain the CD16V protein, a gene (SEQ ID NO: 1) encoding the polypeptide in the extracellular component of CD16V (amino acids 1-208 of GenBank accession number AAH17865.1), with a FLAG sequence (DYKDDDDK, SEQ ID NO: 91) ligated to its C-terminus, was subcloned into a pcDNA3.4 vector (Thermo Fisher Scientific, catalog A14697). The constructed vector was then transfected into ExpiCHO-S cells (Thermo Fisher Scientific, catalog A29133). To obtain the CD16V mutant protein, the amino acid mutations shown in Table 1 (CD16V_K120D, CD16V_K120E, CD16V_K128D, CD16V_K128E, CD16V_K131D, CD16V_K131E, CD16V_K161D, or CD16V_K161E) and genes encoding the extracellular polypeptide of CD16V having a FLAG sequence linked to the C-terminus (SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, or SEQ ID NO: 17) were introduced into the pcDNA3.4 vector. The constructed vector was then transfected into ExpiCHO-S cells. CD16V protein and CD16V mutant proteins were purified from the culture supernatant of ExpiCHO-S cells using a standard method with anti-FLAG® M2 antibody affinity gel (SIGMA-ALDRICH, catalog A2220). Note that the amino acid sequence at positions 1-18 of CD16V in AAH17865.1 is cleaved in the mature form. The positions of the amino acid mutations introduced into the CD16V protein described in this example follow the amino acid numbers registered in 3ay4, and are numbered based on CD16V excluding the amino acid sequence at positions 1-18. Each mutation in the amino acid sequence of each CD16V mutant protein listed in the sequence listing corresponds to the mutation shown in the "AAH17865.1" column of Table 1. 【0171】 Example 3: Preparation of Fc_wt type anti-HER2 antibody or mutant Fc type anti-HER2 antibody In the following study, trastuzumab (Drug Bank accession number DB00072) was used as an anti-HER2 antibody. 【0172】 Antibodies containing the Fc sequence of the wild-type human Igγ1 constant region are collectively called Fc_wt. The expression vector used to produce Fc_wt-type anti-HER2 antibodies was constructed in the following manner: Signal sequence A heavy chain expression vector was constructed by inserting a polynucleotide of the gene encoding the trastuzumab heavy chain variable region (SEQ ID NO: 21), to which the gene encoding TIFF0007875855000002.tif4128 (SEQ ID NO: 19) was attached to the 5' end and the gene encoding the human Igγ1 constant region (SEQ ID NO: 23) was attached to the 3' end, into a pcDNA3.4 vector. Signal sequence A light chain expression vector was constructed by inserting a polynucleotide of the gene encoding the trastuzumab light chain variable region (SEQ ID NO: 27), with the gene encoding TIFF0007875855000003.tif4128 (SEQ ID NO: 25) appended to the 5' end, into a pcDNA3.4 vector. This light chain expression vector will be hereafter referred to as the trastuzumab light chain expression vector. These vectors were co-transfected into ExpiCHO-S cells, and an anti-HER2 antibody containing the wild-type Fc region (hereafter referred to as the Fc_wt type) was prepared from the culture supernatant according to standard procedures. 【0173】 Antibodies containing mutations in the Fc region (hereinafter referred to as "Fc_transmutations," and collectively as mutant Fc antibodies) were prepared. 【0174】 signal sequence A heavy chain expression vector used in the production of Fc_S239K, Fc_S239R, Fc_E294K, and Fc_E294R was constructed by introducing the polynucleotide of the gene encoding the trastuzumab heavy chain variable region (SEQ ID NO: 21) into a pcDNA3.4 vector. This vector had the gene encoding TIFF0007875855000004.tif4128 (SEQ ID NO: 19) attached to the 5' end, and genes encoding the human Igγ1 constant region (SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 29, and SEQ ID NO: 31) with amino acid mutations for substitution with lysine (K) or arginine (R) at S239 or E294 attached to the 3' end. A heavy chain expression vector used in the production of Fc_D265K, Fc_D265R, Fc_E269K, and Fc_E269R was constructed by introducing a polynucleotide of the gene encoding the trastuzumab heavy chain variable region (SEQ ID NO: 39) into a pcDNA3.4 vector. This vector had the gene encoding TIFF0007875855000005.tif4128 (SEQ ID NO: 37) attached to the 5' end, and genes encoding the human Igγ1 constant region (SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, and SEQ ID NO: 47) with amino acid mutations introduced to substitute lysine (K) or arginine (R) at D265 or E269 attached to the 3' end. Each of these expression vectors was co-transfected into ExpiCHO-S cells along with a trastuzumab light chain expression vector, and mutant Fc-type anti-HER2 antibodies were prepared from the culture supernatant according to standard procedures. 【0175】 Example 4: Evaluation of the binding activity of Fc_wt type anti-HER2 antibody and mutant Fc type anti-HER2 antibody to CD16V protein and CD16V mutant protein. The binding activity of the CD16V and CD16V mutant obtained in Example 2 to the Fc_wt type anti-HER2 antibody and mutant Fc type anti-HER2 antibody obtained in Example 3 was evaluated. HER2 protein (Sino Biological, catalog 10004-H08H) was diluted to 2 μg / mL in phosphate-buffered saline (PBS), and 20 μL of the diluted HER2 protein was added to each well of a Maxisorp 384-well clear plate (Thermo Fisher Scientific, catalog 464718) and fixed by incubation at 4°C overnight. After removing the HER2 protein solution, the plate was incubated with 50 μL of PBS containing Blocking One (Nacalai Tesque, catalog 03953-95) at room temperature for 1 hour, and then washed with Tris-buffered saline containing 0.05% Tween 20 (TBS-T, Nippon Gene, catalog 310-07375). Each anti-HER2 antibody obtained in Example 3 was diluted to 4 μg / mL with TBS-T containing 5% Blocking One (hereinafter simply referred to as the diluent), and 20 μL of the diluted antibody was added to each well. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. Each CD16V protein obtained in Example 2 was diluted with the diluent, and a dilution series was prepared with a common ratio of approximately 3 times from the highest concentration of 10 μg / mL, and 20 μL of the diluted CD16V protein was added to each well. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. Next, 20 μL of horseradish peroxidase-labeled anti-FLAG® M2 antibody (SIGMA-ALDRICH, catalog A8592), diluted with the diluent, was added to each well as the detection antibody. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. After adding a TMB + substrate-dye source (DAKO, catalog S1599) and incubating, the reaction was stopped by adding 1M sulfuric acid, and the absorbance at 450 nm and the reference wavelength was measured using Infinite 200 PRO (TECAN). 【0176】 As a result, CD16V_K131D did not bind to Fc_wt, but bound only to Fc_E269R. However, the binding activity between CD16V_K131D and Fc_E269R was lower than the binding activity between Fc_wt and CD16V. Also, Fc_E269R bound to CD16V to a small extent. Both CD16V_K128E and CD16V_K131E were confirmed to bind to mutant Fc, but binding to Fc_wt was only partially confirmed. On the other hand, it was confirmed that CD16V_K128D, CD16V_K120D, CD16V_K120E, CD16V_K161D, and CD16V_K161E did not bind to either Fc_wt or mutant Fc at all, or bound very little (Figure 1). 【0177】 This suggests that positions K128 and K131 in CD16V, and positions E269 and E294 in the Fc region of the antibody, are important amino acid residues for the specific binding activity between the CD16V mutant and the mutant Fc antibody. Therefore, in order to obtain mutant Fc and CD16V mutants that have specific amino acid mutations introduced and bind only specifically to each other, we conducted studies using combinations of these mutations and combinations of these mutations with known mutations that enhance binding activity. 【0178】 Example 5: Preparation of mutant Fc-type anti-HER2 antibody and CD16V mutant protein By combining mutant K128E and K131D or mutant K128E and K131E with CD16V, CD16V_K128E_K131D (hereinafter referred to as CD16V_ED) and CD16V_K128E_K131E (hereinafter referred to as CD16V_EE) were prepared. Specifically, as described in Example 2, an expression vector was constructed by introducing genes (SEQ ID NO: 49 and SEQ ID NO: 51) that have these amino acid mutations and encode the extracellular polypeptide of the CD16V protein linked to a FLAG sequence introduced at the C-terminus into a pcDNA3.4 vector. This expression vector was then transfected into ExpiCHO-S cells. CD16V_ED and CD16V_EE were prepared from their respective culture supernatants in the same manner as in Example 2. 【0179】 It has been reported that inducing N38Q, N74Q, or N169Q mutations in CD16V slightly increases its binding activity to Fc_wt (Journal of Biological Chemistry (2018) 293, pp. 16842-16850). Since CD16V_K131D (hereinafter referred to as CD16V_D) bound only to the Fc_E269R type antibody, CD16V_K131D_N38Q (hereinafter referred to as CD16V_DQ1) and CD16V_K131D_N74Q (hereinafter referred to as CD16V_DQ2) were prepared in which CD16V_D was combined with the N38Q mutation or the N74Q mutation. Specifically, as in Example 2, expression vectors were constructed by introducing genes (SEQ ID NO: 53 and SEQ ID NO: 55) encoding polypeptides of the extracellular domain of the CD16V protein linked to a C-terminal FLAG sequence, in which one of these amino acid mutations was introduced, into a pcDNA3.4 vector. Then, each expression vector was transfected into ExpiCHO-S cells. CD16V_DQ1 and CD16V_DQ2 were prepared from their respective culture supernatants, as in Example 2. 【0180】 Anti-HER2 antibodies of the Fc_E269R_E294K (hereinafter referred to as Fc_RK) type and Fc_E269R_E294R (hereinafter referred to as Fc_RR) type were prepared. Specifically, in the same manner as in Example 3, the signal sequence An expression vector was constructed by introducing a polynucleotide of the gene encoding the trastuzumab heavy chain variable region (SEQ ID NO: 39) into a pcDNA3.4 vector. The gene encoding TIFF0007875855000006.tif4128 (SEQ ID NO: 37) was added to the 5' end, and genes encoding the human Igγ1 constant region (SEQ ID NO: 57 and SEQ ID NO: 59), which had an amino acid mutation to arginine at position E269 and an amino acid mutation to lysine (K) or arginine (R) at position E294, were added to the 3' end. The resulting heavy chain expression vector and the trastuzumab light chain expression vector obtained in Example 3 were co-transfected into ExpiCHO-S cells, and mutant Fc-type anti-HER2 antibodies were prepared from the respective culture supernatants according to standard methods. 【0181】 Example 6: Evaluation of the binding activity of Fc_wt type anti-HER2 antibody and mutant Fc type anti-HER2 antibody to CD16V protein and CD16V mutant protein. The binding activity of CD16V_D obtained in Example 2 and CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2 obtained in Example 5 to the Fc_wt type anti-HER2 antibody obtained in Example 3 or the Fc_RK type anti-HER2 antibody or Fc_RR type anti-HER2 antibody obtained in Example 5 was evaluated in the same manner as in Example 4 (Figure 2). 【0182】 As a result, the Fc_wt type anti-HER2 antibody bound to CD16V, but did not show binding activity to CD16V_D, CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2. On the other hand, the Fc_RK type anti-HER2 antibody or the Fc_RR type anti-HER2 antibody bound to the CD16V mutant but not to CD16V. Therefore, it is clear that introducing a mutation at the E294 position into Fc_E269R increased the binding specificity of the mutant Fc type antibody against the CD16V mutant. Furthermore, the binding activity of CD16V_DQ1 and CD16V_DQ2 to Fc_RK and Fc_RR was almost the same as the binding activity of CD16V to Fc_wt. Therefore, it is clear that CD16V_DQ1 and CD16V_DQ2, in which additional mutations have been introduced into CD16V_D, do not have enhanced Fc_wt binding activity, but only Fc_RR binding activity and Fc_RK binding activity are increased. 【0183】 Example 7: Preparation of anti-EGFR antibodies and anti-EpCAM antibodies of the Fc_wt type and mutant Fc type. To investigate whether the Fc sequence prepared in Example 5 has specific binding activity to CD16V_D obtained in Example 2 and CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2 obtained in Example 5 when used in antibodies other than anti-HER2 antibodies, anti-EGFR antibodies and anti-EpCAM antibodies of the Fc_wt, Fc_RK, and Fc_RR types were prepared. 