Chimeric protein of IL-2 protein and IL-15 protein or variant thereof, fusion protein containing the protein, and genetically engineered cell expressing the protein

Abstract

The present invention relates to a chimeric protein of IL-2 protein and IL-15 protein, a variant thereof, a fusion protein comprising the same, and / or a genetically engineered cell modified to express any one or more thereof. In the present invention, the binding to IL-2Rβ is weakened, the side effects caused by the excessive immune cell activity of IL-2 can be inhibited, and the rapid exhaustion of natural killer cells is prevented, thereby allowing the appropriate level of cell activity and cell survival ability to be maintained for a long time. Accordingly, the present invention can be used for modulating the immune system and treating immune-related diseases, such as cancer, infectious diseases, and autoimmune diseases.

Description

Technical Field

[0001] This invention relates to chimeric proteins of IL-2 and IL-15, variants thereof, fusion proteins containing said chimeric proteins, variants thereof, and / or gene-modified cells expressing at least one thereof. Background Technology

[0002] Interleukin-2 (IL-2), also known as T-cell growth factor (TCGF), is a 15.5 to 16 kDa globular glycoprotein that plays a crucial role in lymphocyte production, survival, and homeostasis. IL-2 has a length of 133 amino acids and forms a quaternary structure consisting of four antiparallel amphiphilic α-helices (Smith, Science 240, 1169-76 (1988); Bazan, Science 257, 410-413 (1992)).

[0003] IL-2 mediates various immune effects by binding to the IL-2 receptor, which comprises three subunits (IL-2R) (i.e., IL-2Rα (CD25), β (CD122), and γ (CD132)). The binding affinity to the IL-2 protein varies depending on the chain of the IL-2 receptor. Specifically, the trimeric IL-2 receptor, composed of α, β, and γ chains, has a high affinity for the IL-2 protein. Compared to the trimeric IL-2 receptor, the dimeric IL-2 receptor, composed of β and γ chains, has a moderate binding affinity (approximately 1 / 100 lower) for the IL-2 protein, while the monomeric IL-2 receptor, composed solely of the α chain, has a low affinity for the IL-2 protein. Both the IL-2 receptor trimer (α, β, γ) and the IL-2 receptor dimer (β, γ) are essential for the activation of IL-2-binding-based cell signaling (Minami et al., Annu Rev Immunol 11, 245-268 (1993)), while the IL-2 receptor monomer (α, CD25) is not essential for cell signaling (Krieg et al., Proc Natl Acad Sci 107, 11906-11 (2010)).

[0004] IL-15 is a 14 kDa to 15 kDa glycoprotein that exists in a membrane-bound form on the cell surface based on its binding to the IL-15 receptor α (IL-15Rα) present on the cell surface. The IL-15 / IL-15Rα complex binds to IL-2Rβ (CD122) and γ (CD132) and shares a cell signaling mechanism with IL-2 (Marek Jakobisiak et al., Cytokine & Growth Factor Reviews 22, 99-108 (2011)).

[0005] IL-2 receptor trimers (α, β, γ) containing the α chain (CD25) are persistently expressed at high levels in FoxP3+CD+4 regulatory T cells. On the other hand, other normal immune effector cells in vivo, such as CD8 T cells and NK cells, are known to express IL-2 receptor dimers (β, γ) at rest and transiently express IL-2 receptor trimers (α, β, γ) when the cells are activated (Fontenot et al., Nature Immunol 6, 1142-51 (2005); H. Asao, Encyclopedia of Endocrine Disease, 60-63 (2004)).

[0006] IL-2 is primarily synthesized by activated T cells, particularly CD4+ helper T cells, which stimulate T cell proliferation and differentiation and induce the production of cytotoxic T lymphocytes (CTLs). IL-2 also induces peripheral blood lymphocytes to differentiate into cytotoxic cells and lymphokine-activated killer (LAK) cells, promotes T cell expression of cytokines and cytolytic molecules, promotes B cell proliferation and differentiation and B cell synthesis of immunoglobulins, and stimulates the production, proliferation, and activation of natural killer (NK) cells (reviewed in, for example, Waldmann, Nat Rev Immunol 6, 595-601 (2009); Olejniczak and Kasprzak, Med Sci Monit 14, RA179-89 (2008); Malek, Annu Rev Immunol 26, 453-79 (2008)).

[0007] In addition, IL-2 is involved in maintaining CD4+CD25+ regulatory T cells (Tregs), also known as suppressor T cells, and inhibits the effector function of T cells and natural killer cells through intercellular contact and the release of immunosuppressive cytokines such as IL-10 or TGF-β (Fontenot et al., Nature Immunol 6, 1142-51 (2005); D'Cruz and Klein, Nature Immunol 6, 1152-59 (2005)).

[0008] IL-2 or IL-15 has a dual function in the immune response, as it increases the population of lymphocytes in the body, enhances the function of these immune cells, and thus mediates the increase and activation of anti-cancer or immune cells, but suppresses anti-tumor immunity mediated by CD8+ T cells and natural killer cells by strongly amplifying regulatory T (Treg) cells that express high-affinity IL-2 receptors. (Brandenburg, S., et al., Eur J Immunol, 2008. 38(6): p. 1643-53; Facciabene, A., et al., Cancer Res, 2012. 72(9): p. 2162-71).

[0009] Patients receiving IL-2-based immunotherapy have experienced serious cardiovascular, pulmonary, renal, hepatic, gastrointestinal, neurological, skin, hematologic, and systemic side effects. Therefore, various IL-2 mutations have been investigated to improve the efficacy of IL-2 therapy and minimize side effects (US 5,229,109 B).

[0010] One approach to addressing the drawbacks of IL-2 or IL-15-based therapies is to prolong the in vivo half-life of IL-2 or IL-15 while selectively activating CD8+ T cells and natural killer cells expressing low-affinity IL-2 receptors. Despite numerous attempts toward this goal, significant results have yet to be achieved (Arenas-Ramirez, N., et al., SciTransl Med, 2016.8(367): p. 367ra166). Studies have shown that continuous stimulation of lymphocytes, particularly natural killer cells, with IL-15 protein actually inhibits immune enhancement and therapeutic efficacy against cancer (Felices et al., JCIInsight, 2018).

[0011] Repeated administration of IL-2 or IL-15 proteins induces low responsiveness of natural killer cells in vivo, thereby reducing their anticancer ability due to deteriorated proliferative capacity and unbalanced activation mechanisms (Frutoso et al, Int J Mol Sci20(18), 4514(2019)). Therefore, continuous improvement of IL-2 variants is needed to minimize low responsiveness of natural killer cells.

[0012] It has been found that natural killer cells modified to express the membrane protein form of IL-15 have higher cell proliferation and survival rates than natural killer cells expressing native soluble IL-15 (Masaru Imamura et al, Blood, 2014, 124(7), 1081-1088). However, the proliferative capacity of natural killer cells modified to express the membrane protein form of IL-15 is unstable, and their cell number depends on the donor; in most cases, only maintenance or a slight increase in cell number is observed.

[0013] Against this backdrop, the inventors have strived to develop a chimeric protein comprising IL-2 and IL-15 sequences that exhibits low affinity and low irritation while prolonging the half-life of IL-2. As a result, the inventors have discovered that chimeric proteins in which a portion of the IL-2 domain is replaced by the IL-15 domain, or variants of chimeric proteins in which a portion of the amino acid sequence of the IL-2Rβ-binding domain of IL-2 is substituted, possess low binding affinity for IL-2Rβ, low STAT5 activity, and excellent inhibitory activity against cancer cell proliferation, without side effects such as pulmonary edema after administration.

[0014] Furthermore, the inventors have discovered that when a chimeric protein including IL-2 and IL-15 sequences is expressed in natural killer cells, it exhibits better proliferation, survival, and activation maintenance capabilities than natural killer cells expressing conventional IL-15 cell membrane proteins, and demonstrates effective tumor cell killing capabilities against various cancers. Based on these findings, this invention was completed.

[0015] Existing technical documents Non-patent literature (Non-patent literature 1) Smith, Science 240, 1169-76 (1988) (Non-patent literature 2) Bazan, Science 257, 410-413 (1992) (Non-patent literature 3) Minami et al., Annu Rev Immunol 11, 245-268 (1993) (Non-patent literature 4) Krieg et al., Proc Natl Acad Sci 107, 11906-11 (2010) (Non-patent literature 5) Marek Jakobisiak et al., Cytokine & Growth Factor Reviews 22, 99-108 (2011) (Non-patent literature 6) Fontenot et al., Nature Immunol 6, 1142-51 (2005) (Non-patent literature 7) H. Asao, Encyclopedia of Endocrine Disease, 60-63 (2004) (Non-patent literature 8) reviewed eg in Waldmann, Nat Rev Immunol 6, 595-601 (2009) (Non-patent literature 9) Olejniczak and Kasprzak, Med Sci Monit 14, RA179-89 (2008) (Non-patent literature 10) Malek, Annu Rev Immunol 26, 453-79 (2008) (Non-patent literature 11) Fontenot et al., Nature Immunol 6, 1142-51 (2005); (Non-patent literature 12) D'Cruz and Klein, Nature Immunol 6, 1152-59 (2005) (Non-patent literature 13) Brandenburg, S., et al., Eur J Immunol, 2008. 38(6):p. 1643-53 (Non-patent literature 14) Arenas-Ramirez, N., et al., Sci Transl Med, 2016. 8(367): p. 367ra166 (Non-patent literature 15) Felices et al., JCI Insight 3(3):e96219 (2018) (Non-patent literature 16) Frutoso et al, Int J Mol Sci 20(18), 4514(2019) (Non-patent literature 17) Masaru Imamura et al, Blood, 2014, 124(7), 1081-1088 Summary of the Invention

[0016] Therefore, one object of the present invention is to provide a chimeric protein or a variant thereof that reduces the excessive activity of natural killer cells, delays cell depletion, and thereby maintains the activity of immune cells for a long time.

[0017] Another object of the present invention is to provide a fusion protein comprising the chimeric protein or a variant thereof.

[0018] Another object of the present invention is to provide a nucleic acid encoding the fusion protein comprising the chimeric protein or a variant thereof.

[0019] Another object of the present invention is to provide a recombinant vector comprising a nucleic acid encoding a fusion protein containing the chimeric protein or a variant thereof.

[0020] Another object of the present invention is to provide a gene-modified cell that expresses the chimeric protein or a variant thereof and / or a fusion protein containing the chimeric protein or a variant thereof.

[0021] Another object of the present invention is to provide a genetically modified immune cell that expresses the chimeric protein or a variant thereof and / or a fusion protein comprising the chimeric protein or a variant thereof.

[0022] Another object of the present invention is to provide a method for producing the chimeric protein or a variant thereof and / or a fusion protein comprising the chimeric protein or a variant thereof, the method comprising: culturing genetically modified cells to express the chimeric protein or a variant thereof and / or a fusion protein comprising the chimeric protein or a variant thereof; and Recover expressed chimeric proteins or variants thereof and / or fusion proteins containing said chimeric proteins or variants thereof.

[0023] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of cancer, the pharmaceutical composition comprising the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cell.

[0024] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of infectious diseases, the pharmaceutical composition comprising the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cell.

[0025] Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of autoimmune diseases, the pharmaceutical composition comprising the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cell.

[0026] Another object of the present invention is to provide a method for preventing or treating cancer using the chimeric protein or variants thereof, the fusion protein and / or the genetically modified cells, its use in preventing or treating cancer, and its use in the preparation of medicaments for preventing or treating cancer.

[0027] Another object of the present invention is to provide a method for preventing or treating infectious diseases using the chimeric protein or variants thereof, the fusion protein and / or the genetically modified cells, its use in preventing or treating infectious diseases, and its use in the preparation of medicaments for preventing or treating infectious diseases.

[0028] Another object of the present invention is to provide a method for preventing or treating autoimmune diseases using the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells, its use in preventing or treating autoimmune diseases, and its use in the preparation of medicaments for preventing or treating autoimmune diseases.

[0029] The present invention provides a chimeric protein comprising IL-2 and IL-15 proteins or a variant thereof.

[0030] The present invention also provides a fusion protein comprising the chimeric protein or a variant thereof.

