Compositions and methods regarding engineered and non-engineered γδ-t cells for treatment of hematological tumors

Abstract

To provide compositions and methods for treatment of hematological tumors with engineered or non-engineered γδ-T cells.SOLUTION: In some embodiments, γδ-T cells comprise a chimeric antigen receptor (CAR) construct. The CAR construct can contain an anti-CD20 binding domain or anti-B cell maturation antigen (BCMA) binding domain, a CD8 hinge and transmembrane domain, a costimulatory domain, a CD3ζ signalling domain, a combination thereof, or all thereof. The CAR construct can contain a domain encoding for a secreted common gamma chain cytokine such as an sIL15 domain.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 62 / 739,822, filed October 1, 2018, the contents of which are incorporated herein in their entirety for all purposes.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The above ASCII copy, created on December 17, 2019, is named ADC-0005-PCT_SL.txt and is 147616 bytes in size. [Background technology]

[0003] Adoptive immune cell therapy has undergone continuous iterations for over 30 years, from early approaches focused on basal lymphokine activation and / or tumor infiltration to more recent strategies such as genetically engineering these immune cells to express engineered antigen receptors, such as chimeric antigen receptors (CARs). While there have been some hints and indications of the curative potential of these approaches, much remains to be done. In particular, successful tumor eradication by CAR-T lymphocytes depends on the persistence and effector function of CAR-T cells, but excess of either can induce graft-versus-host effects in patients. Therefore, the art is testing numerous costimulatory strategies for both T cells and NK cells, particularly αβ T cells, with the aim of balancing efficacy and safety. Notably, given the current lack of understanding regarding the costimulatory requirements of γδ T cells compared to αβ T cells, any actual translation of these various approaches to allogeneic γδ T cells is uncertain at best. See, e.g., Ribot et al., "Searching for 'signal 2': costimulation requirements of γδ T cells," Cell. Mol. Life Sci. (2011) 68:2345-2355.

[0004] Thus, there remains a need for improved strategies to improve cell specificity or selectivity, to improve cell safety, e.g., by reducing or avoiding the graft-versus-host (GVH) effect, to improve cell efficacy, e.g., by avoiding suppression of effector functions, and to improve cell activity and / or survival upon administration to a subject. Methods, cells, compositions, kits, and systems that meet such needs are provided. Summary of the Invention

[0005] Embodiments of the present invention include isolated nucleic acid sequences encoding chimeric antigen receptors (CARs), wherein the CAR comprises a binding domain that specifically binds to a tumor-associated antigen (TAA) expressed on the surface of a hematological tumor cell, the binding domain being, for example, a hinge domain of CD8α, a transmembrane domain of CD8α, a costimulatory signaling region, etc., wherein the costimulatory signaling region is optionally selected from a 4-1BB (CD137) costimulatory signaling region and a CD27 costimulatory signaling region, and a CD3ζ signaling domain.

[0006] Embodiments of the invention further include γδ T cells that are not genetically engineered as described herein, as well as γδ T cells that comprise a nucleic acid encoding a CAR construct as described herein, wherein the γδ T cells functionally express the nucleic acid encoding the CAR on the surface of the γδ T cells. Embodiments of the invention further include a plurality of genetically engineered or non-genetically engineered γδ T cells as described herein. Embodiments of the invention further include methods of generating a γδ T cell or a plurality of γδ T cells as described herein, wherein the method comprises transfecting a γδ T cell(s) with a construct described herein. Embodiments of the invention further include pharmaceutical compositions comprising a pharmaceutically acceptable excipient and a γδ T cell or a plurality of γδ T cells as described herein. Embodiments of the invention further include contacting hematologic tumor cells with an effective tumor cytotoxic amount of γδ T cells as described herein.

[0007] In one aspect, the present invention provides an isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises: (a) a binding domain that specifically binds a tumor-associated antigen (TAA) expressed on the surface of a hematological tumor cell; (b) a hinge domain, such as a CD8α hinge domain; (c) a transmembrane domain, such as a CD8α transmembrane domain; (d) a costimulatory signaling region or a combination of costimulatory signaling regions, optionally wherein the costimulatory signaling region comprises a costimulatory signaling region selected from a 4-1BB (CD137) costimulatory signaling region and a CD27 costimulatory signaling region; and (e) a signaling domain, such as a CD3ζ signaling domain. In some embodiments, the foregoing elements (a)-(e) are encoded in 5' to 3' order on the sense strand of the isolated nucleic acid.

[0008] In some embodiments, the binding domain specifically binds to CD20. In some embodiments, the binding domain selectively binds to, or competes for binding with, an epitope in CD20 bound by an anti-CD20 antibody selected from the group consisting of 3B9, 3H7, 2B7, and 9C11, preferably 3H7. In some embodiments, the binding domain comprises a complementarity-determining region of an anti-CD20 antibody selected from the group consisting of 3B9, 3H7, 2B7, and 9C11, preferably 3H7.

[0009] In some embodiments, the isolated nucleic acid encodes a heavy chain variable region (HCVR) sequence and a light chain variable region (LCVR) sequence, for example, wherein the HCVR sequence and the LCVR sequence are SEQ ID NOs: 99 and 107, respectively, heavy chain complementarity determining region 1, 2, and 3 sequences are SEQ ID NOs: 101, 103, and 105, respectively, and light chain complementarity determining region 1, 2, and 3 sequences are SEQ ID NOs: 109, 111, and 113, respectively, and a heavy chain complementarity determining region 3 (HCDR3) and a light chain CDR3 (LCDR3), wherein the HCDR3 and the LCDR3 are SEQ ID NOs: 345 and 353, 201 and 209, and 249 and 257, and a heavy chain variable region (HCVR) sequence and a light chain variable region (LCVR) sequence, wherein the HCVR sequence and the LCVR sequence are selected from the group consisting of SEQ ID NOs: 339 and 347, 195 and 203, and 243 and 251, and / or a heavy chain complementarity determining region 3 (HCDR3) domain and a light chain CDR3 (LCDR3) domain, wherein wherein the HCDR3 domain comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19, wherein X1 = A, V or T, X2 = K, X3 = D, X4 = P, F or G, X5 = S or H, X6 = Y, X7 = G, X8 = S or H, X9 = G or F, X10 = S or Y, X11 = Y, N or S, X12 = Y, G or H, X13 = G, L or S, X14 = Y, M or D, X15 = Y, D or V, X16 = G, V or absent, X17 = M or absent, X18 = D or absent, X19 = V or absent (sequence number 369), and the LCDR3 domain comprises the amino acid sequence of X1-X2-X3-X4-X5-X6-X7-X8-X9, where X1 = Q, X2 = Q, X3 = R or S, X4 = N, Y or F, X5 = N, D, or Y, X6 = W, X7 = P, X8 = L, and X9 = T (sequence number 370).

[0010] In some embodiments, the isolated nucleic acid encodes a binding domain that specifically binds to CD19 or BCMA. In some embodiments, the binding domain specifically binds to BCMA. In some embodiments, the binding domain selectively binds to, or competes for binding with, an epitope of BCMA bound by an anti-BCMA binding region having a sequence selected from the group consisting of SEQ ID NOs: 27 and 28, SEQ ID NOs: 29 and 30, and SEQ ID NOs: 31 and 32. In some embodiments, the binding domain comprises the complementarity determining regions of an anti-BCMA binding region having a sequence selected from the group consisting of SEQ ID NOs: 27 and 28, SEQ ID NOs: 29 and 30, and SEQ ID NOs: 31 and 32.

[0011] In some embodiments, the CAR comprises a CD8α hinge domain comprising SEQ ID NO: 1 (PTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIY) or SEQ ID NO: 2 (TTTPAPRPPTPAPTIASQPLSLR PEACRPAAGGAVHTRGLDFACDIY). In some embodiments, the CAR comprises a CD8α transmembrane domain comprising SEQ ID NO: 3 (IWAPLAGTCGVLLLSLVITLYC).

[0012] In some embodiments as described herein, the CAR comprises a CD3ζ signaling domain comprising SEQ ID NO: 4 (RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR) or SEQ ID NO: 5 (RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEM GGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR).

[0013] In any one of the foregoing or some embodiments as described herein, the CAR comprises a 4-1BB costimulatory signaling region comprising SEQ ID NO: 6 (KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL) or a CD27 costimulatory signaling region comprising SEQ ID NO: 7 (QRRKYRSNKGESPVEPAEPCH YSCPREEEGSTIPIQEDYRKPEPACSP). In any one of the foregoing or some embodiments as described herein, the isolated nucleic acid encodes a 4-1BB costimulatory signaling region comprising SEQ ID NO: 6 and a CD27 costimulatory signaling region comprising SEQ ID NO: 7.

[0014] In some embodiments as described herein, the isolated nucleic acid encodes a secreted cytokine, or a secreted common gamma chain interleukin, or a secreted common gamma chain interleukin, such as IL-15, preferably wherein the secreted common gamma chain interleukin, such as IL-15, comprises an interleukin polypeptide sequence operably linked to a secretory signal sequence (e.g., a secretory signal of SEQ ID NO: 33 or 49). In some embodiments, the isolated nucleic acid encodes secreted IL-15, preferably wherein the IL-15 comprises the sequence of SEQ ID NO: 34, more preferably wherein the IL-15 comprises the sequence of SEQ ID NO: 34 operably linked to the secretory signal sequence of SEQ ID NO: 33, or wherein the IL-15 comprises the sequence of SEQ ID NO: 34 operably linked to the secretory signal sequence of SEQ ID NO: 49. In some cases, the secreted cytokine, common gamma chain interleukin, and / or IL-15 is carboxy-terminally encoded by a binding region, a hinge and transmembrane domain, a signaling domain, and / or a costimulatory endodomain. In some cases, the secreted cytokine, commonly gamma chain interleukin and / or IL-15, is encoded on the sense strand 3' of the binding region, hinge and transmembrane domain, signaling domain and / or costimulatory endodomain.

[0015] In some embodiments, the nucleic acid encodes a multicistronic linker region configured to facilitate translation of the CAR and the secreted cytokine, general gamma chain cytokine, or IL-15 as separate polypeptides. In some embodiments, the multicistronic linker region encodes a self-cleaving sequence and / or a cleavage polypeptide sequence. Optionally, the self-cleaving sequence is a P2A, F2A, T2A, or E2A self-cleaving sequence. Optionally, the cleavage sequence is a furin cleavage sequence. Optionally, the cleavage sequence (e.g., a furin cleavage sequence) is amino-terminal to the self-cleaving sequence. In some embodiments, the multicistronic linker region encodes an internal ribosome entry site. In some embodiments, the nucleic acid encodes an interleukin or cytokine or an interleukin or cytokine secretion signal amino-terminal to the multicistronic linker region, preferably wherein the multicistronic linker region comprises the sequence of any one of SEQ ID NOs: 43-45, 47, or 52-55, or a combination thereof, or encodes an internal ribosome entry site, e.g., SEQ ID NO: 56 or 60.

[0016] In some embodiments, the binding domain specifically binds to CD20 and the nucleic acid encodes SEQ ID NO: 8, 9, 10, 11, 12, 46, 48, or 57 and 58. In some embodiments, the nucleic acid comprises the sequence of SEQ ID NO: 13, 14, 15, 16, 17, 50, 51, or 59. In some embodiments, the binding domain specifically binds to BCMA and the nucleic acid encodes SEQ ID NO: 35, 36, 37, or 38. In some embodiments, the nucleic acid comprises the sequence of SEQ ID NO: 39, 40, 41, or 42.

[0017] In some aspects, the present invention provides a polypeptide or polypeptides encoded by any one of the aforementioned isolated nucleic acids or as described herein. In some embodiments, the present invention provides a T cell, such as a γδ T cell, comprising any one of the aforementioned polypeptide or polypeptides. In some embodiments, the T cell expresses a functional binding domain as described herein on the surface of the T cell. In some embodiments, the T cell secretes a cytokine, such as a common gamma chain interleukin, such as IL-15.

[0018] In some embodiments, the T cells exhibit in vitro and / or in vivo cytotoxic activity against hematologic tumor cells that exhibit cell surface expression of a tumor-associated antigen (TAA). In some embodiments, the hematologic tumor cytotoxic activity of these, e.g., γδ, T cells is greater than the native level of in vitro and / or in vivo hematologic tumor cytotoxic activity in a control that does not contain a CAR construct, e.g., γδ, T cell. In some embodiments, e.g., γδ, T cells are HLA class I + Hematologic tumor cytotoxic activity against hematologic tumor cells is increased. In some embodiments, the hematologic tumor cytotoxic activity or increased hematologic tumor cytotoxic activity persists for about, at least, or at least about 6 days to 180 days after initial contact with hematologic tumor cells.

[0019] In some embodiments, e.g., γδ T cells proliferate in response to contact with hematological tumor cells that exhibit cell surface expression of a tumor-associated antigen (TAA). In some embodiments, e.g., γδ T cells exhibit increased proliferation in response to contact with hematological tumor cells that exhibit cell surface expression of a tumor-associated antigen (TAA) compared to a control, e.g., γδ T cell, that does not functionally express a nucleic acid encoding a CAR on its surface, e.g., γδ T cell. In some embodiments, e.g., γδ T cells proliferate in a host organism that contains hematological tumor cells that exhibit cell surface expression of a tumor-associated antigen (TAA).

[0020] In some embodiments, the cell proliferation, e.g., of gamma delta T cells, or increased cell proliferation of gamma delta T cells, persists from about, at least, or at least about 6 days to 180 days after initial contact with hematological tumor cells. In some embodiments, for example, the gamma delta T cells express proinflammatory cytokine(s), such as tumor necrosis factor alpha and / or interferon gamma, after contact with hematological tumor cells. In some embodiments, for example, the gamma delta T cells express proinflammatory cytokine(s), such as tumor necrosis factor alpha and / or interferon gamma, after contact with hematological tumor cells, in greater amounts than control T cells that do not functionally express a nucleic acid encoding a CAR on their cell surface.

[0021] In some embodiments, the graft-versus-host response exhibited by, e.g., γδ, T cells is reduced, substantially reduced, substantially absent, or eliminated when the allogeneic host is introduced compared to the graft-versus-host response exhibited by αβ T cells administered to the allogeneic host. In some embodiments, the T cells are γ T cells. In some embodiments, the T cells are δ T cells. In some embodiments, the T cells are γδ T cells. In some embodiments, the T cells are δ1, δ2, δ3, or δ4 T cells, preferably δ2 - δT cells, more preferably δ1δT cells. In some embodiments, the T cells are δ1, δ2, δ3 or δ4γδT cells, preferably δ2 - γδ T cells, more preferably δ1γδ T cells.

[0022] In another aspect, the invention provides a plurality of any one of the foregoing γδ, T cells, etc., or a plurality of γδ, T cells, etc., as described herein. In some embodiments, the plurality is at least about 10 8 For example, γδ T cells, preferably about 10 8 For example, γδ T cells, approximately 10 11In some embodiments, the plurality comprises at least 60%, 80%, or about 60% or 80% to about 90% or 95% δ1, δ2, δ3, or δ4 γδ T cells, preferably δ1 or δ2 γδ T cells, more preferably δ2 - The composition comprises γδ T cells, most preferably δ1 γδ T cells.

[0023] In some embodiments, the invention provides a method of producing, e.g., a γδ T cell as described herein, or a plurality of γδ T cells as described herein, wherein the method comprises transfecting the T cell(s) with a construct comprising an isolated nucleic acid sequence as described herein. Optionally, the method comprises, e.g., gamma, retroviral transduction. Optionally, the method comprises ex vivo expansion of the T cell(s), wherein the ex vivo expansion is performed before and / or after transfection with the isolated nucleic acid sequence. Optionally, the method comprises ex vivo expansion of the T cell(s), wherein the ex vivo expansion is performed before and after transfection with the isolated nucleic acid sequence. Optionally, the method comprises ex vivo expansion of the T cell(s), wherein the ex vivo expansion is performed after transfection with the isolated nucleic acid sequence. In some embodiments, the method comprises producing about 10 T cells that functionally express a CAR described herein within about 30 days of transfection. 8 For example, from γδ T cells, approximately 10 11 This includes generating T cells, e.g., γδ.

[0024] In another aspect, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a γδ, T cell, etc., as described herein.

[0025] In another aspect, the invention provides methods for killing hematologic tumor cells, the methods comprising contacting hematologic tumor cells with an effective tumor cell-killing amount, e.g., of γδ, T cells, a plurality of such cells, and / or a pharmaceutical composition comprising such cells as described herein. In some embodiments, the methods comprise introducing a therapeutically effective amount, e.g., of γδ, T cell(s), or a pharmaceutical composition, into a host organism comprising the hematologic tumor cells. In some embodiments, the methods comprise introducing a therapeutically effective amount, e.g., of γδ, T cell(s), or a pharmaceutical composition, into a host organism comprising the hematologic tumor cells and simultaneously or sequentially elevating general gamma chain cytokine(s).

[0026] In some embodiments, the one or more administration methods for elevating general gamma chain cytokine(s) include administering an effective amount of a general gamma chain cytokine(s) simultaneously with or consecutively with the introduction of, e.g., γδ, T cell(s) to increase the proliferation, cytotoxic activity, persistence, or a combination thereof, of the introduced, e.g., γδ, T cell(s), preferably wherein the method comprises administering IL-2, and more preferably wherein the method comprises administering IL-15. In some embodiments, the one or more administration methods for elevating general gamma chain cytokine(s) include administering an effective amount of a general gamma chain cytokine(s) before and / or after the introduction of T cell(s) to increase the proliferation, cytotoxic activity, persistence, or a combination thereof, of the introduced T cell(s). In some embodiments, the one or more administration methods for elevating general gamma chain cytokine(s) include lymphodepletion before the introduction of T cell(s).

[0027] In some embodiments, the one or more methods of elevating general gamma chain cytokine(s) include secretion of one or more general gamma chain cytokine(s) from the introduced T cell(s). In some embodiments, the method reduces in vivo tumor burden and / or increases mean survival time of the host organism compared to a control organism, wherein the control organism is not treated with the T cell(s) or pharmaceutical composition. In some embodiments, the method is a method of treating cancer in a subject in need of treatment. In some embodiments, the invention provides a pharmaceutical composition described herein for use in treating, e.g., a γδ T cell, a plurality of, e.g., γδ T cell(s), or hematologic tumor cells in a subject in need of treatment.

[0028] In one aspect, the invention provides a method of treating cancer by administering a therapeutically effective amount of γδ T cells, wherein the cancer is hematological tumor cells that exhibit cell surface expression of CD20. Alternatively, the invention provides a method of treating cancer by administering a therapeutically effective amount of γδ T cells, wherein the cancer comprises hematological tumor cells that exhibit cell surface expression of BCMA. In some embodiments, the method comprises administering one or more methods of elevating general gamma chain cytokine(s) simultaneously with or sequentially to the administration of the γδ T cells. In some embodiments, the method comprises administering multiple doses of γδ T cells, wherein the interval between the multiple doses is at least about 1 week, preferably at least about 2, 3, 4, 5, 6, 7, 8, or 12 weeks, and / or no more than once every 6 or 12 months. In some embodiments, the invention provides a pharmaceutical composition for use in any one of the aforementioned therapeutic methods.

[0029] Incorporation by Reference All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was individually and specifically indicated to be incorporated by reference. [Brief explanation of the drawings]

[0030] [Figure 1] Schematic diagrams of chimeric antigen receptor (CAR) embodiments containing one costimulatory signaling endodomain (left) or two costimulatory signaling endodomains (right) are shown. As used herein, costimulatory signaling endodomains are also referred to as costimulation endodomains or costimulatory endodomains. Exemplary costimulatory signaling endodomains useful in exemplary CARs include, but are not limited to, CD28, CD137 (41BB), CD278 (ICOS), CD27, CD134 (OX40), TLR2, and combinations thereof. [Figure 2] The sequences of binding domains that specifically bind to epitopes within CD20 are shown in Figure 2. SEQ ID NOS: 335 to 363, 99, 364, 107, and 365 to 368 are disclosed in order of appearance, respectively. [Figure 3] Figure 1 shows induction of apoptosis of CD20-expressing normal B cells by non-transduced V51 cells and V51 cells transduced with various CD20-specific CAR constructs. [Figure 4] We demonstrate the potent cytotoxic activity of CD20-specific CARγδ T cells against lymphoma cell lines. [Figure 5] Figure 1 shows the cytotoxicity of engineered CAR γδ T cells described herein against Raji cells. Top: Binding domains comprising the CDRs of 3B9, 2B7, 3H7, and 9C11 are tested with a CAR construct encoding a 4-1BB costimulatory endodomain and a CD3ζ signaling domain. [Figure 6]Figure 1 shows the cytotoxicity of engineered CAR γδ T cells described herein against Raji cells. Top: Binding domains containing the CDRs of 3H7 are tested. Bottom: Cytotoxicity of γδ T cells expressing CARs containing the 3H7 binding domain, a CD3ζ signaling domain, and various costimulatory endodomains, as indicated, against Raji cells is shown. The 3H7-CD27z CAR contains a 3H7 binding domain, a CD8α hinge and transmembrane domain, a CD27 costimulatory endodomain, and a CD3ζ signaling domain. The 3H7-5.1 contains a 3H7 binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, and a CD3ζ signaling domain. [Figure 7] Figure 1 shows the results of a cytotoxicity assay involving re-administration of the indicated γδ T cells, with the arrow indicating the time of re-administration. [Figure 8] 1 shows the in vivo efficacy of γδ T cells described herein in a subcutaneous Raji cell NOD scid gamma (NSG) mouse model. [Figure 9] 1 illustrates a table of CD20-specific γδ CAR-T cells within tumors in vivo, demonstrating in vivo expansion of γδ CAR-T cells and tumor elimination. [Figure 10] 1 shows the in vivo expansion of CD20-specific γδ CAR-T cells in CD20+ lymphoma tumors and other organs. [Figure 11] 1 shows the in vivo efficacy of γδ T cells described herein in a disseminated Raji cell NOD scid gamma (NSG) mouse model. [Figure 12] We demonstrate effective treatment of disseminated Raji tumors with CD20-specific γδ CAR-T cells in the SRG-15 mouse model, which expresses human IL-15, without inducing a graft-versus-host (GVH) response. In contrast, CD20-specific αβ CAR-T cells induce a lethal GVH response. [Figure 13] 1 shows the manufacturing process for producing engineered and non-engineered γδ CAR-T cells. [Figure 14]1 shows therapeutic efficacy and durability of CD20 CAR Vδ1 T cells expressing sIL15 in NSG mice subcutaneously implanted with Raji cells followed by re-administration of Raji cells at a different implantation site (day 62). [Figure 15] Figure 1 shows the transduction efficiency of Vδ1 cells with the indicated anti-B cell maturation antigen (BCMA) scFv CAR constructs. BCMA is also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17). [Figure 16] Figure 1 shows the cytotoxic activity of V51 T cells transduced with various anti-BCMA CAR constructs against a panel of multiple myeloma BCMA+ cell lines. The SCABER-Luc cell line is a BCMA-negative control cell line. [Figure 17] Figure 1 shows the cytotoxic activity of V51 T cells transduced with different anti-BCMA CAR constructs against a panel of multiple myeloma and Burkitt's lymphoma BCMA+ cell lines. [Figure 18] 1 shows the efficacy of in vivo treatment with anti-BCMA CARVδ1 T cells against subcutaneously implanted NCI-H929 cells. DETAILED DESCRIPTION OF THE INVENTION

[0031] Definition: For purposes of interpreting this specification, the following definitions shall apply, except that where appropriate, terms used in the singular shall include the plural and vice versa. In the event that any definition set forth conflicts with any document incorporated herein by reference, the definition set forth below shall control. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0032] As used herein, the term "about," when referring to a measurable value such as an amount, temporal duration, and the like, is intended to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and even more preferably ±0.1% from the particular value, as such variations are appropriate for performing the disclosed methods.