【0184】 In the production of Fc_wt, Fc_RK, and Fc_RR type anti-EGFR antibodies, the signal sequence A heavy chain expression vector was prepared by introducing a polynucleotide of the gene encoding the heavy chain variable region of anti-EGFR (cetuximab, Drug Bank accession number DB00002) (SEQ ID NO: 63) into a pcDNA3.4 vector. The gene encoding TIFF0007875855000007.tif4128 (SEQ ID NO: 61) was added to the 5' end, and a gene encoding the human Igγ1 constant region (SEQ ID NO: 23), a gene encoding the human Igγ1 constant region containing Fc_RK (SEQ ID NO: 57), or a gene encoding the human Igγ1 constant region containing Fc_RR (SEQ ID NO: 59) was added to the 3' end. The signal sequence was also prepared. A light chain expression vector was also prepared by introducing a gene encoding the cetuximab light chain region (SEQ ID NO: 67), to which the gene encoding TIFF0007875855000008.tif4128 (SEQ ID NO: 65) was appended to the 5' end, into a pcDNA3.4 vector. 【0185】 In the production of Fc_wt, Fc_RK, and Fc_RR type anti-EpCAM antibodies, the signal sequence A heavy chain expression vector was prepared by introducing a polynucleotide of the gene encoding the heavy chain variable region of anti-EpCAM (edrecolomab, IMGT INN number 7471) (SEQ ID NO: 71) into a pcDNA3.4 vector. The gene encoding TIFF0007875855000009.tif4128 (SEQ ID NO: 69) was added to the 5' end, and a gene encoding the human Igγ1 constant region (SEQ ID NO: 23), a gene encoding the human Igγ1 constant region containing Fc_RK (SEQ ID NO: 57), or a gene encoding the human Igγ1 constant region containing Fc_RR (SEQ ID NO: 59) was added to the 3' end. The signal sequence was also prepared. A light chain expression vector was also prepared by introducing a gene encoding the light chain region of edrecolomab (SEQ ID NO: 75), to which the gene encoding TIFF0007875855000010.tif4128 (SEQ ID NO: 73) was appended to the 5' end, into a pcDNA3.4 vector. 【0186】 Each of these heavy chain expression vectors was co-transfected into ExpiCHO-S cells with either a cetuximab light chain expression vector or an edrecolomab light chain expression vector, and anti-EGFR antibodies and anti-EpCAM antibodies were purified in the same manner as in Example 3. 【0187】 Example 8: Evaluation of the binding activity of Fc_wt type and mutant Fc type anti-EGFR antibodies or anti-EpCAM antibodies against CD16V protein and CD16V mutant protein. The binding activity of CD16V, CD16V_D obtained in Example 2, and CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2 obtained in Example 5, against the Fc_wt, Fc_RK, or Fc_RR type anti-EGFR antibody or anti-EpCAM antibody obtained in Example 7 was evaluated. EGFR protein (Abcam, catalog ab155639) was diluted to 4 μg / mL in PBS, or EpCAM protein (Sino Biological, catalog 10694-H08H) was diluted to 2 μg / mL in PBS. 20 μL of the diluted protein was added to each well of a Maxisorp 384-well clear plate and fixed by incubation at 4°C overnight. The following day, the EGFR protein solution or EpCAM protein solution was removed, and the plate, with 50 μL of PBS containing Blocking One added, was incubated at room temperature for 1 hour, and then washed with TBS-T. Each anti-EGFR antibody obtained in Example 7 was diluted to 2 μg / mL with a diluent, and each anti-EpCAM antibody obtained in the same example was diluted to 10 μg / mL with a diluent. 20 μL of the diluted antibody was added to each well. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. Each CD16V protein obtained in Example 2 or Example 5 was diluted with a diluent to prepare a dilution series with a common ratio of approximately 3 times from the highest concentration of 10 μg / mL. 20 μL of the diluted CD16V protein was added to each well. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. Next, 20 μL of horseradish peroxidase-labeled anti-FLAG® M2 antibody, diluted with a diluent, was added to each well as the detection antibody. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. After adding a TMB+ substrate-dye source and incubating, the reaction was stopped by adding 1M sulfuric acid, and the absorbance at 450 nm and the reference wavelength was measured using Infinite 200 PRO. 【0188】 In the case of anti-EGFR antibodies and anti-EpCAM antibodies, as with anti-HER2 antibodies, Fc_wt type antibodies bound to CD16V but not to the CD16V mutant, while Fc_RK type antibodies and Fc_RR type antibodies did not bind to CD16V but bound only to the CD16V mutant (Figures 3 and 4). These results clearly indicate that Fc_RK and Fc_RR in other antibodies, as well as in anti-HER2 antibodies, specifically bind to the CD16V mutant. 【0189】 Example 9: Evaluation of the binding activity of mutant Fc-type anti-HER2 antibody to CD16V protein and CD16V mutant protein in the presence of excess IgG1 antibody. To confirm the binding activity between the mutant Fc-type anti-HER2 antibody and the CD16V mutant protein in vivo, a competitive assay was performed using the Fc_wt-type antibody. The Fc_wt-type antibody used as the endogenous immunoglobulin mimetic was a human IgG1 antibody against keyhole limpet hemocyanin (KLH), an antigen not present in vivo. This antibody was obtained by standard methods. As in Example 4, the HER2 protein was immobilized on a Maxisorp 384-well clear plate. The following day, after removing the HER2 protein solution, the plate, containing Blocking One and PBS, was incubated at room temperature for 1 hour and then washed with TBS-T. The Fc_wt-type, Fc_RK-type, and Fc_RR-type anti-HER2 antibodies were added to each well and incubated at room temperature for 1 hour. Each CD16V protein and CD16V mutant protein was diluted with a diluent to prepare a dilution series with a common ratio of approximately 3x from the highest concentration of 20 μg / mL. These were then mixed 1:1 with the diluent or an anti-KLH antibody adjusted to 2 mg / mL with the diluent. After washing the plate with TBS-T, 20 μL of these mixed solutions were added and incubated at room temperature for 1 hour. At this time, a common ratio of approximately 3x from the final CD16V protein concentration of 10 μg / mL was present. After washing the plate with TBS-T, 20 μl of anti-FLAG® M2 antibody labeled with horseradish peroxidase, diluted 2,000-fold with the diluent, was added to each well as the detection antibody. After incubation at room temperature for 1 hour, the plate was washed with TBS-T. After adding a TMB+ substrate-dye source and incubating, the reaction was stopped by adding 1 M sulfuric acid, and the absorbance at 450 nm and the reference wavelength was measured using Infinite 200 PRO. 