[0031] The present invention also provides gene-modified cells expressing the chimeric protein or variants thereof and / or fusion proteins comprising the chimeric protein or variants thereof.

[0032] The present invention also provides pharmaceutical compositions for the prevention or treatment of cancer, comprising modified natural killer cells.

[0033] The present invention also provides pharmaceutical compositions for the prevention or treatment of infectious diseases, the pharmaceutical compositions comprising the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells.

[0034] The present invention also provides pharmaceutical compositions for the prevention or treatment of autoimmune diseases, the pharmaceutical compositions comprising the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells.

[0035] The present invention also provides a method for preventing or treating cancer, including administering the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells, its use for preventing or treating cancer, and its use in the preparation of a medicament for preventing or treating cancer.

[0036] The present invention also provides a method for preventing or treating infectious diseases, including administering the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells, its use for preventing or treating infectious diseases, and its use in the preparation of medicaments for preventing or treating infectious diseases.

[0037] The present invention also provides a method for preventing or treating autoimmune diseases, including administering the chimeric protein or a variant thereof, the fusion protein and / or the genetically modified cells, its use for preventing or treating autoimmune diseases, and its use in the preparation of medicaments for preventing or treating autoimmune diseases. Attached Figure Description

[0038] Figure 1a This is a schematic diagram illustrating the IL-2 and / or IL-15 chimeric protein according to the present invention.

[0039] Figure 1b This is a schematic diagram illustrating a fusion protein that includes the IL-15Ra (sushi domain) and the Fc domain.

[0040] Figures 1c to 1f This is an SDS-PAGE gel image showing the prepared and purified fusion proteins GIC-982C1 to GIC-982C15.

[0041] Figure 2 This is a graph illustrating the results of Octet binding assays for Fc-IL2, GIC-982C1 to GIC-982C6, and GIC-982C12 to GIC-982C15.

[0042] Figure 3 This is a graph illustrating the results of the analysis of the STAT-5 cell signaling pathway activity of Fc-IL2, GIC-982C1 to GIC-982C6, and GIC-982C13 to GIC-982C15.

[0043] Figure 4a The structures of the insert gene and vector for genetically modifying natural killer cells to express chimeric proteins including IL-2 and IL-15 are shown (control group: mbIL-15, experimental group: mbIL-2 / IL-15).

[0044] Figure 4b This is a schematic diagram illustrating the natural killer cells that produce chimeric proteins.

[0045] Figure 5 FACS data for genetically modified natural killer cells are shown.

[0046] Figure 6The proliferative capacity of natural killer cells, dependent on culture time, was demonstrated.

[0047] Figure 7 The study demonstrated the in vitro killing ability of genetically modified natural killer cells against tumor cells after treatment. Detailed Implementation

[0048] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as understood by one of ordinary skill in the art. Generally, the nomenclature used herein is well-known and commonly used in the art.

[0049] The inventors are dedicated to developing recombinant cytokines with low affinity and low irritation that can maintain the ability to kill cancer cells while inhibiting the potent cytotoxic side effects of IL-2. Furthermore, by modifying the nonspecificity and potent toxicity of IL-2, a chimeric protein in which some domains of IL-2 are replaced by domains of IL-15 has a weak binding affinity for IL-2Rβ and low intensity of STAT5 activity compared to wild-type IL-2. This has resulted in the development of a novel chimeric protein that exhibits excellent inhibitory activity against cancer cell proliferation without causing the side effects of conventional IL-2, such as pulmonary edema.

[0050] On one hand, the present invention relates to chimeric proteins including IL-2 and IL-15 proteins.

[0051] Chimeric proteins including IL-2 and IL-15 Interleukin-2 (IL-2) is a protein encoded by the human IL-2 gene, also known as "T cell growth factor (TCGF)," and is a 15.5 to 16 kDa globular glycoprotein that plays a crucial role in lymphocyte production, survival, and homeostasis. IL-2 is 133 amino acids in length and forms a quaternary structure consisting of four antiparallel amphiphilic α-helices.

[0052] It has been reported that human IL-2 can have a structure in which a “helix A” domain, an “AB loop” domain, a “helix B” domain, a “BC loop” domain, a “helix C” domain, a “CD loop” domain, and a “helix D” domain are connected in sequence. The “AB loop” domain, the “helix B” domain, and the “CD loop” domain can participate in IL-2Rα binding, and the “helix C” domain can participate in IL-2Rβ binding (Cassell, Current Pharmaceutical Design, 2002, 8, 2171-2183).

[0053] The amino acid or nucleic acid sequences of human IL-2 are shown in Table 1 below: [Table 1] Interleukin-15 (IL-15) is a protein encoded by the human IL-15 gene and is a 14 to 15 kDa glycoprotein. It exists in a membrane-bound form on the cell surface by binding to the IL-15 receptor α (IL-15Rα). The IL-15 / IL-15Rα complex binds to IL-2Rβ (CD122) and γ (CD132) and shares a cell signaling mechanism with IL-2.

[0054] It has been reported that human IL-15 can have a structure in which the “helix A” domain, “AB loop” domain, “helix B” domain, “BC loop” domain, “helix C” domain, “CD loop” domain, and “helix D” domain are connected in sequence, and the “AB loop” domain, “helix B” domain, and “CD loop” domain can participate in IL-15Rα binding (Lowe, Journal of Molecular Biology, 2011, 406, 160-175).

[0055] The sequence of human IL-15 is shown in Table 2 below: [Table 2] IL-2 or IL-15 proteins can increase lymphocyte populations and enhance the function of immune cells in vivo, thus IL-2 or IL-15 can be used to mediate the increase and activation of anticancer agents or immune cells. However, IL-2 or IL-15 has a dual function in immune response because it suppresses antitumor immunity mediated by CD8+ T cells and natural killer cells by strongly expanding regulatory T cells expressing high-affinity IL-2 receptors. In particular, repeated administration of IL-2 or IL-15 proteins can induce low responsiveness of natural killer cells in vivo, thereby reducing proliferative capacity, rapidly exhausting, and diminishing anticancer ability due to an imbalance in activation mechanisms.

[0056] In one embodiment of the present invention, the chimeric protein may include a portion of the IL-2 amino acid sequence and a portion of the IL-15 amino acid sequence.

[0057] In one embodiment of the invention, the chimeric protein may include a portion of the IL-2 amino acid sequence and a portion of the IL-15 amino acid sequence, which are directly linked to each other or indirectly linked via a linker or the like.

[0058] In one embodiment of the invention, the chimeric protein can be generated by replacing a portion of the IL-2 amino acid sequence with the IL-15 amino acid sequence. As used herein, "protein comprising an amino acid sequence replaced by a specific amino acid sequence" may be used interchangeably with "hybrid protein".

[0059] When a portion of the IL-2 amino acid sequence is replaced by the IL-15 amino acid sequence, as in the chimeric protein of the present invention, by controlling the strength of the signal transduction mechanism of the IL-2Rβ (CD122) and γ (CD132) dimer receptors, excessive activity of natural killer cells can be prevented, cell exhaustion can be delayed, and the activity of immune cells can be maintained for a long time.

[0060] In one embodiment of the present invention, at least one domain selected from the group consisting of the “helix A” domain, the “AB ring” domain, the “helix B” domain, the “BC ring” domain, the “helix C” domain, the “CD ring” domain, and the “helix D” domain of IL-2 can be replaced by the “helix A” domain, the “AB ring” domain, the “helix B” domain, the “BC ring” domain, the “helix C” domain, the “CD ring” domain, or the “helix D” domain of IL-5.

[0061] In a preferred embodiment of the present invention, at least one domain selected from the group consisting of the “AB ring” domain, the “spiral B” domain, and the “CD ring” domain of IL-2 can be replaced by the “AB ring” domain, the “spiral B” domain, or the “CD ring” domain of IL-15.

[0062] In a preferred embodiment of the present invention, the “AB loop” domain, “helical B” domain, and “CD loop” domain of IL-2 may be replaced by the “AB loop” domain, “helical B” domain, or “CD loop” domain of IL-15. For example, the amino acid sequence of the chimeric protein or the nucleic acid sequence encoding it may include or consist of the following: the amino acid sequence of SEQ ID NO: 33 shown in Table 3 or the nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 34.

[0063] [Table 3] Chimeric variants including IL-2 and IL-15 proteins On the other hand, the present invention relates to variants of chimeric proteins including IL-2 and IL-15 proteins.

[0064] According to the present invention, a chimeric protein variant in which one or more amino acid sequences of the IL-2Rβ binding domain of IL-2 are mutated further reduces the binding affinity to IL-2Rβ and reduces the activity intensity of STAT5, thereby preventing the overactivation of natural killer cells, delaying cell exhaustion, and maintaining the activity of immune cells for a long time.

[0065] As used herein, the term "variant" includes not only mutations (preferably substitutions, deletions, insertions, etc.) of one or more amino acid residues in the wild-type amino acid sequence, but also truncations of one or more amino acid residues at the N-terminus or C-terminus, and therefore, in this invention, "variant of chimeric protein" is used as a broad concept to include "fragments of chimeric protein variants".

[0066] As used herein, the term “substitution” refers to a modification that involves replacing one or more amino acids with amino acids having similar biochemical properties without causing loss of biological or biochemical function.

[0067] In one embodiment of the invention, the variant of the chimeric protein may include a mutation at at least one of the following positions in the amino acid sequence of SEQ ID NO.: 33: aspartic acid (Asp(D)), leucine (Leu(L)), serine (Ser(S)), asparagine (Asn(N)) at position 77, and valine (Val(V)) at position 80, preferably an amino acid substitution.

[0068] In one embodiment of the invention, the variant of the chimeric protein may include a substitution of at least one amino acid from the amino acid sequence of SEQ ID NO.: 33, namely D73E, L74F, S76D, S76E, S76N, S76K, S76L, N77D, and V80L.

[0069] In one embodiment of the invention, the chimeric protein variant may include a combination of variants selected from the following: D73E / L74F, D73E / S76D, D73E / S76E, D73E / S76N, D73E / S76K, D73E / S76L, D73E / N77D, D73E / V80L, L74F / S76D, L74F / S76E, L74F / S76N, L74F / S76K, L74F / S76L, L74F / N77D, L74F / V80L, S76D / N77D, S76D / V80L, S76E / N77D, S76E / V80L, S76N / N77D, S76N / V80L. , S76K / N77D, S76K / V80L, S76L / N77D, S76L / V80L, D73E / L74F / S76D, D73E / L74F / S76E, D73E / L74F / S76N, D73E / L74F / S76K, D73E / L74F / S76L, D73E / L 74F / N77D, D73E / L74F / V80L, D73E / S76D / N77D, D73E / S76D / V80L, D73E / S7 6E / N77D, D73E / S76E / V80L, D73E / S76N / N77D, D73E / S76N / V80L, D73E / S76K / N77D、D73E / S76K / V80L、D73E / S76L / N77D、D73E / S76L / V80L、D73E / N77D / V80L, S76D / N77D / V80L, S76E / N77D / V80L, S76N / N77D / V80L, S76K / N77D / V 80L, S76L / N77D / V80L, D73E / L74F / S76D / N77D, D73E / L74F / S76E / N77D, D7 3E / L74F / S76N / N77D, D73E / L74F / S76K / N77D, D73E / L74F / S76L / N77D, D73E / L74F / S76D / V80L、D73E / L74F / S76E / V80L、D73E / L74F / S76N / V80L、D73E / L74F / S76K / V80L, D73E / L74F / S76L / V80L, L74F / S76D / N77D / V80L, L74F / S7 6E / N77D / V80L, L74F / S76N / N77D / V80L, L74F / S76K / N77D / V80L, L74F / S76 L / N77D / V80L, D73E / L74F / S76D / N77D / V80L, D73E / L74F / S76E / N77D / V80L,D73E / L74F / S76N / N77D / V80L, D73E / L74F / S76K / N77D / V80L, and D73E / L74F / S76L / N77D / V80L.

[0070] In a preferred embodiment of the invention, the variant of the chimeric protein may include at least one amino acid substitution of D73E, L74F, S76D, N77D and V80L in the amino acid sequence of SEQ ID NO.:33.