[0033] As used herein, the term "γδ T cells (gamma delta T cells)" refers to a subset of T cells that express a distinct T cell receptor (TCR), i.e., γδ TCR, on their surface, which is composed of one γ chain and one δ chain. The term "γδ T cells" includes all subsets of γδ T cells, particularly, but not limited to, Vδ1, Vδ2, and Vδ3 γδ T cells, as well as naive, effector memory, central memory, and terminally differentiated γδ T cells. As a further example, the term "γδ T cells" includes Vδ4, Vδ5, Vδ7, and Vδ8 γδ T cells, as well as Vγ2, Vγ3, Vγ5, Vγ8, Vγ9, Vγ10, and Vγ11 γδ T cells. In some embodiments, γδ T cells are Vδ1 - , Vδ2 - or Vδ1 - and Vδ2 -Compositions and methods for making and using engineered and non-engineered γδ T cells and / or their subtypes include, but are not limited to, those described in US2016 / 0175358, WO2017 / 197347, US9499788, US2018 / 0169147, US9907820, US2018 / 0125889, and US2017 / 0196910, the contents of each of which are incorporated by reference for all purposes, including compositions and methods for making and using engineered and non-engineered γδ T cells and / or their subtypes. The present application further contemplates T cells, or other engineered leukocytes or lymphocytes, that express one gamma chain or one delta chain, optionally in combination with a second polypeptide to form a functional TCR. Such genetically engineered leukocytes or lymphocytes that express one gamma chain or one delta chain may be used in the methods or may be present in the compositions described herein.

[0034] As used herein, the term "T lymphocyte" or "T cell" refers to an immune cell that expresses or has expressed CD3 (CD3+) and a T cell receptor (TCR+). T cells play a central role in cell-mediated immunity. T cells that "express" CD3 and TCR have been genetically engineered to eliminate CD3 and / or TCR cell surface expression.

[0035] As used herein, the term "TCR" or "T cell receptor" refers to dimeric heterologous cell surface signaling proteins that form alpha-beta or gamma-delta receptors, or combinations thereof. αβTCRs recognize antigens presented by MHC molecules, whereas γδTCRs are capable of recognizing antigens independently of MHC presentation.

[0036] The term "MHC" (major histocompatibility complex) refers to a subset of genes that code for cell surface antigen-presenting proteins. In humans, these genes are called human leukocyte antigen (HLA) genes. The abbreviations MHC or HLA are used interchangeably herein.

[0037] As used herein, "activated" refers to a state of T cells that have been sufficiently stimulated to induce detectable cell proliferation. Activation may also be associated with induced cytokine production and detectable effector function. The term "activated T cells" refers, inter alia, to T cells that are undergoing cell division.

[0038] As used herein, the term "antibody" refers to an immunoglobulin molecule that specifically binds to an antigen. Antibodies can be intact immunoglobulins derived from natural or recombinant sources, or can be immunoreactive portions of intact immunoglobulins. Typically, antibodies are tetramers of immunoglobulin molecules. Antibodies of the invention can exist in a variety of forms, including, for example, polyclonal antibodies, monoclonal antibodies, Fv, Fab, and F(ab)2, as well as single-chain antibodies and humanized antibodies (Harlow et al., 1999, In: Using Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY; Harlow et al., 1989, In: Antibodies: A Laboratory Manual, Cold Spring Harbor, NY; Houston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883; Bird et al., 1988, Science 242:423-426).

[0039] The term "antibody fragment" refers to a portion of an intact antibody and refers to the antigen-determining variable region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, scFv antibodies, and multispecific antibodies formed from antibody fragments.

[0040] As used herein, "antibody heavy chain" refers to the larger of the two polypeptide chains present in antibody molecules in their naturally occurring conformation.

[0041] As used herein, "antibody light chain" refers to the smaller of the two polypeptide chains present in antibody molecules in their naturally occurring conformation. Kappa and lambda light chains refer to the two major antibody light chain isotypes.

[0042] As used herein, the term "synthetic antibody" refers to an antibody made using recombinant DNA techniques, such as, for example, antibodies expressed by bacteriophage as described herein. The term should also be construed to mean an antibody made by synthesizing a DNA molecule encoding the antibody, which DNA molecule expresses an antibody protein or amino acid sequence specifying the antibody, wherein the DNA or amino acid sequence is obtained using synthetic DNA or amino acid sequence techniques available and well known in the art.

[0043] As used herein, the term "antigen" or "Ag" is defined as a molecule that elicits an immune response. This immune response may include either antibody production or activation of specific immunocompetent cells, or both. Those skilled in the art will understand that any macromolecule, including proteins or peptides, can serve as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. Thus, those skilled in the art will understand that any DNA containing a nucleotide sequence or partial nucleotide sequence encoding a protein that elicits an immune response encodes an "antigen" as the term is used herein. Furthermore, those skilled in the art will understand that an antigen need not be encoded solely by the full-length nucleotide sequence of a gene. It will be readily apparent that the present invention includes, but is not limited to, the use of partial nucleotide sequences of one or more genes, and that these nucleotide sequences may be arranged in various combinations to elicit a desired immune response. Furthermore, those skilled in the art will understand that an antigen need not be encoded by a "gene" at all. It will be readily apparent that an antigen can be produced, synthesized, or derived from a biological sample. Such biological samples may include, but are not limited to, tissue samples, tumor samples, cells, or biological fluids.

[0044] The term "epitope" includes any protein, lipid, or carbohydrate determinant capable of specific binding to an immunoglobulin or T-cell receptor. Epitopic determinants usually consist of active surface groupings of molecules such as amino acids, lipid, or sugar side chains and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics. The equilibrium dissociation constant (K D ) is 10 -6 ~10 -12 If the antibody is in the range of M, then the antibody is said to specifically bind to the antigen.

[0045] Antibodies 3B9, 9C11, 3H7, 2B7, and 10F2 represent exemplary embodiments of antibodies that specifically recognize CD20. These antibodies, fragments thereof, and their complementarity-determining regions are also described in US 2009 / 0035322 and are designated 3B9-10, 9C11-14, 3H7-6, 2B7-7, and 10F2-13, respectively. As described herein, these antibodies, fragments thereof, and their complementarity-determining regions are useful in generating anti-CD20 chimeric antigen receptor (CAR) constructs and in engineering and using CAR-T cells to treat hematologic tumors that express CD20.

[0046] Binding domains 21587N, 16747P, 16711P, and 16716P represent exemplary embodiments of binding domains that specifically recognize BCMA. These antibodies, fragments thereof, and their complementarity-determining regions are also described in US 16 / 516028, filed July 18, 2019, the contents of which are incorporated by reference in their entirety for all purposes, particularly the binding domains, antibodies, antibody fragments, complementarity-determining regions, polypeptides comprising the complementarity-determining regions, nucleic acids encoding the complementarity-determining regions, and epitope specificity and assays for determining epitope specificity described herein. 21587N, 16747P, 16711P, and 16716P are sometimes referred to as H2aM21587N, H1H16747P, H1H16711P, and H1H16716P, respectively. As described herein, these antibodies, fragments thereof and their complementarity determining regions are useful in generating anti-BCMA chimeric antigen receptor (CAR) constructs and in engineering and using CAR-T cells to treat BCMA-expressing hematologic tumors.

[0047] As used herein, the term "chimeric antigen receptor (CAR)" may refer to, for example, an artificial T cell receptor, T body, single-chain immunoreceptor, chimeric T cell receptor, or chimeric immunoreceptor, and encompasses genetically engineered receptors that transfer artificial specificity to specific immune effector cells. CARs may be used to confer the specificity of a monoclonal antibody to T cells, thereby enabling the generation of large numbers of specific T cells, for example, for use in adoptive cell therapy. In certain embodiments, CARs direct the specificity of cells to, for example, a tumor-associated antigen. In some embodiments, CARs comprise an intracellular activation domain (which enables T cells to be activated upon binding of the targeting moiety to a target cell, such as a targeted tumor cell), a transmembrane domain, and an extracellular domain that can vary in length, and further comprise a disease or disorder-associated, e.g., tumor-antigen-binding region. In certain aspects, CARs comprise a fusion of a single-chain variable fragment (scFv) derived from a monoclonal antibody fused to a CD3-zeta transmembrane domain and endodomain. Other CAR design specificities may be derived from receptor ligands (e.g., peptide receptors) or pattern recognition receptors such as dectin. In certain cases, the spacing of the antigen recognition domain can be altered to reduce activation-induced cell death. In certain cases, the CAR includes domains for adding costimulatory signaling, such as CD3ζ, FcR, CD27, CD28, CD137, DAP10 / 12, and / or OX40, ICOS, TLRs (e.g., TLR2), and the like. In some cases, molecules may be coexpressed with the CAR, such as costimulatory molecules, reporter genes for imaging (e.g., positron emission tomography), gene products that conditionally ablate T cells upon the addition of prodrugs, homing receptors, chemokines, chemokine receptors, cytokines, and cytokine receptors. Furthermore, those skilled in the art will understand that the costimulatory domain need not be encoded solely by the full-length nucleotide sequence of a gene. It is readily apparent that the present invention includes, but is not limited to, the use of partial nucleotide sequences of one or more genes, and that these nucleotide sequences may be arranged in various combinations to elicit a desired immune response.

[0048] As used herein, the term "anti-tumor effect" refers to a biological effect that may be manifested by a reduction in tumor volume, a reduction in tumor cell number, a reduction in the number of metastases, an increase in life expectancy, or an improvement in various physiological symptoms associated with a cancerous condition. An "anti-tumor effect" may also be manifested by the ability of the peptides, polynucleotides, cells, and antibodies of the present invention to prevent the development of tumors in the first place.

[0049] The term "autoantigen" as used herein refers to any self-antigen that is mistakenly recognized as foreign by the immune system. Autoantigens include, but are not limited to, cellular proteins, phosphorylated proteins, cell surface proteins, cellular lipids, nucleic acids, and glycoproteins, including cell surface receptors.

[0050] As used herein, the term "autologous" is intended to refer to any material that originates from the same individual into which it is subsequently reintroduced.

[0051] As used herein, the term "allogeneic" is intended to refer to any material derived from an animal that is subsequently introduced into another animal of the same species.

[0052] The term "therapeutically effective amount" refers to an amount of a composition that elicits the biological or medical response of a tissue, system, or subject that is desired by a researcher, veterinarian, physician, or other clinician. The term "therapeutically effective amount" includes an amount of a composition that, when administered, is sufficient to prevent the onset of, or to alleviate to some extent, one or more of the signs or symptoms of, the disease or disorder (e.g., blood cancer) being treated. The therapeutically effective amount will vary depending on the composition, the disease and its severity, and the age, weight, etc., of the subject being treated.

[0053] As used herein, the term "treating" a disease means reducing the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject.

[0054] Administration "in combination with" one or more other therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order.

[0055] As used herein, the term "pharmaceutically acceptable" refers to a material, such as, but not limited to, a salt, carrier, or diluent, that does not abrogate the biological activity or properties of the compound and that is relatively non-toxic. In other words, the material may be administered to an individual without causing undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is included.

[0056] "Encode" refers to the inherent property of a particular sequence of nucleotides in a polynucleotide, such as a gene, cDNA, or mRNA, to serve as a template for synthesizing other polymers and macromolecules in a biological process, either having a defined sequence of nucleotides (i.e., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of the mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, which is the nucleotide sequence identical to that of the mRNA and usually provided in a sequence listing, and the non-coding strand, which is used as a template for transcription of the gene or cDNA, can be said to encode the protein or other product of that gene or cDNA.

[0057] "Isolated" means changed or removed from its natural state. For example, a nucleic acid or peptide naturally present in a living animal is not "isolated," but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is "isolated." An isolated nucleic acid or protein can exist in a substantially purified form, or it can exist in a non-native environment, such as, for example, a host cell.

[0058] Unless otherwise specified, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. Nucleotide sequences that encode proteins and RNA may contain introns.

[0059] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal amenable to the methods described herein. In certain non-limiting embodiments, the patient, subject, or individual is a human.

[0060] As used herein, the term "specifically binds" with respect to antibodies refers to an antibody that recognizes a specific antigen but does not substantially recognize or bind to other molecules in a sample. For example, an antibody that specifically binds to an antigen from one species may also bind to that antigen from one or more other species. However, such cross-species reactivity does not in itself change the classification of the antibody according to its characteristics. In another example, an antibody that specifically binds to an antigen may bind to various alleles of the antigen. However, such cross-reactivity does not in itself change the classification of the antibody according to its characteristics. In some cases, the terms "specific binding" or "specifically binding" may be used in reference to the interaction of an antibody, protein, or peptide with a second chemical species, but this interaction depends on the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody is meant to recognize and bind to a specific protein structure rather than a general protein. If an antibody is specific for epitope "A," then in a reaction containing labeled "A" and the antibody, the presence of a molecule containing epitope A (or free, unlabeled A) will reduce the amount of labeled A that binds to the antibody.

[0061] In some embodiments, specific binding is at least about 1x10 -8 M or less (e.g., K D(The smaller the , the stronger the binding). Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. Furthermore, multispecific antibodies that bind to a first antigen and one or more additional antigens, or bispecific antibodies that bind to two different regions of an antigen, are nevertheless considered to be "specifically binding" antibodies as used herein.

[0062] Hematological cancers are cancers that originate in the blood or bone marrow. Examples of hematological (or hematopoietic) cancers include acute leukemia (such as acute lymphocytic leukemia, acute myeloid leukemia, acute myelogenous leukemia, and myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroleukemia), chronic leukemia (chronic myeloid (granulocytic) leukemia, chronic myelogenous leukemia, and chronic lymphocytic leukemia), polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma (low-grade and high-grade), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia, and myelodysplasia. In a preferred embodiment, the hematological cancer expresses or overexpresses CD20. In a preferred embodiment, the hematological cancer expresses or overexpresses B-cell maturation antigen (BCMA), also known as tumor necrosis factor receptor superfamily member 17 (TNFRSF17).

[0063] An "expression cassette" refers to a nucleic acid comprising expression control sequences operably linked to a nucleic acid encoding a transcript or polypeptide to be expressed. An expression cassette contains sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. An expression cassette can be a component of a vector such as a cosmid, a plasmid (e.g., naked in a liposome or contained in a liposome), or a virus (e.g., a lentivirus, retrovirus, adenovirus, and adeno-associated virus). An expression cassette can be present in a host cell, such as a γδ T cell.

[0064] Ranges: Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values ​​within that range. For example, description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within the range such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0065] Chimeric antigen receptor constructs: Aspects of the invention include nucleic acids encoding CARs, as well as constructs and vectors comprising such nucleic acids. In some cases, the nucleic acid is a component of, e.g., a heterologous expression cassette. In some embodiments, the nucleic acid is a component of, e.g., a heterologous retroviral vector. In some embodiments, the nucleic acid is a component of, e.g., a heterologous αβ or γδ T cell, preferably a γδ T cell. In some embodiments, the nucleic acid is a component of, e.g., a heterologous γ + T cells and / or delta + In some embodiments, the nucleic acid is a heterologous, α - T cells and / or β - It is a component of T cells.

[0066] Described herein are nucleic acids encoding a CAR binding domain that specifically binds to a tumor-associated antigen (TAA) expressed on the surface of hematologic tumor cells. Exemplary TAAs include CD19, CD20, and BCMA. In some embodiments, the binding domain is a CD19-binding domain, such as the CD19-binding domain described in U.S. Patent No. 9,540,445, the contents of which are incorporated by reference in their entirety and for all purposes, particularly the binding domain, antibody, antibody fragment, complementarity-determining region, polypeptide comprising the complementarity-determining region, nucleic acid encoding the complementarity-determining region, and epitope specificity and assays for determining epitope specificity described herein. In some embodiments, the binding domain is a CD20-binding domain, such as the CD20-binding domain described in U.S. Patent Application Publication No. 2009 / 0035322, the contents of which are incorporated by reference in their entirety and for all purposes, particularly the binding domain, antibody, antibody fragment, complementarity-determining region, polypeptide comprising the complementarity-determining region, nucleic acid encoding the complementarity-determining region, and epitope specificity and assays for determining epitope specificity described herein. In some embodiments, the binding domain is a BCMA-binding domain, such as the BCMA-binding domain described in WO 2018 / 133877 or the BCMA-binding domain described in US 16 / 516028, filed July 18, 2019, the contents of each of which are incorporated by reference in their entirety and for all purposes, particularly the binding domain, antibody, antibody fragment, complementarity determining region, polypeptide comprising the complementarity determining region, nucleic acid encoding the complementarity determining region, and epitope specificity and assays for determining epitope specificity described herein. Typically, the region encoding the binding domain is 5' to a linker region (e.g., a region encoding the CD8α hinge domain).

[0067] In some embodiments, the binding domain binds to an antigen as expressed as a full-length functional polypeptide on the surface of a cell. In some embodiments, the binding domain binds to an antigen as presented in an MHC:antigen complex. In some embodiments, the binding domain binds to an antigen in an HLA-restricted manner. Binding domains that exhibit specificity for MHC:antigen complexes are described, for example, in WO / 2016 / 199140 and WO / 2016 / 199141, the contents of each of which are incorporated by reference in their entirety and for all purposes, particularly the binding domains, antibodies, antibody fragments, complementarity determining regions, polypeptides comprising the complementarity determining regions, nucleic acids encoding the complementarity determining regions, and epitope specificity and assays for determining epitope specificity described therein.

[0068] Exemplary CD20 binding domains include, but are not limited to, 3B9, 3H7, 2B7, 9C11, or 10F2, or a binding domain that selectively binds to an epitope within CD20 bound by 3B9, 3H7, 2B7, or 9C11, or 3H7, or that competes for binding with 3B9, 3H7, 2B7, 9C11, or 10F2, or 3B9, 3H7, 2B7, or 9C11, or 3H7. Additionally or alternatively, the CD20 binding domain may comprise a complementarity determining region of an anti-CD20 antibody selected from the group consisting of 3B9, 3H7, 2B7, 9C11, and 10F2, selected from the group consisting of 3B9, 3H7, 2B7, and 9C11, or including a complementarity determining region of an anti-CD20 antibody selected from the group consisting of 3H7. The present disclosure also contemplates CD20, CD19 and BCMA binding domains that compete for binding with the sequences provided herein.

[0069] Using known methods, it is possible to determine whether a CD20 binding domain binds to the same epitope as a reference antibody or binding domain, or competes for binding with the reference antibody or binding domain. For example, to determine whether a test antibody binds to the same epitope as a reference binding domain, the reference binding domain can be bound to CD20 under saturating conditions. The ability of the test binding domain to bind to the CD20 molecule can then be evaluated. If the test binding domain is able to bind to CD20 following saturation binding with the reference binding domain, it can be concluded that the test binding domain binds to a different epitope than the reference binding domain. On the other hand, if the test binding domain is unable to bind to CD20 following saturation binding with the reference binding domain, the test binding domain may bind to the same epitope as the epitope bound by the reference binding domain.

[0070] When a binding domain competes for binding with a reference binding domain, the binding methodology described above is performed in two ways. In the first way, the reference binding domain is allowed to bind to CD20 under saturating conditions, followed by assessing the binding of the test binding domain to the CD20 molecule. In the second way, the test binding domain is allowed to bind to CD20 molecules under saturating conditions, followed by assessing the binding of the reference binding domain to the CD20 molecule. If, in both ways, only the first (saturating) binding domain is able to bind to the CD20 molecule, it is concluded that the test and reference binding domains compete for binding to CD20. As will be understood by those skilled in the art, a binding domain that competes for binding with a reference binding domain may not necessarily bind to the same epitope as the reference binding domain, but may sterically block binding of the reference binding domain by binding to an overlapping or adjacent epitope. The above-described methods for determining competition and epitope binding with an anti-CD20 binding domain can similarly be applied to anti-CD19 binding domains and anti-BCMA binding domains.

[0071] Two binding domains bind to the same or overlapping epitopes if each competitively inhibits (blocks) the binding of the other to the antigen. That is, a 1-fold, 5-fold, 10-fold, 20-fold, or 100-fold excess of one binding domain inhibits the binding of the other by at least 50%, e.g., 75%, 90%, or even 99%, as measured in a competitive binding assay (see, e.g., Junghans et al., Cancer Res. 1990 50:1495-1502). Alternatively, two binding domains have the same epitope if essentially all amino acid mutations in the antigen that reduce or eliminate binding of one binding domain also reduce or eliminate binding of the other. Two binding domains have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one binding domain also reduce or eliminate binding of the other.

[0072] Further routine experiments (e.g., peptide mutations and binding analysis) can be performed to ascertain whether the observed lack of binding of the test binding domain is indeed due to binding to the same epitope as the reference binding domain, or whether steric hindrance (or other phenomena) is responsible for the observed lack of binding. These types of experiments can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry, or any other quantitative or qualitative binding assay available in the art.

[0073] The present disclosure provides antibodies and CARs that have "substantial identity" or "substantial similarity" to the sequences provided herein in the CDR or framework regions. The terms "substantial identity" or "substantially identical," when referring to a nucleic acid or fragment thereof, indicate that when optimally aligned with another nucleic acid (or the complementary strand of another nucleic acid), there is, for example, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% nucleotide sequence identity as measured by any known sequence identity algorithm, such as FASTA, BLAST, or GAP, as described below. A nucleic acid molecule having substantial identity to a reference nucleic acid molecule can, in certain instances, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0074] When applied to polypeptides, the term "substantial similarity" or "substantially similar" means that two peptide sequences share at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% sequence identity when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights. In some embodiments, residue positions that are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is the replacement of an amino acid residue with another amino acid residue having a side chain (R group) with similar chemical properties (such as charge or hydrophobicity). Generally, conservative amino acid substitutions do not substantially alter the functional properties of a protein. Where two or more amino acid sequences differ from each other by conservative substitutions, the percent or degree of identity may be adjusted upwards to restore the conservative nature of the substitution. Methods for making this adjustment are well known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24:307-331, which is incorporated herein by reference. Examples of amino acid groups having side chains of similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine; 2) aliphatic hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartic acid and glutamic acid; and 7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine.Instead, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix, as disclosed in Gonnet et al. (1992) Science 256:1443 45, incorporated herein by reference. A "moderately conservative" substitution is any change that has a non-negative value in the PAM250 log-likelihood matrix.

