KR20190095942A - Anti-human CXCR3 Antibody for the Treatment of Vitiligo - Google Patents

Abstract

Provided are methods and uses of anti-CXCR3 antibodies for treating CXCR3-associated disorders such as vitiligo. In certain embodiments, the anti-CXCR3 antibody is a humanized anti-human CXCR3 antibody with enhanced effector function against cells expressing CXCR3 on the surface.

Description

Anti-human CXCR3 Antibody for the Treatment of Vitiligo

The present invention relates to antibodies and methods of using the antibodies for treating disorders associated with CXCR3 signaling, such as vitiligo.

Vitiligo is a palliative autoimmune skin disease in which autoreactive CD8 ⁺ T cells kill melanocytes in the epidermis leading to patchy depigmentation. This is one of the most common autoimmune diseases occurring in 1% of the world's population, but there are currently no FDA-approved treatments for vitiligo.

CXC motif chemokine receptor 3 (CXCR3) is a chemokine receptor expressed primarily on antigenic experience (memory), effector and activating T cells, and these subsets of T cells are their primary ligands: CXCL9 (MIG), CXCL10 (IP-10), And recruitment to tissue inflammation sites in response to CXCL11 (I-TAC). CXCR3 and CXCL10 are expressed in human T1D patients (Uno et al., Endocr J 57: 991-96 (2010); Roep et al., Clin Exp Immunol 159: 338-43 (2003); Tanaka et al., Diabetes 58: 2285-2291 (2009)). In such patients, CXCL10 is expressed in remaining insulin-producing beta cells in the islets. CXCR3 is expressed on invading T cells surrounding the islet. Similar expression patterns have been reproduced in non-obese diabetic (NOD) mice, a mouse model of type 1 diabetes (Morimoto et al., J Immunol 173: 7017-24 (2004)); Li et al., World J Gastroenterol 11 (30): 4750-2 (2005); Sarkar et al., Diabetes 61 (2): 436-46 (2012).

CXCR3 is expressed by dermal CD3 ⁺ lymphocytes and plasmacytoid dendritic cells and its chemokine ligands, CXCL10 and CXCL9, are upregulated in psoriasis lesions (Rottman et al., Lab Invest 81 (3): 335-47 ( 2001), Chen et al., Arch Dermatol Res 302 (2): 113-23 (2010). CXCR3 is also expressed on infiltrating T cells present in certain types of inflammatory tissue, while CXCL9, CXCL10 and CXCL11 are often produced by local cells in inflammatory lesions.

Upregulation of CXCR3 has been associated with various autoimmune disorders. CXCR3 expression is upregulated upon activation by antigen (usually absent in naive T cells). CXCR3 recruits these cells (including T helper 1 (Th1) cells) to tissue inflammation sites in response to their primary ligands. Beta cells in the islets of Langerhans express CXCL9 and CXCL10 (Frigerio at al., Nat Med 8: 1414-20 (2002)), and T cells infiltrating the pancreas express CXCR3 (documents). Christen et al., J Immunol 171: 6838-45 (2003); Van Halteren at al., Diabetologia 48: 75-82 (2005); Uno et al., Endocr J 57: 991- 96 (2010); Roep et al., Clin Exp Immunol 159: 338-43 (2003); Tanaka et al., Diabetes 58: 2285-2291 (2009); Sarkar et al., Diabetes 61 (2): 436-46 (2012). US Patent No. 8,865,870 to Youd et al. Discloses anti-CXCR3 antibodies.

Given the prevalence of vitiligo and other disorders involving CXCR3, there is a need for additional methods of targeting CXCR3 signaling, for example to reduce or treat the progression of disorders such as vitiligo in a patient.

Provided herein are methods of treating CXCR3-associated diseases and disorders such as vitiligo by administering a humanized antibody or antigen binding fragment thereof that specifically binds CXCR3. In certain embodiments, the anti-CXCR3 antibodies provided for the methods and uses herein have the ability to induce depletion of CXCR3-expressing cells, or are engineered to enhance the ability to induce depletion of CXCR3-expressing cells, thereby enhancing CXCR3. It can treat associated diseases and disorders such as vitiligo.

In some embodiments, a humanized antibody or antigen binding fragment thereof provided for the methods and uses herein has a germinality of at least 0.885 (when comparing all residues of the VH and VL chain except the D-region residues of the VH). score) or a germline score of at least 0.950 (only when comparing residues of the framework region as determined by IMTG). In some embodiments, a humanized antibody or antigen binding fragment thereof provided herein has a KD of at least 1 × 10 ⁻⁹ M. In some embodiments, a humanized antibody or antigen binding fragment thereof provided herein has a kd of less than 7 × 10 ⁻⁵ 1 / Ms. In some embodiments, a humanized antibody or antigen binding fragment thereof provided herein has a germline score of at least 0.885 (when comparing all residues of the VH and VL chain except for the D-region residues of VH), or only as determined by IMTG Comparing the residues of the framework region) with a germline score of at least 0.950 and a KD of at least 1 × 10 ⁻⁹ M and at least 0.885 (when comparing all residues of the VH and VL chain except for the D-region residues of VH) A germline score, or a germline score of at least 0.950 (only when comparing residues of the framework region as determined by IMTG). In some embodiments, a humanized antibody or antigen binding fragment thereof provided herein has a germline score of at least 0.885 (when comparing all residues of the VH and VL chain except for the D-region residues of VH), or only as determined by IMTG Comparing the residues of the framework regions) with a germline score of at least 0.950, a KD of at least 1 × 10 ⁻⁹ M, and a kd of less than 7 × 10 ⁻⁵ 1 / Ms.