[0071] In a preferred embodiment of the present invention, the amino acid sequence or nucleic acid sequence of the chimeric protein variant may include or consist of the following: amino acid sequences of SEQ ID NO.: 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73 or 75, or nucleic acid sequences encoding the chimeric protein variant of SEQ ID NO: 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 70, 72 or 74, as shown in Table 4 below.

[0072] [Table 4] *The underlined portion of the amino acid sequence indicates the mutation location.

[0073] This includes chimeric proteins containing IL-2 and IL-15 proteins, or fusion proteins of their variants. On the other hand, the present invention relates to fusion proteins including chimeric proteins or variants thereof.

[0074] As used herein, the term "fusion protein" refers to a protein that, in addition to a chimeric protein comprising IL-2 and IL-15 proteins or variants thereof, includes another fusion domain or amino acid sequence.

[0075] As used herein, the term "fusion protein" refers to a protein in which a chimeric protein or a variant thereof is fused with an additional peptide sequence (fusion domain).

[0076] As used herein, the term "fusion domain" refers to an additional domain or portion that can be directly or indirectly linked to and incorporated into a chimeric protein or a variant thereof according to the invention, and fusion domain is used to denote functional and non-functional polypeptides.

[0077] In this invention, fusion domains can be fused to confer expression characteristics, such as secretion from cells, expression on cell surfaces, anchoring to cell membranes or intracellular localization; adding substrates or other recognition sequences for post-translational modifications; or altering tissue localization, tissue rejection or other ADME characteristics; or adding proteins or peptides with another function, but not limited thereto.

[0078] In this invention, the fusion domain may be, for example, a transmembrane domain, a hinge domain, an intracellular signal transduction domain, an immunomodulatory domain, a localization domain, an immune co-stimulatory factor / receptor, a cytokine, a growth factor, an albumin-binding domain, an Fc domain, a transferrin fusion domain, albumin, PEG, hyaluronic acid, or other therapeutic peptides, but is not limited thereto, and may include any domain (or portion) that is not decisive for the expression or activity of chimeric proteins or variants thereof, including the IL-2 and IL-15 proteins of this invention.

[0079] In this invention, in order to immobilize the fusion protein expressed in modified natural killer cells onto the cell membrane or to express it on the cell surface, a fusion domain comprising a membrane protein or a transmembrane domain may be used.

[0080] In this invention, "membrane protein" refers to a protein that is inserted into or attached to the surface of a membrane, including a lipid bilayer, and includes "integrated membrane proteins" that are inserted into the lipid bilayer, and "peripheral membrane proteins" that are attached to the surface of the lipid bilayer. An "integrated membrane protein" is a transmembrane "membrane protein".

[0081] As used herein, the term "integrated membrane protein" is a protein that penetrates the cell membrane and includes a "transmembrane domain" that penetrates the membrane, an "intracellular domain" located within the cell, and an "extracellular domain" exposed outside the cell. The number of transmembrane domains varies among membrane proteins. Since the lipid bilayer is composed of highly hydrophobic lipids, most transmembrane domains through this region are composed of hydrophobic amino acids. Almost all transmembrane domains are composed of α-helical structures. In membrane proteins with multiple transmembrane domains, the transmembrane domains are arranged in a loop to form a cylindrical structure. This cylindrical structure opens and closes in response to specific signals and is therefore used as a channel for transporting specific ions or biological substances. Very few membrane proteins have transmembrane domains that are β-sheet structures. The cylindrical structure formed by arranging multiple domains in a β-sheet structure in a circular pattern is called a "β-barrel."

[0082] In this invention, in the case of "peripheral membrane proteins," lipoproteins are formed through covalent bonds between protein molecules and lipid molecules, and hydrophobic bonds are formed between the lipid molecules of the lipoprotein and the lipid monolayer of the cell membrane, thereby anchoring the protein to the surface of the cell membrane. Peripheral membrane proteins are, for example, GPI (glycosylphosphatidylinositol)-anchored proteins. The fatty acid of the GPI is inserted into the lipid monolayer, and its phosphate group on the other side is covalently linked to the protein, so that the GPI anchors the protein to the surface of the lipid bilayer.

[0083] In this invention, examples of membrane proteins include receptors, ligands, immunoglobulins, blood group glycoproteins, or combinations thereof. Membrane proteins may be selected from the group consisting of: IL-15 receptor α (IL-15Rα), CD8α, CD4, CD3ε, CD3γ, CD3δ, CD3ζ, CD28, CD137, FcεRIγ, T cell receptors (TCRs, e.g., TCRα and / or TCRβ), nicotinic acetylcholine receptors, GABA receptors, and fragments thereof, but are not limited thereto. Specific examples of immunoglobulins include IgG, IgA, IgM, IgE, IgD, and combinations thereof. Specific examples of blood group glycoproteins include, but are not limited to, blood group glycoprotein A, blood group glycoprotein D, and combinations thereof.

[0084] In this invention, in addition to the transmembrane domain, the fusion protein may also include a hinge domain or a sushi domain. As used herein, the term "hinge domain" refers to a sequence of amino acids present between the transmembrane domain and the extracellular domain of a membrane-anchored protein. In this invention, the domains present in the fusion protein may be combinations of domains from the same protein, or combinations of domains from different proteins, forming a chimeric protein.

[0085] In one embodiment of the invention, in addition to the IL-2 and IL-15 chimeric protein or variants thereof according to the invention, the fusion protein may include the IL-15 receptor α (IL-15Rα) protein or fragments thereof.

[0086] In this invention, the IL-15 receptor α (IL-15Rα) protein fragment may include a “sushi domain”, which is the shortest region of the receptor with IL-15 binding activity.

[0087] In this invention, the IL-15 receptor α (IL-15Rα) protein fragment may include a "sushi domain" as the shortest region of the receptor having IL-15 binding activity and a "transmembrane domain" as the cell membrane permeable region of the IL-15 receptor α. In this invention, the fusion protein may be a fusion protein in which the IL-15 receptor α (IL-15Rα) protein or a fragment thereof is fused with a chimeric protein or a variant thereof.

[0088] In this invention, the sequences of human IL-15 receptor α (IL-15Rα) and its fragments are listed, for example, in Table 5 below, but are not limited thereto.

[0089] [Table 5] In this invention, the chimeric protein or a variant thereof can be directly or indirectly (e.g., via a linker) linked to the IL-15 receptor α (IL-15Rα) protein. In one embodiment of the invention, the fusion protein may comprise a structure of formula (I) or formula (II): N'-X-[L1] n -Y-C'(Formula (I)) N'-Y-[L1] n -X-C'(Equation (II)) Where N' is the N-terminus of the fusion protein, and C' is the C-terminus of the fusion protein. X is a chimeric protein or a variant thereof that includes IL-2 and IL-15 proteins. Y is the IL-15 receptor α (IL-15Rα) protein or a fragment thereof. L1 is a connector, and n is an integer of 0 or greater, preferably 0 or 1.

[0090] In this invention, fragments of the IL-15Rα protein may include at least one of the IL15Rα membrane protein domain and the IL15Rα sushi domain. For example, the IL15Rα membrane protein domain may include or consist of the amino acid sequence shown in SEQ ID NO: 79, and the IL15Rα sushi domain may include or consist of the amino acid sequence shown in SEQ ID NO: 81, but is not limited thereto.

[0091] In this invention, the connector can be any of the various connectors known in the art. For example, the connector can include, but is not limited to, a hinged structural domain between a transmembrane structural domain and an extracellular structural domain, a flexible connector, a rigid connector, etc.

[0092] More specifically, a linker refers to an amino acid sequence of sufficient length to allow the protein to form appropriate secondary and tertiary structures. In some embodiments, the linker is a peptide linker comprising at least one, but less than 100 amino acids, for example, 2 to 60 amino acids, preferably 10 to 40 amino acids, more preferably 15 to 40 amino acids, even more preferably 19 to 30 amino acids, and most preferably 20 to 26 amino acids. In some embodiments, the linker has 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acid residues.

[0093] In this invention, the linker is a CD4-derived hinge, a CD8α-derived hinge, a CD28-derived hinge, a CD34-derived hinge, an IgG1-derived hinge, an IgG2-derived hinge, an IgG3-derived hinge, an IgG4-derived hinge, PLrigid, a 2aa GS linker, a 6aa [GS]x linker, a 10aa [Gs]x linker, a 10aa flexible protein domain linker, an 8aa protein domain linker, a flexible linker 2x (GGGS), a flexible linker 2x (GGGGS), a 13-amino acid linker [GGGS GGGGS GGGS], a splitting fluorophore linker, a Freiburg standard, 15 Flexible glycine-serine protein domain linkers, Freiburg standard, short linkers (Gly-Gly-Ser-Gly), medium linkers (Gly-Gly-Ser-Gly) x2, long linkers (Gly-Gly-Ser-Gly) x3, glycine linkers, (HL5)2 peptide helical linkers, rigid; domain separation of fusion proteins, glycine-serine linkers (GSGGS), glycine-serine linkers (GSSGS), (G2S)3 linkers, SEG linkers, GSAT linkers, Z-EGFR-1907_short linkers, Z-EGFR-1907_medium linkers, Z-EGFR-1907_SEG-linkers, (Gly4Ser)3 flexible peptide linkers or (SSSSG)x2 serine-glycine linkers or Whitlow 218 linkers (GSTGSGSKPGSGEGSTKG), but not limited to these.

[0094] In this invention, in addition to the adapters for the fusion of the chimeric protein or its variants with the functional domains according to the invention, the adapters can be any adapters that 1) perform additional functions such as improving biological activity, increasing expression yield and improving pharmacokinetic characteristics, 2) do not negatively affect the expression, secretion or functional activity of each domain of the fusion protein, and 3) do not exhibit immunogenicity.

[0095] In this invention, preferably, the linker is QSFGLLDPK (CD3-derived hinge amino acid sequence, SEQ ID NO: 83) or a variant thereof, LSEGDKVKMDSRIQVLSRGVNQT (CD4-derived hinge amino acid sequence, SEQ ID NO: 84) or a variant thereof, KPTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY (CD8α-derived hinge amino acid sequence, SEQ ID NO: 85) or a variant thereof, IEVMYPPPYLDNERSNGTIIHVKGKHLCPSPLFPGPSKP (CD28-derived hinge amino acid sequence, SEQ ID NO: 86) or a variant thereof, ELPTQGTFSNVSTNVS (CD34-derived hinge amino acid sequence, SEQ ID NO: 87) or a variant thereof, (GGGGS)n (where n is an integer greater than or equal to 1, SEQ ID NO: 88) or GSTGSGSKPGSGEGSTKG (Whitlow 218 linker, SEQ ID NO: 88). NO: 89), but not limited to this.

[0096] In this invention, preferably, the connector may include, but is not limited to, the sequence represented by SEQ ID NO: 87, 88 or 89.

[0097] In this invention, the fusion protein may include the Fc domain of an immunoglobulin.

[0098] The Fc domain of an immunoglobulin may include the heavy chain constant region 2 (CH2) and heavy chain constant region 3 (CH3). The Fc domain may not include the variable regions of the heavy and light chains, or the light chain constant region 1 (CH1). The immunoglobulin may be IgG, IgA, IgE, IgD, or IgM, preferably IgG4.

[0099] Furthermore, the Fc domain of an immunoglobulin can be not only a wild-type Fc domain but also an Fc domain variant. Additionally, as used herein, the term "Fc domain variant" can have a glycosylation pattern different from that of the wild-type Fc domain, or it can have an increased glycosylation pattern, a decreased glycosylation pattern, or a deglycosylated form compared to the wild-type Fc domain. The Fc domain of an immunoglobulin can include glycosylated Fc domains. Fc domains or variants can be sialylated, fucosylated, or glycosylated, the degree of which is controlled by culture conditions or host gene manipulation.

[0100] Furthermore, the glycans of the immunoglobulin Fc domain can be modified using conventional methods such as chemical methods, enzymatic methods, and genetic engineering methods using microorganisms. Additionally, Fc domain variants can be mixed with the Fc regions of immunoglobulins IgG, IgA, IgE, IgD, or IgM.

[0101] In this invention, when the fusion protein includes an Fc domain, the chimeric protein and / or its variants according to the invention may be attached to the N'-terminus and / or C'-terminus of the Fc domain, and another fusion domain may be attached to a terminus different from the terminus to which the chimeric protein or its variants are attached.