[0075] Sequence identity and / or similarity of polypeptides is typically measured using sequence analysis software. Protein analysis software matches similar sequences, which is used to measure similarity by assigning various substitutions, deletions, and other modifications, including conservative amino acid substitutions. For example, GCG software includes programs such as GAP and BESTFIT, which can be used with default parameters, to measure sequence identity or identity between homologous polypeptides from various species of organisms, or between closely related polypeptides, or between a wild-type protein and its mutant protein. See, for example, GCG Version 6.1. Polypeptide sequences can also be compared using FASTA with default or recommended parameters, i.e., the GCG Version 6.1 program. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the optimal overlap regions between the query and search sequences (Pearson (2000) supra). Sequences can be compared using the Smith-Waterman homology search algorithm, using an affine gap search with a gap opening penalty of 12, a gap extension penalty of 2, and a BLOSUM matrix of 62. Another preferred algorithm for comparing the sequences disclosed herein to databases containing a large number of sequences from various organisms is the computer program BLAST, particularly BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and (1997) Nucleic Acids Res. 25:3389-3402, the contents of each of which are incorporated by reference.

[0076] Provided herein are anti-CD20, anti-BCMA, or anti-CD19 CARs comprising variants of any of the HCVR, LCVR, and / or CDR amino acid sequences disclosed herein with one or more substitutions (e.g., conservative substitutions). For example, the disclosure includes anti-CD20 CARs having HCVR, LCVR, and / or CDR amino acid sequences with, for example, 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 amino acid substitution compared to any of the HCVR, LCVR, and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, or LCDR3) amino acid sequences disclosed herein. For example, an anti-CD20 CAR may include 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitution (such as a conservative amino acid substitution) compared to any of the HCVR, LCVR, and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3) amino acid sequences disclosed herein.

[0077] Similarly, the disclosure includes anti-BCMA CARs having HCVR, LCVR and / or CDR amino acid sequences with, for example, 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 amino acid substitution compared to any of the HCVR, LCVR and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3) amino acid sequences disclosed herein. For example, an anti-BCMA CAR may include 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitution (such as a conservative amino acid substitution) compared to any of the HCVR, LCVR, and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3) amino acid sequences disclosed herein.

[0078] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain having a heavy chain complementarity determining region 3 (HCDR3) and a light chain CDR3 (LCDR3), wherein the HCDR3 and LCDR3 are selected from the group consisting of SEQ ID NOs: 345 (AKDPSYGSGSYHSYYGMDV) and 353 (QQRFNWPLT), 201 (VKDFHYGSGSNYGMDV) and 209 (QQSNDWPLT), and 249 (TKDGSYGHFYSGLDV) and 257 (QQRYYWPLT).

[0079] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain having a heavy chain variable region (HCVR) sequence and a light chain variable region (LCVR) sequence, wherein the HCVR sequence and the LCVR sequence are set forth in SEQ ID NOs: 339 (EEQLVESGGDLVQPGRSLRLSCAASGFTFHDYTMH WVRQAPGKGLEWVSGISWNSGSLGYADSVKGRFTISRDNAKKSLYLQMNSLRAEDTALYYCAKDPSYGSGSYHSYYGMDVWGQGTTVTVSS) and 347 (EIVLTQSPATLSLSPGE RATLSCWASQSISRYLVWYQQKCGQAPRLLIYEASKRATGIPVRFSGSGSGTDFTLTISSLESEDFAVYYCQQRFNWPLTFGGGTKVEIK), 195 (EVQLAESGGDLVQSGRSLRLSCAAS GITFHDYAMHWVRQPPGKGLEWVSGISWNSDYIGYADSVKGRFTISRDNAKKSLYLQMNSLRPDDTALYYCVKDFHYGSGSNYGMDVWGQGTTVTVSP) and 203 (EIVMTQSPATL SMSPGERATLSCRASQSVSRNLAWYQQKVGQAPRLLISGASTRATGIPARFSGSGSGTEFTLTINSLQSEDFAVYYCQQSNDWPLTFGQGTRLEIK), 243 (VQLVESGGGLVQPGR SLRLSCAASGFTFYDYAMHWVRQAPGKGLEWVSGISWNSDTIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCTKDGSYGHFYSGLDVWGQGTTVTVSS) and 251 (EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYVASNRATGIPARFSGSGSGTDFTLTISSLEPDDFAVYYCQQRYYWPLTFGGGTKVEIK).

[0080] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain having a heavy chain complementarity determining region 3 (HCDR3) domain and a light chain CDR3 (LCDR3) domain, wherein the HCDR3 domain comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19, where X1=A, V or T, X2=K, X3=D, X4=P, F or G, X5=S or H, X6=Y, X7=G, X8=S or H, X9=G or F, X10=Y, X11=Y, X12=Y, X13=Y, X14=Y, X15=Y, X16=Y, X17=Y, X18=Y, X19=Y, X20=Y, X21=Y, X22=Y, X23=Y, X24=Y, X25=Y, X26=Y, X27=Y, X28=Y, X29=Y, X30=Y, X31=Y, X32=Y, X33=Y, X34=Y, X35=Y, X36=Y, X37=Y, X38=Y, X39=Y, X40=Y, X41=Y, X42=Y, X43=Y, X44=Y, X45=Y, X46=Y, X47=Y, X48=Y, X49=Y, X50=Y, X51=Y, X52=Y, X53=Y, X54=Y, X55=Y, X56=Y, X57=Y, X58=Y, X59=Y, X60=Y, X61=Y, X62=Y, X63 =S or Y, X11 =Y, N or S, X12 =Y, G or H, X13 =G, L or S, X14 =Y, M or D, X15 =Y, D or V, X16 =G, V or absent, X17 =M or absent, X18 =D or absent, X19 =V or absent (SEQ ID NO: 369), and the LCDR3 domain comprises the amino acid sequence of X1-X2-X3-X4-X5-X6-X7-X8-X9, where X1 =Q, X2 =Q, X3 =R or S, X4 =N, Y or F, X5 =N, D, or Y, X6 =W, X7 =P, X8 =L, X9 =T (SEQ ID NO: 370).

[0081] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain having a heavy chain variable region (HCVR) sequence and a light chain variable region (LCVR) sequence, wherein the HCVR sequence and the LCVR sequence are SEQ ID NOs: 99 (EVQLVESGGGLVQPGRSLRLSCAASGFTFYDYAMHWVRQAPGKGLEWVSGISWNSGYIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDNSYGKFYYGLDVWGQGTTVTVSS) and 107 (EIVMTQSPATLSVSPGERTTLSCRASQSVSSNLAWYLQKPGQAPR LLIYGASTRATGIPARFSGSGSGTEFILTISSLQSEDFAVYYCQQYNNWPITFGQGTRLEIK).

[0082] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain that binds to the same epitope as, competes with, or is the anti-CD20 binding domain having heavy chain complementarity determining regions (HCDRs) and light chain complementarity determining regions (LCDRs), wherein the HCDR and LCDR sequences are the HCDR sequence of SEQ ID NO: 99 and the LCDR sequence of SEQ ID NO: 107, respectively.

[0083] In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain that binds to the same epitope as, competes with, or is an anti-CD20 binding domain having an HCDR1 that is or comprises SEQ ID NO: 101 (GFTFYDYA), an HCDR2 that is or comprises SEQ ID NO: 103 (ISWNSGYI), and / or an HCDR3 that is or comprises SEQ ID NO: 105 (AKDNSYGKFYYGLDV). In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain that binds to the same epitope as, competes with, or is an anti-CD20 binding domain having an LCDR1 that is or comprises SEQ ID NO: 109 (QSVSSN), an LCDR2 that is or comprises SEQ ID NO: 111 (GAS), and / or an LCDR3 that is or comprises SEQ ID NO: 113 (QQYNNWPIT). In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain that binds to the same epitope as, competes with, or is an anti-CD20 binding domain having an HCDR1 that is or comprises SEQ ID NO: 101, an HCDR2 that is or comprises SEQ ID NO: 103, an HCDR3 that is or comprises SEQ ID NO: 105, an LCDR1 that is or comprises SEQ ID NO: 109, an LCDR2 that is or comprises SEQ ID NO: 111, and an LCDR3 that is or comprises SEQ ID NO: 113. In some embodiments, the isolated nucleic acid encodes an anti-CD20 binding domain having an HCDR1 that is or comprises SEQ ID NO: 101, an HCDR2 that is or comprises SEQ ID NO: 103, an HCDR3 that is SEQ ID NO: 105, an LCDR1 that is or comprises SEQ ID NO: 109, an LCDR2 that is or comprises SEQ ID NO: 111, and an LCDR3 that comprises SEQ ID NO: 113.

[0084] Exemplary BCMA binding domains include, but are not limited to, binding domains that selectively bind to an epitope within BCMA that is bound by, or competes for binding with, the BCMA binding domain described in WO2018 / 133877 or the BCMA binding domain described in US 16 / 516028, filed July 18, 2019. Alternatively or additionally, the BCMA binding domain may comprise the complementarity determining regions of an anti-BCMA antibody selected from the group consisting of the anti-BCMA antibodies or chimeric antigen receptors described in WO2018 / 133877 and the anti-BCMA antibodies or chimeric antigen receptors described in US 16 / 516028, filed July 18, 2019.

[0085] Exemplary BMCA binding domains include, but are not limited to, binding domains that selectively bind to or compete for binding with an epitope in BCMA bound by anti-BCMA-CAR16716P, anti-BCMA-CAR16747P, and / or anti-BCMA-CAR21587N. Alternatively or additionally, the BCMA binding domain may comprise a complementarity determining region of an anti-BCMA CAR selected from the group consisting of anti-BCMA-CAR16716P, anti-BCMA-CAR16747P, and anti-BCMA-CAR21587N.

[0086] In some embodiments, the isolated nucleic acid encodes an anti-BCMA binding domain having a heavy chain complementarity determining region 3 (HCDR3) and a light chain CDR3 (LCDR3), wherein the HCDR3 and LCDR3 are selected from the group consisting of SEQ ID NO:21 (RAGDNWNWFDP) and SEQ ID NO:22 (QQAKSVPFT), SEQ ID NO:23 (EGGNYGMDV) and SEQ ID NO:24 (QQANSFPPT), and SEQ ID NO:25 (FAEYCGGNICYYYGMDV) and SEQ ID NO:26 (QQCGGSPWT).

[0087] In some embodiments, the isolated nucleic acid encodes an anti-BCMA binding domain having a heavy chain variable region (HCVR) sequence and a light chain variable region (LCVR) sequence, wherein the HCVR sequence and the LCVR sequence are identical to those of the 16716P binding domain HCVR SEQ ID NO: 27 (MSVPTQVLGLLLLWLTDARCEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYVMSWVRQAPGKGLEWVSAIIGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNS LRAEDTAVYYCAKRAGDNWNWFDPWGQGTLVTV) and 16716P binding domain LCVR SEQ ID NO: 28 (DIQMTQSPSSVSASLGDRVTITCRASQGISSWLAWYQRKPGKAPKLLIYAASSLQSGVPSRFSGSGSGADFTLTISSLQPEDFATYYCQQAKSVPFTFGPGTKVDIK), 16747P binding domain HCVR SEQ ID NO: 29 (MSVPTQVLGLLLLWLTDARCQVQLVESGGGLV KPGGSLRLSCAASGFTFSDYYISWIRQAPGKGLEWVSYISSSGSSIKYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGNYGMDVWGQGTTVTV) and 16747P binding domain LCVR SEQ ID NO: 30 (DIQMTQSPSSVSASVGDRVTITCRASQGINNWLVWYQQKPGKAPKLLIYAATSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPTFGQGTKLEIK), 21587N binding domain HCVR sequence number 31 (MSVPTQVLGLLLLWLTDARCQVQLQESGPGLVKPSETLSLTCTVSGGSINYYYWNWIRQPPGKGLEWIGYISYSGNTNYNPSLKSRVTISVATSRNQFSLTLSSVTAADTAVYYCARFAEYCGGNICYYYGMDVWGQGTTVTV), and 21587N binding domain LCVR sequence number 32 (EIVLTQSPGTLSLSPGERATFSCRASQSVGSSFLAWYQQKPGQAPRRLMYGASNRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQCGGSPWTFGQGTKVEIK).

[0088] Provided herein are anti-BCMA CARs comprising variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein having one or more substitutions (such as conservative substitutions). For example, the disclosure includes anti-BCMA CARs having HCVR, LCVR and / or CDR amino acid sequences with, for example, 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 amino acid substitution compared to any of the HCVR, LCVR and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, or LCDR3) amino acid sequences disclosed herein. For example, an anti-BCMA CAR may include 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 amino acid substitution (such as a conservative amino acid substitution) compared to any of the HCVR, LCVR, and / or CDR (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 or LCDR3) amino acid sequences disclosed herein.

[0089] Exemplary binding domains described herein typically comprise, from amino-terminus to carboxy-terminus, a heavy chain region followed by a light chain region (VH-VL). Where a particular order of VH and VL regions in a binding domain is explicitly or implicitly described, the disclosure is also understood to describe alternative embodiments in which the order of VH and VL regions is reversed, for example, in a CAR comprising an scFV or scFv binding domain. Thus, a description of a VH-VL order also describes alternative VL-VH orders, for example, in a CAR comprising an scFV or scFv binding domain. Furthermore, a description of a VL-VH order also describes alternative VH-VL orders, for example, in a CAR comprising an scFV or scFv binding domain.

[0090] Generally, the nucleic acids encoding the CARs described herein include an extracellular linker portion that encodes a peptide linker that connects the binding domain to the transmembrane domain. Exemplary linker portions include, but are not limited to, a linker portion encoding a CD8α hinge domain, e.g., SEQ ID NO: 1 (PTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY) or SEQ ID NO: 2 (TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY). Typically, the region encoding the peptide linker (e.g., the CD8α hinge domain) is 3' to the region encoding the binding domain and 5' to the region encoding the transmembrane domain.

[0091] The nucleic acids encoding the CARs described herein comprise a transmembrane domain. The transmembrane domain can link an extracellular antigen-binding domain, e.g., a hinge, to one or more intracellular signaling components. For example, the transmembrane domain can link an antigen-binding domain, e.g., a hinge, to a CD3ζ signaling domain and, optionally, one or two costimulatory endodomains. Exemplary transmembrane domains include, but are not limited to, the CD8α transmembrane domain, e.g., SEQ ID NO: 3 (IWAPLAGTCGVLLLSLVITLYC). Typically, the region encoding the transmembrane domain (e.g., the CD8α transmembrane domain) is 3' to a region encoding a peptide linker (e.g., the CD8α hinge domain) and 5' to a region encoding one or more cytoplasmic domains.

[0092] In some embodiments, the isolated nucleic acid encodes a cytoplasmic region comprising one or more cytoplasmic domains. The region encoding the cytoplasmic region is typically 3' to the region encoding the transmembrane domain. The cytoplasmic domain is typically a signaling domain that provides an activation signal for γδ T cell proliferation, cytotoxic activity, and / or proinflammatory cytokine expression (e.g., TNF-α or IFNγ). An exemplary cytoplasmic domain is the CD3ζ signaling domain. In some embodiments, the CD3ζ signaling domain is or comprises SEQ ID NO: 4 (RVKFSRSADAPAYQQGQNQLYNELNLGR REEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR). In some embodiments, the CD3 zeta signaling domain is or comprises SEQ ID NO: 5 (RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDV LDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR). In some embodiments, the cytoplasmic region comprises multiple (e.g., 2, 3, 4, 5, or 6) signaling domains, such as multiple (e.g., 2, 3, 4, 5, or 6) CD3 zeta signaling domains, each independently selected from SEQ ID NOs: 4 and 5. In some embodiments, the cytoplasmic region comprises multiple (e.g., 2, 3, 4, 5, or 6) non-CD3 zeta signaling domains and a CD3 zeta signaling domain. In some embodiments, the cytoplasmic region comprises a non-CD3 zeta signaling domain and multiple (e.g., 2, 3, 4, 5, or 6) CD3 zeta signaling domains.

[0093] The cytoplasmic region may comprise one or more costimulatory endodomains. The region encoding one or more costimulatory endodomains may be 5' or 3' to the region encoding the signaling domain. In some embodiments, the region encoding one or more costimulatory endodomains may be 5' to the region encoding the signaling domain. In some embodiments, the region encoding one or more costimulatory endodomains is 5' to the region encoding the signaling domain, and an additional region encoding one or more costimulatory endodomains is 3' to the signaling domain. Exemplary costimulatory endodomains include, but are not limited to, CD28, CD137 (4-1BB), CD278 (ICOS), CD27, CD134 (OX40), Dap10, Dap12, DNAm-1, 2B4, a SLAM domain, and a TLR2 costimulatory endodomain, and combinations thereof.

[0094] In some embodiments, the construct encodes at least one 4-1BB costimulatory endodomain and, optionally, a second costimulatory endodomain selected from 4-1BB, 2B4, ICOS, CD28, and CD27 costimulatory endodomains. In some embodiments, the construct encodes at least two 4-1BB costimulatory endodomains or two 4-1BB costimulatory endodomains in combination with one, two, three, or four or more costimulatory endodomains selected from 4-1BB, ICOS, CD28, and CD27. In some embodiments, the 4-1BB costimulatory endodomain comprises SEQ ID NO: 6 (KRGRKKLLYIFKQPFMRPVQTT QEEDGCSCRFPEEEEGGCEL).

[0095] In some embodiments, the construct encodes one CD27 costimulatory endodomain and, optionally, a second costimulatory endodomain selected from 4-1BB, ICOS, CD28, and CD27 costimulatory endodomains. In some embodiments, the construct encodes a CD27 costimulatory endodomain and a 4-1BB costimulatory endodomain. In some embodiments, the construct encodes two CD27 costimulatory endodomains. In some embodiments, the CD27 costimulatory endodomain comprises SEQ ID NO: 7 (QRRKYRSNKGESPVEPAEPCHYSCPREEEGSTIPIQED YRKPEPACSP).

[0096] In some embodiments, the construct encodes a secretion signal, e.g., SEQ ID NO: 33 (MALPVTALLLPLALLLHAARP), operably linked to promote secretion of a C-terminal polypeptide, such as a cytokine that supports activation, cytotoxicity, and / or persistence of T cells (e.g., CAR-T cells). In some embodiments, the construct encodes a secretion signal, e.g., SEQ ID NO: 33, operably linked to promote secretion of a general gamma chain cytokine, such as IL-15, or an active fragment thereof, e.g., SEQ ID NO: 34 (NWVNVISDLKKIED LIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS). Exemplary general gamma chain cytokines include IL-2 and IL-15. In some embodiments, the general gamma chain cytokine is selected from IL-2, IL-7, and IL-15. In some embodiments, the general gamma chain cytokine is IL-15. IL-15 sequences, including codon-optimized nucleic acid sequences encoding sIL15, are disclosed herein and in WO2007 / 037780.

[0097] In some embodiments, the construct encodes one or more multicistronic linker regions, for example, between the signaling domain and / or costimulatory endodomain and the operably linked secretion signal to promote cytokine secretion. A multicistronic linker region is a region of a polypeptide or RNA sequence that facilitates the production of multiple distinct polypeptides from a single transcription product. In some embodiments, the multicistronic linker region encodes a cleavage sequence. Suitable cleavage sequences include self-cleaving sequences, such as P2A, F2A, E2A, or T2A cleavage sequences, and / or sequences that are cleaved by endogenous proteases, such as furin.

[0098] In some embodiments, the cleavage sequence is a P2A cleavage sequence. In some embodiments, the cleavage sequence is a furin cleavage sequence. In some embodiments, the cleavage sequence is a P2A and furin cleavage sequence. In some embodiments, the cleavage sequence is a P2A cleavage sequence of SEQ ID NO: 43 (SGSGATNFSLLKQAGDVEENPGP). In some embodiments, the cleavage sequence is a furin cleavage sequence of SEQ ID NO: 44 (RAKR). In some embodiments, the cleavage sequence is a P2A + furin cleavage sequence of SEQ ID NO: 45 (RAKRSGSGATNFSLLKQAG DVEENPGP).

[0099] In some embodiments, the cleavage sequence is or comprises the P2A cleavage sequence of SEQ ID NO: 52 (ATNFSLLKQAGDVEENPGP). In some embodiments, the cleavage sequence is or comprises the F2A cleavage sequence of SEQ ID NO: 53 (VKQTLNNFDLLKLAGDVESNPGP). In some embodiments, the cleavage sequence is or comprises the E2A cleavage sequence of SEQ ID NO: 54 (QCTNYALLKLAGDVESNPGP). In some embodiments, the cleavage sequence is or comprises the T2A cleavage sequence of SEQ ID NO: 55 (EGRSLLTCGDVEENPGP). In certain embodiments, multiple self-cleavage sequences may be encoded at the carboxy terminus of the signaling and / or costimulatory domain and the amino terminus of the encoded secreted cytokine (e.g., a general gamma chain cytokine such as IL-15), wherein the multiple self-cleavage sequences are preferably independently selected from the group consisting of a P2A cleavage sequence, a T2A cleavage sequence, an E2A cleavage sequence, and an F2A cleavage sequence. In certain aspects, one or more self-cleaving sequences and one or more sequences cleaved by an endogenous protease are encoded in the constructs described herein, hi certain embodiments, the endogenous protease recognition site is encoded amino-terminally to the self-cleaving sequence.

[0100] In some embodiments, the multicistronic linker region encodes an internal ribosome entry site. An exemplary internal ribosome entry site is encoded by SEQ ID NO: 56().

[0101] Another exemplary internal ribosome entry site is encoded by SEQ ID NO: 60 (AGCAGGTTTCCCCAACTGACACAAAACGTGCAACTTGAAACTCCGCCTGGTCTTTCCAGGTCTAGAGGGGTAACACTTTGTACTGCGTTTGGCTCCACGCTCGATCCACTGGCGAGTGTTAGTAACAGCACTGTTGCTTCGTAGCGGAGCATGACGGCCGTGGGAACTCCTCCTTGGTAACAAGGACCCACGGGGCCAAAAGCCACGCCCACACGGGCCCGTCATGTGTGCAACCCCAGCACGGCGACTTTACTGCGAAACCCACTTTAAAGTGACATTGAAACTGGTACCCACACACTGGTGACAGGCTAAGGATGCCCTTCAGGTACCCCGAGGTAACACGCGACACTCGGGATCTGAGAAGGGGACTGGGGCTTCTATAAAAGCGCTCGGTTTAAAAAGCTTCTATGCCTGAATAGGTGACCGGAGGTCGGCACCTTTCCTTTGCAATTACTGACCAC).