In certain embodiments, humanized CXCR3 antibodies are provided wherein certain light chain variable regions are paired with specific heavy chain variable regions. In certain embodiments, humanized CXCR3 antibodies provided herein comprise variant human IgG1 Fc regions that confer enhanced effector function on cells expressing human CXCR3 on the surface.

In a first aspect, provided herein is a humanized anti-human CXCR3 antibody or pharmaceutical composition thereof for use in a method of depleting CXCR3 expressing cells in a subject, wherein the humanized anti-human CXCR3 antibody comprises a heavy chain variable region (VH). Light chain (LC) having a heavy chain (HC) having and a light chain variable region (VL). In some embodiments, CD4 + T-cells are depleted. In some embodiments, CD8 + T-cells are depleted. In some embodiments, CD4 + and CD8 + T cells are depleted. In some embodiments, CD4 + memory T-cells are depleted. In some embodiments, CD8 + memory T-cells are depleted. In some embodiments, CD4 + memory T-cells and CD8 + memory T-cells are depleted. In one embodiment, the subject has a T-cell-mediated autoimmune disease. In yet another embodiment, the subject has vitiligo.

In one embodiment of the first aspect, the VH and VL of the humanized anti-human CXCR3 antibodies provided herein comprise the amino acid sequences of the sequence pairs shown in Table 1 , and the HC comprises the amino acid sequence of any one of the following SEQ ID NOs: Further comprises human IgG1 Fc regions: 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11.

TABLE 1

In another embodiment of the first aspect, the VH of the humanized anti-human CXCR3 antibody provided herein comprises the amino acid sequence of SEQ ID NO: 20 and the VL comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the humanized human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2, or SEQ ID NO: 9, or SEQ ID NO: 10, or SEQ ID NO: 11.

In another embodiment of the first aspect, the VH of the humanized anti-human CXCR3 antibody provided herein comprises the amino acid sequence of SEQ ID NO: 18 and the VL comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, the antibody comprises a human IgG1 Fc region. In some embodiments, the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2, or SEQ ID NO: 9, or SEQ ID NO: 10, or SEQ ID NO: 11.

In another embodiment of the first aspect, the HC and LC of the humanized anti-human CXCR3 antibodies provided herein comprise the amino acid sequences of the sequence number pairs shown in Table 2 .

TABLE 2

In another embodiment of the first aspect, the HC of the humanized anti-human CXCR3 antibody provided herein comprises a human IgG1 Fc region having a reduced fucose content. In some embodiments, provided humanized anti-human CXCR3 antibodies are produced in host cells cultured in medium containing glycation inhibitors. In one embodiment, the glycation inhibitor is kifunensin.

In a second aspect, provided herein is a humanized anti-human CXCR3 antibody or pharmaceutical composition thereof for use in a method of treating a T-cell-mediated autoimmune disease, wherein the humanized anti-human CXCR3 antibody comprises a heavy chain variable region ( Heavy chain (HC) with VH) and light chain (LC) with light chain variable region (VL). In one embodiment, the T-cell-mediated disease is vitiligo.

In a third aspect, provided herein is a method of treating a T-cell-mediated autoimmune disease, wherein the method comprises a heavy chain (HC) and a light chain variable region (VL) having a heavy chain variable region (VH) in a subject in need thereof. Administering a humanized anti-human CXCR3 antibody comprising a light chain (LC). In some embodiments, the T-cell-mediated autoimmune disease is vitiligo.

In a fourth aspect, provided herein is a method of treating a T-cell-mediated autoimmune disease, wherein the method comprises a heavy chain (HC) and a light chain variable region (VL) having a heavy chain variable region (VH) in a subject in need thereof. Providing instructions for administering a humanized anti-human CXCR3 antibody comprising a light chain (LC) having In some embodiments, the T-cell-mediated autoimmune disease is vitiligo.

In a fifth aspect, provided herein is a kit for treating a T-cell-mediated autoimmune disease, comprising a heavy chain (HC) having a heavy chain variable region (VH) and a light chain (LC) having a light chain variable region (VL). Humanized anti-human CXCR3 antibodies comprising, and instructions for administering humanized anti-human CXCR3 to human subjects in need thereof.

In some embodiments of the first, second, third, fourth, and fifth aspects provided herein, the VH and VL of the humanized anti-human CXCR3 antibody comprise the amino acid sequences of the sequence pairs shown in Table 1 and are HC Comprises a human IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11. In another embodiment, the VH comprises the amino acid sequence of SEQ ID NO: 20 and the VL comprises the amino acid sequence of SEQ ID NO: 24. In another embodiment, the VH comprises the amino acid sequence of SEQ ID NO: 20, the VL comprises the amino acid sequence of SEQ ID NO: 24, and the human IgG1 Fc region is SEQ ID NO: 2, or SEQ ID NO: 9, or SEQ ID NO: 10, or The amino acid sequence of SEQ ID NO: 11.

In other embodiments of the second, third, fourth, fifth, sixth, and seventh aspects provided herein, the VH comprises the amino acid sequence of SEQ ID NO: 18 and the VL comprises the amino acid sequence of SEQ ID NO: 22. do. In some embodiments, the humanized human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2, or SEQ ID NO: 9, or SEQ ID NO: 10, or SEQ ID NO: 11.