【0190】 As a result, the binding activity between CD16V and Fc_wt-type anti-HER2 antibody in the presence of anti-KLH antibody was very low at the highest concentration of CD16V (10 μg / mL), and the absorbance when reacting with CD16V at 3 μg / mL or less was at the same level as the background. On the other hand, the binding activity between Fc_RK-type and Fc_RR-type anti-HER2 antibodies and each CD16V variant (CD16V_D, CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2) in the presence of anti-KLH antibody decreased, but remained within a certain range of decrease (Figure 5). Since the majority of immunoglobulins in serum is IgG1, this suggests that mutant Fc-type antibodies and CD16V variants show binding at lower concentrations than Fc_wt and CD16V in serum. 【0191】 Example 10: Establishment of KHYG-1 cells expressing CD16V It is known that the CD16V sequence has both a mature form (SEQ ID NO: 78) and an immature form (GenBank accession number: AAH17865.1, SEQ ID NO: 90). The gene encoding the immature form (SEQ ID NO: 89) was inserted into the multi-cloning site of the pLVSIN-CMV Pur vector (Takara Bio, catalog 6183). Similarly, the genes encoding the amino acid sequences (SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88) of the CD16V mutants (CD16V_D, CD16V_ED, CD16V_EE, CD16V_DQ1, CD16V_DQ2) (SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88) (SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87) were introduced into the pLVSIN-CMV Pur vector to construct vectors for each CD16V mutant. Lenti-X® 293T cell line (Takara Bio, catalog 632180) was transfected with a high-titer lentivirus packaging mix (Takara Bio, catalog 6194) along with lipofectamine 2000 (Thermo Fisher Scientific, catalog 11668027), and the resulting culture supernatant was collected. PEG-it virus precipitate solution (5×) (System Biosciences, catalog LV810A-1) was added to each culture supernatant, and the cells were incubated overnight at 4°C to concentrate lentiviral vectors expressing CD16V or CD16V mutants. KHYG-1 cells (JCRB Cell Bank, JCRB0156) were infected with a lentiviral vector, and transgenic cells were selected using puromycin to prepare KHYG-1 cells expressing the CD16V gene or the CD16V mutant gene, resulting in KHYG-1 (CD16V / KHYG-1) cells expressing the CD16V gene or the CD16V mutant. 【0192】 The expression levels of CD16V or CD16V mutant expressed in prepared CD16V / KHYG-1 cells or CD16V mutant / KHYG-1 cells were measured by flow cytometry. A phycoerythrin-labeled anti-human CD16 antibody (clone: ​​3G8, Biolegend, catalog 302008) was added to each KHYG-1 cell suspended in a STAIN BUFFER (BD Bioscience, catalog 554656) and incubated on ice for 20 minutes. After washing the cells three times with the STAIN BUFFER, 7-AAD (BD Bioscience, catalog 559925) was added and incubated in the dark for 15 minutes. The fluorescence intensity of phycoerythrin was then measured in the 7-AAD-negative viable cell fraction using a FACS array (BD Bioscience). FlowJo (BD Bioscience) was used for analysis. As a result, all established cells expressed CD16V or the CD16V mutant at an expression rate of 75% or higher (Figure 6). 【0193】 Example 11: Evaluation of antibody ADCC activity using KHYG-1 cells expressing CD16V To evaluate the ADCC activity of CD16V / KHYG-1 cells or CD16V mutant / KHYG-1 cells, 10,000 HER2-positive SK-BR-3 cells and 100,000 CD16V / KHYG-1 cells or CD16V mutant / KHYG-1 cells, stained with calcein AM solution (Dojindo Laboratories, C396) in a 1:10 ratio, were incubated for 4 hours in 200 μL of medium in a 96-well plate (CORNING, catalog 353077) in the presence of each anti-HER2 antibody diluted in a 4-fold dilution series starting from a maximum concentration of 4 μg / mL. The fluorescence intensity derived from calcein AM in 100 μL of the supernatant was measured using FlexStation 3 (Molecular Devices) to observe the viability of SK-BR-3 cells. Cytotoxic activity was calculated using the following formula. In the formula, T-MAX is the value when 1% TritonX-100 (SIGMA-ALDRICH, catalog 30-5140) is added to SK-BR-3 cells, and T-Spon is the value when only culture medium is added. Cytotoxic activity (%) (lysis rate (%)) = 100 × (data - (T-Spon)) / ((T-MAX) - (T-Spon)) 【0194】 As a result, CD16V / KHYG-1 cells showed concentration-dependent cytotoxicity against SK-BR-3 cells only with the Fc_wt type anti-HER2 antibody. Cytotoxicity was extremely low with the Fc_RK type anti-HER2 antibody and the Fc_RR type anti-HER2 antibody. On the other hand, the Fc_wt type anti-HER2 antibody showed no cytotoxic activity or very low cytotoxic activity in all CD16V mutant / KHYG-1 cells evaluated. Only the Fc_RK type anti-HER2 antibody and the Fc_RR type anti-HER2 antibody showed cytotoxic activity in all CD16V mutant / KHYG-1 cells evaluated (Figure 7). Therefore, it is clear that CD16V mutants and Fc mutants possess specific binding activity, and that the combination exhibits specificity in ADCC induction. 【0195】 Example 12: Evaluation of ADCC activity in the presence of human serum using KHYG-1 cells expressing CD16V. To identify the effect of human serum on the ADCC activity of CD16V / KHYG-1 cells or CD16V mutant / KHYG-1 cells, cytotoxicity against SK-BR-3 cells was evaluated in the same manner as in Example 11, using 1 μg / mL of each anti-HER2 antibody added to a medium containing 5% fetal bovine serum (FBS) or 5% human serum. The complement activity of each serum was thermally inactivated to avoid interference with the evaluation of ADCC activity due to the induction of complement-dependent cytotoxicity. 【0196】 As a result, the cytotoxicity of CD16V / KHYG-1 against SK-BR-3 in the case of Fc_wt type anti-HER2 antibodies was reduced in the presence of human serum compared to FBS. On the other hand, the cytotoxicity of CD16V mutant / KHYG-1 against SK-BR-3 in the case of Fc_RK type anti-HER2 antibodies in the presence of human serum was almost the same as in the presence of FBS (Figure 8). Similarly, the cytotoxicity of CD16V mutant / KHYG-1 against SK-BR-3 in the case of Fc_RR type anti-HER2 antibodies in the presence of human serum was almost the same as in the presence of FBS, except for CD16V_EE / KHYG-1 (Figure 8). 