[0102] Further suitable internal ribosome entry sites include, but are not limited to, those described in Nucleic Acids Res. 2010 Jan;38(Database issue):D131-6. doi:10.1093 / nar / gkp981. Epub 2009 Nov. 16, those described in iresite.org, those described in WO2018 / 215787, the sequence described in GenBank accession No. KP019382.1 and the IRES element disclosed in GenBank accession No. LT727339.1.

[0103] Additional multicistronic linker regions, such as self-cleaving and IRES elements, are disclosed in US2018 / 0360992 and US8865467.

[0104] In some embodiments, the isolated nucleic acid is SEQ ID NO: 8 (MSVPTQVLGLLLLWLTDARCEIVMTQSPATLSVSPGERTTLSCRASQSVSSNLAWYLQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFILTISSLQSEDFAVYYCQQYNNWPITFGQGTRLEIKGGGGSGGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFYDYAMHWVRQAPGKGLEWVSGISWNSGYIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDNSYGKFYYGLDVWGQGTTVTVSSTTTPAPRPPTPAPTIASQPL SLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPAEPCHYSCPREEEGSTIPIQEDYRKPEPACSPRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding the 3H7-CD8-CD27z polypeptide, which in order contains the following domains: a 3H7-binding domain, a CD8α hinge and transmembrane domain, a CD27 costimulatory endodomain, and a CD3ζ signaling domain.

[0105] In some embodiments, the isolated nucleic acid is SEQ ID NO: 9 (MSVPTQVLGLLLLWLTDARCEIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGTSTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPLTFGGGTKVEIKGGGGSGGGGSGGGGEVQLVESGGGLVQPGRSLRLSCVASGFTFNDYAMHWVRQAPGKGLEWVSVISWNSDSIGYADSVKGRFTISRDNAKNSLYLQMHSLRAEDTALYYCAKDNHYGSGSYYYYQYGMDVWGQGTTVTVSSTTTPAPRPPTPAPT IASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding the 3B9-CD8-BBz polypeptide, which in order contains the following domains: a 3B9 binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, and a CD3ζ signaling domain.

[0106] In some embodiments, the isolated nucleic acid is SEQ ID NO: 10 (MSVPTQVLGLLLLWLTDARCEIVMTQSPATLSVSPGERTTLSCRASQSVSSNLAWYLQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFILTISSLQSEDFAVYYCQQYNNWPITFGQGTRLEIKGGGGSGGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFYDYAMHWVRQAPGKGLEWVSGISWNSGYIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDNSYGKFYYGLDVWGQGTTVTVSSTTTPAPRPPTPAPTI ASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding the 3H7-CD8-BBz polypeptide, which in turn contains the following domains: a 3H7-binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, and a CD3ζ signaling domain.

[0107] In some embodiments, the isolated nucleic acid is SEQ ID NO: 11 (MSVPTQVLGLLLLWLTDARCEIVLTQSPATLSLSPGERAALSCRASQSVSNYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPLTFGGGTKVEIRGGGGSGGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFRDYTMHWVRQGPGKGLEWVSGISWNSDYIGYADSVKGRFTISRDNAKNSLYLQMNSLRVEDTALYYCAKLSGTYRDYFYGVDVWGQGTTVTVSSTTTPAPRPPTPAPTI ASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding the 2B7-CD8-BBz polypeptide, which in order contains the following domains: 2B7 binding domain, CD8α hinge and transmembrane domain, 4-1BB costimulatory endodomain, and CD3ζ signaling domain.

[0108] In some embodiments, the isolated nucleic acid is SEQ ID NO: 12 (MSVPTQVLGLLLLWLTDARCEIVVTQSPATLSLSPGERATLSCRTSQTTTSYLAWYRQKPGQAPRLLIYDASNRAAGIPARFSGSGSGTDFTLTINSLEPEDFAVYYCQLRTNWITFGQGTRLEIKGGGGSGGGGSGGGGQVQLVESGGDSVKPGGSLRLSCAASGFTFSDSYMTWIRQAPGKGLEWVSFISSSGSTIYYADSVKGRFTISRDNVKKSLYLQMNRLRAEDTAVYYCAREEPGNYVYYGMDVWGQGTTVTVSSTTTPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding the 9C11-CD8-BBz polypeptide, which in order contains the following domains: 9C11 binding domain, CD8α hinge and transmembrane domain, 4-1BB costimulatory endodomain, and CD3ζ signaling domain.

[0109] In some embodiments, the isolated nucleic acid is SEQ ID NO: 20 (MSVPTQVLGLLLLWLTDARCE IVMTQSPATLSVSPGERTTLS CRASQSVSSNLAWYLQKPGQA PRLLIYGASTRATGIPARFSG SGSGTEFILTISSLQSEDFAV YYCQQYNNWPITFGQGTRLEI KGGGGSGGGGSGGGGEVQLVE SGGGLVQPGRSLRLSCAASGF TFYDYAMHWVRQAPGKGLEWV SGISWNSGYIGYADSVKGRFT ISRDNAKNSLYLQMNSLRAED TALYYCAKDNSYGKFYYGLDV WGQGTTVTVSSTTTPAPRPPT PAPTIASQPLSLRPEACRPAA GGAVHTRGLDFACDIYIWAPL AGTCGVLLLSLVITLYCRVKF SRSADAPAYQQGQNQLYNELN LGRREEYDVLDKRRGRDPEMG GKPQRRKNPQEGLYNELQKDK MAEAYSEIGMKGERRRGKGHD GLYQGLSTATKDTYDALHMQA LPPR), which encodes the 3H7-CD3z polypeptide, which contains, in order, the following domains: a 3H7 binding domain, a CD8α hinge and transmembrane domain, and a CD3ζ signaling domain.

[0110]

[0111]

[0112]

[0113]

[0114]

[0115] In some embodiments, the isolated nucleic acid comprises a codon-optimized sequence encoding a CD8α hinge region. Exemplary codon-optimized CD8α hinge region nucleic acid sequences include, but are not limited to, SEQ ID NO: 18 (ACCACCACCCCTGCACCAAGGCCCCCGACTCCCGCGCCCACCATCGCGTCA CAGCCTCTTAGCCTGCGACCGGAAGCATGCAGACCAGCTGCCGGGGGGGCCGTGCATACGAGAGGTTTGGACTTCGCCTGCGAT). In some embodiments, the CD8α hinge region is encoded by the following SEQ ID NO: 19 (ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGAT).

[0116] In some embodiments, the isolated nucleic acid encodes the 3B9 binding domain and comprises the following sequence encoding the CD8 alpha hinge domain SEQ ID NO: 18. In some embodiments, the isolated nucleic acid encodes the 2B7 binding domain and comprises the following sequence encoding the CD8 alpha hinge domain SEQ ID NO: 18. In some embodiments, the isolated nucleic acid encodes the 9C11 binding domain and comprises the following sequence encoding the CD8 alpha hinge domain SEQ ID NO: 18. In some embodiments, the isolated nucleic acid encodes the 3H7 binding domain and comprises the following sequence encoding the CD8 alpha hinge domain SEQ ID NO: 19.

[0117] In some embodiments, the isolated nucleic acid is SEQ ID NO: 35 (MSVPTQVLGLLLLWLTDARCEVQLVESGGGLVQPGGSLRLSCAASGFTFSSYVMSWVRQAPGKGLEWVSAIIGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKRAGDNWNWFDPWGQGTLVTVSSGGGGSGGGGSGGGGDIQMTQSPSSVSASLGDRVTITCRASQGISSWLAWYQRKPGKAPKLLIYAASSLQSGVPSRFSGSGSGADFTLTISSLQPEDFATYYCQQAKSVPFTFGPGTKVDIKTTTPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding an anti-BCMA-CAR polypeptide containing, in order, the following domains: 16716P binding domain, CD8α hinge and transmembrane domain, 4-1BB costimulatory endodomain and CD3ζ signaling domain.

[0118] In some embodiments, the isolated nucleic acid is SEQ ID NO: 36 (MSVPTQVLGLLLLWLTDARCQVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYISWIRQAPGKGLEWVSYISSSGSSIKYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCAREGGNYGMDVWGQGTTVTVSSGGGGSGGGGSGGGGDIQMTQSPSSVSASVGDRVTITCRASQGINNWLVWYQQKPGKAPKLLIYAATSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPTFGQGTKLEIKTTTPAPRPPTPAPTIASQ PLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding an anti-BCMA-CAR polypeptide containing, in order, the following domains: 16747P binding domain, CD8α hinge and transmembrane domain, 4-1BB costimulatory endodomain and CD3ζ signaling domain.

[0119] In some embodiments, the isolated nucleic acid is SEQ ID NO: 37 (MSVPTQVLGLLLLWLTDARCQVQLQESGPGLVKPSETLSLTCTVSGGSINYYYWNWIRQPPGKGLEWIGYISYSGNTNYNPSLKSRVTISVATSRNQFSLTLSSVTAADTAVYYCARFAEYCGGNICYYYGMDVWGQGTTVTVSSGGGGSGGGGSGGGGEIVLTQSPGTLSLSPGERATFSCRASQSVGSSFLAWYQQKPGQAPRRLMYGASNRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQCGGSPWTFGQGTKVEIKTTTPAPRPPTPAPT IASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR), encoding an anti-BCMA-CAR polypeptide containing, in order, the following domains: 21587N binding domain, CD8α hinge and transmembrane domain, 4-1BB costimulatory endodomain, and CD3ζ signaling domain.

[0120] In some embodiments, the isolated nucleic acid encodes SEQ ID NO: 38(*), an anti-BCMA-CAR polypeptide comprising the following domains, in order: a 16747P binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, a CD3ζ signaling domain, a furin+P2A cleavage domain, a secretion signal, and a sIL15 domain.

[0121]

[0122]

[0123]

[0124]

[0125] In some embodiments, the isolated nucleic acid encodes SEQ ID NO: 46(*), an anti-CD20 CAR polypeptide comprising, in order, the following domains: a 3H7 binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, a CD3ζ signaling domain, a P2A cleavage domain (GSGATNFSLLKQAGDVEENPGP, SEQ ID NO: 47), a secretion signal, and a sIL15 domain.

[0126] In some embodiments, the isolated nucleic acid encodes an anti-CD20 CAR polypeptide comprising, in order, SEQ ID NO: 48(*), the following domains: a 3H7 binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, a CD3ζ signaling domain, a P2A cleavage domain of SEQ ID NO: 47, a secretion signal of SEQ ID NO: 49 (MRISKPHLRSISIQCYLCLLLNSHFLTEAGIHVFILGCFSAGLPKTEA), and a sIL15 domain.

[0127]

[0128]

[0129] In some embodiments, the isolated nucleic acid is SEQ ID NO: 57 (MSVPTQVLGLLLLWLTDARCEIVMTQSPATLSVSPGERTTLSCRASQSVSSNLAWYLQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFILTISSLQSEDFAVYYCQQYNNWPITFGQGTRLEIKGGGGSGGGGSGGGGEVQLVESGGGLVQPGRSLRLSCAASGFTFYDYAMHWVRQAPGKGLEWVSGISWNSGYIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDNSYGKFYYGLDVWGQGTTVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLY IFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR*), an anti-CD20 CAR polypeptide comprising, in order, the following domains: a 3H7 binding domain, a CD8α hinge and transmembrane domain, a 4-1BB costimulatory endodomain, and a CD3ζ signaling domain, but via an internal ribosome entry site (e.g., the site encoded by SEQ ID NO: 56) in the region encoding SEQ ID NO: 57, this isolated nucleic acid is encoded as SEQ ID NO: 58 (MALPVTALLLPLALLLHAARPNWVNVISDLKKIEDLIQSMHIDATLYTESDVHP SCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS*), further encoding the secretion signal of SEQ ID NO: 33 and the sIL15 domain.

[0130]

[0131] In some embodiments, the isolated nucleic acid is a linear nucleic acid. In some embodiments, the isolated nucleic acid is a circular nucleic acid. In some embodiments, the isolated nucleic acid is a vector, such as a plasmid vector, an adenoviral vector, an adeno-associated viral vector, a viral vector, a retroviral vector, or a lentiviral vector. In some embodiments, the isolated nucleic acid, or a contiguous portion of the isolated nucleic acid, e.g., comprising a binding domain, a transmembrane domain, and one or more signaling and / or costimulatory endodomains, is integrated into the genome of a host cell, such as a host γδ T cell. In an exemplary embodiment, the isolated nucleic acid is a retroviral vector.

[0132] γδT cells: An embodiment of the invention includes a γδ T cell that functionally expresses an isolated nucleic acid described herein, thereby expressing a CAR on the surface of the γδ T cell.

[0133] Embodiments of the invention may alternatively or additionally include γδ T-cells that have in vitro or in vivo cytotoxic activity against hematologic tumor cells that exhibit cell surface expression of a tumor-associated antigen (TAA). In some cases, the cytotoxic activity is innate. In some cases, the cytotoxicity is at least partially, significantly (>about 25%), or entirely due to the presence of a CAR construct having a binding domain that specifically binds to a TAA expressed on the surface of the hematologic tumor cells. In some cases, the γδ T-cells exhibit hematologic tumor cytotoxic activity that is greater than the innate level of in vitro and / or in vivo hematologic tumor cytotoxic activity of control γδ T-cells. In some cases, the control γδ T-cells do not comprise a CAR construct. In some cases, the control γδ T-cells comprise a CAR construct lacking a binding domain described herein, a hinge region described herein, a transmembrane domain described herein, a signaling domain described herein, and / or a costimulatory endodomain described herein.

[0134] In some cases, the cytotoxicity is at least partially, significantly (>about 25%), or entirely due to the presence of a CAR construct having a binding domain that specifically binds to CD20 or an epitope within CD20. In some cases, the γδ T cells functionally express a CD20-specific CAR encoded by an isolated nucleic acid described herein.

[0135] In some embodiments, the γδ T cells described herein may exhibit HLA-restricted (e.g., HLA class I-restricted) cytotoxicity. In other embodiments, most (>50%), substantially all (>90%), or all cytotoxic activity is not HLA-restricted (e.g., HLA class I-restricted). HLA-restricted cytotoxic activity can be assessed by comparing in vitro cytotoxicity against HLA (e.g., HLA class I) (null) tumor cell lines with in vitro cytotoxicity against HLA+ (e.g., HLA class I+) tumor cell lines. In some embodiments, HLA-restricted cytotoxic activity is at least partially, significantly (>25%), or completely provided by the use of a T cell receptor-like binding domain. T cell receptor-like binding domains are binding domains that specifically recognize antigens when presented on the surface of cells in complex with MHC molecules. T cell receptor-like binding domains are further described, for example, in WO2016 / 199141.

[0136] The γδ T cells described herein can exhibit potent and / or sustained hematologic tumor cytotoxic activity. In some cases, the hematologic tumor cytotoxic activity can persist for at least about 6 to 120 days, or at least about 6 to 180 days, from initial contact with hematologic tumor cells. In some cases, the hematologic tumor cytotoxic activity of the γδ T cells described herein or their progeny can persist for at least about 6 to 120 days, or at least about 6 to 180 days, from initial contact with hematologic tumor cells or from administration of the γδ T cells described herein. This sustained hematologic tumor cytotoxic activity can be demonstrated in vitro, in vivo, or both in vitro and in vivo.

[0137] Embodiments of the present invention may alternatively or additionally include γδ T cells that proliferate in response to contact with cells that exhibit cell surface expression or overexpression of a tumor-associated antigen (TAA). The cells that exhibit cell surface expression or overexpression of a tumor-associated antigen (TAA) may be normal blood cells, such as normal B cells. The cells that exhibit cell surface expression or overexpression of a tumor-associated antigen (TAA) may be hematologic tumor cells. In some cases, the proliferation is an innate activity. In some cases, the proliferation is at least partially, significantly (>about 20% or >about 25%), or entirely due to the presence of a CAR construct having a binding domain that specifically binds to a TAA expressed on the surface of a blood cell or hematologic tumor cell. In some cases, the γδ T cells exhibit higher levels of proliferation in vitro and / or in vivo compared to control γδ T cells. In some cases, the control γδ T cells do not comprise a CAR construct. In some cases, the control γδ T-cell comprises a CAR construct lacking a binding domain described herein, a hinge region described herein, a transmembrane domain described herein, a signaling domain described herein, and / or a costimulatory endodomain described herein.

[0138] In some cases, proliferation is at least partially, significantly (>about 20% or >about 25%), or entirely due to the presence of a CAR construct having a binding domain that specifically binds to CD20 or an epitope within CD20. In some cases, γδ T cells that exhibit proliferation in response to contact with blood cells or hematological tumor cells that exhibit cell surface expression of CD20 functionally express a CD20-specific CAR encoded by an isolated nucleic acid described herein.

[0139] The γδ T-cells described herein can exhibit robust and / or sustained proliferation in a host organism comprising blood cells or hematologic tumor cells that exhibit cell surface expression or overexpression of a tumor-associated antigen (TAA). In some cases, proliferation can persist for at least about 6 to 120 days, or at least about 6 to 180 days, from initial contact with the hematologic tumor cells or from the date of administration of the γδ T-cells to the host organism. In some cases, proliferation of the γδ T-cells described herein or their progeny in a host organism comprising blood cells or hematologic tumor cells that exhibit cell surface expression or overexpression of a tumor-associated antigen (TAA) can persist for at least about 6 to 120 days, or at least about 6 to 180 days, from initial contact with the blood cells or hematologic tumor cells or from the date of initial administration of the γδ T-cells to the host organism. In some cases, proliferation in the host organism is at least partially, significantly (>about 20% or >about 25%), or entirely due to the presence of a CAR construct having a binding domain that specifically binds to CD20 or an epitope within CD20. In some cases, γδ T cells exhibiting proliferation in a host organism containing blood cells or hematological tumor cells that exhibit cell surface expression of CD20 functionally express a CD20-specific CAR encoded by an isolated nucleic acid described herein.

[0140] In some embodiments, a γδ T-cell described herein expresses or persistently expresses a pro-inflammatory cytokine, such as tumor necrosis factor alpha or interferon gamma, after contact with blood cells or hematologic tumor cells. In some embodiments, a γδ T-cell described herein or its progeny expresses or persistently expresses a pro-inflammatory cytokine, such as tumor necrosis factor alpha or interferon gamma, after contact with blood cells or hematologic tumor cells, such as in a host organism that contains the blood cells or hematologic tumor cells.

[0141] In some embodiments, γδ T cells or pharmaceutical compositions comprising γδ T cells, when introduced into an allogeneic host, exhibit substantially no or no graft-versus-host response. In some embodiments, γδ T cells or pharmaceutical compositions comprising γδ T cells, when introduced into an allogeneic host, exhibit a clinically acceptable level of graft-versus-host response. In some embodiments, a clinically acceptable level is an amount of graft-versus-host response that does not require cessation of γδ T cell therapy to achieve therapeutically effective treatment. In some embodiments, a clinically acceptable level of graft-versus-host response (GvHD) is an acute response less severe than Grade C according to the applicable IBMTR grading scale. The severity of acute graft-versus-host response is determined by assessing the degree of skin, liver, and gastrointestinal involvement. The stages of involvement in individual organs are combined to create an overall grade that has prognostic significance. Grade I (A) GvHD is considered mild disease, Grade II (B) GvHD is moderate, Grade III (C) is severe, and Grade IV (D) is life-threatening. The IBMTR grading system defines the severity of acute GvHD as follows (Rowlings et al., Br J Haematol 1997;97:855): Grade A - No liver or gastrointestinal involvement, only stage 1 skin involvement (maculopapular rash on <25% of the body) Grade B - Stage 2 skin lesions, Stage 1 to 2 intestinal or liver lesions Grade C - Stage 3 disease in any organ system (generalized erythroderma, bilirubin 6.1-15.0 mg / dL, diarrhea 1500-2000 mL / day) • Grade D - Stage 4 involvement of any organ system (generalized erythroderma with blister formation, bilirubin >15 mg / dL, diarrhea >2000 mL / day or pain or ileus). See also Tables 1 and 2 in Schoemans et al., Bone Marrow Transplantation volume 53, pages 1401-1415 (2018), which also discloses criteria for assessing and grading acute GvHD.

[0142] In some embodiments, the γδ T cells, or pharmaceutical compositions comprising γδ T cells, exhibit a reduced or substantially reduced graft-versus-host response when introduced into an allogeneic host compared to the graft-versus-host response exhibited by control αβ T cells, or a control pharmaceutical composition comprising control αβ T cells, administered to the allogeneic host. Optionally, the control αβ T cells are allogeneic, non-genetically engineered control αβ T cells. Optionally, the control αβ T cells do not comprise a CAR or do not comprise the same CAR as the reference γδ T cells.

[0143] The γδ T cells described herein can be δ1, δ2, δ3, or δ4 γδ T cells, or a combination thereof. In some cases, the γδ T cells are mostly (>50%), substantially (>90%), essentially all, or entirely δ2 - In some cases, the γδ T cells are mostly (>50%), substantially (>90%), essentially all, or entirely δ1 γδ T cells.

[0144] γδ T cells can be obtained from allogeneic or autologous donors. γδ T cells can be partially or completely purified or unpurified and expanded ex vivo. Methods and compositions for ex vivo expansion include, but are not limited to, those described in WO2017 / 197347. Expansion can be performed before or after, or both before and after, the CAR construct is introduced into the γδ T cell(s).

[0145] The γδ T cells described herein can be stored, for example, cryopreserved, for use in adoptive cell transfer.

[0146] Methods for inhibiting or killing tumor cells One or more non-engineered γδ T cell populations, genetically engineered γδ T cell populations, and / or mixtures thereof having cytotoxic activity against hematologic tumor cells can be administered to a subject in any order or simultaneously. When administered simultaneously, multiple non-engineered γδ T cell populations, genetically engineered γδ T cell populations, and / or mixtures thereof of the present invention can be provided in a single, unified form, such as an intravenous injection, or in multiple forms, such as multiple intravenous infusions, subcutaneous injections, or tablets. Non-engineered γδ T cell populations, genetically engineered γδ T cell populations, and / or mixtures thereof of the present invention can be packaged together or separately in a single package or multiple packages. One or all of the non-engineered γδ T cell populations, genetically engineered γδ T cell populations, and / or mixtures thereof of the present invention can be given in multiple doses. When not administered simultaneously, the timing between multiple doses can vary by about one week, one month, two months, three months, four months, five months, six months, or up to about one year. In some cases, the non-engineered enriched γδ T cell population, the genetically engineered enriched γδ T cell population, and / or a mixture thereof of the present invention may be expanded in vivo within the subject's body after administration to the subject. One or more non-engineered γδ T cell populations, one or more genetically engineered γδ T cell populations, and / or a mixture thereof can be frozen to provide cells for multiple treatments with the same cell preparation. One or more non-engineered γδ T cell populations, one or more genetically engineered γδ T cell populations, and / or a mixture thereof of the present disclosure, as well as pharmaceutical compositions comprising the same, can be packaged as a kit. The kit may include instructions (e.g., written instructions) for using the non-engineered γδ T cell population, the genetically engineered γδ T cell population, and / or a mixture thereof, as well as compositions comprising the same.