[0102] In this invention, for example, the fusion protein may include an IL-2 / IL-15 chimeric protein or a variant thereof, or the sushi domain of the IL-15 receptor α.

[0103] For example, fusion proteins can have the structure of formula (III) or formula (IV).

[0104] In implementation, the fusion protein may have the structure of formula (III) or formula (IV): N'-X-[L1] l -Y-[L2] n -[H] m -Fc structural domain-C' (Equation (III)) N'-[H] m -Fc domain-[L1] l -Y-[L2] n -XC' (Formula (IV)) in N' is the N-terminus of the fusion protein, and C' is the C-terminus of the fusion protein. X is a chimeric protein or a variant thereof that includes IL-2 and IL-15 proteins; Y is the IL-15 receptor α (IL-15Rα) protein or a fragment thereof; L1 and L2 are connectors; H is the Fc hinge domain of immunoglobulin; and l, n, and m are each an independent integer of 0 or greater, preferably 0 or 1.

[0105] In this invention, the Fc hinge domain can be a hinge domain derived from IgG, IgA, IgE, IGD or IgM, preferably a hinge domain of IgG4, but is not limited thereto.

[0106] In this invention, for example, when the fusion protein is presented on the cell membrane surface, it may additionally include membrane proteins or transmembrane domains.

[0107] In this invention, the IL-15Rα protein fragment may include at least one of the IL-15Rα membrane protein domain and the IL-15Rα sushi domain.

[0108] In this invention, the fusion protein may further include an immunoregulatory domain or an intracellular signal transduction domain.

[0109] In this invention, the immunomodulatory domain or intracellular signal transduction domain is a domain located in the cytoplasmic direction of a membrane anchoring protein, and refers to the site that activates or inhibits the immune response when the target antigen binds to the extracellular domain.

[0110] In this invention, the immunomodulatory domain or intracellular signal transduction domain can be an intracellular signal transduction domain capable of activating natural killer cells to an appropriate level, and is, for example, derived from the following intracellular signal transduction domains: IL-15Rα, CD3, CD28, CD40L, ICOS, OX40, 4-1BB, TNFR2, DAP10, 2B4, CD3ζ, CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, LFA-1 (lymphocyte function-associated antigen-1), CD2, CD7, LIGHT, NKG2C and / or B7-H3, but is not limited thereto.

[0111] In this invention, the fusion protein can polymerize into a dimer, trimer, or high-polymer form depending on the linker, fusion domain, etc. In one embodiment of the invention, the fusion protein is used in a dimer form through the hinge region of the fusion protein including the Fc domain, but is not limited thereto.

[0112] As in embodiments of the present invention, the fusion protein may include or be composed of an amino acid sequence selected from, for example, the amino acid sequences shown in SEQ ID NO: 90 to 133, but is not limited thereto.

[0113] As in embodiments of the present invention, the fusion protein may include, or be composed of, the amino acid sequence of SEQ ID NO:185, but is not limited thereto.

[0114] Nucleic acid and recombinant vector On the other hand, the present invention relates to nucleic acids encoding chimeric proteins or variants thereof, or fusion proteins.

[0115] As used herein, “nucleic acid” can exist in cellular form, cell lysate, or in a partially purified or substantially pure form. When referring to nucleic acids, “isolated” or “substantially pure” means nucleic acids that have been purified by standard techniques, including alkali / SDS treatment, CsCl banding, column chromatography, agarose gel electrophoresis, and other techniques known in the art, and are therefore isolated from other cellular components or other contaminants (e.g., other cellular nucleic acids or proteins). The nucleic acids of this invention can be DNA or RNA.

[0116] In this invention, the nucleic acid encoding the chimeric protein may include or be composed of the nucleic acid sequence of SEQ ID NO: 34.

[0117] In this invention, the nucleic acid encoding the variant of the chimeric protein may include or consist of the following nucleic acid sequences: SEQ ID NO: 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72 or 74.

[0118] In this invention, in addition to the amino acid sequence of the chimeric protein or its variant protein according to the invention or the nucleic acid sequence encoding it, the nucleic acid sequence encoding the fusion protein can be readily designed based on the amino acid sequence of the further included linker or fusion domain.

[0119] In this invention, the nucleic acid encoding the fusion protein may include or consist of the following: nucleic acid sequences selected from SEQ ID NO:135 to 178, but are not limited thereto.

[0120] In this invention, the nucleic acid encoding the fusion protein may include or be composed of the nucleic acid sequence of SEQ ID NO: 186, but is not limited thereto.

[0121] On the other hand, the present invention relates to recombinant vectors containing the nucleic acids of the present invention.

[0122] In this invention, the recombinant vector can be selected without limitation from vectors known in the art, as long as it can induce protein expression of nucleic acids encoding peptides. For example, when *Escherichia coli* (… E. coliWhen yeast is used as a host, vectors including the T7 series (T7A1, T7A2, T7A3, etc.), lac, lacUV5, and temperature-dependent vectors (including λphoA, phoB, rmB, tac, trc, trp, or 1PL promoters) can be used. When yeast is used as a host, vectors containing ADH1, AOX1, GAL1, GAL10, PGK, or TDH3 promoters can be used, and when Bacillus is used as a host, vectors including the P2 promoter can be used. These are provided only as representative embodiments, and those skilled in the art can choose any vector suitable from a variety of vectors known in the art, other than those including promoters, as long as it is suitable for the host as a vector including a promoter for inducing the expression of the interferon λ variant according to the invention.

[0123] As used herein, the term "vector" refers to a DNA product containing a DNA sequence operatively linked to a suitable regulatory sequence capable of expressing that DNA in a suitable host. Vectors can be plasmids, phage particles, or simple potential genomic inserts. When transformed into a suitable host, vectors can be replicated or perform functions independently of the host genome, or some may integrate with the genome. Plasmids are currently the most commonly used form of vector, and therefore the terms "plasmid" and "vector" are often used interchangeably. However, this invention covers other forms of vectors known in the art or having the same functions as those known in the art. Protein expression vectors used in *E. coli* include: the pET family vectors from Novagen, Inc. (USA); the pBAD family vectors from Invitrogen Corp. (USA); the PHCE or pCOLD vectors from Takara Bio Inc. (Japan); and the pACE family vectors from GenoFocus Inc. (Korea). In *Bacillus subtilis* (… Bacillus subtilis In genomics, the target gene can be inserted into a specific part of the genome to achieve protein expression, or the pHT family of vectors from MoBiTech (Germany) can be used. Even in fungi and yeast, protein expression is possible using genome insertion or self-replicating vectors. Plant protein expression vectors utilizing the T-DNA system, such as those from Agrobacterium tumefaciens (…), can be used. Agrobacterium tumefaciens ) or Agrobacterium rhizogenes ( Agrobacterium rhizogenes Typical expression vectors used for expression in mammalian cell cultures are based on, for example, pRK5 (EP 307, 247), pSV16B (WO91 / 08291), and pVL1392 (Pharmingen).

[0124] As used herein, the term "expression control sequence" refers to the DNA sequence necessary for the expression of a coding sequence that is operatively linked to a specific host organism. Such control sequences include promoters for performing transcription, operon sequences for controlling such transcription, sequences encoding appropriate mRNA ribosome binding sites, and sequences controlling the termination of transcription and translation. For example, control sequences suitable for prokaryotes include promoters, optional operon sequences, and ribosome binding sites. Control sequences suitable for eukaryotic cells include promoters, polyadenylation signals, and enhancers. The promoter is the most influential factor on gene expression levels in plasmids. SRα promoters, cytomegalovirus-derived promoters, and the like are preferred promoters for high expression.

[0125] To express the DNA sequence of the present invention, the vector can use any of a variety of expression control sequences. Useful expression control sequences include, for example, early and late promoters of SV40 or adenovirus, lac system, trp system, TAC or TRC system, T3 and T7 promoters, major operon and promoter regions of bacteriophage λ, control regions encoding fd proteins, promoters of 3-phosphoglycerate kinase or other diol lyases, promoters of phosphatases such as Pho5, promoters of yeast α-mating type systems, and other sequences and various combinations thereof having known conformations and inducible activities for controlling gene expression in prokaryotic or eukaryotic cells or viruses. The T7 RNA polymerase promoter φ10 can be used to express proteins in *E. coli*.

[0126] When a nucleic acid sequence is aligned with another nucleic acid sequence based on its functional relationship, it is said to be "operably linked" to it. This can be one or more genes and one or more control sequences linked in such a way that the gene can be expressed when a suitable molecule (e.g., a transcription activator protein) is linked to one or more control sequences. For example, when expressed as a pre-protein involved in polypeptide secretion, the DNA of the pre-sequence or secretion leader sequence is operably linked to the DNA of the polypeptide; when a promoter or enhancer affects the transcription of a sequence, the promoter or enhancer is operably linked to the coding sequence; or when a ribosome binding site affects the transcription of a coding sequence, the ribosome binding site is operably linked to the coding sequence; or when the ribosome binding site is positioned to facilitate translation, the ribosome binding site is operably linked to the coding sequence. Generally, the term "operably linked" means that the linked DNA sequence is in contact with it, and the secretion leader sequence is in contact with it and is present in the reading frame. However, the enhancer does not need to be in contact with it. The ligation of these sequences is achieved by joining (ligation) at a convenient restriction enzyme cleavage site. When such a site is not available, synthetic oligonucleotide adaptors or linkers according to conventional methods are used.

[0127] As used herein, the term "expression vector" generally refers to a recombinant vector into which a foreign DNA fragment has been inserted, and typically means a double-stranded DNA fragment. Here, "foreign DNA" means foreign DNA that is not naturally present in the host cell. Once the expression vector is present in the host cell, it can replicate independently of the host chromosomal DNA and can produce several copies of the vector and its inserted (foreign) DNA.

[0128] As is well known in the art, in order to increase the expression level of a transfected gene in recombinant cells, the gene should be operatively linked to a transcriptional or translational expression control sequence that functions in a selected expression host. Preferably, the expression control sequence and the corresponding gene are contained in a single expression vector containing bacterial selection markers and origins of replication. When the expression host is a eukaryotic cell, the expression vector should further include useful expression markers from the eukaryotic expression host.

[0129] Cells expressing chimeric proteins including IL-2 and IL-15, their variants, and / or gene-modified proteins including such chimeric proteins or their variants On the other hand, the present invention relates to cells expressing gene-modified chimeric proteins, variants thereof, and / or fusion proteins including the chimeric proteins and variants thereof according to the present invention.

[0130] In this invention, genetically modified cells can be produced without restriction by any method, as long as the method is modified to express chimeric proteins, their variants, and / or fusion proteins.

[0131] In this invention, preferably, the modified natural killer cells can be generated by introducing nucleic acids encoding chimeric proteins, their variants, and / or fusion proteins, or recombinant vectors containing them, into host cells.

[0132] In this invention, host cells can refer to cells used for expression, in which genes, recombinant vectors, etc., have been introduced to produce proteins. Host cells can be used without limitation, as long as they are cells capable of expressing chimeric proteins or their variants or fusion proteins, and eukaryotic cells are preferred, yeast cells, insect cells, or animal cells are more preferred, and animal cells are most preferred. For example, host cells can be CHO cell lines or HEK cell lines primarily used for expressing fusion proteins, but are not limited thereto.

[0133] A variety of expression host / vector combinations can be used to express chimeric proteins or their variants or fusion proteins. Expression vectors suitable for eukaryotic hosts include, for example, expression control sequences from SV40, bovine papillomavirus, adenovirus, adeno-associated virus, cytomegalovirus, and retroviruses. Expression vectors suitable for bacterial hosts include bacterial plasmids, such as those obtained from *E. coli*, such as pBlueScript, pGEX2T, pUC vector, col E1, pCR1, pBR322, pMB9 and their derivatives; plasmids with a broad host range, such as RP4; phage DNA, such as various phage λ derivatives like λgt10, λgt11, and NM989; and other DNA phages such as M13 and filamentous single-stranded DNA phages. Expression vectors useful for yeast cells include 2μ plasmids and their derivatives. The vector used for insect cells is pVL 941.

[0134] Recombinant vectors can be introduced into host cells through methods such as transformation or transfection. As used herein, the term "transformation" means introducing DNA into a host and making the DNA replicable using extrachromosomal factors or chromosomal integration. As used herein, the term "transfection" means that the expression vector is accommodated by the host cell, regardless of whether any coding sequence is actually expressed.