In other embodiments of the first, second, third, fourth, and fifth aspects provided herein, the HC and LC of the antibody comprise the amino acid sequences of the sequence pairs shown in Table 2

In other embodiments of the first, second, third, fourth, and fifth aspects provided herein, the HC of the humanized anti-human CXCR3 antibody provided herein comprises a human IgG1 Fc region with a reduced fucose content. do. In some embodiments, provided humanized anti-human CXCR3 antibodies are produced in host cells cultured in medium containing glycation inhibitors. In some embodiments, the glycation inhibitor is kifunensin.

1 is a series of six graphs showing the amount of various T cell subsets in the blood after treatment with antibodies as follows: "Hamster CXCR3-173" (Hamster anti-mouse CXCR3), "CXCR3 mIgG2a Dab" (effector function) Hamster CXCR3-173 engineered to replace a mouse IgG2a constant region with a mutation to remove, "CXCR3 mIgG2a WT" (hamster CXCR3-173 engineered to replace a mouse wild-type IgG2a constant region), "CXCR3 mIgG1-agly" ( Hamster CXCR3-173 engineered to replace the mouse IgG1 constant region with the N297G mutation), and "untreated" (representing an untreated control).
FIG. 2A is a series of six graphs showing the binding of various Fc-engineered versions of CXCR3-173 by recombinant mouse FcγRI (mFcRI) as measured by surface plasmon resonance (Biacore) binding assays. BMP5, anti-BMP5 mIgG1 isotype control; mIgG1 agly, CXCR3-173 engineered to replace mouse IgG1 constant region with N297G mutations; mIgG2a WT, CXCR3-173 engineered to replace the mouse wild type IgG2a constant region; mIgG2a Dab, CXCR3-173 engineered to replace mouse IgG2a constant region with mutations to eliminate effector function; mIgG3, CXCR3-173 engineered to replace mouse wild-type IgG3 constant region; Hamster CXCR3, parent hamster mAb CXCR3-173.
FIG. 2B is a series of six graphs showing the binding of various Fc-engineered versions of CXCR3-173 by recombinant mouse FcγRIIb (mFcRIIb) as measured by the Biacore binding assay. The antibody name is the same as for FIG. 2A.
FIG. 2C is a series of six graphs showing the binding of various Fc-engineered versions of CXCR3-173 by recombinant mouse FcγRIII (mFcRIII) as measured by the Biacore binding assay. The antibody name is the same as for FIG. 2A.
FIG. 2D is a series of six graphs showing the binding of various Fc-engineered versions of CXCR3-173 by recombinant mouse FcγRIV (mFcRIV) as measured by the Biacore binding assay. The various antibody names are the same as for FIG. 2A.
3A is a table summarizing structure-effector functional properties for anti-human CXCR3 antibodies comprising engineered human IgG1 constant regions.
3B shows antibody-dependent cytotoxicity (ADCC) -mediated in vitro of CHO-human CXCR3 target cells by various anti-human CXCR3 antibodies at the indicated concentrations and effector: target (E: T) ratios of 5: 1. Bar graph depicting dissolution. Effector cells are natural killer ((NK) cells from a single donor. IgG, human IgG1 isotype control.Anti-human CXCR3 mAb tested was clone 4 (CXCR3 CL4), clone 12 (CXCR3 CL12), clone 82 (CXCR3). CL82), clone 135 (CXCR3 CL135), 53Hu37, and 53Hu37 engineered Fc variants M1, M2, and M3 (as described in Figure 3A) Kif, Kifunensine treatment.ALEM, Alemtuzumab.
3C is a bar graph depicting ADCC-mediated lysis in vitro of CHO-human CXCR3 target cells by various antibodies at the indicated concentrations and effector: target cell (E: T) ratios of 3: 1. Effector cells are from NK-like cell line (NK92-CD16V). IgG, human IgG1 isotype control. The anti-human CXCR3 mAb tested was 53Hu37 and engineered Fc variant M1 (as in FIG. 3A). CXCR3 CL4, anti-human CXCR3 mAb clone 4. Kif, kifunensine-treated. ALEM is alemtuzumab.
FIG. 4 shows the indicated affinity, KD, and BIAu37 for binding to human FcγRIIa (rhFcγRIIa), human FcγRIII-F158 (rhFcγRIII-F158), human FcγRIII-V158 (rhFcγRIII-V158), and mouse FcγRIV (rmFcγRIV). A table summarizing the data. M1-M3 are as described in FIG . 3A , which is kif defucosylated 53Hu37.
FIG. 5A is a series of six graphs depicting depletion of the indicated T-cell subset in vivo in cynomolgus monkeys treated with the indicated antibodies administered at a dose of 2 mg / kg body weight. M1: S239D / I332E variant of 53H37; Kif: defucosylated 53H37; Veh: vehicle control. N = 8. for the CXCR3 antibody group N = 6. for the vehicle group
FIG. 5B shows combined pharmacokinetic data evaluating the concentration of the indicated antibody in serum of cynomolgus monkeys pretreated with the indicated amount of time using a single dose of the indicated antibody administered at a dose of 2 mg / kg body weight . Is a graph. The anti-human CXCR3 mAbs tested were 53 Hu37, kifunensine-treated 53 Hu37 (53 Hu37 kif), and 53 Hu37 M1 variants (53 Hu37 M1).
6 is a table showing the sequence numbers and the corresponding sequences.

DETAILED DESCRIPTION Specific exemplary embodiments according to the present invention will now be described in detail, which specific examples are illustrated in the accompanying drawings.

Provided herein are methods and uses for a humanized antibody or antigen binding fragment thereof that specifically binds human CXCR3. In certain embodiments, an anti-CXCR3 antibody provided herein has been engineered to have the ability to induce depletion of CXCR3-expressing cells or to enhance the ability to induce depletion of CXCR3-expressing cells, thereby resulting in CXCR3-associated diseases and disorders. Treat. In some embodiments, a therapy for treating vitiligo by targeting CXCR3 is disclosed.