[0147] In some cases, the method for treating a hematological cancer comprises administering to a subject a therapeutically effective amount of a non-engineered γδ T cell population, a genetically engineered γδ T cell population, and / or a mixture thereof, wherein administering treats the hematological cancer. In some embodiments, the therapeutically effective amount of a non-engineered γδ T cell population, a genetically engineered γδ T cell population, and / or a mixture thereof is administered for at least about 10 seconds, 30 seconds, 1 minute, 10 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, 24 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, or 1 year. In some embodiments, the therapeutically effective amount of a non-engineered γδ T cell population, a genetically engineered γδ T cell population, and / or a mixture thereof is administered for at least 1 week. In some embodiments, a therapeutically effective amount of a non-engineered γδ T cell population, an engineered γδ T cell population and / or a mixture thereof is administered for at least two weeks.

[0148] The non-genetically engineered γδ T cell populations, genetically engineered γδ T cell populations, and / or mixtures thereof described herein can be administered before, during, or after the onset of a disease or symptom, and the timing of administration of a pharmaceutical composition comprising a γδ T cell population can vary. For example, the γδ T cell populations can be used as prophylactic agents and can be administered continuously to subjects exhibiting symptoms or a tendency toward a disease to reduce the likelihood of the disease or symptom occurring. The initial administration can be via any practical route, such as by any route described herein using any formulation described herein. In some embodiments, the γδ T cell populations of the present disclosure are administered intravenously. One or more doses of the γδ T cell populations can be administered as soon as practicable after the onset of a hematological cancer, for a period of time necessary to treat the immune disease, for example, from about 24 hours to about 48 hours, from about 48 hours to about 1 week, from about 1 week to about 2 weeks, from about 2 weeks to about 1 month, or from about 1 month to about 3 months. In some embodiments, one or more doses of the γδ T cell population may be administered years after the onset of cancer, and before and after other treatments.

[0149] In some embodiments, the γδ T cell population is administered simultaneously or sequentially with one or more methods for elevating common gamma chain cytokine(s). As used herein, "one or more methods for elevating common gamma chain cytokine(s)" refers to a method or combination of methods for altering the physiological state of a subject such that the level of at least one common gamma chain cytokine is elevated in the subject. In some embodiments, the method elevates the level of one or more common gamma chain cytokine(s) selected from the group consisting of IL-2, IL-7, and IL-15, preferably wherein the method elevates the level of IL-15 in the subject. In some embodiments, the method comprises lymphodepletion. In some embodiments, the method comprises administering one or more common gamma chain cytokine(s) to the subject. Optionally, IL-2, IL-7, and / or IL-15, preferably IL-15, are administered. In some embodiments, the method comprises secreting the common gamma chain cytokine(s), such as from the administered γδ T cells. In some cases, IL-2, IL-7 and / or IL-15, preferably IL-15, is secreted.

[0150] In some embodiments, the one or more administration methods for increasing general gamma chain cytokine(s) include lymphodepletion before introducing γδ T cell(s). In some embodiments, the one or more administration methods for increasing general gamma chain cytokine(s) include administering an effective amount of general gamma chain cytokine(s) simultaneously with or consecutively with the introduction of γδ T cell(s) to increase the proliferation, cytotoxic activity, persistence, or a combination thereof, of the introduced γδ T cell(s), preferably wherein the method comprises administering IL-2 or one or more mimetics thereof, more preferably wherein the method comprises administering IL-15 or one or more mimetics thereof. The administration of the general gamma chain cytokine(s) may increase the proliferation, cytotoxic activity, persistence, or a combination thereof, of the introduced γδ T cell(s) before and / or after introducing γδ T cell(s). Exemplary amounts of IL-15 include, but are not limited to, between 0.01 and 10 μg / kg / dose for IL-15 every 24 hours. Exemplary amounts of IL-2 include about 3×10 6 and approximately 22 x 10 6 For example, the dosing regimen for IL2 in RCC is 600,000 International Units / kg (0.037 mg / kg) IV every 8 hours infused over 15 minutes for up to 14 doses.

[0151] In some embodiments, the one or more administration methods that elevate general gamma chain cytokine(s) comprise lymphodepletion prior to administration of γδ T-cell(s), prior to simultaneous administration with the introduced γδ T-cell(s), or prior to sequential administration of general gamma chain cytokine(s) effective to increase proliferation, cytotoxic activity, persistence, or a combination thereof, of the introduced γδ T-cell(s). [Example]

[0152] Example 1 1x106 Human PBMCs (PBMCs) at 100 μg / mL were activated with engineered cell culture medium on pre-coated anti-Vδ1 antibodies D1-08 or D1-35 in 24-well plates (Costar) in the presence of IL-2 (100 U / mL) for 5 days (see WO2017 / 197347). On day 5, the cell cultures were transduced with γ-retroviral constructs encoding chimeric antigen receptors (2B7-5.1, SEQ ID NO: 11; 3B9-5.1, SEQ ID NO: 9; 3H7-5.1, SEQ ID NO: 10; and 9C11-5.1, SEQ ID NO: 12) in the presence of retronectin. On day 6, the cells were returned to engineered cell culture medium and further expanded with feeding and IL-2 supplementation as needed. On days 17, 18, or 19, the cells were harvested and depleted of remaining αβ T cells using an AutoMACS® kit (Miltenyi Biotec). The purity and transduction efficiency of the γδ cell population were assessed by FACS. In parallel, untransduced cell cultures were grown in the same manner without the addition of retroviral supernatant. As shown in Figure 3, untransduced expanded Vδ1 cells from multiple donors were not cytotoxic to normal B cells from allogeneic donors. Transduction of CD20CAR into Vδ1 cells conferred potent cytotoxicity to these cells against normal B cells. Cytotoxicity was measured as % Annexin V+ cells by flow cytometry in a 4-hour assay.

[0153] Example 2 V51 cells were activated, transduced, and expanded in the same manner as described above. The 3H7 CAR construct, SEQ ID NO: 10, was used to demonstrate cytotoxicity against two CD20+ cell lines, Daudi and Raji. As shown in Figure 4, introduction of the CAR increased the innate cytotoxicity of non-engineered V51 cells.

[0154] Example 3 V51 cells were activated, transduced, and expanded in the same manner as described above. Four different constructs (SEQ ID NOs: 9, 10, 11, and 12) were introduced into V51 cells during expansion and tested against Raji-Luc cells. Cytotoxicity was measured by total luminescence measurement after 18 hours of co-culture at various E / T ratios and addition of the luminescent substrate D-luciferin (Perkin Elmer). As shown in FIG. 5, anti-CD20CAR cells containing the 4-1BB costimulatory endodomain described herein exhibited potent cytotoxic activity against Raji cells.

[0155] Example 4 CAR constructs were generated with several different domains and a CD20 binding domain (3H7-CD3z, SEQ ID NO: 20, 3H7-5.1, SEQ ID NO: 10, and 3H7-CD27z, SEQ ID NO: 8). CAR constructs were transfected as described above, and cytotoxicity was tested against Raji-Luc cells (18-hour cytotoxicity at various E / T ratios) as described in the previous example. Figure 6 shows potent cytotoxic activity against Raji cells bearing various signaling and / or costimulatory endodomain(s).

[0156] Example 5 Various CAR constructs were introduced into expanded V51 cells and tested in a long-term cytotoxicity assay (serial killing) by target cell rechallenge using an IncuCyte® instrument. Briefly, Raji cells were labeled with NucRed reagent, and the total fluorescence of the cells was recorded over time. Without the addition of any cytokines, cells were co-cultured in growth medium at an E / T ratio of 3 for 72 hours. After 72 hours, the cultures were rechallenged with another dose of Raji cells and monitored for killing. This procedure was repeated for up to 144 hours in cultures from which Raji cells had been removed. In Figure 7, 3H7-ICOSz is a construct in which the 4-1BB costimulatory endodomain was replaced with the ICOS endodomain (WLTKKKYSSSVHDPNGEYMFMRAVNTAKKSRLTDVTL (SEQ ID NO: 354)).

[0157] Example 6 Raji cells were subcutaneously implanted into NSG mice (Jackson Labs). Tumors grew to approximately 100 mm 3 Once the animals reached a size of 5 × 10, they were cultured in a 5 × 10 -well culture to compare the efficacy of various costimulatory endodomains ("co-stim" or "costim") in vivo. 6 Vδ1CD20 CAR cells were used to treat these tumors. Animals were simultaneously administered IL-2 (60,000 U / dose) three times a week until the end of the study. As shown in Figure 8, the tested constructs demonstrated potent in vivo efficacy in treating hematologic tumors in NSG mice. Without wishing to be bound by theory, it is hypothesized that the optimized CAR constructs, 3H7-5.1, 3H7-CD3z, and 3H7-CD27z, exhibit superior in vivo tumor control, proliferation, activation, persistence, and / or cytotoxicity compared to non-optimized CAR constructs.

[0158] Example 7 Raji cells were subcutaneously implanted into NSG mice (Jackson Labs). Tumors grew to approximately 100 mm 3 Once the animals reached 100 μg / ml, they were transferred to 5 × 10 6 Animals were treated with Vδ1 CD20 CAR+ cells. On days 2 and 6, tumors and various other organs were extracted and digested, and the resulting cell suspensions were analyzed for (A) the presence of γδ T cells and Raji cells (Figure 9) and (B) γδ cell proliferation as evidenced by CellTrace Violet dye dilution (Figure 10). Animals were simultaneously administered IL-2 (60,000 U / dose) three times a week until day 6. As shown in Figure 9, the transferred γδ T cells robustly expanded in the intratumoral environment and promoted a significant decrease in the ratio of tumor cells to γδ T cells from days 2 to 6. As shown in Figure 10, γδ T cells robustly and preferentially proliferated in the intratumoral space.

[0159] Example 8 NSG mice were inoculated with Raji-Luc cells (0.5 ml / animal). On day 4, animals were inoculated with 8.7 x 10 6 Vδ1CAR+ cells (SEQ ID NO: 10) or 6.8x10 transduced with the same construct5 Subjects with hematological cancers were treated with αβ T cells. Animal survival was monitored over a 140-day period. All animals received three doses of IL-2 (60,000 / animal) on days 0, 1, and 2 (FIG. 11). As shown in FIG. 11, administration of γδ T cells as described herein increased survival time in subjects with hematological cancers.

[0160] Example 9 SRG-15 mice expressing human IL-15 (Herndler-Brandstetter et al., PNAS, 2017) were transfected with Raji-Luc cells (0.5x10 6 On day 4, animals were inoculated with 20.2 x 10 6 of Vδ1CAR+ cells (SEQ ID NO: 10) or 1.9x10 transduced with the same construct 6 The animals were treated with αβ T cells. Animal survival was monitored for 70 days ( FIG. 12 ). As shown in FIG. 12 , the transferred γδ T cells did not induce a GvHD response. In contrast, the transferred αβ T cells did induce a GvHD response.

[0161] Example 10 Inject Raji cells (1 x 10) into the right hind flank of NSG mice. 6 / animal) was subcutaneously inoculated. The tumor volume was approximately 100 mm 3 Once the mice reached 100 μg / ml, they were randomized to receive 5x10 CAR-encoding CD20CAR or CARCD20 and soluble IL-15. 6 On day 62, four animals in the CD20+sIL15CAR T group that did not have measurable tumors were treated with 1x10 IL-2 in the contralateral (left) flank. Animals were simultaneously administered IL-2 (60000U / dose, Peprotech, 3 times per week) until the end of the study. 6 The animals were then re-administered subcutaneously with γδ CAR-T cells carrying a heterologous soluble IL-15-encoding nucleic acid construct. As shown in Figure 14, administration of γδ CAR-T cells carrying a heterologous soluble IL-15-encoding nucleic acid construct resulted in a durable anti-tumor effect lasting for more than 60 days (e.g., 60 to 110 days).

[0162] Example 11 1x10 cells in growth medium in 24-well plates (Costar) precoated with anti-Vδ1 antibody D1-08 or D1-35 for 5 days in the presence of IL-2 (100 U / mL). 6 Human PBMCs were activated at 1000kJ / mL. On day 5, cell cultures were transduced with a gamma-retroviral construct encoding the BCMA chimeric antigen receptor (SEQ ID NOs: 35-38) in the presence of retronectin. On day 6, cells were returned to growth medium and further expanded by feeding and IL-2 supplementation as needed. On days 17, 18, or 19, cells were harvested and depleted of remaining αβ T cells using an AutoMACS® kit (Miltenyi Biotec). The purity and transduction efficiency of the γδ cell population were assessed by FACS (Figure 15). Briefly, CAR-T cells were stained by incubating cells with 1 μg / mL of soluble recombinant biotinylated BCMA (Acro Biosystems). Binding detection was performed using streptavidin-BV421 at the manufacturer's recommended dilution of 1:500. In parallel, untransduced cell cultures were grown in the same manner without the addition of retroviral supernatant. The expanded cells were tested in an in vitro cytotoxicity assay of BCMA-positive cell lines. As shown in Figures 16 and 17, untransduced expanded Vδ1 cells induced some degree of cytotoxicity against multiple myeloma and Burkitt's lymphoma cell lines, which are known to express BCMA to varying degrees. This cytotoxicity was enhanced by introducing a BCMA CAR construct. Cytotoxicity was measured by total luminescence measurement in 96-well plates after adding the luminescent substrate D-luciferin (Perkin Elmer) after 18 hours of coculture at the indicated E / T ratio. A BCMA-negative Scaber cell line was used as a control.

[0163] Example 12 NCI-H929 multiple myeloma cells (1x10 6 ( / animal) were subcutaneously implanted into NSG mice (Jackson Labs). Tumors grew to approximately 200 mm 3Once the animals reached a size of 5 x 10, they were cultured at 5 x 10 to compare the in vivo efficacy of the 16716P and 16747P scFv-derived CAR constructs. 6 Anti-BCMA CAR+ cells were used to treat tumors in vivo. Animals were simultaneously administered IL-2 (13,000 IU / dose, Proleukin®) three times a week until the end of the study. The results are shown in Figure 18. As shown in Figure 19, anti-BCMA CAR+ cells demonstrated potent in vivo tumor burden control.