[0135] It should be understood that not all vectors and expression control sequences function identically when expressing the DNA sequences of the present invention. Similarly, not all hosts function identically within the same expression system. However, those skilled in the art will be able to make appropriate selections from a variety of vectors, expression control sequences, and hosts without departing from the scope of the present invention, without incurring excessive experimental burden. For example, the selection of a vector should take into account the host, as the vector should replicate within the host. The number of replications of the vector, the ability to control the number of replications, and the expression of other proteins encoded by the respective vector, such as antibiotic markers, should also be considered. Many factors should be considered when selecting an expression control sequence. For example, the relative strength, controllability, and compatibility with the DNA sequences of the present invention, particularly aspects related to possible secondary structures, should be considered. Factors such as the selected vector, the toxicity of the product encoded by the DNA sequences of the present invention, secretion characteristics, the ability to precisely fold proteins, culture and fermentation factors, and the ease of purifying the product encoded by the DNA sequences of the present invention from the host can be considered when selecting a single-cell host. Within these factors, those skilled in the art can select various vector / expression control sequence / host combinations capable of expressing the DNA sequences of the present invention in fermentation or large-scale animal cultures. As a screening method for cloning protein cDNA through expression cloning, methods such as binding, panning, and membrane emulsification can be applied.

[0136] Genes and recombinant vectors can be introduced into host cells using various methods known in the art. The chimeric protein or its variants or fusion protein according to the invention can be directly introduced into the genome of a host cell and exist as a factor on the chromosome. It will be apparent to those skilled in the art that even when the gene is inserted into the host cell's genome chromosome, it will have the same function as when the recombinant vector is introduced into the host cell.

[0137] Methods for producing chimeric proteins comprising IL-2 and IL-15 proteins or variants thereof and / or fusion proteins comprising such chimeric proteins or variants thereof. On the other hand, the present invention relates to methods for producing chimeric proteins or variants thereof and / or fusion proteins.

[0138] The chimeric proteins or variants and / or fusion proteins according to the present invention can be produced by various protein production methods known in the art.

[0139] In this invention, chimeric proteins or their variants and / or fusion proteins can be produced using genetically modified cells.

[0140] For example, the production method includes: culturing genetically modified cells to express chimeric proteins or variants and / or fusion proteins thereof; and recovering the expressed chimeric proteins or variants and / or fusion proteins thereof.

[0141] In this invention, chimeric proteins or their variants, protein complexes or fusion proteins can also be produced in a cell-free protein synthesis system.

[0142] Cell-free protein synthesis systems involve adding substrates or enzymes to cell lysates or extracts and synthesizing proteins in vitro using the key elements necessary for protein synthesis (ATP (adenosine triphosphate), amino acids, etc.). Cell-free protein synthesis systems advantageously overcome the drawbacks of conventional protein production methods using cells. Furthermore, cell-free protein synthesis allows for high-rate protein synthesis without any cell culture. This is achieved by extracting only the intracellular mechanisms and factors associated with protein production from the cells and artificially repeating the protein synthesis process while excluding extracellular physiological control mechanisms. This enables the rapid, large-scale production of target proteins. Compared to cell culture processes that express proteins within the space of the cell membrane and cell wall, it is a completely open system without physical barriers and allows for flexible modification of protein synthesis conditions for various research applications. Moreover, intracellular protein synthesis systems present major problems in terms of production and yield when the produced proteins are cytotoxic, whereas cell-free protein synthesis systems can produce peptides or proteins without these issues.

[0143] In this invention, in addition to examples of host cells used for protein expression and production, immune cells and the like can be used as host cells and modified to express chimeric proteins, variants thereof, and / or fusion proteins containing such chimeric proteins or variants according to the invention. In this case, the function or characteristics of the host cell (e.g., immune cell) can be modified or improved by expressing chimeric proteins, variants thereof, and / or fusion proteins containing such chimeric proteins or variants thereof according to the invention.

[0144] Specifically, in one embodiment of the present invention, in order to develop genetically modified natural killer cells that maintain appropriate levels of cell activity and cell viability for a longer period of time compared to original or previously reported modified natural killer cells, it was found that when a chimeric protein including IL-2 and IL-15 is expressed in natural killer cells, it has a higher expression yield than other proteins, inhibits side effects caused by excessive immune cell activity of IL-2, prevents rapid depletion of natural killer cells, and maintains significantly higher levels of cell activity and cell viability for a significantly longer period of time. Furthermore, it was found that the genetically modified natural killer cells exhibit excellent cancer cell killing ability without side effects such as pulmonary edema.

[0145] Therefore, on the other hand, the present invention relates to a modified immune cell that expresses a chimeric protein comprising IL-2 and IL-15 proteins, variants thereof, and / or a fusion protein comprising the chimeric protein or a variant thereof.

[0146] Modified immune cells expressing chimeric proteins including IL-2 and IL-15, their variants, and / or fusion proteins containing such chimeric proteins or their variants. As used herein, the term "modified immune cell expressing a chimeric protein comprising IL-2 and IL-15 proteins, its variants, and / or a fusion protein comprising the chimeric protein, its variants" refers to a genetically modified immune cell expressing a chimeric protein, its variants, or a fusion protein comprising the chimeric protein, its variants, according to the invention.

[0147] The immune cells according to the present invention can be genetically modified to express chimeric proteins, their variants and / or fusion proteins.

[0148] In this invention, immune cells are preferably animal-derived immune cells, more preferably human-derived immune cells, and may include, but are not limited to, B cells, T cells (e.g., cytotoxic T cells, effector T cells, and helper T cells), natural killer cells (NK cells), NKT cells, dendritic cells, and cells capable of differentiating into immune cells, such as hematopoietic stem cells, iPSCs, and adult stem cells.

[0149] In this invention, the most preferred immune cells are natural killer cells.

[0150] In this invention, "natural killer cells" ("NK cells") refer to a type of cytotoxic lymphocyte in the immune system. NK cells provide a rapid response to virus-infected cells and respond to transformed cells. Typically, immune cells detect peptides from pathogens, provided by the major histocompatibility complex (MHC) molecule on the surface of infected cells, thus triggering cytokine release and leading to cell lysis or cell death. However, NK cells are unique because they have the ability to recognize stress cells regardless of the presence of pathogen-derived peptides on the MHC molecule. These cells are called "natural killer cells" based on the initial concept that they can kill their targets without prior activation. NK cells are known to be large granular lymphocytes (LGLs) that differentiate and mature in the bone marrow and then enter the circulatory system from the bone marrow.

[0151] In some embodiments, the immune cells are mammalian immune cells. Examples of “mammalian” or “mammal” include primates (e.g., humans), canines, felines, rodents, pigs, ruminants, etc. Specific examples include humans, dogs, cats, horses, cattle, sheep, goats, rabbits, guinea pigs, rats, and mice. In some embodiments, the mammalian immune cells are human-derived immune cells.

[0152] In this invention, chimeric proteins, variants and / or fusion proteins expressed in immune cells can be secreted extracellularly, expressed on the cell membrane, and / or expressed on the cell surface.

[0153] In this invention, signal sequences for transporting chimeric proteins, their variants, and / or fusion proteins may be included.

[0154] In this invention, the signal sequence (signal peptide or leader sequence) can be located at the N-terminus of the fusion protein. The function of the signal sequence is to move the fusion protein to the desired location. For example, the signal sequence can be transported to a transport channel such as the endoplasmic reticulum or a transporter, and then cleaved at the residues of the signal sequence, but is not limited thereto.

[0155] In this invention, the signal sequence (signal peptide or leader sequence) can be located at the N-terminus of the fusion protein. The function of the signal sequence is to move the fusion protein to the desired location. For example, the signal sequence can be transported to a transport channel such as the endoplasmic reticulum or a transporter, and can then be cleaved at residues of the signal sequence, but is not limited thereto.

[0156] In this invention, chimeric proteins, their variants, and / or fusion proteins are more preferably immobilized on the cell membrane of immune cells or expressed on the cell surface.

[0157] In this invention, chimeric proteins, their variants, and / or fusion proteins can be directly or indirectly (e.g., via ionic, nonionic, or covalent bonds) bound (conjugated; fused) to the surface of immune cells (e.g., on the surface of immune cells or within the cell membrane) using various linkers known in the art (see reference [Hermanson, G., Bioconjugate Techniques, Academic Press 1996]).

[0158] In this invention, the fusion protein is preferably a fusion protein having the structure of formula (I) or formula (II), as can be seen from the examples, but is not limited thereto.

[0159] In this invention, the fusion protein may exist on the cell membrane of immune cells, but is not limited thereto.

[0160] The genetically modified immune cells of the present invention can maintain appropriate levels of immune cell activity and cell survival capacity for a long time, and therefore cell immunotherapy agents using the modified immune cells can be used to prevent or treat various diseases.

[0161] Therefore, in another respect, the present invention relates to the use of genetically modified immune cells for the prevention or treatment of diseases.

[0162] This invention relates to pharmaceutical compositions comprising genetically modified immune cells for the prevention or treatment of diseases.

[0163] This invention relates to methods for preventing or treating diseases, including administering genetically modified immune cells.

[0164] This invention relates to the use of genetically modified immune cells in the preparation of medicaments for the prevention or treatment of diseases.

[0165] In this invention, any disease can be applied without limitation, as long as it is reported to use immune cells, preferably natural killer cells, for prevention or treatment, and examples include, but are not limited to, cancer, infectious diseases and autoimmune diseases.

[0166] In this document, "cancer" has the same meaning as "tumor," and the compositions according to the present invention for the prevention or treatment of disease can be applied to all types of cancer, including solid cancers and leukemia. Unlike leukemia, solid cancers refer to cancers that form masses in organs and cancers that occur in most organs. In this invention, cancer can be selected from the group consisting of: colon cancer, melanoma, stomach cancer, liver cancer, lung cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, cervical cancer, bladder cancer, kidney cancer, gallbladder cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumors, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, and lymphoma.

[0167] The pharmaceutical compositions according to the present invention for the prevention or treatment of diseases may contain only a pharmaceutically effective amount of genetically modified immune cells, or, in addition to the said compound, may contain at least one pharmaceutically acceptable loading agent, excipient, or diluent. The term "pharmaceutical effective amount" refers to an amount sufficient to prevent, improve, and treat disease symptoms.

[0168] As used herein, the term "pharmaceutically acceptable" means that when administered to a human, it is physiologically acceptable and will not cause common allergic reactions such as gastrointestinal disturbances or dizziness or similar reactions. Examples of loading agents, excipients, and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. Furthermore, the composition may further comprise fillers, anti-aggregating agents, lubricants, humectants, flavoring agents, emulsifiers, and preservatives.

[0169] The compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained, or delayed release of the active ingredient upon administration to mammals. The formulation may be in the form of powder, granules, tablets, emulsion, syrup, aerosol, soft or hard gelatin capsules, sterile injectable solutions, or sterile powder.

[0170] Furthermore, the compositions according to the invention can be administered via any of a variety of routes of administration, including intravenous, transdermal, subcutaneous, intramuscular, or oral administration. The dosage of the active ingredient can be appropriately selected based on various factors such as the route of administration, the patient's age, sex, weight, and the severity of the patient's disease. Moreover, the compositions according to the invention for the prevention or treatment of disease can be administered in combination with known therapeutic agents or compounds that have the effect of preventing, improving, or treating the target disease.

[0171] In this invention, specific amino acid and nucleotide sequences have been described; however, it will be apparent to those skilled in the art that amino acid sequences substantially identical to those of the enzymes implemented in this invention, and nucleotide sequences encoding them, fall within the scope of this invention. "Substantially identical" includes cases where the amino acid or nucleotide sequences have a very high degree of homology, and means shared structural features unrelated to sequence homology, or proteins having the same function as those used in this invention. Proteins with partially deleted sequences other than those constituting the subject matter of this invention, or fragments of the nucleotide sequences encoding such proteins, also fall within the scope of this invention. Therefore, this invention includes all amino acid or nucleotide sequences having the same function as those used in this invention, regardless of fragment length.