【0197】 Therefore, these results indicate that human serum interferes with the interaction between Fc_wt type antibodies and CD16V, but Fc_RK type antibodies or Fc_RR type antibodies and human serum IgG compete less for ADCC-inducing activity against CD16V variant / KHYG-1. 【0198】 Example 13: Evaluation of cytotoxic activity of T cells expressing CD16V CAR CAR-T cells are known to be one of the most potent effector cells against cancer cells in cancer immunotherapy. To broaden the applications of the CD16V mutant and Fc mutant combination, we expressed a chimeric receptor (CD16V CAR or CD16V mutant CAR) in which the extracellular domain of CD16V or the CD16V mutant is fused with a signaling domain in primary T cells, and evaluated the cytotoxic activity of these T cells expressing CD16V CAR or the CD16V mutant CAR (CD16V / CAR-T or CD16V mutant / CAR-T) against cancer cells. 【0199】 The gene (SEQ ID NO: 92) encoding the amino acid sequence of CD16V CAR, i.e., a fusion protein of the extracellular domain of CD16V and the signaling domains of CD3ζ and CD137 via the CD8a hinge and CD8 transmembrane domain, and the signaling sequence The gene encoding TIFF0007875855000011.tif4128, with the gene appended to its 5' end, was inserted into the multi-cloning site of the pLVSIN-EF1α Neo Vector (Takara Bio, catalog 6184). Similarly, the signal sequence was inserted. Each gene encoding the amino acid sequence of the CAR of each CD16V mutant (CD16V_D, CD16V_ED, CD16V_EE, CD16V_DQ1, and CD16V_DQ2), with the gene encoding TIFF0007875855000012.tif4128 appended to the 5' end (SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, and SEQ ID NO: 102), was inserted into the multi-cloning site of the pLVSIN-EF1α Neo Vector. Using these vectors, lentiviral vectors expressing CD16V or CD16V mutants were prepared in the same manner as in Example 10. 【0200】 CD16V / CAR-T and CD16V mutant / CAR-T cells were prepared by infecting human T cells purified from human peripheral blood mononuclear cells (Lonza, catalog CC-2702) with a lentiviral vector using a pan-T cell isolation kit (Miltenyi Biotec, catalog 130-096-535). The expression rate of CD16V or each CD16V mutant in T cells exceeded 60%. To evaluate the cytotoxic activity of CD16V / CAR-T or CD16V mutant / CAR-T cells, 10,000 SK-BR-3 cells and 150,000 CD16V / CAR-T or CD16V mutant / CAR-T cells, stained with calcein AM solution in a 1:15 ratio, were incubated for 4 hours in 200 μL of medium in the presence of each anti-HER2 antibody diluted in a 4-fold dilution series from a peak concentration of 16 μg / mL. As in Example 11, the fluorescence intensity derived from calcein AM in 100 μL of supernatant was measured, and the cytotoxic activity was calculated. 【0201】 As a result, CD16V / CAR-T showed concentration-dependent cytotoxicity against SK-BR-3 cells only with the Fc_wt type anti-HER2 antibody. Cytotoxicity was extremely low with the Fc_RK type and Fc_RR type anti-HER2 antibodies. On the other hand, the Fc_wt type anti-HER2 antibody either did not induce cytotoxic activity or induced very low cytotoxic activity in all CD16V mutant / CAR-T cells. Only the Fc_RK type and Fc_RR type anti-HER2 antibodies induced cytotoxic activity in all CD16V mutant / CAR-T cells (Figure 9). Therefore, it is clear that the combination of CD16V mutant and Fc mutant maintains specificity when applied to a chimeric protein similar to CAR. 【0202】 The Fcγ receptor variants and Fc region variants of the present invention do not inherently bind to endogenous immunoglobulins or endogenous Fcγ receptors, but the Fcγ receptor variants and Fc region variants specifically bind to each other. Therefore, these combinations are expected to enable immunotherapy without endogenous molecules reducing drug efficacy. 【0203】 Equivalents Those skilled in the art will recognize, or confirm through routine experiments, many equivalents of the specific embodiments and methods described herein. Such equivalents are intended to be included in the appended claims. 【0204】 [Sequence Listing Free Text] The artificial sequences are described in the numbered headings of the sequence list below. <223> It is described below. Specifically, the nucleotide sequence indicated by SEQ ID NO: 1 in the sequence listing is a nucleotide sequence that codes for a protein in which the FLAG sequence is ligated to the C-terminus of the extracellular segment of the CD16V sequence. The amino acid sequence indicated by SEQ ID NO: 2 is the amino acid sequence coded by SEQ ID NO: 1. The nucleotide sequences indicated by SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, and SEQ ID NO: 17 are nucleotide sequences that code for a protein in which the FLAG sequence is ligated to the C-terminus of the protein and a mutation has been introduced into the extracellular segment of the CD16V sequence. The amino acid sequences shown in SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, and SEQ ID NO: 18 are the amino acid sequences encoded by SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, and SEQ ID NO: 17, respectively. SEQ ID NO: 19 and SEQ ID NO: 37 are nucleotide sequences that encode signal sequences ligated to the N-terminus of the gene encoding the heavy chain variable region of trastuzumab. SEQ ID NO: 21 and SEQ ID NO: 39 are nucleotide sequences encoding the heavy chain variable region of trastuzumab, and SEQ ID NO: 23 is a nucleotide sequence encoding the human Igγ1 constant region.SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 38, and SEQ ID NO: 40 are amino acid sequences encoded by SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 37, and SEQ ID NO: 39, respectively. SEQ ID NO: 27 is a nucleotide sequence encoding the light chain region of trastuzumab, and SEQ ID NO: 28 is the amino acid sequence encoded by SEQ ID NO: 27. SEQ ID NO: 25 is a nucleotide sequence encoding a signal sequence ligated to the 5' end of the light chain region of trastuzumab, and SEQ ID NO: 26 is the amino acid sequence encoded by SEQ ID NO: 25. SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, and SEQ ID NO: 35 are nucleotide sequences encoding proteins in which mutations have been introduced into the gene encoding the human Igγ1 constant region, while SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, and SEQ ID NO: 36 are amino acid sequences encoded by SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, and SEQ ID NO: 35, respectively. The nucleotide sequences indicated by SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, and SEQ ID NO: 47 are nucleotide sequences encoding a protein in which an amino acid mutation has been introduced into the gene encoding the human Igγ1 constant region, while SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, and SEQ ID NO: 48 are the amino acid sequences encoded by SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, and SEQ ID NO: 47, respectively.The nucleotide sequences shown in SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, and SEQ ID NO: 55 are nucleotide sequences that encode a protein in which a FLAG sequence is ligated to the C-terminus of the protein and a mutation has been introduced into the extracellular segment of the CD16V sequence. The amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, and SEQ ID NO: 56 are the amino acid sequences encoded by SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, and SEQ ID NO: 55, respectively. The nucleotide sequences shown in SEQ ID NO: 57 and SEQ ID NO: 59 are nucleotide sequences that encode a protein in which a mutation has been introduced into the human Igγ1 constant region. SEQ ID NO: 58 and SEQ ID NO: 60 are the amino acid sequences encoded by SEQ ID NO: 57 and SEQ ID NO: 59. SEQ ID NO: 61 is a nucleotide sequence encoding a signal sequence ligated to the N-terminus of the gene encoding the heavy chain variable region of cetuximab, and SEQ ID NO: 65 is a nucleotide sequence encoding a signal sequence ligated to the N-terminus of the gene encoding the light chain region of cetuximab. SEQ ID NO: 62 and SEQ ID NO: 66 are the amino acid sequences encoded by SEQ ID NO: 61 and SEQ ID NO: 65, respectively. SEQ ID NO: 63 and SEQ ID NO: 67 are nucleotide sequences encoding the heavy chain variable region and light chain region of cetuximab, respectively, and SEQ ID NO: 64 and SEQ ID NO: 68 are the amino acid sequences encoded by SEQ ID NO: 63 and SEQ ID NO: 67, respectively. SEQ ID NO: 69 is a nucleotide sequence that encodes a signal sequence ligated to the N-terminus of the gene encoding the heavy chain variable region of edrecolomab, and SEQ ID NO: 73 is a nucleotide sequence that encodes a signal sequence ligated to the N-terminus of the gene encoding the light chain region of edrecolomab.SEQ ID NO: 70 and SEQ ID NO: 74 are amino acid sequences encoded by SEQ ID NO: 69 and SEQ ID NO: 73, respectively. SEQ ID NO: 71 and SEQ ID NO: 75 are nucleotide sequences encoding the heavy chain variable region and light chain region of edrecolomab, respectively, and SEQ ID NO: 72 and SEQ ID NO: 76 are amino acid sequences encoded by SEQ ID NO: 71 and SEQ ID NO: 75, respectively. SEQ ID NO: 77 is a nucleotide sequence encoding the mature CD16V sequence, and SEQ ID NO: 78 is the amino acid sequence encoded by SEQ ID NO: 77. SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, and SEQ ID NO: 87 are nucleotide sequences encoding the sequence of the CD16V mutant with the introduced mutation, and SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, and SEQ ID NO: 88 are amino acid sequences encoded by SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, and SEQ ID NO: 87, respectively. SEQ ID NO: 89 is a nucleotide sequence encoding the immature CD16V sequence, and SEQ ID NO: 90 is the amino acid sequence encoded by SEQ ID NO: 89. SEQ ID NO: 91 is the amino acid sequence of the FLAG. The nucleotide sequence shown in SEQ ID NO: 92 is a nucleotide sequence encoding a fusion protein of the extracellular partial protein of CD16V and the signal domains of CD3ζ and CD137 via the CD8a hinge and CD8 transmembrane domain, with the signal sequence ligated to the N-terminus. The amino acid sequence shown in SEQ ID NO: 93 is the amino acid sequence encoded by SEQ ID NO: 92.The nucleotide sequences shown in SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, and SEQ ID NO: 102 are nucleotide sequences encoding a fusion protein of a mutated extracellular partial protein of CD16V and the signal domains of CD3ζ and CD137 via the CD8a hinge and CD8 transmembrane domain, with the signal sequence ligated to the N-terminus. The amino acid sequences shown in SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, and SEQ ID NO: 103 are the amino acid sequences encoded by SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, and SEQ ID NO: 102, respectively. SEQ ID NO: 104 is the amino acid sequence of the signal sequence ligated to the N-terminus of the CD16V CAR protein or the CD16V mutant CAR protein.

Claims

[Claim 1] A polypeptide comprising a modified Fc region of IgG, The modified Fc region comprises an amino acid sequence that is non-natural and has 90% or more amino acid sequence identity with SEQ ID NO: 24, and at least one amino acid mutation compared to the Fc region of wild-type or natural IgG. The at least one amino acid mutation is (i) A mutation from glutamate to arginine (E269R mutation) at the position corresponding to position 269 of SEQ ID NO: 24, according to EU index number assignment. (ii) A mutation from glutamate to arginine at position 294 of SEQ ID NO: 24 according to EU index number assignment (E294R mutation), and (iii) A mutation from glutamate to lysine (E294K mutation) at the position corresponding to position 294 of SEQ ID NO: 24, as assigned by the EU index number. Selected from the group consisting of, The polypeptide does not have binding activity to wild-type or native Fcγ receptors, and can bind to non-native Fcγ receptors that have at least one amino acid mutation compared to wild-type or native Fcγ receptors, and The non-natural Fcγ receptor contains an amino acid sequence that has more than 90% amino acid sequence identity with SEQ ID NO: 78, and The at least one amino acid mutation is (i) A mutation from lysine to aspartic acid at position 131 of SEQ ID NO: 78 (K131D mutation), (ii) A mutation from lysine to glutamate at position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) A mutation from lysine to glutamate at position 131 of SEQ ID NO: 78 (K131E mutation) Selected from the group