[0164] The foregoing merely illustrates the principles of the present invention. It will be appreciated by those skilled in the art that, although not explicitly described or shown herein, they will be able to devise various configurations which embody the principles of the present invention and are within its spirit and scope. Furthermore, all examples and conditional language recited herein are intended primarily to aid the reader's understanding of the principles of the present invention and the concepts contributed by the inventors to further this technology, and should not be construed as being limited to such specifically recited examples and conditions. Furthermore, all statements herein reciting principles, aspects, and specific examples thereof are intended to encompass both structural and functional equivalents. Furthermore, such equivalents are intended to include both currently known equivalents and equivalents developed in the future, i.e., any elements developed to perform the same function, regardless of structure. Therefore, the scope of the present invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the present invention are embodied by the appended claims. SEQUENCE LISTING <110> ADICET BIO, INC. <120> COMPOSITIONS AND METHODS REGARDING ENGINEERED AND NON- ENGINEERED GAMMA DELTA-T CELLS FOR TREATMENT OF HEMATOLOGICAL TUMORS <130> ADC-0005-PCT <140> PCT / US2019 / 054132 <141> 2019-10-01 <150> 62 / 739,822 <151> 2018-10-01 <160> 370 <170> PatentIn version 3.5 <210> 1 <211> 39 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 1 Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro 1 5 10 15 Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu 20 25 30 Asp Phe Ala Cys Asp Ile Tyr 35 <210> 2 <211> 47 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 2 Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 1 5 10 15 Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 20 25 30 Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr 35 40 45 <210> 3 <211> 22 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 3 Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu 1 5 10 15 Val Ile Thr Leu Tyr Cys 20 <210> 4 <211> 113 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 4 Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 1 5 10 15 Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 20 25 30 Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 35 40 45 Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln 50 55 60 Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu 65 70 75 80 Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr 85 90 95 Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro 100 105 110 Arg <210> 5 <211> 112 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 5 Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 1 5 10 15 Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 20 25 30 Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 35 40 45 Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys 50 55 60 Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg 65 70 75 80 Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala 85 90 95 Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 100 105 110 <210> 6 <211> 42 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 6 Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 1 5 10 15 Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 20 25 30 Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 35 40 <210> 7 <211> 48 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 7 Gln Arg Arg Lys Tyr Arg Ser Asn Lys Gly Glu Ser Pro Val Glu Pro 1 5 10 15 Ala Glu Pro Cys His Tyr Ser Cys Pro Arg Glu Glu Glu Gly Ser Thr 20 25 30 Ile Pro Ile Gln Glu Asp Tyr Arg Lys Pro Glu Pro Ala Cys Ser Pro 35 40 45 <210> 8 <211> 493 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 8 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Gln Arg Arg Lys 325 330 335 Tyr Arg Ser Asn Lys Gly Glu Ser Pro Val Glu Pro Ala Glu Pro Cys 340 345 350 His Tyr Ser Cys Pro Arg Glu Glu Glu Gly Ser Thr Ile Pro Ile Gln 355 360 365 Glu Asp Tyr Arg Lys Pro Glu Pro Ala Cys Ser Pro Arg Val Lys Phe 370 375 380 Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 385 390 395 400 Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 405 410 415 Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg 420 425 430 Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 435 440 445 Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 450 455 460 Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 465 470 475 480 Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 485 490 <210> 9 <211> 492 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 9 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Thr Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asn Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile 180 185 190 Ser Trp Asn Ser Asp Ser Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 His Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn His Tyr Gly Ser Gly Ser Tyr Tyr Tyr Tyr Gln Tyr Gly Met Asp 245 250 255 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro 260 265 270 Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu 275 280 285 Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His 290 295 300 Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu 305 310 315 320 Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr 325 330 335 Cys Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe 340 345 350 Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg 355 360 365 Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser 370 375 380 Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr 385 390 395 400 Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys 405 410 415 Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys 420 425 430 Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala 435 440 445 Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys 450 455 460 Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr 465 470 475 480 Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 485 490 <210> 10 <211> 487 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 10 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg 325 330 335 Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln 340 345 350 Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu 355 360 365 Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala 370 375 380 Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu 385 390 395 400 Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp 405 410 415 Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly 420 425 430 Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu 435 440 445 Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu 450 455 460 Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His 465 470 475 480 Met Gln Ala Leu Pro Pro Arg 485 <210> 11 <211> 488 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 11 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Leu Ser Pro Gly Glu Arg Ala Ala Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95 Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser 100 105 110 Asn Trp Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Arg Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Asp Tyr Thr Met His 165 170 175 Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Asp Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Val Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Leu 225 230 235 240 Ser Gly Thr Tyr Arg Asp Tyr Phe Tyr Gly Val Asp Val Trp Gly Gln 245 250 255 Gly Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro 260 265 270 Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro 275 280 285 Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu 290 295 300 Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys 305 310 315 320 Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly 325 330 335 Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val 340 345 350 Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu 355 360 365 Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp 370 375 380 Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn 385 390 395 400 Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg 405 410 415 Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu 420 425 430 Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 435 440 445 Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 450 455 460 Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 465 470 475 480 His Met Gln Ala Leu Pro Pro Arg 485 <210> 12 <211> 486 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 12 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Val Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Thr Ser Gln Thr 35 40 45 Thr Thr Ser Tyr Leu Ala Trp Tyr Arg Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Asp Ala Ser Asn Arg Ala Ala Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn 85 90 95 Ser Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Leu Arg Thr 100 105 110 Asn Trp Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly Gly 115 120 125 Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Gln Val Gln Leu 130 135 140 Val Glu Ser Gly Gly Asp Ser Val Lys Pro Gly Gly Ser Leu Arg Leu 145 150 155 160 Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser Tyr Met Thr Trp 165 170 175 Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Phe Ile Ser 180 185 190 Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe 195 200 205 Thr Ile Ser Arg Asp Asn Val Lys Lys Ser Leu Tyr Leu Gln Met Asn 210 215 220 Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Glu 225 230 235 240 Pro Gly Asn Tyr Val Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr 245 250 255 Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr 260 265 270 Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala 275 280 285 Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe 290 295 300 Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val 305 310 315 320 Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys 325 330 335 Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr 340 345 350 Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu 355 360 365 Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro 370 375 380 Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly 385 390 395 400 Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro 405 410 415 Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu 420 425 430 Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile 435 440 445 Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr 450 455 460 Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met 465 470 475 480 Gln Ala Leu Pro Pro Arg 485 <210> 13 <211> 1482 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 13 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 120 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 180 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 300 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 360 gggacacggc tggagattaa aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc aactggtgga gtctggggga ggcttggtac agcctggcag gtccctgaga 480 ctctcctgtg cagcctctgg attcaccttt tatgattatg ccatgcactg ggtccggcaa 540 gctccaggga agggcctgga gtgggtctca ggtattagtt ggaatagtgg ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagctgag gacacggcct tgtattactg tgcaaaagat 720 aacagctatg gaaagttcta ctacggtttg gacgtctggg gccaagggac cacggtcacc 780 gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg 840 cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg 900 agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg 960 gtccttctcc tgtcactggt tatcaccctt tactgccaac gacgcaagta ccgctccaat 1020 aaaggagagt caccagtaga acccgccgaa ccttgtcact attcatgtcc acgcgaagag 1080 gagggttcaa cgatccctat tcaggaagat tacagaaagc cggaacctgc ttgtagcccc 1140 agagtgaagt tcagccgcag cgccgacgcc cctgcctacc agcagggcca gaaccagctg 1200 tataacgagc tgaacctggg caggcgggag gaatacgacg tgctggacaa gcgcagaggc 1260 cgggaccctg agatgggcgg caagccccag aggcggaaga acccccagga aggcctgtat 1320 aacgaactgc agaaagacaa gatggccgag gcctacagcg agatcggcat gaagggcgag 1380 cggcgacgcg gcaagggcca cgacggcctg taccagggcc tgtccaccgc caccaaggac 1440 acctacgacg ccctgcacat gcaggccctg cctccccgtt ag 1482 <210> 14 <211> 1464 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 14 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 120 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 180 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 300 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 360 gggacacggc tggagattaa aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc aactggtgga gtctggggga ggcttggtac agcctggcag gtccctgaga 480 ctctcctgtg cagcctctgg attcaccttt tatgattatg ccatgcactg ggtccggcaa 540 gctccaggga agggcctgga gtgggtctca ggtattagtt ggaatagtgg ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagctgag gacacggcct tgtattactg tgcaaaagat 720 aacagctatg gaaagttcta ctacggtttg gacgtctggg gccaagggac cacggtcacc 780 gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg 840 cagcccctgt ccctgcgcc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg 900 agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg 960 gtccttctcc tgtcactggt tatcaccctt tactgcaaac ggggcagaaa gaaactcctg 1020 tatatattca aacaaccatt tatgagacca gtacaaacta ctcaagagga agatggctgt 1080 agctgccgat ttccagaaga agaagaagga ggatgtgaac tgagagtgaa gttcagcagg 1140 agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200 ggacgaagag aggagtacga tgttttggac aagagacgtg gccgggaccc tgagatgggg 1260 ggaaagccgc agagaaggaa gaaccctcag gaaggcctgt acaatgaact gcagaaagat 1320 aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1380 cacgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac 1440 atgcaggccc tgccccctcg ctaa 1464 <210> 15 <211> 1479 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 15 atgagcgttc caacccaagt tctgggactg cttctgctct ggttgactga cgctaggtgc 60 gaaatagtaa tgacccaatc cccagccact ctctccgtta gcccaggtga aagagccact 120 cttagttgca gggctagtca atccgtatct agcaacctgg cctggtacca gcaaaagccc 180 ggacaagcgc cgcggttgtt gatctatggg acgagcacac gagctacggg tattccggcc 240 aggttctcag ggtctggctc cggaaccgaa tttacattga cgatcagtag tctgcaatca 300 gaggatttcg ccgtttacta ttgccaacag tacaataatt ggccgctcac attcggggga 360 ggaaccaagg tcgagattaa gggaggtggg ggtagtgggg gcggggggtc aggaggtgga 420 ggagaggtac agttggtaga aagcggcggg gggttggttc aacctggacg gagtctgaga 480 ttgtcttgcg tggcttccgg ctttactttc aatgattacg ccatgcactg ggtacgccag 540 gcgcctggaa agggtctgga gtgggtttcc gtgatatcct ggaatagtga tagtataggc 600 tatgccgata gtgtaaaagg aaggtttaca atctctaggg ataacgctaa gaacagcctg 660 taccttcaaa tgcatagtct ccgggctgag gacacagcct tgtactattg tgctaaggac 720 aatcattatg gaagcgggtc atattattac tatcaatatg ggatggatgt gtggggtcag 780 ggaacgaccg ttacggtatc ctcaaccacc acccctgcac caaggccccc gactcccgcg 840 cccaccatcg cgtcacagcc tcttagcctg cgaccggaag catgcagacc agctgccggg 900 ggggccgtgc atacgagagg tttggacttc gcctgcgata tctacatctg ggcgcccttg 960 gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caaacggggc 1020 agaagaaac tcctgtatat attcaaacaa ccatttatga gaccagtaca aactactcaa 1080 gaggaagatg gctgtagctg ccgatttcca gaagaagaag aaggaggatg tgaactgaga 1140 gtgaagttca gcaggagcgc agacgcccc gcgtaccagc agggccagaa ccagctctat 1200 aacgagctca atctaggacg aagagaggag tacgatgttt tggacaagag acgtggccgg 1260 gaccctgaga tgggggaaa gccgcagaga aggagaacc ctcaggaagg cctgtacaat 1320 1380 cggaggggca aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc 1440 tacgacgccc ttcacatgca ggccctgccc cctcgctaa 1479 <210> 16 <211> 1467 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 16 atgtccgtac ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccgcc 120 ctctcctgca gggccagtca gagtgttagc aactacttag cctggtacca acagaaacct 180 ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc 240 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct 300 gaagattttg cagtttatta ctgtcagcag cgtagcaact ggccgctcac tttcggcgga 360 gggaccaagg tggagatcag aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc agctggtgga gtctggggga ggcttggtac agcctggcag gtccctgcga 480 ctctcctgtg cagcctctgg attcaccttt cgagattata ccatgcactg ggtccggcaa 540 ggtccaggga agggcctgga atgggtctca ggtattagtt ggaatagtga ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagttgag gacacggcct tgtattactg tgcaaagctc 720 agtgggacct acagggacta cttctacgga gtggacgtct ggggccaagg gaccacggtc 780 accgtctcct caaccaccac ccctgcacca aggcccccga ctcccgcgcc caccatcgcg 840 tcacagcctc ttagcctgcg accggaagca tgcagaccag ctgccggggg ggccgtgcat 900 acgagaggtt tggacttcgc ctgcgatatc tacatctggg cgcccttggc cgggacttgt 960 ggggtccttc tcctgtcact ggtatcacc cttactgca aacggggcag aaagaaactc 1020 ctgtatatat tcaaacaacc attatgaga ccagtacaaa ctactcaaga ggaagatggc 1080 tgtagctgcc gatttccaga agaagaagaa ggaggatgtg aactgagagt gaagttcagc 1140 aggagcgcag acgcccccgc gtaccagcag ggccagaacc agctctataa cgagctcaat 1200 ctaggacgaa gagaggagta cgatgttttg gacaagagac gtggccggga ccctgagatg 1260 gggggaaagc cgcagagaag gaagaaccct caggaaggcc tgtacaatga actgcagaaa 1320 gataagatgg cggaggccta cagtgagatt gggatgaaag gcgagcgccg gaggggcaag 1380 gggcacgatg gcctttacca gggtctcagt acagccacca aggacaccta cgacgccctt 1440 cacatgcagg ccctgccccc tcgctaa 1467 <210> 17 <211> 1461 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 17 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaattgtgg tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 120 ctctcctgca ggaccagtca gactactacc agctacttag cctggtaccg acagaaacct 180 ggccaggctc ccaggctcct catctatgat gcatccaaca gggccgctgg catcccagcc 240 aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcaacag cctggagcct 300 gaagattttg cagtttatta ctgtcagctg cgtaccaact ggatcacctt cggccaaggg 360 acacgactgg agattaaagg tggaggtgga tctggaggag gaggatccgg tggaggaggt 420 caggtgcagc tggtggagtc tgggggagac tcggtcaagc ctggagggtc cctgagactc 480 tcctgtgcag cctctggatt caccttcagt gactcctaca tgacttggat ccgccaggct 540 ccagggaagg ggctggagatg ggtttcattc attagtagta gtggaagtac catatattat 600 gcagactctg tgaagggccg attcaccatt tccagggaca acgtcaagaa gtcattgtat 660 720 ccaggaaact acgtctatta cggtatggac gtctggggcc aagggaccac ggtcaccgtc 780 tcctcaacca ccacccctgc accaaggccc ccgactcccg cgcccaccat cgcgtcacag 840 cctcttagcc tgcgaccgga agcatgcaga ccagctgccg ggggggccgt gcatacgaga 900 ggtttggact tcgcctgcga tatctacatc tgggcgccct tggccgggac ttgtggggtc 960 cttctcctgt cactggttat caccctttac tgcaaacggg gcaagaagaa actcctgtat 1020 atattcaaac aaccatttat gagaccagta caaactactc aagagaaga tggctgtagc 1080 tgccgatttc cagaagaaga agaaggagga tgtgaactga gagtgaagtt cagcaggagc 1140 gcagacgccc ccgcgtacca gcagggccag aaccagctct ataacgagct caatctagga 1200 cgaagagagg agtacgatgt tttggacaag agacgtggcc gggaccctga gatgggggga 1260 aagccgcaga gaaggaagaa ccctcaggaa ggcctgtaca atgaactgca gaaagataag 1320 atggcggagg cctacagtga gattgggatg aaaggcgagc gccggagggg caaggggcac 1380 gatggccttt accagggtct cagtacagcc accaaggaca cctacgacgc ccttcacatg 1440 caggccctgc cccctcgcta a 1461 <210> 18 <211> 135 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 18 accaccaccc ctgcaccaag gcccccgact cccgcgccca ccatcgcgtc acagcctctt 60 agcctgcgac cggaagcatg cagaccagct gccggggggg ccgtgcatac gagaggtttg 120 gacttcgcct gcgat 135 <210> 19 <211> 135 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 19 accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg 60 tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg 120 gacttcgcct gtgat 135 <210> 20 <211> 445 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 20 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Arg Val Lys Phe 325 330 335 Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 340 345 350 Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 355 360 365 Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg 370 375 380 Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met 385 390 395 400 Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly 405 410 415 Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp 420 425 430 Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 435 440 445 <210> 21 <211> 11 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 21 Arg Ala Gly Asp Asn Trp Asn Trp Phe Asp Pro 1 5 10 <210> 22 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 22 Gln Gln Ala Lys Ser Val Pro Phe Thr 1 5 <210> 23 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 23 Glu Gly Gly Asn Tyr Gly Met Asp Val 1 5 <210> 24 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 24 Gln Gln Ala Asn Ser Phe Pro Pro Thr 1 5 <210> 25 <211> 17 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 25 Phe Ala Glu Tyr Cys Gly Gly Asn Ile Cys Tyr Tyr Tyr Gly Met Asp 1 5 10 15 Val <210> 26 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 26 Gln Gln Cys Gly Gly Ser Pro Trp Thr 1 5 <210> 27 <211> 138 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 27 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Ser Tyr Val Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Lys Arg Ala Gly Asp Asn Trp Asn Trp Phe Asp 115 120 125 Pro Trp Gly Gln Gly Thr Leu Val Thr Val 130 135 <210> 28 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 28 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Leu Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Arg Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Ala Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Lys Ser Val Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 105 <210> 29 <211> 136 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 29 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Asp Tyr Tyr Ile Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Ser Ile Lys Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 85 90 95 Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Glu Gly Gly Asn Tyr Gly Met Asp Val Trp 115 120 125 Gly Gln Gly Thr Thr Val Thr Val 130 135 <210> 30 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 30 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Asn Asn Trp 20 25 30 Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ala Ala Thr Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 31 <211> 143 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 31 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val 20 25 30 Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser 35 40 45 Ile Asn Tyr Tyr Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly 50 55 60 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asn Thr Asn Tyr Asn 65 70 75 80 Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Ala Thr Ser Arg Asn 85 90 95 Gln Phe Ser Leu Thr Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Phe Ala Glu Tyr Cys Gly Gly Asn Ile Cys Tyr 115 120 125 Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val 130 135 140 <210> 32 <211> 108 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 32 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Phe Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser 20 25 30 Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Leu 35 40 45 Met Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Cys Gly Gly Ser Pro 85 90 95 Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <210> 33 <211> 21 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 33 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro 20 <210> 34 <211> 114 <212> PRT <213> Homo sapiens <400> 34 Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile 1 5 10 15 Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His 20 25 30 Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln 35 40 45 Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu 50 55 60 Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val 65 70 75 80 Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile 85 90 95 Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn 100 105 110 Thr Ser <210> 35 <211> 485 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 35 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Ser Tyr Val Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Gly Ser Thr Tyr Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 85 90 95 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Lys Arg Ala Gly Asp Asn Trp Asn Trp Phe Asp 115 120 125 Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly 130 135 140 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Asp Ile Gln Met Thr Gln 145 150 155 160 Ser Pro Ser Ser Val Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Thr 165 170 175 Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala Trp Tyr Gln Arg 180 185 190 Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu 195 200 205 Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Ala Asp 210 215 220 Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 225 230 235 240 Tyr Cys Gln Gln Ala Lys Ser Val Pro Phe Thr Phe Gly Pro Gly Thr 245 250 255 Lys Val Asp Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro 260 265 270 Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys 275 280 285 Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala 290 295 300 Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu 305 310 315 320 Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys 325 330 335 Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr 340 345 350 Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly 355 360 365 Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 370 375 380 Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 385 390 395 400 Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 405 410 415 Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr 420 425 430 Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly 435 440 445 Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln 450 455 460 Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln 465 470 475 480 Ala Leu Pro Pro Arg 485 <210> 36 <211> 483 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 36 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Asp Tyr Tyr Ile Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Ser Ile Lys Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 85 90 95 Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Glu Gly Gly Asn Tyr Gly Met Asp Val Trp 115 120 125 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Asp Ile Gln Met Thr Gln Ser Pro 145 150 155 160 Ser Ser Val Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg 165 170 175 Ala Ser Gln Gly Ile Asn Asn Trp Leu Val Trp Tyr Gln Gln Lys Pro 180 185 190 Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Thr Ser Leu Gln Ser 195 200 205 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 210 215 220 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 225 230 235 240 Gln Gln Ala Asn Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu 245 250 255 Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro 260 265 270 Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro 275 280 285 Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp 290 295 300 Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 305 310 315 320 Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu 325 330 335 Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu 340 345 350 Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Gly Gly Cys 355 360 365 Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 370 375 380 Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 385 390 395 400 Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 405 410 415 Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu 420 425 430 Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys 435 440 445 Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu 450 455 460 Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu 465 470 475 480 Pro Pro Arg <210> 37 <211> 491 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 37 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val 20 25 30 Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser 35 40 45 Ile Asn Tyr Tyr Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly 50 55 60 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asn Thr Asn Tyr Asn 65 70 75 80 Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Ala Thr Ser Arg Asn 85 90 95 Gln Phe Ser Leu Thr Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val 100 105 110 Tyr Tyr Cys Ala Arg Phe Ala Glu Tyr Cys Gly Gly Asn Ile Cys Tyr 115 120 125 Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 130 135 140 Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu 145 150 155 160 Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly Glu 165 170 175 Arg Ala Thr Phe Ser Cys Arg Ala Ser Gln Ser Val Gly Ser Ser Phe 180 185 190 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Arg Leu Met 195 200 205 Tyr Gly Ala Ser Asn Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 210 215 220 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 225 230 235 240 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Cys Gly Gly Ser Pro Trp 245 250 255 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Thr Thr Thr Pro Ala 260 265 270 Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser 275 280 285 Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr 290 295 300 Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala 305 310 315 320 Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys 325 330 335 Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 340 345 350 Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 355 360 365 Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg 370 375 380 Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 385 390 395 400 Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 405 410 415 Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn 420 425 430 Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu 435 440 445 Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly 450 455 460 His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr 465 470 475 480 Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 485 490 <210> 38 <211> 645 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 38 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val 20 25 30 Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 35 40 45 Phe Ser Asp Tyr Tyr Ile Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly 50 55 60 Leu Glu Trp Val Ser Tyr Ile Ser Ser Ser Gly Ser Ser Ile Lys Tyr 65 70 75 80 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys 85 90 95 Asn Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 100 105 110 Val Tyr Tyr Cys Ala Arg Glu Gly Gly Asn Tyr Gly Met Asp Val Trp 115 120 125 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly 130 135 140 Gly Gly Gly Ser Gly Gly Gly Gly Asp Ile Gln Met Thr Gln Ser Pro 145 150 155 160 Ser Ser Val Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg 165 170 175 Ala Ser Gln Gly Ile Asn Asn Trp Leu Val Trp Tyr Gln Gln Lys Pro 180 185 190 Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Thr Ser Leu Gln Ser 195 200 205 Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr 210 215 220 Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys 225 230 235 240 Gln Gln Ala Asn Ser Phe Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu 245 250 255 Glu Ile Lys Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro 260 265 270 Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro 275 280 285 Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp 290 295 300 Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 305 310 315 320 Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg Lys Lys Leu Leu 325 330 335 Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu 340 345 350 Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys 355 360 365 Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 370 375 380 Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 385 390 395 400 Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 405 410 415 Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu 420 425 430 Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys 435 440 445 Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu 450 455 460 Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu 465 470 475 480 Pro Pro Arg Arg Ala Lys Arg Ser Gly Ser Gly Ala Thr Asn Phe Ser 485 490 495 Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala 500 505 510 Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala 515 520 525 Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu 530 535 540 Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser 545 550 555 560 Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu 565 570 575 Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp 580 585 590 Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn 595 600 605 Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu 610 615 620 Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met 625,630,635,640 Phe Ile Asn Thr Ser 645 <210> 39 <211> 1458 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide” <400> 39 atgagcgtgc ctacccaggt gctgggactg ctgctgctgt ggctgacaga cgcaaggtgc 60 gaggtgcagc tggtggagtc cggaggagga ctggtgcagc caggaggatc cctgaggctg 120 tcttgcgccg ccagcggctt cacctttagc tcctacgtga tgtcctgggt gcgccaggca 180 cctggcaagg gactggagtg ggtgtctgcc atcatcggct ctggcggcag cacatactat 240 gccgacagcg tgaagggccg gttcaccatc tccagagata actctaagaa tacactgtat 300 ctgcagatga acagcctgag ggcagaggac accgccgtgt actattgcgc caagagagcc 360 ggcgacaact ggaattggtt tgatccatgg ggccaggca ccctggtgac agtgtcttagc 420 ggaggaggag gatctggagg aggaggaagc ggcggaggag gcgacatcca gatgacacag 480 tcccatcct ctgtgagcgc ctccctgggc gataggtga ccatcacatg tcgcgcctct 540 caggcatca gctcctggct ggcatgtac cagaggaagc caggcaggc ccctaagctg 600 ctgatctatg cagcatctag cctgcagagc ggagtgcctt cccggttctc tggaagcgga 660 tccggagcag actttaccct gatactcc tctctgcagc cagaggattt cgccacctac 720 tattgtcagc aggccaagtc cgtgccattc accttggcc ccggcacaaa ggtggatatc 780 aagaccacca cccctgcacc aaggccccg actcccgcgc ccaccaccacgc gtcacagcct 840 cttagcctgc gaccggaagc atgcagacca gctgccgggg gggccgtgca tacgagaggt 900 ttggacttcg cctgcgatat ctacatctgg gcgcccttgg ccgggacttg tggggtcctt 960 1020. ctcctgtcac tggttatcac cctttactgc aaacggggca gaaagaact cctgtatata ttcaaacaac catttatgag accagtacaa actactcaag aggaagatgg ctgtagctgc cgatttccag aagaagaaga aggaggatgt gaactgagag tgaagttcag caggagcgca gacgccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaag 1260 ccgcagagaa ggaagaaccc tcaggaaggc ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa ggcgagcgcc ggaggggcaa ggggcacgat 1380. ggcctttacc agggtctcag tacagccacc aaggacacct acgacgccct tcacatgcag 1440 gccctgcccc ctcgctaa 1458 <210> 40 <211> 1452 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 40 atgagcgtgc ctacccaggt gctgggactg ctgctgctgt ggctgacaga cgcaaggtgc 60 caggtgcagc tggtggagag cggaggagga ctggtgaagc caggaggaag cctgaggctg 120 tcctgcgccg cctctggctt cacctttagc gactactata tctcctggat caggcaggca 180 cctggcaagg gactggagtg ggtgtcctac atcagctcct ctggcagctc catcaagtat 240 gccgactctg tgaagggccg gttcaccatc tccagagata acgccaagaa ttctctgtac 300 ctgcagatga acagcctgcg ggccgaggac acagccgtgt actattgcgc cagagagggc 360 ggcaattatg gcatggacgt gtggggccag ggcaccacag tgaccgtgtc tagcggcggc 420 ggcggctctg gaggaggagg aagcggcgga ggaggcgaca tccagatgac acagagccca 480 tccagcgtga gcgccagcgt gggcgatagg gtgaccatca catgtcgcgc ctcccagggc 540 atcaacaatt ggctggtgtg gtaccagcag aagccaggca aggcccccaa gctgctgatc 600 tatgcagcca cctccctgca gtctggagtg cctagccggt tctccggatc tggaagcgga 660 accgacttta ccctgacaat cagctccctg cagccagagg attttgccac atactattgt 720 cagcaggcca actctcttcc ccctaccttt ggccagggca caaagctgga gatcaagacc 780 accacccctg caccaaggcc cccgactccc gcgcccacca tcgcgtcaca gcctcttagc 840 ctgcgaccgg aagcatgcag accagctgcc ggtggggcgg tgcatacgag aggtttggac 900 ttcgcctgcg atatctacat ctgggcgccc ttggccggga cttgtggggt ccttctcctg 960 tcactggtta tcacccttta ctgcaaacgg ggcaagaa aactcctgta tatattcaaa 1020 caaccattta tgagaccagt acaaactact caagagaag atggctgtag ctgccgattt 1080 ccagaagaag aagaaggagg atgtgaactg agagtgaagt tcagcaggag cgcagacgcc 1140 cccgcgtacc agcagggcca gaaccagctc tataacgagc tcaatctagg acgaagagag 1200 gagtacgatg ttttggacaa gagacgtggc cgggaccctg agatgggggg aaagccgcag 1260 agaaggaaga accctcagga aggcctgtac aatgaactgc agaaagataa gatggcggag 1320 gcctacagtg agattgggat gaaaggcgag cgccggaggg gcaaggggca cgatggcctt 1380 taccagggtc tcagtacagc caccaaggac acctacgacg cccttcacat gcaggccctg 1440 ccccctcgct aa 1452 <210> 41 <211> 1476 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 41 atgagcgtgc ctacccaggt gctgggactg ctgctgctgt ggctgacaga cgcaaggtgc 60 caggtgcagc tgcaggagtc tggccctggc ctggtgaagc catccgagac cctgtctctg 120 acctgcacag tgagcggcgg ctccatcaat tactattact ggaactggat caggcagcca 180 cctggcaagg gactggagtg gatcggctac atcagctatt ccggcaacac caattacaac 240 ccttctctga agagcagggt gaccatcagc gtggccacat cccgcaatca gttcagcctg 300 acactgagct ccgtgaccgc agcagacaca gccgtgtatt actgcgcaag gtttgcagag 360 tactgcggag gcaacatctg ttattactat ggcatggacg tgtggggcca gggcaccaca 420 gtgaccgtgt ctagcggcgg cggcggctct ggaggaggag gaagcggagg aggaggagag 480 atcgtgctga cccagtcccc aggcacactg tctctgagcc ctggagagag ggccacattc 540 tcttgtcgcg cctcccagtc tgtgggctcc tcttttctgg cctggtacca gcagaagcca 600 ggacaggcac cacggagact gatgtatgga gcatccaata gggcaaccgg aatcccagac 660 agattcagcg gctccggctc tgcacagac ttcaccctga caatcagcag actggagcca 720 gaggacttcg ccgtgtacta ttgccagcag tgtggatt cccatggac ctttggccag 780 ggaacaagg tggagatcaa gaccaccacc cctgcaccaa ggccccgac tcccgcgccc 840 accatcgcgt cacagcctct tagcctgcga ccggaagcat gcagaccagc tgccgggggg 900 gccgtgcata cgagaggttt ggactcgcc tgcgatatct acatctgggc gcccttggcc 960 gggacttgtg gggtccttct cctgtcactg gttatcacc tttactgca acggggcaga 1020 aagaaactcc tgtatattat caacaacca tttatgagac cagtacaac tactcagag 1080 gaagatggct gtagctgccg atttccagaa gagagaag gaggatgtga actgagagtg 1140 aagttcagca ggagcgcaga cgcccccgcg taccagcagg