[0172] Example The present invention will be described in more detail below with reference to the following embodiments. However, it will be apparent to those skilled in the art that the following embodiments are for illustrative purposes only and should not be construed as limiting the scope of the invention.

[0173] Example 1. Selection of IL-2 / IL-15 fusion protein candidates with reduced binding affinity for IL-2R (IL-2Rβ) compared to IL-2 and Fc-IL-2. Example 1-1: Generation of a fusion protein including IL-2 / IL-15 chimeric protein, IL-15Rα, and Fc domain To generate a fusion protein comprising a chimeric protein or a variant thereof, the fusion protein comprising an Fc domain, an IL-15Rα (sushi domain), and IL-2 / IL-15, and nucleic acid sequences encoding amino acid sequences of SEQ ID NO: 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, or 75, and SEQ ID NO: 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, or 74, were synthesized via the gBlock gene fragment service of Integrated DNA Technologies and then cloned into the pcDNA3.4 vector (Figure 1B).

[0174] The resulting fusion protein has the following formula: N'-[hinge domain]-[Fc domain]-[connector]-[IL15Rα / sh]-[connector]-[IL-2 / IL-15 chimeric protein or its variant]-C'; or N'-[IL-2 / IL-15 chimeric protein or its variants]-[connector]-[IL15Rα / sh]-[connector]-[hinge domain]-[Fc domain]-C' The generated vectors were introduced into CHO cells (Expi-CHO™) to express each fusion protein. After vector introduction, cultures were collected for 5 days at 37°C, 125 RPM, and 8% CO2, and the culture medium was purified. The purified fusion proteins, the names of the contained IL-2 / IL-15 chimeric proteins or their variants, and their amino acid and nucleic acid sequence numbers are shown in Tables 6 and 7 below: [Table 6] Fusion protein (IL-2 / IL-15 variant) (mutation) amino acid sequence SEQ IDNO: GIC-982C1(IL-2 / IL-15) GSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGL PSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSL SLSLGGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGGGGSGGGGSAPTSSSTKKTQLQ LEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 90 GIC-982C2(IL-2 / IL-15v1)(D73E) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRELISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 91 GIC-982C3(IL-2 / IL-15v2)(L74F) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 92 GIC-982C4(IL-2 / IL-15v3)(S76D) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 93 GIC-982C5(IL-2 / IL-15v4)(N77D) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISDINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 94 GIC-982C6(IL-2 / IL-15v5)(V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 95 GIC-982C7(IL-2 / IL-15v6)(L74F、S76D、N77D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 96 GIC-982C8(IL-2 / IL-15v7)(D73E、S76D、N77D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRELIDDINLIVLELKGSGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 97 GIC-982C9(IL-2 / IL-15v8)(D73E、L74F、N77D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPREFISDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 98 GIC-982C10(IL-2 / IL-15v9)(D73E、L74F、S76D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPREFIDNILIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 99 GIC-982C11(IL-2 / IL-15v10)(D73E、L74F、S76D、N77D) GSAESKYGPPCPPAPEAAGGPSVFLFPPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPREFIDNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 100 GIC-982C12(IL-2 / IL-15v11)(D73E、L74F、S76D、N77D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPREFIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 101 GIC-982C13(IL-2 / IL-15v6)(S76E) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIENINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 102 GIC-982C14(IL-2 / IL-15v7)(S76N) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLINNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 103 GIC-982C15(IL-2 / IL-15v8)(S76K) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIKNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 104 GIC-982C16(S76L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLILNINVIVLELKGSGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 105 GIC-982C17(L74F、S76D) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 106 GIC-982C18(S76D、N77D) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDDINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 107 GIC-982C19(S76D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 108 GIC-982C20(L74F、S76D、N77D) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDDINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 109 GIC-982C21(S76D、N77D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 110 GIC-982C22(L74F、S76D、V80L) GSAESKYGPPCPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLGGGGSGGGSIGSITCPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGSGGGSGGGSGAPTSSTTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLT 111 GIC-982N1 APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 112 GIC-982N2(D73E) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRELISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 113 GIC-982N3(L74F) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRDFISNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 114 GIC-982N4(S76D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRDLIDNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 115 GIC-982N5(N77D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISDINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 116 GIC-982N6(V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLISNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 117 GIC-982N7(L74F、S76D、N77D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 118 GIC-982N8(D73E、S76D、N77D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRELIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 119 GIC-982N9(D73E、L74F、N77D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPREFISDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 120 GIC-982N10(D73E、L74F、S76D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPREFIDNILILLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 121 GIC-982N11(D73E、L74F、S76D、N77D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPREFIDDINVIVLELKGSGNNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 122 GIC-982N12(D73E、L74F、S76D、N77D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPREFIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 123 GIC-982N13(S76E) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIENINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 124 GIC-982N14(S76N) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLINNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 125 GIC-982N15(S76K) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIKNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 126 GIC-982N16(S76L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRDLILNINVIVLELKGSGNNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 127 GIC-982N17(L74F、S76D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDNINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 128 GIC-982N18(S76D、N77D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTMKCFLLELQVISLESKNFHLRPRDLIDDINVIVLELKGSGNNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 129 GIC-982N19(S76D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 130 GIC-982N20(L74F、S76D、N77D) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDDINVIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 131 GIC-982N21(S76D、N77D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDLIDDINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 132 GIC-982N22(L74F、S76D、V80L) APTSSSTKKTQLQLEHLLLDLQMILNGSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESKNFHLRPRDFIDNINLIVLELKGSNGNVTESGCKECEELEEKNIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSGGGGSITCPPPMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRGGGGSGGGGSGSAESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDQLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVLHEALHNHYTQKSLSLSLG 133 Fc-IL2 AESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT 134 [Table 7] Fusion protein (IL-2 / IL-15 variant) (variant) Nucleic acid sequence SEQ ID NO: GIC-982C1 (IL-2 / IL-15) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 135 GIC-982C2 (IL-2 / IL-15v1) (D73E) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGCTGATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 136 GIC-982C3 (IL-2 / IL-15v2) (L74F) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 137 GIC-982C4 (IL-2 / IL-15v3) (S76D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 138 GIC-982C5 (IL-2 / IL-15v4) (N77D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 139 GIC-982C6(IL-2 / IL-15v5)(V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 140 GIC-982C7(IL-2 / IL-15v6)(L74F、S76D、N77D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 141 GIC-982C8(IL-2 / IL-15v7)(D73E、S76D、N77D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGCTGATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 142 GIC-982C9(IL-2 / IL-15v8)(D73E, L74F, N77D, V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCTCCGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 143 GIC-982C10(IL-2 / IL-15v9)(D73E、L74F、S76D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACAACATCAACCTGATCGTCCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 144 GIC-982C11(IL-2 / IL-15v10)(D73E、L74F、S76D、N77D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 145 GIC-982C12(IL-2 / IL-15v11)(D73E、L74F、S76D、N77D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 146 GIC-982C13(IL-2 / IL-15v6)(S76E) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGAAAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 147 GIC-982C14(IL-2 / IL-15v7)(S76N) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCAACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 148 GIC-982C15(IL-2 / IL-15v8)(S76K) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCAAGAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 149 GIC-982C16(S76L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCCTCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 150 GIC-982C17(L74F、S76D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 151 GIC-982C18(S76D, N77D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 152 GIC-982C19(S76D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 153 GIC-982C20(L74F、S76D、N77D) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 154 GIC-982C21(S76D、N77D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 155 GIC-982C22(L74F、S76D、V80L) GGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGCGGTGGCGGAGGATCTGGCGGAGGCGGATCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACC 156 GIC-982N1 GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 157 GIC-982N2(D73E) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGCTGATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 158 GIC-982N3(L74F) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCTCCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 159 GIC-982N4(S76D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 160 GIC-982N5(N77D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 161 GIC-982N6(V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 162 GIC-982N7(L74F、S76D、N77D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 163 GIC-982N8(D73E、S76D、N77D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGCTGATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 164 GIC-982N9(D73E、L74F、N77D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCTCCGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 165 GIC-982N10(D73E、L74F、S76D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACAACATCAACCTGATCGTCCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 166 GIC-982N11(D73E、L74F、S76D、N77D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 167 GIC-982N12(D73E、L74F、S76D、N77D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCAAGAGAGTTCATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 168 GIC-982N13(S76E) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGAAAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 169 GIC-982N14(S76N) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCAACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 170 GIC-982N15(S76K) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCAAGAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 171 GIC-982N16(S76L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGTTGGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCCTCAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 172 GIC-982N17(L74F、S76D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACAACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 173 GIC-982N18(S76D, N77D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 174 GIC-982N19(S76D、V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 175 GIC-982N20(L74F、S76D、N77D) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACGACATCAACGTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 176 GIC-982N21(S76D, N77D, V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCGACGACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 177 GIC-982N22(L74F, S76D, V80L) GCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAACGGCTCCATGCACATCGACGCTACCCTGTACACCGAGTCCGACGTGCACCCTTCCTGTAAAGTGACCGCCATGAAGTGCTTTCTGCTCGAGCTGCAAGTGATCTCCCTGGAATCCAAGAACTTCCACCTGAGGCCTCGGGACTTCATCGACAACATCAACCTGATCGTGCTGGAACTGAAGGGCTCCAACGGCAACGTGACCGAGTCTGGCTGTAAAGAGTGCGAGGAACTGGAAGAGAAGAACATCGTCGAGTTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCTCCACACTGACCGGCGGCGGAGGAAGCGGTGGCGGCGGTAGCGGAGGTGGTGGTTCTGGTGGTGGCGGTTCTATTACATGCCCTCCTCCAATGTCCGTGGAACACGCCGACATCTGGGTCAAGTCCTACAGCCTGTACTCCAGAGAGCGGTACATCTGCAACTCCGGCTTCAAGAGAAAGGCCGGCACCTCTAGCCTGACCGAGTGCGTGCTGAACAAGGCCACCAATGTGGCCCACTGGACAACCCCTAGCCTGAAGTGTATTAGAGGTGGCGGAGGATCTGGCGGAGGCGGATCTGGATCCGCCGAGTCTAAGTACGGCCCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCCGGAGGGCCCAGTGTGTTTCTGTTCCCTCCAAAGCCTAAGGACCAGCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGACCCTGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCTGTGCTGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGC 178 Fc-IL2 GCCGAGTCTAAGTACGGACCTCCTTGTCCTCCATGTCCTGCTCCAGAAGCTGCTGGCGGCCCTTCCGTGTTTCTGTTCCCTCCAAAGCCTAAGGACACCCTGATGATCTCTCGGACCCCTGAAGTGACCTGCGTGGTGGTGGATGTGTCCCAAGAGGATCCCGAGGTGCAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAACGCCAAGACCAAGCCTAGAGAGGAACAGTTCAACTCCACCTACAGAGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGGCCTGCCTTCCAGCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTTTACACCCTGCCTCCAAGCCAAGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAATGGGAGAGCAATGGCCAGCCTGAGAACAACTACAAGACCACACCTCCTGTGCTGGACTCCGACGGCTCCTTCTTTCTGTACTCCCGCCTGACCGTGGACAAGTCCAGATGGCAAGAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCTCTGTCCCTTGGAGGTGGCGGAGGATCTGCTCCTACCTCCTCCAGCACCAAGAAAACCCAGCTGCAGTTGGAGCATCTGCTGCTGGACCTGCAGATGATCCTGAATGGCATCAACAATTACAAGAACCCCAAGCTGACCCGGATGCTGACCTTCAAGTTCTACATGCCCAAGAAGGCCACCGAGCTGAAACATCTGCAGTGCCTGGAAGAGGAACTGAAGCCCCTGGAAGAAGTGCTGAATCTGGCCCAGTCCAAGAACTTCCACCTGAGGCCTCGGGACCTGATCTCCAACATCAACGTGATCGTGCTCGAGCTGAAGGGCTCCGAGACAACCTTCATGTGCGAGTACGCCGACGAGACAGCTACCATCGTGGAATTTCTGAACCGGTGGATCACCTTCTGCCAGTCCATCATCAGCACCCTGACC 179 GIC-982C1 was purified to GIC-982C15 using an open column containing protein A resin. The protein A resin from Repligen was washed and equilibrated with PBS and protein A binding buffer from Thermo. The supernatant, filtered through a 0.22 μm filter, was then reacted with the protein A resin. The protein A resin reacted in the PBS-washed open column was collected, and the column was washed with protein A binding buffer at a volume 10 times the resin volume. The binding buffer was then added to a collection tube at a volume 1 / 10 that of elution buffer, followed by elution and collection with Thermo's IgG elution buffer. The collected fusion protein was replaced with PBS buffer.