consisting of, Polypeptide. [Claim 2] (a) The wild-type or natural Fcγ receptor is wild-type or natural CD16A, and the non-natural Fcγ receptor containing at least one amino acid mutation is a non-natural CD16A containing at least one amino acid mutation. (b) The wild-type or natural CD16A contains the amino acid sequence shown in SEQ ID NO: 78, (c) The non-natural CD16A containing at least one amino acid mutation contains either or both of the K131D mutation and the K128E mutation, (d) The non-natural CD16A containing at least one amino acid mutation contains either or both of the K131E mutation and the K128E mutation, (e) a non-natural CD16A containing at least one amino acid mutation contains the K131D mutation and further contains at least one mutation selected from (f) an asparagine-to-glutamine mutation at the position corresponding to position 38 of SEQ ID NO: 78 (N38Q mutation) and (g) an asparagine-to-glutamine mutation at the position corresponding to position 74 of SEQ ID NO: 78 (N74Q mutation), and / or (h) Non-natural CD16A containing at least one amino acid mutation contains the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88, The polypeptide according to claim 1. [Claim 3] (a) The polypeptide comprises a modified Fc region of human Igγ1 containing the amino acid sequence of SEQ ID NO: 24, The modified Fc region is (i) A mutation from glutamate to arginine (E269R mutation) at the position corresponding to position 269 of SEQ ID NO: 24 according to the EU index number assignment, (ii) At least one mutation selected from (a) a glutamate-to-arginine mutation at the position corresponding to position 294 of SEQ ID NO: 24 according to EU index number assignment (E294R mutation) and (b) a glutamate-to-lysine mutation at the position corresponding to position 294 of SEQ ID NO: 24 according to EU index number assignment (E294K mutation) Including, and / or (b) The polypeptide is an antibody, and / or (c) The polypeptide is an antibody that binds to a cancer antigen. The polypeptide according to claim 1 or 2. [Claim 4] A polypeptide according to any one of claims 1 to 3 for use in a method of treating or preventing a patient's disease or disorder using immunotherapy, The method includes the step of administering to the patient the polypeptide and cells expressing a non-natural Fcγ receptor having at least one amino acid mutation compared to the wild-type or natural Fcγ receptor, The polypeptide is capable of binding to the non-native Fcγ receptor, which contains at least one amino acid mutation. [Claim 5] (a) The cells are human immune cells selected from the group consisting of T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. (b) The cells are derived from stem cells selected from the group consisting of pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells, and / or The pluripotent stem cells are either induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). (c) The cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene, and / or The cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein that is covalently bound either directly or via a linker sequence to at least one portion of an HLA-1α chain selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. (d) The cells contain, through genetic manipulation, human leukocyte antigen (HLA) class II related genes selected from the group consisting of regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, and / or (e) The cell contains one or more polynucleotides encoding a single-chain fusion HLA class II protein or an HLA class II protein, A polypeptide for use according to claim 4. [Claim 6] The polypeptide for use according to claim 4 or 5, wherein the method is a method for treating or preventing cancer. [Claim 7] A pharmaceutical composition comprising a polypeptide according to any one of claims 1 to 3 and a pharmaceutically acceptable excipient. [Claim 8] A pharmaceutical composition according to claim 7 for use in combination with cells for immunotherapy, wherein the cells express a non-natural Fcγ receptor comprising at least one amino acid mutation compared to a wild-type or native Fcγ receptor, and the polypeptide is capable of binding to the non-natural Fcγ receptor comprising at least one amino acid mutation. (a) The cell is a human immune cell selected from the group consisting of T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. (b) The cells are derived from stem cells selected from the group consisting of pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells, and / or The pluripotent stem cells are either induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). (c) The cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene, and / or The cell further comprises a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein that is covalently bound either directly or via a linker sequence to at least one portion of an HLA-1α chain selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. (d) The cells contain genetically modified human leukocyte antigen (HLA) class II related genes selected from the group consisting of regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, and / or (e) The cell contains one or more polynucleotides encoding a single-chain fusion HLA class II protein or an HLA class II protein, The pharmaceutical composition according to claim 7. [Claim 9] A kit for treating or preventing a patient's disease or disability using immunotherapy, (i) a polypeptide according to any one of claims 1 to 3, and (ii) a cell expressing a non-native Fcγ receptor having at least one amino acid mutation compared to the wild-type or native Fcγ receptor, A kit comprising a polypeptide capable of binding to a non-native Fcγ receptor containing at least one amino acid mutation. [Claim 10] (a) The cells are human immune cells selected from the group consisting of T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. (b) The cells are derived from stem cells selected from the group consisting of pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells, and / or The pluripotent stem cells are either induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). (c) The cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene, and / or The cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein that is covalently bound either directly or via a linker sequence to at least one portion of an HLA-1α chain selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. (d) The cells contain, through genetic manipulation, human leukocyte antigen (HLA) class II related genes selected from the group consisting of regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, and / or (e) The cell contains one or more polynucleotides encoding a single-chain fusion HLA class II protein or an HLA class II protein, The kit according to claim 9. [Claim 11] A cell expressing a non-natural CD16A comprising an amino acid sequence having 90% or more amino acid sequence identity with SEQ ID NO: 78, and at least one amino acid mutation compared to wild-type or natural CD16A, The at least one amino acid mutation is selected from the group consisting of (i) a mutation from lysine to aspartate at the position corresponding to position 131 of SEQ ID NO: 78 (K131D mutation), (ii) a mutation from lysine to glutamate at the position corresponding to position 128 of SEQ ID NO: 78 (K128E mutation), and (iii) a mutation from lysine to glutamate at the position corresponding to position 131 of SEQ ID NO: 78 (K131E mutation). The non-natural CD16A does not have binding activity to polypeptides containing the Fc region of wild-type or natural IgG, and can bind to polypeptides containing the modified Fc region of IgG, and The modified Fc region includes an amino acid sequence that is non-natural and has 90% or more amino acid sequence identity with SEQ ID NO: 24, and at least one amino acid mutation compared to the Fc region of wild-type or natural IgG, and The at least one amino acid mutation is (i) A mutation from glutamate to arginine (E269R mutation) at the position corresponding to position 269 of SEQ ID NO: 24, according to EU index number assignment. (ii) A mutation from glutamate to arginine at position 294 of SEQ ID NO: 24 according to EU index number assignment (E294R mutation), and (iii) A mutation from glutamate to lysine (E294K mutation) at the position corresponding to position 294 of SEQ ID NO: 24, as assigned by the EU index number. Selected from the group consisting of, cell. [Claim 12] (a) The CD16A containing at least one amino acid mutation contains either or both of the K131D mutation and the K128E mutation, (b) The CD16A containing at least one amino acid mutation contains either or both of the K131E mutation and the K128E mutation, (c) The CD16A containing at least one amino acid mutation contains the K131D mutation, (d) SEQ ID NO: A mutation from asparagine to glutamine at position 38 of 78 (N38Q mutation) and (e) SEQ ID NO: Mutation from asparagine to glutamine at position 74 of 78 (N74Q mutation) Further including at least one more selected mutation, or (f) The CD16A containing at least one amino acid mutation contains the amino acid sequence shown in SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, or SEQ ID NO: 88, The cell according to claim 11. [Claim 13] (a) The cells are human immune cells selected from the group consisting of T cells, macrophages, dendritic cells, NKT cells, NK cells, microglia, osteoclasts, granulocytes, monocytes, and innate immune cells. (b) The cells are derived from stem cells selected from the group consisting of pluripotent stem cells, hematopoietic stem cells, adult stem cells, fetal stem cells, mesenchymal stem cells, postpartum stem cells, multipotent stem cells, and embryonic germ cells, and / or The pluripotent stem cells are either induced pluripotent stem cells (iPS cells) or embryonic stem cells (ES cells). (c) The cells contain genetically modified disruption in the β-2 microglobulin (B2M) gene, and / or The cells further comprise a polynucleotide capable of encoding a single-chain fusion human leukocyte antigen (HLA) class I protein, which includes at least one portion of a B2M protein that is covalently bound either directly or via a linker sequence to at least one portion of an HLA-1α chain selected from the group consisting of HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. (d) The cells contain, through genetic manipulation, human leukocyte antigen (HLA) class II related genes selected from the group consisting of regulatory factor X-related ankyrin-containing protein (RFXANK), regulatory factor 5 (RFX5), regulatory factor X-related protein (RFXAP), class II transactivator (CIITA), HLA-DPA (α chain), HLA-DPB (β chain), HLA-DQA, HLA-DQB, HLA-DRA, HLA-DRB, HLA-DMA, HLA-DMB, HLA-DOA, and HLA-DOB, and / or (e) The cell contains one or more polynucleotides encoding a single-chain fusion HLA class II protein or an HLA class II protein, The cell according to claim 11 or 12. [Claim 14] A pharmaceutical composition comprising cells and a pharmaceutically acceptable excipient according to any one of claims 11 to 13. [Claim 15] A pharmaceutical composition according to claim 14, for use in combination with a polypeptide comprising a modified Fc region of IgG for immunotherapy, The modified Fc region is non-natural and contains at least one amino acid mutation compared to the Fc region of wild-type or natural IgG, and The polypeptide does not have binding activity to wild-type or natural CD16A, and can bind to non-natural CD16A containing at least one amino acid mutation expressed by the cell. (a) The polypeptide comprises a modified Fc region of human Igγ1 containing the amino acid sequence of SEQ ID NO: 24, The modified Fc region is (i) A mutation from glutamate to arginine (E269R mutation) at the position corresponding to position 269 of SEQ ID NO: 24 according to the EU index number assignment, (ii) At least one mutation selected from (a) a glutamate-to-arginine mutation at the position corresponding to position 294 of SEQ ID NO: 24 according to EU index number assignment (E294R mutation) and (b) a glutamate-to-lysine mutation at the position corresponding to position 294 of SEQ ID NO: 24 according to EU index number assignment (E294K mutation) Including, and / or (b) The polypeptide is an antibody, and / or (c) The polypeptide is an antibody that binds to a cancer antigen. Pharmaceutical composition. [Claim 16] A pharmaceutical composition according to claim 14 or 15, for the treatment of cancer.

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