gccagaacca gctctataac 1200 gagctcaatc taggacgaag agaggagtac gatgttttgg acagagacg tggccgggac 1260 cctgagatgg ggggaaagcc gcagagaagg aagaaccctc aggaaggcct gtacaatgaa 1320 ctgcagaaag ataagatggc ggaggcctac agtgagattg ggatgaaagg cgagcgccgg 1380 aggggcaagg ggcacgatgg cctttaccag ggtctcagta cagccaccaa ggacacctac 1440 gacgcccttc acatgcaggc cctgccccct cgctaa 1476 <210> 42 <211> 1938 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 42 atgagcgtgc ctacccaggt gctgggactg ctgctgctgt ggctgacaga cgcaaggtgc 60 caggtgcagc tggtggagag cggaggagga ctggtgaagc caggaggaag cctgaggctg 120 tcctgcgccg cctctggctt cacctttagc gactactata tctcctggat caggcaggca 180 cctggcaagg gactggagtg ggtgtcctac atcagctcct ctggcagctc catcaagtat 240 gccgactctg tgaagggccg gttcaccatc tccagagata acgccaagaa ttctctgtac 300 ctgcagatga acagcctgcg ggccgaggac acagccgtgt actattgcgc cagagagggc 360 ggcaattatg gcatggacgt gtggggccag ggcaccacag tgaccgtgtc tagcggcggc 420 ggcggctctg gaggaggagg aagcggcgga ggaggcgaca tccagatgac acagagccca 480 tccagcgtga gcgccagcgt gggcgatagg gtgaccatca catgtcgcgc ctcccagggc 540 atcaacaatt ggctggtgtg gtaccagcag aagccaggca aggcccccaa gctgctgatc 600 tatgcagcca cctccctgca gtctggagtg cctagccggt tctccggatc tggaagcgga 660 accgacttta ccctgacaat cagctccctg cagccagagg attttgccac atactattgt 720 cagcaggcca actccttccc ccctaccttt ggccagggca caaagctgga gatcaagacc 780 accacccctg caccaaggcc cccgactccc gcgcccacca tcgcgtcaca gcctcttagc 840 ctgcgaccgg aagcatgcag accagctgcc ggtggggcgg tgcatacgag aggtttggac 900 ttcgcctgcg atatctacat ctgggcgccc ttggccggga cttgtggggt cttctcctg 960 tcactggtta tcacccttta ctgcaaacgg ggcagaaaga aactcctgta tatattcaaa 1020 caaccattta tgagaccagt acaaactact caagaggaag atggctgtag ctgccgattt 1080 ccagaagaag aagaaggagg atgtgaactg agagtgaagt tcagcaggag cgcagacgcc 1140 cccgcgtacc agcagggcca gaaccagctc tataacgagc tcaatctagg acgaagagag 1200 gagtacgatg ttttggacaa gagacgtggc cgggaccctg agatgggggg aaagccgcag 1260 agaaggaaga accctcagga aggcctgtac aatgaactgc agaaagataa gatggcggag 1320 gcctacagtg agattgggat gaaaggcgag cgccggaggg gcaaggggca cgatggcctt 1380 taccagggtc tcagtacagc caccaaggac acctacgacg cccttcacat gcaggccctg 1440 ccccctcgcc gcgcgaagcg atcaggcagc ggggcgacaa atttcagcct tctgaaacaa 1500 gcaggcgacg tggaagaaaa ccccggtcca atggccttac cagtgaccgc cttgctcctg 1560 ccgctggcct tgctgctcca cgccgccagg ccgaactggg tgaatgtaat aagtgatttg 1620 aaaaaaattg aagatcttat tcaatctatg catattgatg ctactttata tacggaaagt 1680 gatgttcacc ccagttgcaa agtaacagca atgaagtgct ttctcttgga gttacaagtt 1740 atttcacttg agtccggaga tgcaagtatt catgatacag tagaaaatct gatcatccta 1800 gcaaacaaca gtttgtcttc obedient gtaacagaat ctggatgcaa agaatgtgag 1860 gaactggagg aaaaaaatat taaagaattt ttgcagagtt ttgtanot tgtccaaatg 1920 ttcatcaaca cttcttga 1938 <210> 43 <211> 23 <212> domestic worker <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 43 Ser Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp 1 5 10 15 Val Glu Glu Asn Pro Gly Pro 20 <210> 44 <211> 4 <212> PRT <213> Unknown <220> <221> source <223> / note="Description of Unknown: furin cleavage sequence" <400> 44 Arg Ala Lys Arg 1 <210> 45 <211> 27 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 45 Arg Ala Lys Arg Ser Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys 1 5 10 15 Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro 20 25 <210> 46 <211> 644 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 46 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg 325 330 335 Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln 340 345 350 Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu 355 360 365 Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala 370 375 380 Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu 385 390 395 400 Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp 405 410 415 Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly 420 425 430 Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu 435 440 445 Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu 450 455 460 Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His 465 470 475 480 Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Ala Thr Asn Phe Ser Leu 485 490 495 Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala Leu 500 505 510 Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu His Ala Ala 515 520 525 Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp 530 535 540 Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp 545 550 555 560 Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu 565 570 575 Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr 580 585 590 Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly 595 600 605 Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys 610 615 620 Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe 625 630 635 640 Ile Asn Thr Ser <210> 47 <211> 22 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 47 Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val 1 5 10 15 Glu Glu Asn Pro Gly Pro 20 <210> 48 <211> 671 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 48 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg 325 330 335 Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln 340 345 350 Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu 355 360 365 Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala 370 375 380 Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu 385 390 395 400 Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp 405 410 415 Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly 420 425 430 Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu 435 440 445 Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu 450 455 460 Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His 465 470 475 480 Met Gln Ala Leu Pro Pro Arg Gly Ser Gly Ala Thr Asn Phe Ser Leu 485 490 495 Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Met Arg Ile 500 505 510 Ser Lys Pro His Leu Arg Ser Ile Ser Ile Gln Cys Tyr Leu Cys Leu 515 520 525 Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly Ile His Val Phe Ile 530 535 540 Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys Thr Glu Ala Asn Trp Val 545 550 555 560 Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile Gln Ser Met 565 570 575 His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His Pro Ser Cys 580 585 590 Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln Val Ile Ser 595 600 605 Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu Asn Leu Ile 610 615 620 Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val Thr Glu Ser 625 630 635 640 Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe 645 650 655 Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn Thr Ser 660 665 670 <210> 49 <211> 48 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 49 Met Arg Ile Ser Lys Pro His Leu Arg Ser Ile Ser Ile Gln Cys Tyr 1 5 10 15 Leu Cys Leu Leu Leu Asn Ser His Phe Leu Thr Glu Ala Gly Ile His 20 25 30 Val Phe Ile Leu Gly Cys Phe Ser Ala Gly Leu Pro Lys Thr Glu Ala 35 40 45 <210> 50 <211> 1935 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 50 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 120 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 180 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 300 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 360 gggacacggc tggagattaa aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc aactggtgga gtctggggga ggcttggtac agcctggcag gtccctgaga 480 ctctcctgtg cagcctctgg attcaccttt tatgattatg ccatgcactg ggtccggcaa 540 gctccaggga agggcctgga gtgggtctca ggtattagtt ggaatagtgg ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagctgag gacacggcct tgtattactg tgcaaaagat 720 aacagctatg gaaagttcta ctacggtttg gacgtctggg gccaagggac cacggtcacc 780 gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg 840 cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg 900 aggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg 960 gtccttctcc tgtcactggt tatcaccctt tactgcaaac ggggcagaaa gaaactcctg 1020 tatatattca aaaaccatt tatgagacca gtacaacta ctcaagagga agatggctgt 1080 1140 agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200 ggacgaag aggagtacga tgttttggac aagacgtg gccgggaccc tgagatgggg 1260 ggaaagccgc agaaaggaa gaaccctcag gaagcctgt acaatgaact gcagaaagat 1320 aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1440. ccgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac atgcaggccc tgccccctcg cggtagcggg gctacgaact tctcccttct taaacaagcg ggagacgtgg aagaaaatcc cggacctatg gccttaccag tgaccgcctt gctcctgccg 1560 ctggccttgc tgctccacgc cgccaggccg aactgggtga atgtaataag tgatttgaaa aaaattgaag atcttattca atctatgcat attgatgcta ctttatatac ggaaagtgat gttcacccca gttgcaaagt aacagcaatg aagtgctttc tcttggagtt acaagttatt tcacttgagt ccggagatgc aagtattcat gatacagtag aaaatctgat catcctagca aacaacagtt tgtcttctaa tgggaatgta acagaatctg gatgcaaaga atgtgaggaa ctggaggaa aaaatattaa agaatttttg cagagttttg tacatattgt ccaaatgttc atcaacactt from <210> 51 <211> 2016 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 51 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 120 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 180 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 300 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 360 gggacacggc tggagattaa aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc aactggtgga gtctggggga ggcttggtac agcctggcag gtccctgaga 480 ctctcctgtg cagcctctgg attcaccttt tatgattatg ccatgcactg ggtccggcaa 540 gctccaggga agggcctgga gtgggtctca ggtattagtt ggaatagtgg ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagctgag gacacggcct tgtattactg tgcaaaagat 720 aacagctatg gaaagttcta ctacggtttg gacgtctggg gccaagggac cacggtcacc 780 gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg 840 cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg 900 agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg 960 gtccttctcc tgtcactggt tatcaccctt tactgcaaac ggggcagaaa gaaactcctg 1020 tatatattca aaaaccatt tatgagacca gtacaacta ctcaagagga agatggctgt 1080 1140 agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200 ggacgaag aggagtacga tgttttggac aagacgtg gccgggaccc tgagatgggg 1260 ggaaagccgc agaaaggaa gaaccctcag gaagcctgt acaatgaact gcagaaagat 1320 aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1380 cacgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac 1440 atgcaggccc tgccccctcg cggtagcggg gctacgaact tctcccttct taaaagcg 1500 ggagacgtgg aagaaaatcc cggacctatg agaatttcga aaccacattt gagaagtatt 1560 tccatccagt gctacttgtg tttacttcta aacagtcatt ttctaactga agctggcatt 1620 catgtcttca ttttggggctg tttcagtgca gggcttccta aaacagaagc caactgggtg 1680 aatgtaataa gtgatttgaa aaaaattgaa gatcttattc aatctatgca tattgatgct 1740 actttatata cggaaagtga tgttcacccc agttgcaaag taacagcaat gaagtgcttt 1800 ctcttggagt tacaagttat ttcacttgag tccggagatg caagtattca tgatacagta 1860 gaaaatctga tcatcctagc aaacaacagt ttgtcttcta atgggaatgt aacagaatct 1920 ggatgcaaag aatgtgagga actggaggaa aaaaatatta aagaatttt gcagagtttt 1980 gtacatattg tccaaatgtt catcaacact tcttga 2016 <210> 52 <211> 19 <212> PRT <213> Porcine teschovirus 1 <400> 52 Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn 1 5 10 15 Pro Gly Pro <210> 53 <211> 23 <212> PRT <213> Foot-and-mouth disease virus <400> 53 Val Lys Gln Thr Leu Asn Asn Phe Asp Leu Leu Lys Leu Ala Gly Asp 1 5 10 15 Val Glu Ser Asn Pro Gly Pro 20 <210> 54 <211> 20 <212> PRT <213> Equine rhinitis A virus <400> 54 Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly Asp Val Glu Ser 1 5 10 15 Asn Pro Gly Pro 20 <210> 55 <211> 17 <212> PRT <213> Thosea asigna virus <400> 55 Glu Gly Arg Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly 1 5 10 15 Pro <210> 56 <211> 553 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 56 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat 60 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct 120 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg 180 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc 240 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg 300 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg 360 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga 420 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt 480 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttccttttgaa 540 aaacacgatg ata 553 <210> 57 <211> 487 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide” <400> 57 Met Ser Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 1 5 10 15 Asp Ala Arg Cys Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser 20 25 30 Val Ser Pro Gly Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Asn Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro 50 55 60 Arg Leu Leu Ile Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala 65 70 75 80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser 85 90 95 Ser Leu Gln Ser Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn 100 105 110 Asn Trp Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Gly 115 120 125 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Glu Val Gln 130 135 140 Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg Ser Leu Arg 145 150 155 160 Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr Ala Met His 165 170 175 Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile 180 185 190 Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val Lys Gly Arg 195 200 205 Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met 210 215 220 Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys Ala Lys Asp 225 230 235 240 Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp Gly Gln Gly 245 250 255 Thr Thr Val Thr Val Ser Ser Thr Thr Thr Pro Ala Pro Arg Pro Pro 260 265 270 Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu 275 280 285 Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 290 295 300 Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly 305 310 315 320 Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Lys Arg Gly Arg 325 330 335 Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln 340 345 350 Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu 355 360 365 Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala 370 375 380 Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu 385 390 395 400 Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp 405 410 415 Pro Glu Met Gly Gly Lys Pro Gln Arg Arg Lys Asn Pro Gln Glu Gly 420 425 430 Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu 435 440 445 Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu 450 455 460 Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His 465 470 475 480 Met Gln Ala Leu Pro Pro Arg 485 <210> 58 <211> 135 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 58 Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 1 5 10 15 His Ala Ala Arg Pro Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys 20 25 30 Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr 35 40 45 Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe 50 55 60 Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile 65 70 75 80 His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser 85 90 95 Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu 100 105 110 Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His Ile Val 115 120 125 Gln Met Phe Ile Asn Thr Ser 130 135 <210> 59 <211> 2489 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 59 atgtccgtgc ctacccaggt gctgggcctg ctgctgctgt ggctgaccga cgccagatgc 60 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 120 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 180 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 240 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 300 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 360 gggacacggc tggagattaa aggtggaggt ggatctggag gaggaggatc cggtggagga 420 ggtgaagtgc aactggtgga gtctggggga ggcttggtac agcctggcag gtccctgaga 480 ctctcctgtg cagcctctgg attcaccttt tatgattatg ccatgcactg ggtccggcaa 540 gctccaggga agggcctgga gtgggtctca ggtattagtt ggaatagtgg ttacataggc 600 tatgcggact ctgtgaaggg ccgattcacc atctccagag acaacgccaa gaactccctg 660 tatctgcaaa tgaacagtct gagagctgag gacacggcct tgtattactg tgcaaaagat 720 aacagctatg gaaagttcta ctacggtttg gacgtctggg gccaagggac cacggtcacc 780 gtctcctcaa ccacgacgcc agcgccgcga ccaccaacac cggcgcccac catcgcgtcg 840 cagcccctgt ccctgcgcc agaggcgtgc cggccagcgg cggggggcgc agtgcacacg 900 agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg gacttgtggg 960 gtccttctcc tgtcactggt tatcaccctt tactgcaaac ggggcagaaa gaaactcctg 1020 tatatattca aacaaccatt tatgagacca gtacaaacta ctcaagagga agatggctgt 1080 agctgccgat ttccagaaga agaagaagga ggatgtgaac tgagagtgaa gttcagcagg 1140 agcgcagacg cccccgcgta ccagcagggc cagaaccagc tctataacga gctcaatcta 1200 ggacgaag aggagtacga tgttttggac aagacgtg gccgggaccc tgagatgggg 1260 ggaaagccgc agaaaggaa gaaccctcag gaagcctgt acaatgaact gcagaaagat 1320 aagatggcgg aggcctacag tgagattggg atgaaaggcg agcgccggag gggcaagggg 1380 cacgatggcc tttaccaggg tctcagtaca gccaccaagg acacctacga cgcccttcac 1440 atgcaggccc tgccccctcg ctagagtact gcggccgcta cgtaaattcc gcccctctcc 1500 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 1560 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 1620 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 1680 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 1740 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 1800 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 1860 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 1920 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 1980 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt 2040 tttcctttga aaaacacgat gatattaatt aagccaccgc catggcctta ccagtgaccg 2100 ccttgctcct gccgctggcc ttgctgctcc acgccgccag gccgaactgg gtgaatgtaa 2160 taagtgattt gaaaaaatt gaagaatctta ttcaatctat gcatattgat gctactttat 2220 atacggaaag tgatgttcac cccagttgca aagtaacagc aatgaagtgc tttctcttgg 2280 agttacaagt tatttcactt gagtccggag atgcaagtat tcatgataca gtagaaaatc 2340 tgatcatcct agcaacaac agttgtctt ctaatgggaa tgtaacagaa tctggatgca 2400 aagaatgtga ggaacggag gaaaaaaata ttaagaatt tttgcagagt ttgtacata 2460 ttgtccaaat gttcatcaac acttcttga 2489 <210> 60 <211> 461 <212> DNA <213> Foot-and-mouth disease virus A <400> 60 agcaggtttc cccaacctgac acaaacgtg caactgaa ctccgcctgg tctttccagg 60 tacagggg tacactttg tactgcgttt ggctccacgc talltacactt ggcgagtgtt 120 agtaacagca ctgttgctc gtagcggagc atgacggccg tgggaactcc tccttggtaa 180 caaggacccca cggggccaaa agccacgccc acacggggcc gtcatgtgtg caaccccagc 240 acggcgactt tactgcgaaa cccactttaa agtgacattg aaactggtac ccacacactg 300 gtgacaggct aaggatgccc ttcaggtacc ccgaggtaac acgcgacact cgggatctga 360 gaaggggact ggggcttcta taaaagcgct cggtttaaaa agcttctatg cctgaatagg 420 tgaccggagg tcggcacctt tccttgcaa ttactgacca c 461 <210> 61 <400> 61 000 <210> 62 <400> 62 000 <210> 63 <400> 63 000 <210> 64 <400> 64 000 <210> 65 <400> 65 000 <210> 66 <400> 66 000 <210> 67 <400> 67 000 <210> 68 <400> 68 000 <210> 69 <400> 69 000 <210> 70 <400> 70 000 <210> 71 <400> 71 000 <210> 72 <400> 72 000 <210> 73 <400> 73 000 <210> 74 <400> 74 000 <210> 75 <400> 75 000 <210> 76 <400> 76 000 <210> 77 <400> 77 000 <210> 78 <400> 78 000 <210> 79 <400> 79 000 <210> 80 <400> 80 000 <210> 81 <400> 81 000 <210> 82 <400> 82 000 <210> 83 <400> 83 000 <210> 84 <400> 84 000 <210> 85 <400> 85 000 <210> 86 <400> 86 000 <210> 87 <400> 87 000 <210> 88 <400> 88 000 <210> 89 <400> 89 000 <210> 90 <400> 90 000 <210> 91 <400> 91 000 <210> 92 <400> 92 000 <210> 93 <400> 93 000 <210> 94 <400> 94 000 <210> 95 <400> 95 000 <210> 96 <400> 96 000 <210> 97 <400> 97 000 <210> 98 <400> 98 000 <210> 99 <211> 122 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 99 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Gly Tyr Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asp Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 100 <400> 100 000 <210> 101 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 101 Gly Phe Thr Phe Tyr Asp Tyr Ala 1 5 <210> 102 <400> 102 000 <210> 103 <211> 8 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 103 Ile Ser Trp Asn Ser Gly Tyr Ile 1 5 <210> 104 <400> 104 000 <210> 105 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 105 Ala Lys Asp Asn Ser Tyr Gly Lys Phe Tyr Tyr Gly Leu Asp Val 1 5 10 15 <210> 106 <400> 106 000 <210> 107 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 107 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Thr Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30 Leu Ala Trp Tyr Leu Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Ile Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Ile 85 90 95 Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 108 <400> 108 000 <210> 109 <211> 6 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 109 Gln Ser Val Ser Ser Asn 1 5 <210> 110 <400> 110 000 <210> 111 <211> 3 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 111 Gly Ala Ser 1 <210> 112 <400> 112 000 <210> 113 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 113 Gln Gln Tyr Asn Asn Trp Pro Ile Thr 1 5 <210> 114 <400> 114 000 <210> 115 <400> 115 000 <210> 116 <400> 116 000 <210> 117 <400> 117 000 <210> 118 <400> 118 000 <210> 119 <400> 119 000 <210> 120 <400> 120 000 <210> 121 <400> 121 000 <210> 122 <400> 122 000 <210> 123 <400> 123 000 <210> 124 <400> 124 000 <210> 125 <400> 125 000 <210> 126 <400> 126 000 <210> 127 <400> 127 000 <210> 128 <400> 128 000 <210> 129 <400> 129 000 <210> 130 <400> 130 000 <210> 131 <400> 131 000 <210> 132 <400> 132 000 <210> 133 <400> 133 000 <210> 134 <400> 134 000 <210> 135 <400> 135 000 <210> 136 <400> 136 000 <210> 137 <400> 137 000 <210> 138 <400> 138 000 <210> 139 <400> 139 000 <210> 140 <400> 140 000 <210> 141 <400> 141 000 <210> 142 <400> 142 000 <210> 143 <400> 143 000 <210> 144 <400> 144 000 <210> 145 <400> 145 000 <210> 146 <400> 146 000 <210> 147 <400> 147 000 <210> 148 <400> 148 000 <210> 149 <400> 149 000 <210> 150 <400> 150 000 <210> 151 <400> 151 000 <210> 152 <400> 152 000 <210> 153 <400> 153 000 <210> 154 <400> 154 000 <210> 155 <400> 155 000 <210> 156 <400> 156 000 <210> 157 <400> 157 000 <210> 158 <400> 158 000 <210> 159 <400> 159 000 <210> 160 <400> 160 000 <210> 161 <400> 161 000 <210> 162 <400> 162 000 <210> 163 <400> 163 000 <210> 164 <400> 164 000 <210> 165 <400> 165 000 <210> 166 <400> 166 000 <210> 167 <400> 167 000 <210> 168 <400> 168 000 <210> 169 <400> 169 000 <210> 170 <400> 170 000 <210> 171 <400> 171 000 <210> 172 <400> 172 000 <210> 173 <400> 173 000 <210> 174 <400> 174 000 <210> 175 <400> 175 000 <210> 176 <400> 176 000 <210> 177 <400> 177 000 <210> 178 <400> 178 000 <210> 179 <400> 179 000 <210> 180 <400> 180 000 <210> 181 <400> 181 000 <210> 182 <400> 182 000 <210> 183 <400> 183 000 <210> 184 <400> 184 000 <210> 185 <400> 185 000 <210> 186 <400> 186 000 <210> 187 <400> 187 000 <210> 188 <400> 188 000 <210> 189 <400> 189 000 <210> 190 <400> 190 000 <210> 191 <400> 191 000 <210> 192 <400> 192 000 <210> 193 <400> 193 000 <210> 194 <400> 194 000 <210> 195 <211> 123 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 195 Glu Val Gln Leu Ala Glu Ser Gly Gly Asp Leu Val Gln Ser Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Thr Phe His Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Tyr Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Asp Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Val Lys Asp Phe His Tyr Gly Ser Gly Ser Asn Tyr Gly Met Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Pro 115 120 <210> 196 <400> 196 000 <210> 197 <400> 197 000 <210> 198 <400> 198 000 <210> 199 <400> 199 000 <210> 200 <400> 200 000 <210> 201 <211> 16 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 201 Val Lys Asp Phe His Tyr Gly Ser Gly Ser Asn Tyr Gly Met Asp Val 1 5 10 15 <210> 202 <400> 202 000 <210> 203 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 203 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Met Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Val Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Ser Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Asn Asp Trp Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 204 <400> 204 000 <210> 205 <400> 205 000 <210> 206 <400> 206 000 <210> 207 <400> 207 000 <210> 208 <400> 208 000 <210> 209 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 209 Gln Gln Ser Asn Asp Trp Pro Leu Thr 1 5 <210> 210 <400> 210 000 <210> 211 <400> 211 000 <210> 212 <400> 212 000 <210> 213 <400> 213 000 <210> 214 <400> 214 000 <210> 215 <400> 215 000 <210> 216 <400> 216 000 <210> 217 <400> 217 000 <210> 218 <400> 218 000 <210> 219 <400> 219 000 <210> 220 <400> 220 000 <210> 221 <400> 221 000 <210> 222 <400> 222 000 <210> 223 <400> 223 000 <210> 224 <400> 224 000 <210> 225 <400> 225 000 <210> 226 <400> 226 000 <210> 227 <400> 227 000 <210> 228 <400> 228 000 <210> 229 <400> 229 000 <210> 230 <400> 230 000 <210> 231 <400> 231 000 <210> 232 <400> 232 000 <210> 233 <400> 233 000 <210> 234 <400> 234 000 <210> 235 <400> 235 000 <210> 236 <400> 236 000 <210> 237 <400> 237 000 <210> 238 <400> 238 000 <210> 239 <400> 239 000 <210> 240 <400> 240 000 <210> 241 <400> 241 000 <210> 242 <400> 242 000 <210> 243 <211> 122 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 243 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Tyr Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Thr Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Thr Lys Asp Gly Ser Tyr Gly His Phe Tyr Ser Gly Leu Asp Val Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 244 <400> 244 000 <210> 245 <400> 245 000 <210> 246 <400> 246 000 <210> 247 <400> 247 000 <210> 248 <400> 248 000 <210> 249 <211> 15 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 249 Thr Lys Asp Gly Ser Tyr Gly His Phe Tyr Ser Gly Leu Asp Val 1 5 10 15 <210> 250 <400> 250 000 <210> 251 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 251 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Val Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Asp Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Tyr Tyr Trp Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 252 <400> 252 000 <210> 253 <400> 253 000 <210> 254 <400> 254 000 <210> 255 <400> 255 000 <210> 256 <400> 256 000 <210> 257 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 257 Gln Gln Arg Tyr Tyr Trp Pro Leu Thr 1 5 <210> 258 <400> 258 000 <210> 259 <400> 259 000 <210> 260 <400> 260 000 <210> 261 <400> 261 000 <210> 262 <400> 262 000 <210> 263 <400> 263 000 <210> 264 <400> 264 000 <210> 265 <400> 265 000 <210> 266 <400> 266 000 <210> 267 <400> 267 000 <210> 268 <400> 268 000 <210> 269 <400> 269 000 <210> 270 <400> 270 000 <210> 271 <400> 271 000 <210> 272 <400> 272 000 <210> 273 <400> 273 000 <210> 274 <400> 274 000 <210> 275 <400> 275 000 <210> 276 <400> 276 000 <210> 277 <400> 277 000 <210> 278 <400> 278 000 <210> 279 <400> 279 000 <210> 280 <400> 280 000 <210> 281 <400> 281 000 <210> 282 <400> 282 000 <210> 283 <400> 283 000 <210> 284 <400> 284 000 <210> 285 <400> 285 000 <210> 286 <400> 286 000 <210> 287 <400> 287 000 <210> 288 <400> 288 000 <210> 289 <400> 289 000 <210> 290 <400> 290 000 <210> 291 <400> 291 000 <210> 292 <400> 292 000 <210> 293 <400> 293 000 <210> 294 <400> 294 000 <210> 295 <400> 295 000 <210> 296 <400> 296 000 <210> 297 <400> 297 000 <210> 298 <400> 298 000 <210> 299 <400> 299 000 <210> 300 <400> 300 000 <210> 301 <400> 301 000 <210> 302 <400> 302 000 <210> 303 <400> 303 000 <210> 304 <400> 304 000 <210> 305 <400> 305 000 <210> 306 <400> 306 000 <210> 307 <400> 307 000 <210> 308 <400> 308 000 <210> 309 <400> 309 000 <210> 310 <400> 310 000 <210> 311 <400> 311 000 <210> 312 <400> 312 000 <210> 313 <400> 313 000 <210> 314 <400> 314 000 <210> 315 <400> 315 000 <210> 316 <400> 316 000 <210> 317 <400> 317 000 <210> 318 <400> 318 000 <210> 319 <400> 319 000 <210> 320 <400> 320 000 <210> 321 <400> 321 000 <210> 322 <400> 322 000 <210> 323 <400> 323 000 <210> 324 <400> 324 000 <210> 325 <400> 325 000 <210> 326 <400> 326 000 <210> 327 <400> 327 000 <210> 328 <400> 328 000 <210> 329 <400> 329 000 <210> 330 <400> 330 000 <210> 331 <400> 331 000 <210> 332 <400> 332 000 <210> 333 <400> 333 000 <210> 334 <400> 334 000 <210> 335 <400> 335 000 <210> 336 <400> 336 000 <210> 337 <400> 337 000 <210> 338 <400> 338 000 <210> 339 <211> 126 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 339 Glu Glu Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asp Tyr 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Gly Ser Leu Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Lys Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asp Pro Ser Tyr Gly Ser Gly Ser Tyr His Ser Tyr Tyr Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 340 <400> 340 000 <210> 341 <400> 341 000 <210> 342 <400> 342 000 <210> 343 <400> 343 000 <210> 344 <400> 344 000 <210> 345 <211> 19 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 345 Ala Lys Asp Pro Ser Tyr Gly Ser Gly Ser Tyr His Ser Tyr Tyr Gly 1 5 10 15 Met Asp Val <210> 346 <400> 346 000 <210> 347 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 347 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Trp Ala Ser Gln Ser Ile Ser Arg Tyr 20 25 30 Leu Val Trp Tyr Gln Gln Lys Cys Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Glu Ala Ser Lys Arg Ala Thr Gly Ile Pro Val Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Phe Asn Trp Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 348 <400> 348 000 <210> 349 <400> 349 000 <210> 350 <400> 350 000 <210> 351 <400> 351 000 <210> 352 <400> 352 000 <210> 353 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <400> 353 Gln Gln Arg Phe Asn Trp Pro Leu Thr 1 5 <210> 354 <211> 37 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 354 Trp Leu Thr Lys Lys Lys Tyr Ser Ser Ser Val His Asp Pro Asn Gly 1 5 10 15 Glu Tyr Met Phe Met Arg Ala Val Asn Thr Ala Lys Lys Ser Arg Leu 20 25 30 Thr Asp Val Thr Leu 35 <210> 355 <211> 382 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 355 gaagtacagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60 tcctgtgtag cctctggatt cacctttaat gattatgcca tgcactgggt ccggcaagct 120 ccagggaagg gcctggaatg ggtctcagtt attagttgga atagtgatag cataggctat 180 gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240 ctgcaaatgc acagtctgag agctgaggac acggccttgt attactgtgc aaaagataat 300 cactatggtt cggggagtta ttactactac caatacggta tggacgtctg gggccaaggg 360 accacggtca ccgtctcctc ag 382 <210> 356 <211> 127 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 356 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asn Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Trp Asn Ser Asp Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met His Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asp Asn His Tyr Gly Ser Gly Ser Tyr Tyr Tyr Tyr Gln Tyr 100 105 110 Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 <210> 357 <211> 322 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 357 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacca gcagaaacct 120 ggccaggctc cccgactcct catctatggt acatccacca gggccactgg tatcccagcc 180 aggttcagtg gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240 gaagattttg cagtttatta ctgtcaacaa tataataact ggccgctcac tttcggcgga 300 gggaccaagg tggagatcaa ac 322 <210> 358 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 358 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Thr Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 359 <211> 367 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 359 caggtgcagc tggtggagtc tgggggagac tcggtcaagc ctggagggtc cctgagactc 60 tcctgtgcag cctctggatt caccttcagt gactcctaca tgacttggat ccgccaggct 120 ccagggaagg ggctggagtg ggtttcattc attagtagta gtggaagtac catatattat 180 gcagactctg tgaagggccg attcaccatt tccagggaca acgtcaagaa gtcattgtat 240 ctgcagatga acagactgag agccgaggac acggccgtgt attactgtgc gagagaagaa 300 ccaggaaact acgtctatta cggtatggac gtctggggcc aagggaccac ggtcaccgtc 360 tcctcag 367 <210> 360 <211> 122 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 360 Gln Val Gln Leu Val Glu Ser Gly Gly Asp Ser Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Ser 20 25 30 Tyr Met Thr Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Phe Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Val Lys Lys Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Arg Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Glu Pro Gly Asn Tyr Val Tyr Tyr Gly Met Asp Val Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 361 <211> 319 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 361 gaaattgtgg tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc 60 ctctcctgca ggaccagtca gactactacc agctacttag cctggtaccg acagaaacct 120 ggccaggctc ccaggctcct catctgat gcatccaaca gggccgctgg catcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactca ccatcacag cctagagcct 240 gaagattttg cagtttatta ctgtcagctg cgtaccaact ggatcacctt cggccaaggg 300 Acacgactgg National 319 <210> 362 <211> 106 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide” <400> 362 Glu Ile Val Val Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Served as Cys Arg Thr Served as Gln Thr Thr 20 25 30 Leu Wing Trp Tyr Arg Gln Lys Pro Gly Gln Wing Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Ala Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Leu Arg Thr Asn Trp Ile Thr 85 90 95 Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 <210> 363 <211> 367 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 363 gaagtgcaac tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60 tcctgtgcag cctctggatt caccttttat gattatgcca tgcactgggt ccggcaagct 120 ccagggaagg gcctggagtg ggtctcaggt attagttgga atagtggtta cataggctat 180 gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240 ctgcaaatga acagtctgag agctgaggac acggccttgt attactgtgc aaaagataac 300 agctatggaa agttctacta cggtttggac gtctggggcc aagggaccac ggtcaccgtc 360 tcctcag 367 <210> 364 <211> 322 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 364 gaaatagtga tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagaaccacc 60 ctctcctgca gggccagtca gagtgttagc agcaacttag cctggtacct tcagaaacct 120 ggccaggctc ccaggctcct catctatggt gcatccacca gggccactgg tatcccagcc 180 aggttcagtg gcagtgggtc tgggacagag ttcattctca ccatcagcag cctgcagtct 240 gaagattttg cagtttatta ctgtcagcag tataataact ggccgatcac cttcggccaa 300 gggacacggc tggagattaa ac 322 <210> 365 <211> 370 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 365 gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgcgactc 60 tcctgtgcag cctctggatt cacctttcga gattatacca tgcactgggt ccggcaaggt 120 ccagggaagg gcctggaatg ggtctcaggt attagttgga atagtgatta cataggctat 180 gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat 240 ctgcaaatga acagtctgag agttgaggac acggccttgt attactgtgc aaagctcagt 300 gggacctaca gggactactt ctacggagtg gacgtctggg gccaagggac cacggtcacc 360 gtctcctcag 370 <210> 366 <211> 123 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide" <400> 366 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Asp Tyr 20 25 30 Thr Met His Trp Val Arg Gln Gly Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Asp Tyr Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Leu Ser Gly Thr Tyr Arg Asp Tyr Phe Tyr Gly Val Asp Val 100 105 110 Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 367 <211> 322 <212> DNA <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polynucleotide" <400> 367 gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccgcc 60 ctctcctgca gggccagtca gagtgttagc aactacttag cctggtacca acagaaacct 120 ggccaggctc ccaggctcct catctgat gcatccaaca gggccactgg catcccagcc 180 aggttcagtg gcagtgggtc tgggacagac ttcactca ccatcagcag cctagagcct 240 gaagattttg cagtttatta ctgtcagcag cgtagcact ggccgctcac ttcggcgga 300 scream scream ac 322 <210> 368 <211> 107 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic polypeptide” <400> 368 Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Leu Ser Gly 1 5 10 15 Glu Arg Only Leu Only Cys Arg Only Only On Gln Only Only on Asn Tyr 20 25 30 Leu Wing Trp Tyr Gln Gln Lys Pro Gly Gln Wing Pro Arg Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 65 70 75 80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Arg 100 105 <210> 369 <211> 19 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <220> <221> VARIANT <222> (1)..(1) <223> / replace="Val" or "Thr" <220> <221> VARIANT <222> (4)..(4) <223> / replace="Phe" or "Gly" <220> <221> VARIANT <222> (5)..(5) <223> / replace="His" <220> <221> VARIANT <222> (8)..(8) <223> / replace="His" <220> <221> VARIANT <222> (9)..(9) <223> / replace="Phe" <220> <221> VARIANT <222> (10)..(10) <223> / replace="Tyr" <220> <221> VARIANT <222> (11)..(11) <223> / replace="Asn" or "Ser" <220> <221> VARIANT <222> (12)..(12) <223> / replace="Gly" or "His" <220> <221> VARIANT <222> (13)..(13) <223> / replace="Leu" or "Ser" <220> <221> VARIANT <222> (14)..(14) <223> / replace="Met" or "Asp" <220> <221> VARIANT <222> (15)..(15) <223> / replace="Asp" or "Val" <220> <221> VARIANT <222> (16)..(16) <223> / replace="Val" or " " <220> <221> VARIANT <222> (17)..(17) <223> / replace=" " <220> <221> VARIANT <222> (18)..(18) <223> / replace=" " <220> <221> VARIANT <222> (19)..(19) <223> / replace=" " <220> <221> SITE <222> (1)..(19) <223> / note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 369 Ala Lys Asp Pro Ser Tyr Gly Ser Gly Ser Tyr Tyr Gly Tyr Tyr Gly 1 5 10 15 Met Asp Val <210> 370 <211> 9 <212> PRT <213> Artificial Sequence <220> <221> source <223> / note="Description of Artificial Sequence: Synthetic peptide" <220> <221> VARIANT <222> (3)..(3) <223> / replace="Ser" <220> <221> VARIANT <222> (4)..(4) <223> / replace="Tyr" or "Phe" <220> <221> VARIANT <222> (5)..(5) <223> / replace="Asp" or "Tyr" <220> <221> SITE <222> (1)..(9) <223> / note="Variant residues given in the sequence have no preference with respect to those in the annotations for variant positions" <400> 370 Gln Gln Arg Asn Asn Trp Pro Leu Thr 1 5