[0175] At this point, the isolated and purified fusion protein was subjected to SDS-PAGE under reducing (R) or non-reducing (NR) conditions and confirmed by Coomassie brilliant blue staining (Figures 1C to 1F).

[0176] Examples 1-2: Measurement of IL-2Rβ binding affinity using Octet binding assay To determine the binding affinity of GIC-982C1 to GIC-982C6 and GIC-982C12 to GIC-982C15 for IL-2Rβ, Octet binding assays were performed using a Ni-NTA sensor chip (ForteBio 18-5101). Fc-IL2 was selected as a control for the analysis. First, the sensor chip was activated by immersion in distilled water for 5 minutes. The sensor was baselined in PBS for 3 minutes and then immersed in a solution containing 6 µg / ml IL-2Rβ-his (Acro CD2-H5221) to bind to the sensor. The baseline was then held in PBS for 3 minutes and then immersed in serially diluted 1 / 2 doses of the test substance from 2,000 nM to 31.25 nM for 5 minutes to bind to IL-2Rβ. PBS buffer without the test substance was used as a blank for the analysis at this point. The results were then transferred back to PBS and dissociated for 10 minutes. All steps were performed at room temperature, with the plate shaken at 1,000 RPM. Experimental results obtained using ForteBio data analysis software are shown in Table 8 below. Figure 2 As shown.

[0177] The binding affinity was confirmed using Octet. The results showed that GIC-982C1 to GIC-982C6 and GIC-982C12 to GIC-982C15 exhibited relatively higher dissociation constants (Kd) than Fc-IL2, which implies a weakened binding affinity for IL-2Rβ.

[0178] [Table 8] Examples 1-3: Confirmation of the role of IL-2 receptor (IL-2R)-mediated STAT5 mechanism activity using STAT5 signal transduction assays. To evaluate STAT5 activity in GIC-982C1 to GIC-982C6 and GIC-982C12 to GIC-982C15, HEK-Blue CD122 / 132 cells (InvivoGen hkb-il2bg) expressing IL-2Rβ and a common γ chain were cultured, and STAT5 activity was analyzed. IL2-Fc was selected as a control group for analysis.

[0179] HEK-Blue CD122 / 132 cells were cultured in DMEM [(Gibco, Cat No. 10569010) + 10% FBS (Gibco, Cat No. 26140079) + penicillin / streptomycin (Gibco, Cat No. 15140122) + Normocin (InvivoGen, Ant-Nr-2) + 1XHEK-Blue™ Selection (InvivoGen, Hb-Sel) + 1 µg / mL puromycin (InvivoGen, Ant-Pr-1)] and screened for activity using antibiotic-free DMEM [(Gibco, Cat No. 10569010) + 10% FBS (Gibco, Cat No. 26140079) + penicillin / streptomycin (Gibco, Cat No. 15140122)]. Cultured cells were treated with trypsin-EDTA, collected, and washed with added culture medium. The collected HEK-Blue CD122 / 132 cells were centrifuged at 300 x g for 5 minutes, the supernatant was removed, and the residue was resuspended in fresh culture medium. The HEK-Blue CD122 / 132 cells were then counted and diluted with culture medium to adjust the cell number to 2.5 x 10⁻⁶ cells / year. 5 Cells / mL. The control group protein was diluted to the highest concentration used in the assay (based on 100 ng / mL IL-2), loaded onto plates, and serially diluted 1 / 10. 20 mL of the diluted protein was loaded onto a flat-bottomed plate, and 180 mL (45,000 cells) of the pre-prepared cells were added. The plates were incubated at 37°C for 24 hours in the presence of 5% CO2.

[0180] To prepare the QUANTI-Blue™ solution (InvivoGen, rep-qbs) for detection, 750 mL of QB reagent and QB buffer were added to 73.5 mL of distilled water, vortexed to mix thoroughly, and incubated at room temperature for 10 minutes. 20 mL of each supernatant from plates cultured for 24 hours was loaded onto a new plate, and 180 mL of the previously prepared QUANTI-Blue™ solution was added. The plates were then incubated at 37°C in the dark for 1 hour. The absorbance measured at 630 nm using a microplate reader is shown below. Figure 3 In, and from this EC 50 As shown in Table 9.

[0181] As a result, GIC-982C1 to GIC-982C5 and GIC-982C13 to GIC-982C15 were found to have lower STAT5 activity than Fc-IL2 and were substances that could maintain appropriate levels of cell activity while preventing cell overactivity.

[0182] [Table 9] Examples 1-4: Determination of the anticancer efficacy and reduction of side effects of IL-2 / IL-15 fusion protein in a mouse in vivo anticancer model To determine the anticancer efficacy of the IL-2 / IL-15 chimeric protein and its ability to reduce the side effect of pulmonary edema caused by excessive immune activation, 5 x 10 g of the protein were administered subcutaneously to Balb / c mice. 5 A mouse model was established using CT26 (a colon cancer cell line) cells / head. The volume of the generated tumor was measured using calipers, and when the volume was approximately 70 mm... 3 Mice were divided into groups. Then, IL2-Fc, GIC-982C1, GIC-982C4, and GIC-982C15 were administered intraperitoneally to mice at a dose of 10 mg / kg twice weekly for two weeks, for a total of four administrations. Tumor volume was measured on days 1, 4, 7, 11, 14, 16, and 18 after administration to determine anticancer efficacy, and the results are shown in Table 10.

[0183] In addition, the condition of the mice was observed to determine if any side effects occurred due to the administration. Adverse reactions in deceased subjects were visually assessed, and autopsies were performed on deceased subjects to examine the condition of internal organs. To determine the occurrence of side effects due to the administration, the removed lungs were weighed, dried for 2 days, and then weighed again to calculate the wet / dry ratio. The results are shown in Table 11.

[0184] As a result, under a high dose of 10 mpk, Fc-IL2 caused lung death due to pulmonary edema induced by immune hyperactivity, while GIC-982C1, GIC-982C4, and GIC-982C15 exhibited excellent inhibitory activity against cancer cell proliferation. Furthermore, Fc-IL2 showed increased wet lung weight, while the wet / dry ratio of GIC-982C1, GIC-982C4, and GIC-982C15 was the same as that of the carrier, indicating that pulmonary edema did not occur.

[0185] [Table 10] [Table 11] Example 2. Confirmation of the generation and effects of immune cells modified to express IL-2 / IL-15 fusion protein. Example 2-1: Plasmid Construction Two cloning vectors were generated using retroviral vectors (Biovec pharma, SinVec(K)-GFP-BSD, Vec-033), which contain the amino acid sequences of SEQ ID NO: 183 and 185 and the nucleic acid sequences of SEQ ID NO: 184 and 186 as shown in Table 12 below, and their schematic diagram is shown in Figure 4A.

[0186] A schematic diagram of immune cells expressing the IL-2 / IL-15 fusion protein vector is shown in Figure 4B, and the expressed fusion protein is as follows: N'-[IL-2 / IL-15 chimeric protein]-[linker]-[IL15Rα]-C' [Table 12] Example 2-2: Retrovirus Production and Concentration To generate BaEV pseudotyped retroviruses, 6.0 x 10⁻⁶ cells were prepared the day before transfection. 6293Vec-BaEV cell lines (Biovec pharma, Vec-006) were cultured in T75 flasks at 37°C in a 5% CO2 incubator for 24 hours. When the cell density reached 80%, 15 µg of plasmid encoding the chimeric protein was transfected using lipofectamine 2000 (Invitrogen, 11668500). Forty-eight hours after transfection, the culture medium containing retroviruses was harvested, centrifuged at 500 xg at 4°C for 10 min, and passed through a 0.45 µm PVDF membrane filter (MILLIPORE, SE1M003M00) to remove cell debris. The culture medium containing the cell-debrided retroviruses was mixed with Retro-X concentrator (Clontech, 631456) reagent at a 3:1 ratio (virus medium: Retro-X concentrator reagent) for retrovirus concentration and reacted under refrigeration at 4°C. After 18 hours of reaction, the retrovirus pellet obtained by centrifugation at 1,500 × g and 4 °C for 45 min was resuspended in Opti-MEM medium (Gibco, 11058021) and concentrated. The concentrated retrovirus was stored at -80 °C.

[0187] Examples 2-3: Preparation of natural killer cells derived from human peripheral blood mononuclear cells (PBMCs) Blood collected from different healthy donors was centrifuged at 400 xg for 30 min using a Ficoll-Hypaque concentration gradient, and PBMCs were isolated from the erythrocyte sedimentation rate (ESR) layer. The isolated PBMCs were counted using an ADAM-MC2 automated cell counter (NanoEnTek), transferred to new tubes to obtain CD3(-) cells, and then centrifuged at 350 xg for 10 min at 4°C. After centrifugation, the supernatant was removed, and cells were centrifuged at 1 x 10⁻⁶ cells per 1 x 10⁻⁶ cells. 7 Each cell was dispensed with 80 mL of CliniMACS buffer (20% human serum albumin, EDTA 2 mM) and 22 mL of CD3 magnetic beads (Miltenyi biotech, 130-050-101) to suspend the cell pellet, and incubated at 4°C in the dark for 15 minutes. After the reaction, 10 mL of CliniMACS buffer was dispensed for washing, centrifuged at 350 x g at 4°C for 10 minutes, and then each 1 x 10⁻⁶ cells were washed. 8 Each cell was dispensed with 0.5 mL of CliniMACS buffer to suspend the cell pellet.

[0188] Flow 3 mL of CliniMACS buffer into an LD column (Miltenyi Biotec, 130-042-901) and soak, then flow cell suspension into the column to obtain CD3(-) cells that pass through the column. Wash with 10 mL of CliniMACS buffer and centrifuge the cell pellet at 350 x g and 4 °C for 10 min. Resuspend the cell pellet in CliniMACS buffer and measure the cell count using an automated cell counter. Then, every 1 x 10⁻⁶ cells... 7 Each cell was dispensed with 80 mL of CliniMACS buffer and 22 mL of CD56 magnetic beads (Miltenyi biotech, 130-050-401), the cell pellet was resuspended, and the reaction was carried out at 4°C in the dark for 15 minutes. For washing, 10 mL of CliniMACS buffer was dispensed and centrifuged at 350 x g at 4°C for 10 minutes, and then each 1 x 10⁻⁶ cell pellet was centrifuged. 8 Each cell was dispensed with 0.5 mL of CliniMACS buffer to suspend the cell pellet.

[0189] Flow 3 mL of CliniMACS buffer onto an LS column (Miltenyi Biotec, 130-042-401) and soak, allowing the cell suspension to flow. Then, detach the column from the magnetic holder, add 5 mL of CliniMACS buffer, pressurize with the stopcock, and obtain CD3(-)CD56(+) natural killer cells in a new tube. Centrifuge the obtained cells at 350 x g and 4 °C for 10 min. After centrifugation, resuspend the cell pellet in CliniMACS buffer and measure the cell count using an automated cell counter. Count the cells at 10 x 10⁻⁶ cells per cell line. 6 / mL to 20 x 10 6 / mL of cells were suspended in CryoStor CS10 cryopreservative (Biolife solution, 210102). The suspended cells were dispensed into 1 mL vials, initially frozen once using a cell freezing container (Corning, CLS432001), and then transferred to an LN2 container for secondary freezing.