Antibodies

As used herein, the term “antibody” refers to four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, and multimers thereof (eg, IgM Refers to an immunoglobulin molecule comprising Each heavy chain has a heavy chain variable region (V _H) Or abbreviated VH) and heavy chain constant region (C _H or CH). The heavy chain constant region comprises three domains, C _H 1, C _H 2 and C _H 3. The Fc portion of the heavy chain includes C _H 2 and C _H 3.

Each light chain comprises a light chain variable region (abbreviated as V _L ) and a light chain constant region (C _L or CL). The light chain constant region contains one domain (C _L 1).

The V _H and V _L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDRs), interspersed with more conserved regions called framework regions (FR). Each V _H and V _L consists of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

As used herein, the term “antigen binding fragment” of an antibody refers to any naturally occurring, enzyme obtainable, synthetic or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. It includes. Antigen binding fragments of an antibody can be prepared using any suitable standard technique, such as, for example, proteolytic cleavage or recombinant genetic engineering techniques involving manipulation and expression of DNA encoding antibody variable domains and optionally antibody constant domains. Can be derived from an antibody molecule. Non-limiting examples of antigen binding moieties include: (i) Fab fragments; (ii) F (ab ') ₂ fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; And (vii) a minimal recognition unit comprising an amino acid residue that mimics the hypervariable region of the antibody (eg, an isolated complementarity determining region (CDR)). Other engineered molecules such as bispecific, trispecific, tetraspecific antibodies, and minibodies are also included within the expression "antigen binding fragment".

In certain embodiments, the CXCR3 antibody or antigen binding fragment comprises at least one antigen binding domain. In some embodiments, the antibody or fragment is multispecific and comprises two or more (eg, two, three, four, five or more) antigen binding domains such that the antibody or fragment has high affinity for CXCR3 and at least one other It may bind to an antigen or may bind to two or more CXCR3 molecules at the same or different epitopes. An antigen binding moiety may comprise one or more fragments of an antibody that retain the ability to specifically bind an antigen. Such fragments may comprise heavy and / or light chain variable regions from the parent antibody or from variants of the parent antibody.

As used herein, the term “antigen” refers to a binding site or epitope recognized by an antibody or antigen binding fragment thereof.

An “epitope” or “antigen determinant” is the portion of an antigen molecule responsible for specific interactions with the antigen binding domain of an antibody.

As used herein, an “bind” in connection with an antibody or antigen-binding fragment thereof refers to an antibody that forms one or more non-covalent bonds with its cognate antigen by noncovalent interactions between the antibody and the antibody combining sites of the antigen. Or the ability of an antigen binding fragment. The antigen may be an isolated antigen or may be presented in association with another entity, such as in the context of a polypeptide on the cell surface.

As used herein, the term “specifically binds” at least about 1 × 10 ⁻⁶ M, 1 × 10 ⁻⁷ M, 1 × 10 ⁻⁸ M, 1 × 10 ⁻⁹ M, 1 × 10 Refers to the ability of an antibody or antigen-binding fragment thereof to bind antigen at ^-10 M, 1 x 10 ^-11 M, 1 x 10 ^-12 M, or more Kd. In certain embodiments, the term refers to the ability of an antibody or antigen-binding fragment thereof to bind an antigen with an affinity at least twice higher than the affinity for a nonspecific antigen. However, it will be understood that the antibody or antigen-binding fragment thereof may specifically bind to two or more antigens (eg, human and cynomolgus CXCR3) with which the sequence is associated. Nonspecific binding usually has low affinity and medium to high binding capacity. If desired, nonspecific binding can be reduced by substantially changing the binding conditions without substantially affecting specific binding. These conditions are known in the art and those skilled in the art using routine techniques can select appropriate conditions. Conditions are usually defined in terms of the concentration of the antibody, the ionic strength of the solution, the temperature, the time considered for binding, and the concentration of blocking molecules such as serum albumin and milk casein.

Affinity constants can be determined by kinetic standard methods for antibody response, such as immunoassays (eg ELISA), or surface plasmon resonance (SPR). Instruments and methods for real-time detection and monitoring of binding rates are known and commercially available (eg, Biacore 2000, Biacore Ab (Swept Uppsala), and GE Healthcare Life Sciences).

As used herein, “complementarity determining region” or “CDR” refers to one of a plurality of portions within each variable region of an antibody or antigen binding fragment that together form an antigen binding site of the antibody. Each variable region domain contains three CDRs called CDR1, CDR2 and CDR3. Thus, the variable heavy chain domain (VH) comprises CDR-H1, CDR-H2 and CDR-H3 and the variable light domain (VL) comprises CDR-L1, CDR-L2, and CDR-L3. The three CDRs are non-contiguous along the linear amino acid sequence but are in close proximity within the folded polypeptide. CDRs are located in a loop connecting the parallel strands of the beta sheet of the variable domain.

As used herein, the term “framework (FR) amino acid residues” refers to amino acids within the framework region of the Ig chain. As used herein, the term “framework region” or “FR region” includes amino acid residues that are part of a variable region but not part of a CDR. Thus, the variable region framework is between about 100 and 120 amino acids in length, but includes only amino acids external to the CDRs.