Claims

1. A δ1γδ T cell, a) A nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR is in the order 5' to 3': (i) A binding domain that specifically binds to CD20 expressed on the surface of blood cells; (ii) CD8α hinge domain containing Sequence ID No. 2 (TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDY); (iii) CD8α transmembrane domain containing Sequence ID No. 3 (IWAPLAGTCCGVLLLLSLVITLYC); (iv) The 4-1BB costimulatory signaling region containing SEQ ID NO: 6 (KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL); and (v) A nucleic acid encoding a CAR, comprising a CD3ζ signaling domain containing Sequence ID No. 4 (RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRRGKGHDGLYQGLSTAKDTYDALHMQALPPR), Includes, or b) A polypeptide comprising a CAR containing an amino acid sequence encoded by the nucleic acid described in a) above, δ1γδT cells, wherein the δ1γδT cells functionally express the nucleic acid-encoded CAR on the binding domain of the polypeptide or on the surface of the δ1γδT cells.

2. The binding domain is the following complementarity determination region (CDR): CDRH1 containing the sequence of sequence number 101; CDRH2 containing the sequence of sequence number 103; CDRH3 containing the sequence of sequence number 105; CDRL1 containing the sequence of sequence number 109; CDRL2 containing the sequence of sequence number 111; and CDRL3 containing the sequence of sequence number 113 The δ1γδT cell according to claim 1, comprising:

3. The δ1γδT cell according to claim 1, wherein the binding domain comprises a heavy chain variable region (HCVR) sequence containing the sequence of SEQ ID NO: 99 and a light chain variable region (LCVR) sequence containing the sequence of SEQ ID NO:

107.

4. The δ1γδT cell according to claim 1, wherein the nucleic acid encodes a polypeptide containing the sequence of SEQ ID NO:

10.

5. The δ1γδT cell according to claim 1, wherein the nucleic acid comprises the sequence of sequence number 14.

6. The δ1γδT cell according to any one of claims 1 to 5, wherein the nucleic acid further encodes secreted IL-15.

7. The secreted IL-15 is operably linked to a secretion signal sequence containing SEQ ID NO: 33 (MALPVTALLLPLAALLHAARP) and SEQ ID NO: 34 (NWVNVISDLKKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIIILANNSLSLSSNGNVTESGCKECEELEEEKNIKEFLQSFVHIVQMFI The δ1γδT cell according to claim 6, comprising the sequence of NTS.

8. The δ1γδT cell according to claim 6 or 7, wherein the nucleic acid further encodes a P2A cleavage sequence including SEQ ID NO: 47 (GSGATNFSLLKQAGDVEENPGP) at the amino terminus of the secreted IL-15.

9. The δ1γδT cell according to claim 1, wherein the nucleic acid encodes a polypeptide containing the sequence of SEQ ID NO:

46.

10. The δ1γδT cell according to claim 1, wherein the nucleic acid comprises the sequence of SEQ ID NO:

50.

11. A δ1γδ T cell, a) A nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR is in the order 5' to 3': (i) A binding domain that specifically binds to CD20 expressed on the surface of blood cells, the following CDR: CDRH1 containing the sequence of sequence number 101; CDRH2 containing the sequence of sequence number 103; CDRH3 containing the sequence of sequence number 105; CDRL1 containing the sequence of sequence number 109; CDRL2 containing the sequence of sequence number 111; and CDRL3 containing the sequence of sequence number 113 A binding domain including; (ii) CD8α hinge domain containing the sequence of Sequence ID No. 2; (iii) CD8α transmembrane domain containing the sequence of Sequence ID No. 3; (iv) The 4-1BB costimulatory signaling region containing the sequence of Sequence ID No. 6; and (v) A nucleic acid encoding a CAR, containing a CD3ζ signaling domain containing the sequence of Sequence ID No. 4 Includes, or b) A polypeptide comprising a CAR containing an amino acid sequence encoded by the nucleic acid described in a) above, δ1γδT cells, wherein the δ1γδT cells functionally express the nucleic acid-encoded CAR on the binding domain of the polypeptide or on the surface of the δ1γδT cells.

12. The δ1γδT cells according to any one of claims 1 to 11, wherein the δ1γδT cells proliferate in response to contact with cells exhibiting cell surface expression or overexpression of CD20, and the cells exhibiting cell surface expression of CD20 are normal blood cells.

13. The δ1γδT cells according to any one of claims 1 to 11, wherein the δ1γδT cells proliferate in response to contact with cells exhibiting cell surface expression or overexpression of CD20, and the cells exhibiting cell surface expression or overexpression of CD20 are hematological tumor cells.

14. A cell population comprising a plurality of δ1γδT cells according to any one of claims 1 to 13.

15. The cell population according to claim 14, comprising at least 10⁸ δ¹γδT cells.

16. The cell population according to claim 15, comprising 108 δ1γδT cells to 1011 δ1γδT cells.

17. The cell population according to any one of claims 14 to 16, comprising at least 60% δ1γδT cells.

18. The cell population according to claim 17, comprising 60% to 95% δ1γδT cells.

19. A pharmaceutical composition comprising a therapeutically effective amount of δ1γδT cells according to any one of claims 1 to 13, or a cell population according to any one of claims 14 to 18, and a pharmaceutically acceptable excipient.

20. A δ1γδT cell according to any one of claims 1 to 13, a cell population according to any one of claims 14 to 18, or a pharmaceutical composition according to claim 19, for use in the treatment of a target hematological cancer.

21. A δ1γδ T cell, a population of δ1γδ T cells, or a pharmaceutical composition for use according to claim 20, wherein the blood cancer is lymphoma.

22. A δ1γδT cell according to any one of claims 1 to 13, a cell population according to any one of claims 14 to 18, or a pharmaceutical composition according to claim 19, for use in killing B cells.

23. A δ1γδT cell, a population of δ1γδT cells, or a pharmaceutical composition for use according to claim 22, wherein the B cells are hematological tumor cells.

24. A δ1γδT cell, a δ1γδT cell population, or a pharmaceutical composition for use according to any one of claims 20 to 23, wherein the treatment comprises multiple administrations of δ1γδT cells, a δ1γδT cell population, or a pharmaceutical composition, with an interval of at least one week between such multiple administrations.

25. A δ1γδT cell, a population of δ1γδT cells, or a pharmaceutical composition for use according to claim 24, wherein multiple administrations of δ1γδT cells are performed at intervals of six months or less.

26. δ1γδT cells, δ1γδT cell population, or pharmaceutical composition for use according to any one of claims 20 to 24, wherein the treatment further comprises introducing one or more agents to a subject that increase common gamma chain cytokines, either concurrently or sequentially with the δ1γδT cells, δ1γδT cell population, or pharmaceutical composition.

27. ​​δ1γδT cells, δ1γδT cell population, or pharmaceutical composition for use according to any one of claims 20 to 24, wherein the treatment further comprises introducing one or more agents that increase common gamma chain cytokines to a subject before and / or after the introduction of the δ1γδT cells, δ1γδT cell population, or pharmaceutical composition.

28. A δ1γδT cell, a population of δ1γδT cells, or a pharmaceutical composition for use according to claim 26 or 27, wherein one or more agents that increase the common gamma chain cytokine(s) deplete lymphocytes before the introduction of δ1γδT cells(s).

29. A δ1γδT cell, a population of δ1γδT cells, or a pharmaceutical composition for use according to claim 26 or 27, wherein one or more agents that increase the common gamma chain cytokines induce the secretion of one or more common gamma chain cytokines from the introduced δ1γδT cells.

30. A δ1γδT cell, a population of δ1γδT cells, or a pharmaceutical composition for use according to any one of claims 26 to 29, wherein the common gamma chain cytokine(s) are IL-12 and / or IL-15.

31. A method for producing a δ1γδT cell according to any one of claims 1 to 13 or a cell population according to any one of claims 14 to 18, wherein the method comprises transfecting a δ1γδT cell (or more) with the nucleic acid according to any one of claims 1 to 13 by gamma retrovirus transduction.

32. The method according to claim 31, comprising the proliferation of the δ1γδT cells (or more) in ex vivo, wherein the proliferation in ex vivo is carried out before and / or after the transfection of the nucleic acid.

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