[0190] Examples 2-4: Generation of natural killer cells expressing chimeric proteins including IL-2 and IL-15 To culture the isolated natural killer cells from Examples 2-3, 100 mL of CD335 (NKp46)-biotin and 100 mL of CD2-biotin, included in the NK cell activation / expansion kit (Miltenyi Biotec, 130-094-483), were dispensed into new tubes, mixed, and then combined with 500 mL of antibiotin MACSI beads (MACSIBeads). 300 mL of CliniMACS buffer was added to the same tube, and the mixture was reacted at 4°C for 2 hours at 4.0 rpm using a microtube spinneret (Thermo Scientific, Hulamixer, 15920D). The frozen natural killer cells from Examples 2-3 were thawed, and cell counts were measured. The previously prepared beads were washed with 1 mL of CliniMACS buffer, based on 5 mL beads / 1 x 10⁶ cells / day. 6 Cells were suspended in CN5-101 medium (CTS™ NK-Xpander™ medium, 1X supplement (Gibco, A5019001), 5% human AB serum (Milan Analytica AG, #000084) and 50 nM GI-101 (GI-inovation, 8.2 mg / mL)), seeded in 6-well plates, and cultured at 37°C for 4 days in the presence of 5% CO2.

[0191] To infect the retroviruses generated in Examples 2-2, untreated 12-well plates were coated with 10 µg / mL of reverse transcriptase protein (Takara, T100B) according to the manufacturer's instructions. 300 mL of retrovirus equivalent to 1 to 10 MOI was added to each well, and the plates were centrifuged at 2,000 x g at 32°C for 2 hours to adsorb the retroviruses onto the protein-coated plates. The supernatant was removed, and the plates were washed once with 1 mL of DPBS (WELGENE, LB001-02). On day 4 after the start of culture, the number of natural killer cells was measured and counted at 0.8 x 10⁻⁶. 6 Cells were suspended at a density of 100 cells / mL in CN5-101 medium. 1 mL of the result was seeded into each well of a plate coated with the prepared natural killer cells, and then further seeded with 0.6 mL of CN5-101 medium. The result was centrifuged at 1,000 x g at 32°C for 15 min to ligate the retrovirus and natural killer cells together, and cultured at 37°C for 3 days in the presence of 5% CO2. 6 The cells were seeded at a density of 10 cells / mL and then passaged.

[0192] Specifically, on the third day after transduction and culture, the expression of the chimeric protein was detected by flow cytometry at a concentration of 0.5 x 10⁻⁶ cells / mL. 6 Cells were suspended at a density of 0.5 x 10⁶ cells / mL and cultured in CN5-101 medium containing GI-101 and CN5 medium without GI-101 (CTS™ NK-Xpander™ medium, 1X supplement (Gibco, A5019001), 5% human AB serum (Milan Analytica AG, #000084)). Cell counts were then measured under both CN5-101 and CN5 medium conditions, at a density of 0.5 x 10⁶ cells / mL. 6 Cells were passaged at a density of 1 cell / mL for 5 mL of cells.

[0193] Examples 2-5: Analysis of the expression of chimeric proteins containing IL-2 and IL-15 Retroviral transduction was induced, natural killer cells were obtained on day 3, and 0.2 x 10⁻⁶ cells were counted using an automated cell counter. 6 Cells were seeded in 96-well plates. 250 µl of FACS buffer (DPBS containing 2% FBS) was added, and the cells were centrifuged at 350 x g for 5 minutes. After centrifugation, the supernatant was removed, and the cell pellet was resuspended in FITC-labeled anti-CD56 antibody (Biolegend, 362546), BV650-labeled anti-IL-15Rα antibody (BD, 747701), and a LIVE / DEAD-fixable purple dye sample (Invitrogen, L34955A), and incubated at 4°C for 30 minutes. Then, 150 µl of FACS buffer was added, and the cells were centrifuged at 350 x g for 5 minutes at 4°C. The supernatant was removed, and the cell pellet was resuspended in 200–250 µl of FACS buffer and seeded onto FACS tubes. The expression of IL-15Rα, which constitutes the chimeric protein, was measured using flow cytometry (FACSymphony™ A3 Cell Analyzer, BD).

[0194] As a result, from Figure 5 It can be seen that natural killer cells expressing membrane-bound chimeric proteins containing IL-2 and IL-15 (mbIL-2 / IL-15) on the cell membrane surface showed an expression efficiency of 88%, which was higher than the 25.1% of membrane-bound IL-15 protein (mbIL-15).

[0195] Examples 2-6: Comparison of in vitro cell counting and cell viability On day 3 post-transduction, the cultured natural killer cells were cultured at the same cell density and quantity in CN5 medium for 19 days, and then in CN5-101 medium supplemented with GI-101 for 25 days. On days 0, 4, 7, 10, 13, 16, 19, 22, and 25, cells adhering to the bottom of the flask were removed with a scraper (SPL, 90020), and aspirated and decomposed using a pipette to obtain samples for cell counting. The cell density (x 10⁻¹⁰) of the obtained cells was measured using an automated cell counter. 6 Cell counts were performed using cell density per mL, cell viability (%), and dead cell count. Total cell counts on days 0 and 4 were calculated by multiplying the cell density of live cells by the total volume of the cell culture medium (mL), and then multiplied by 0.5 x 10⁻⁶. 6 Natural killer cells were suspended in 5 mL of water at a cell density of 1 cell / mL and passaged. Therefore, the total cell count on day 7 was calculated as follows: Predicted total cell count = viable cell density (x 10) 6 (cells / mL) / seeding cell density of the previous culture day (0.5 x 10⁻⁶) 6 (cells / mL) x Total cell count from the previous culture day (x 10) 6 (cells) Cell fold expansion was calculated using both the calculated total cell count and the predicted total cell count. Cell fold expansion was calculated by dividing the total cell count or predicted total cell count for each culture day by the total cell count on day 0 of culture. Cell proliferation was stimulated in the presence of GI-101 (a cell proliferation-promoting protein), or cultured in the absence of GI-101, and the results were shown in… Figure 6 middle.

[0196] As a result, from Figure 6 It can be seen that, compared with untransformed normal natural killer cells (UTD) and natural killer cells expressing membrane-bound IL-15 protein (mbIL-15), natural killer cells expressing membrane-bound chimeric proteins containing IL-2 and IL-15 (mbIL-2 / IL-15) have better proliferation and survival capabilities in the presence or absence of GI-101 (a cell proliferation-promoting protein).

[0197] Example 3. This demonstrated the improved anticancer efficacy and side effects of the IL-2 / IL-15 fusion protein and natural killer cells expressing this fusion protein compared to the control group. Example 3-1: Confirmation of in vitro cell-killing efficacy using various cancer cell lines Target tumor cell lines (SKOV3, HCT116, A549) used for in vitro cytotoxic activity evaluation were infected with a lentivirus (Satorious, 4475) containing GFP and a puromycin resistance gene, and cultured in R10 medium (10% fetal bovine serum, 1% penicillin-streptomycin) containing 0.5 g / mL puromycin. Only transduced tumor cells were selected and cultured. The day before in vitro cytotoxic activity evaluation, the target tumor cell lines were cultured at 3 x 10⁻⁶ cells / mL. 4 1 cell / mL (A549 and HCT116 cell lines) or 5 x 10 4 The target tumor cell line was suspended at a density of 100 µl / mL (SKOV3 cell line) in R10 medium, and 100 µl of the target tumor cell line was seeded into each well of a 96-well plate (Corning, 3799) and cultured at 37°C in the presence of 5% CO2 for 16 hours.

[0198] Natural killer cells expressing mbIL-2 / IL-15, generated in Examples 2-4, were thawed, washed, and seeded into 96-well plates containing target tumor cells prepared the previous day at an E / T (effectant to target) ratio of 10:1 or 3:1. The plates were imaged every 4 hours for 96 hours using an Incucyte® S3 device (Sartorius) mounted in a cell culture incubator. Positive target tumor cells exhibiting GFP fluorescence were then counted and analyzed from the stored images.

[0199] As a result, from Figure 7 It can be seen that, compared with UTD and mbIL-15, natural killer cells expressing mbIL-2 / IL-15 have high cell killing ability in the three types of tumor cells.

[0200] Industrial applicability According to the present invention, natural killer cells modified to express chimeric proteins comprising IL-2 and IL-15, their variants, or fusion proteins containing such chimeric proteins or their variants, suppress the side effects caused by excessive immune cell activity due to IL-2 and prevent the rapid depletion of natural killer cells, thereby maintaining appropriate levels of cell activity and cell viability for a prolonged period. Therefore, based on chimeric proteins comprising IL-2 and IL-15, which have excellent innate immune function, the present invention provides protein complexes comprising such chimeric proteins, fusion proteins comprising such chimeric proteins, and natural killer cells modified to express such proteins.

[0201] Although specific configurations of the invention have been described in detail, those skilled in the art will understand that this detailed description is provided for illustrative purposes as a preferred embodiment and should not be construed as limiting the scope of the invention. Therefore, the essential scope of the invention is defined by the appended claims and their equivalents.

[0202] Sequence List Free Text Electronic files have been attached.

Claims

1. A chimeric protein or a variant thereof, wherein the chimeric protein or the variant thereof is obtained by replacing the AB loop domain, helical B domain and CD loop domain of IL-2 with the AB loop domain, helical B domain and CD loop domain of IL-15, respectively.

2. The chimeric protein or a variant thereof according to claim 1, wherein, The chimeric protein or a variant thereof includes the amino acid sequence of SEQ ID NO:

33.

3. The chimeric protein or a variant thereof according to claim 1, wherein, The chimeric protein or its variants comprise a substitution at at least one amino acid position selected from the group consisting of amino acids at positions 73, 74, 76, 77, and 80 of the amino acid sequence of SEQ ID NO:

33.

4. The chimeric protein or a variant thereof according to claim 1, wherein, The variant includes at least one amino acid substitution selected from the group consisting of D73E, L74F, S76D, S76E, S76N, S76K, S76L, N77D and V80L of the amino acid sequence of SEQ ID NO:

33.

5. The chimeric protein or a variant thereof according to claim 1, wherein, The variant includes at least one amino acid substitution selected from the group consisting of D73E, L74F, S76D, N77D and V80L in the amino acid sequence of SEQ ID NO:

33.

6. The chimeric protein or a variant thereof according to claim 1, wherein, The variants include amino acid sequences of SEQ ID NO: 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73 or 75.

7. A fusion protein comprising the chimeric protein of claim 1 or a variant thereof.

8. The fusion protein according to claim 7 further comprises IL-15 receptor α protein or a fragment thereof.

9. The fusion protein according to claim 8, wherein, The fragment of the IL-15 receptor α protein includes the sushi domain of the IL-15 receptor α protein.

10. The fusion protein according to claim 7 further includes an Fc domain.

11. A nucleic acid encoding a chimeric protein according to claim 1 or a variant thereof, or a fusion protein comprising the chimeric protein or a variant thereof.

12. A vector comprising the nucleic acid according to claim 11.

13. A host cell, said host cell having the nucleic acid according to claim 11 or the vector according to claim 12 introduced therein.

14. A genetically modified cell that expresses the chimeric protein or a variant thereof according to claim 1, or a fusion protein comprising the chimeric protein or a variant thereof.

15. The gene-modified cell according to claim 14, wherein, The genetically modified cells are natural killer cells.

16. The genetically modified cell according to claim 14, wherein, The fusion protein includes membrane proteins.

17. The gene-modified cell according to claim 16, wherein, The membrane proteins are selected from the group consisting of: IL-15 receptor α (IL-15Rα), CD8α, CD4, CD3ε, CD3γ, CD3δ, CD3ζ, CD28, CD137, FcεRIγ, T cell receptor, nicotinic acetylcholine receptor, GABA receptor and fragments thereof.

18. The genetically modified cell according to claim 14, wherein, The fusion protein has the structure of either formula (I) or formula (II): N'-X-[L1] n -Y-C’(Formula (I)) N'-Y-[L1] n -X-C’(Formula (II)) Wherein, X is a chimeric protein including IL-2 and IL-15 proteins or a variant thereof; Y is the IL-15 receptor α (IL-15Rα) protein or a fragment thereof; L1 is a connector; and n is an integer of 0 or greater.

19. A pharmaceutical composition for the prevention or treatment of cancer, infectious diseases, or autoimmune diseases, comprising at least one of the following: The chimeric protein or a variant thereof according to claim 1; The fusion protein according to claim 7; and The gene-modified cell according to claim 14.

20. A method for producing a chimeric protein or a variant thereof according to claim 1, or a fusion protein comprising said chimeric protein or a variant thereof. The method includes culturing host cells according to claim 13 to produce a chimeric protein or a variant thereof, or a fusion protein comprising the chimeric protein or a variant thereof.

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