As used herein, the term "% identity" or "percent identity" means that when comparing two sequences over a particular region, the two sequences have a specified number of identical residues at the same position. The terms "% similarity" or "percent similarity" have a similar meaning, but in addition to the number of identical amino acids between two sequences, it is also contemplated that the amino acids are not identical and are conservatively substituted. The percent identity can be determined using known computer algorithms such as the BLASTP, BLASTN, and FASTA programs (Altschul, SF, et al., J Mol). Biol 215: 403 (1990)), which is described, for example, in Pearson et al., Proc . Natl Acad Sci USA 85: 2444 (1988)] as the default parameter. For example, identity can be determined using the BLAST function of the National Center for Biotechnology Information (NCBI) database.

In certain embodiments, the antibody provided herein is a humanized antibody. A “humanized antibody” binds to a desired antigen and has one or more CDRs from a non-human species (eg, a mouse antibody) and has at least some portion of a framework region and / or constant domain from a human immunoglobulin molecule. It is an antibody molecule having a. Known human Ig sequences are described, for example, in ncbi.nlm.nih.gov/entrez-/query.fcgi; atcc.org/phage/hdb.html; sciquest.com; abcam.com; antibodyresource.com/onlinecomp.html; And Kabat et al., Sequences of Proteins of Immunological Interest, US Dept. Health (1983). Imported human sequences reduce or enhance immunogenicity or reduce binding, affinity, on-rate, off-rate, affinity, specificity, half-life, or any other suitable feature. Or can be used in the alteration, as is known in the art. Methods recognized in the art for antibody humanization are described in Jones et al., Nature 321: 522 (1986); Verhoeyen et al., Science 239: 1534 (1988); Sims et al., J Immunol 151: 2296 (1993); Chothia and Lesk, J Mol Biol 196: 901 (1987); Carter et al., Proc Natl Acad Sci USA 89: 4285 (1992); Presta et al., J Immunol 151: 2623 (1993); US Patent No. 5,589,205; US Patent No. 5,565,332; US Patent No. 6,180,370; US Patent No. 6,632,927; US Patent No. 7,241,877; US Patent No. 7,244,615; US Patent No. 7,244,832; US Patent No. 7,262,050; And US Patent Publication No. 2004/0236078, filed April 30, 2004, which are hereby incorporated by reference in their entirety.

In certain embodiments, certain framework residues in the humanized antibodies provided herein have been substituted with corresponding residues from the CDR donor antibody to alter, eg, improve, antigen binding. Substituted with framework residues from anti-human CXCR3 antibodies. Such framework substitutions were identified by modeling the interaction of CDR and framework residues to identify framework residues important for antigen binding and sequence comparisons to identify specific framework residues at specific locations. In some embodiments, humanized antibody variants of the invention were prepared using 4D humanization. For methods used in 4D humanization, see International Publication Nos. 2009/032661 (eg, paragraphs [0037]-[0044], incorporated herein by reference in its entirety). Briefly, 4D humanization may include: a) building a 3D model of the variable domain to be humanized; b) identifying flexible residues in variable domains using molecular dynamics simulations of 3D models of the domains; c) identifying the closest human germline by comparing the molecular dynamics trajectory of the 3D model with the molecular dynamics trajectories of the 49 human germline; And d) mutating flexible residues that are not part of the CDRs to their human germline counterparts (identified in step (c)).

In some embodiments, humanized CXCR3 antibodies, or antigen binding fragments thereof, comprising the VH and VL sequences set forth in Table 1 are provided.

In some embodiments, humanized CXCR3 antibodies, or antigen binding fragments thereof, are provided comprising the heavy (HC) and light chain (LC) sequences disclosed in Table 2 .

Antibody Effector  Function / depletion active

In certain embodiments, the anti-CXCR3 antibodies disclosed herein have been engineered to have the ability to induce depletion of CXCR3-expressing cells or to enhance the ability to induce depletion of CXCR3-expressing cells, thereby resulting in CXCR3-associated diseases and disorders. It can be cured. CXCR3-expressing cells that may be depleted by the antibodies disclosed herein may include CD4 ⁺ T cells and / or CD8 ⁺ T cells. CXCR3-expressing cells that may be depleted by the antibodies disclosed herein may include CD4 ⁺ memory T cells and / or CD8 ⁺ memory T cells. As used herein, “depletion” with respect to CXCR3 ⁺ cells (ie, cells expressing CXCR3 on the taxosome surface) refers to the removal of such cells from the cell population. Reference to depletion includes complete or partial depletion. Depletion may also be permanent or temporary, and may be of varying degrees in size and / or location. Depletion can be a result of cell death, such as by apoptosis or necrosis. Depletion can be performed by any method known in the art (eg, flow cytometry, immune tissue, before or after exposure to an antibody or antigen binding fragment provided herein, or in the absence and presence of an antibody or antigen binding fragment provided herein. Chemistry, etc.) can be used to measure the number of CXCR3 ⁺ cells in a population. After exposure to the antibody or antigen binding fragment provided herein, the CXCR3 ⁺ cells are at least or approximately 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more.

In certain embodiments, the humanized anti-human CXCR3 antibody is an enhanced effector for cells expressing human CXCR3 on the surface compared to a corresponding humanized anti-human CXCR3 antibody comprising a wild type Fc region, eg, a wild type human IgG1 Fc. Indicates a function. As used herein, “enhanced effector function” refers to antibody-dependent cytotoxicity (ADCC), complement, to a suitable target cell, compared to a reference antibody having the same antigen specificity and wild-type human IgG1 Fc region under the same conditions. -Measurably increased ability of an antibody to induce any one or more of -mediated cytotoxicity (CDC), or antibody-dependent cell-mediated phagocytosis (ADCP). In certain embodiments, the reference antibody comprises a variant human Fc region. In certain embodiments, the effector function is ADCC, ADCP, or CDC, or any combination thereof. In certain embodiments, the effector function is ADCC, or CDC, or both ADCC and CDC. In certain embodiments, the effector function is ADCC. In certain embodiments, the effector function is CDC. In certain embodiments, the effector function is both ADCC and CDC. In certain embodiments, the effector function is ADCP.

As used herein, “variant human IgG1 Fc region” refers to a human IgG1 Fc region that has been engineered or modified to include one or more amino acid mutations or amino acid modifications as compared to wild type human IgG1 Fc. In certain embodiments, the wild type human IgG1 Fc region comprises the following amino acid sequence:

In certain embodiments, the variant human IgG1 Fc region comprises at least one of the following amino acid substitutions: G236A, S239D, S267E, H268F, S324T, I332E (Eu numbering), or any combination thereof. In certain embodiments, the variant human IgG1 Fc region comprises at least one of the following sets of amino acid substitutions: S239D / I332E, G236A / S267E / H268F / S324T / I332E, and S239D / H268F / S324T / I332E (Eu numbering). In certain embodiments, the variant human IgG1 Fc region comprises amino acid substitution S239D / I332E. In other embodiments, the variant human IgG1 Fc region comprises amino acid substitutions G236A / S267E / H268F / S324T / I332E. In another embodiment, the variant human IgG1 Fc region comprises amino acid substitutions S239D / H268F / S324T / I332E.

For example, in certain embodiments, the variant human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 3, 4, 5, 6, 7 or 8.

In certain embodiments, the variant human IgG1 Fc region comprises a sequence that is at least 90% identical to any one or more of SEQ ID NOs: 3-8, provided that in each case the indicated amino acid substitutions are maintained. In various embodiments, the variant human IgG1 Fc region is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95 with any one or more of SEQ ID NOs: 3-8 % Identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical sequences, provided that in each case, the specified amino acid substitutions are maintained.

In certain embodiments, the variant human IgG1 Fc region comprises at least one of the following sets of amino acid substitutions: S239D / I332E, G236A / S267E / H268F / S324T / I332E, and S239D / H268F / S324T / I332E (Eu numbering). For example, in certain embodiments, the variant human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 9, 10, or 11.

In certain embodiments, at least one amino acid substitution is S239D / I332E (Eu numbering). In another embodiment, the at least one amino acid substitution is G236A / S267E / H268F / S324T / I332E (Eu numbering). In certain embodiments, the at least one amino acid substitution is S239D / H268F / S324T / I332E (Eu numbering).

In certain embodiments, the variant human IgG1 Fc region of a humanized anti-CXCR3 antibody provided herein comprises a sequence that is at least 90% identical to any one of SEQ ID NOs: 9-11, provided that in each case the specified amino acid substitutions and Improved effector functionality is maintained. In various embodiments, the variant human IgG1 Fc region is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95 with any one or more of SEQ ID NOs: 9-11 % Identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical sequences, provided that in each case, the specified amino acid substitutions are maintained.

CDR Variant :

In addition to the foregoing embodiments, provided herein are anti-CXCR3 antibodies comprising six CDRs, wherein the VH is

(i) SEQ ID NO: 12, SEQ ID NO: 35, and SEQ ID NO: 14;

(ii) SEQ ID NO: 12, SEQ ID NO: 35, and SEQ ID NO: 45;

(iii) SEQ ID NO: 12, SEQ ID NO: 36, and SEQ ID NO: 45;

(iv) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 14;

(v) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 45;

(vi) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 46

(vii) SEQ ID NO: 12, SEQ ID NO: 38, and SEQ ID NO: 14;

(viii) SEQ ID NO: 12, SEQ ID NO: 38, and SEQ ID NO: 45;

(ix) SEQ ID NO: 12, SEQ ID NO: 39, and SEQ ID NO: 14;

(x) SEQ ID NO: 12, SEQ ID NO: 39, and SEQ ID NO: 47;

(xi) SEQ ID NO: 12, SEQ ID NO: 40, and SEQ ID NO: 14;

(xii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 45

(xiii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 46;

(xiv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 48;

(xv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 49;

(xvi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 50;

(xvii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 51;

(xviii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 52;

(xix) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 53;

(xx) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 54;

(xxi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 55;

(xxii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 56;

(xxiii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 57;

(xxiv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 58;

(xxv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 59;

(xxvi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 60;

(xxvii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 61;

(xxviii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 62;

(xxix) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 63;

(xxx) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 64;

(xxxi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 65;

(xxxii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 66;

(xxxiii) SEQ ID NO: 34, SEQ ID NO: 13, and SEQ ID NO: 14;

(xxxiv) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 14;

(xxxv) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 46;

(xxxvi) SEQ ID NO: 12, SEQ ID NO: 42, and SEQ ID NO: 14;

(xxxvii) SEQ ID NO: 12, SEQ ID NO: 42, and SEQ ID NO: 45;

(xxxviii) SEQ ID NO: 12, SEQ ID NO: 43, and SEQ ID NO: 14;

(xxxix) SEQ ID NO: 12, SEQ ID NO: 44, and SEQ ID NO: 14;

(xl) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 14; or

(xli) comprises a CDR having the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 68;

VL

(i) SEQ ID NO: 69, SEQ ID NO: 16, and SEQ ID NO: 17;

(ii) SEQ ID NO: 70, SEQ ID NO: 16, and SEQ ID NO: 17;

(iii) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 17;

(iv) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 75;

(v) SEQ ID NO: 72, SEQ ID NO: 16, and SEQ ID NO: 17;

(vi) SEQ ID NO: 73, SEQ ID NO: 16, and SEQ ID NO: 17;

(vii) SEQ ID NO: 74, SEQ ID NO: 16, and SEQ ID NO: 17;

(viii) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 75;

(ix) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 76;

(x) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 77;

(xi) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 78; or

(xii) CDRs having the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 79.

In addition to the foregoing embodiments, provided herein are anti-CXCR3 antibodies comprising six CDRs, wherein the VH is

(i) SEQ ID NO: 12, SEQ ID NO: 35, and SEQ ID NO: 14;

(ii) SEQ ID NO: 12, SEQ ID NO: 35, and SEQ ID NO: 45;

(iii) SEQ ID NO: 12, SEQ ID NO: 36, and SEQ ID NO: 45;

(iv) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 14;

(v) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 45;

(vi) SEQ ID NO: 12, SEQ ID NO: 37, and SEQ ID NO: 46;

(vii) SEQ ID NO: 12, SEQ ID NO: 38, and SEQ ID NO: 14;

(viii) SEQ ID NO: 12, SEQ ID NO: 38, and SEQ ID NO: 45;

(ix) SEQ ID NO: 12, SEQ ID NO: 39, and SEQ ID NO: 14;

(x) SEQ ID NO: 12, SEQ ID NO: 39, and SEQ ID NO: 47; or

(xi) comprises a CDR having the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 40, and SEQ ID NO: 14;

VL

(i) SEQ ID NO: 69, SEQ ID NO: 16, and SEQ ID NO: 17;

(ii) SEQ ID NO: 70, SEQ ID NO: 16, and SEQ ID NO: 17;

(iii) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 17;

(iv) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 100;

(v) SEQ ID NO: 72, SEQ ID NO: 16, and SEQ ID NO: 17;

(vi) SEQ ID NO: 73, SEQ ID NO: 16, and SEQ ID NO: 17;

(vii) SEQ ID NO: 74, SEQ ID NO: 16, and SEQ ID NO: 17;

(viii) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 75;

(ix) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 76;

(x) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 77;

(xi) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 78; or

(xii) CDRs having the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 79.

In addition to the foregoing embodiments, provided herein are anti-CXCR3 antibodies comprising six CDRs, wherein the VH is

(i) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 45

(ii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 46;

(iii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 48;

(iv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 49;

(v) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 50;

(vi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 51;

(vii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 52;

(viii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 53;

(ix) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 54;

(x) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 55; or

(xi) comprises a CDR having the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 56;

VL

(i) SEQ ID NO: 69, SEQ ID NO: 16, and SEQ ID NO: 17;

(ii) SEQ ID NO: 70, SEQ ID NO: 16, and SEQ ID NO: 17;

(iii) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 17;

(iv) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 75;

(v) SEQ ID NO: 72, SEQ ID NO: 16, and SEQ ID NO: 17;

(vi) SEQ ID NO: 73, SEQ ID NO: 16, and SEQ ID NO: 17;

(vii) SEQ ID NO: 74, SEQ ID NO: 16, and SEQ ID NO: 17;

(viii) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 75;

(ix) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 76;

(x) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 77;

(xi) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 78; or

(xii) CDRs having the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 79.

In addition to the foregoing embodiments, provided herein are anti-CXCR3 antibodies comprising six CDRs, wherein the VH is

(i) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 57;

(ii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 58;

(iii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 59;

(iv) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 60;

(v) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 61;

(vi) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 62;

(vii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 63;

(viii) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 64;

(ix) SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 65; or

(x) comprises a CDR having the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 66;

VL

(i) SEQ ID NO: 69, SEQ ID NO: 16, and SEQ ID NO: 17;

(ii) SEQ ID NO: 70, SEQ ID NO: 16, and SEQ ID NO: 17;

(iii) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 17;

(iv) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 75;

(v) SEQ ID NO: 72, SEQ ID NO: 16, and SEQ ID NO: 17;

(vi) SEQ ID NO: 73, SEQ ID NO: 16, and SEQ ID NO: 17;

(vii) SEQ ID NO: 74, SEQ ID NO: 16, and SEQ ID NO: 17;

(viii) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 75;

(ix) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 76;

(x) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 77;

(xi) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 78; or

(xii) CDRs having the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 79.

In addition to the foregoing embodiments, provided herein are anti-CXCR3 antibodies comprising six CDRs, wherein the VH is

(i) SEQ ID NO: 34, SEQ ID NO: 13, and SEQ ID NO: 14;

(ii) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 14;

(iii) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 71;

(iv) SEQ ID NO: 12, SEQ ID NO: 42, and SEQ ID NO: 14;

(v) SEQ ID NO: 12, SEQ ID NO: 42, and SEQ ID NO: 70;

(vi) SEQ ID NO: 12, SEQ ID NO: 43, and SEQ ID NO: 14;

(vii) SEQ ID NO: 12, SEQ ID NO: 44, and SEQ ID NO: 14;

(viii) SEQ ID NO: 12, SEQ ID NO: 41, and SEQ ID NO: 14; or

(ix) a CDR having the amino acid sequence of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 68;

VL

(i) SEQ ID NO: 69, SEQ ID NO: 16, and SEQ ID NO: 17

(ii) SEQ ID NO: 70, SEQ ID NO: 16, and SEQ ID NO: 17;

(iii) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 17;

(iv) SEQ ID NO: 71, SEQ ID NO: 16, and SEQ ID NO: 75;

(v) SEQ ID NO: 72, SEQ ID NO: 16, and SEQ ID NO: 17;

(vi) SEQ ID NO: 73, SEQ ID NO: 16, and SEQ ID NO: 17;

(vii) SEQ ID NO: 74, SEQ ID NO: 16, and SEQ ID NO: 17;

(viii) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 75;

(ix) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 76;

(x) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 77;

(xi) SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 78; or

(xii) CDRs having the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 79.

In addition to the above embodiments,

And the amino acid sequence of SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, or SEQ ID NO: 90 Provided herein are anti-CXCR3 antibodies comprising VH.

In addition to the above embodiments,

Comprising a VH comprising the amino acid sequence of SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, or SEQ ID NO: 101 Anti-CXCR3 antibodies are provided herein.

In addition to the above embodiments,

A VH comprising an amino acid sequence selected from SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, or SEQ ID NO: 111 Provided herein are anti-CXCR3 antibodies.

In addition to the above embodiments,

Comprising the amino acid sequence of SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118, SEQ ID NO: 119, SEQ ID NO: 120, SEQ ID NO: 121, or SEQ ID NO: 122 Provided herein are anti-CXCR3 antibodies comprising VH.

In addition to the above embodiments,

Amino acids of SEQ ID NO: 123, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, or SEQ ID NO: 134 Provided herein are anti-CXCR3 antibodies comprising a VL comprising a sequence.

In certain embodiments, the anti-human CXCR3 antibodies comprise VH / VL pairs, each comprising an amino acid sequence illustrated in Table 3 . (The indicated change (s) relate to the corresponding 53Hu37 VH or VL sequence (SEQ ID NO: 20, and SEQ ID NO: 24), respectively):

TABLE 3

Neutralizing antibodies

In certain embodiments, the humanized anti-human CXCR3 antibodies provided herein are CXCR3 neutralizing antibodies. In certain exemplary embodiments, the CXCR3 antibody has neutralizing activity in addition to enhanced effector function. The combined effect of CXCR3 neutralization and CXCR3 ⁺ cell depletion may be advantageous whenever it is desirable to reduce or eliminate CXCR3-mediated effects, eg recruitment of T cells.

"CXCR3 neutralizing antibodies" bind to CXCR and block the activity of receptors such as typical physiological and genetic responses resulting from CXCR3 ligands that bind to CXCR3. The neutralizing activity may be complete (100% neutralizing) or partial, such as about 10, 20, 30, 40, 50, 60, 70, 80, 90, 95 (or any percentage in between) or more neutralization, and the antibody Will depend on various factors known to those skilled in the art, such as concentration, affinity, and epitope, and specific assays used to assess neutralizing activity. The neutralizing activity of CXCR3 neutralizing antibodies can be indicated by assays that measure, for example, inhibition of ligand binding, GTP binding, calcium migration, cell chemotaxis, and / or receptor internalization. Many assays for determining the activity of neutralizing antibodies, particularly CXCR3 neutralizing antibodies, are known to those skilled in the art and can be readily modified to determine if a particular antibody is neutralizing.

For example, in some embodiments, the neutralizing activity of an anti-CXCR3 antibody is substantially in a chemotaxis assay as described in the package insert for an antibody produced by clone 49801 and sold by R & D Systems® (Cat. No. MAB160). Can be evaluated. Neutralization dose 50 (ND ₅₀ ) is defined as the concentration of antibody required to produce half of the maximum inhibition of cell surface CXCR3-mediated recombinant human CXCL11 (rhCXCL11) response in a reactive cell line, at a specific rhCXCL11 concentration. To determine the ability of antibodies to block rhCXCL11 induced chemotaxis of BaF / 3 cells transfected with hCXCR3, 7 ng / mL of rhCXCL11 was added to the lower compartment of a 96-well chemotaxis chamber (NeuroProbe, Cabin Zone, MD). do. The chemotaxis chamber is then assembled using a PVP-free polycarbonate filter (pore size of 5 μm). Passaging dilutions of antibody (eg, 0.001 to 10000 μg / mL) and 0.25 × 10 ⁶ cells / well are added to the upper wells of the chamber. After 3 hours of incubation at 37 ° C. in a 5% CO ₂ -humidified incubator, the chamber is disassembled and the cells transferred to the lower chamber are transferred to a working plate and quantified using, for example, Resazurin fluorescence. .

Colvin et al., Mol Cell Biol 26: 5838-49 (2006) describe, in certain embodiments, additional assays that can be used to determine the neutralizing activity of neutralizing anti-CXCR3 antibodies. Briefly, 300-19 cells, a murine pre-B-cell leukemia cell line functionally expressing CXCR4, can be used. After transfection, the strain may functionally express other chemokine receptors, such as human CXCR3 (see, eg, paragraphs 201 through 209 of US Patent Publication No. 2010/0061983, which is incorporated herein by reference). 300-19 cells expressing human CXCR3 can be grown in complete RPMI medium containing 10% fetal bovine serum (FBS). To assess the binding of CXCR3 ligand to CXCR3 in the presence of candidate neutralizing CXCR3 antibodies, in 150 μL total volume of binding buffer (0.5% BSA, 5 mM MgCl ₂ , 1 mM CaCl ₂ , 50 mM HEPES, pH 7.4). 400,000 CXCR3 / 300-19 cells are placed in a 96 well tissue culture plate. A total of 0.04 nM of ¹²⁵ I labeled CXCL10 (New England Nuclear, Boston, Mass.) Or CXCL11 (Amersham Biosciences, Piscataway, NJ) and 5 x 10 ⁶ nM to 500 nM of unlabeled CXCL10 or CXCL11 (Peprotech, Rocky Hill, NJ) can be added to the cells and incubated for 90 minutes with shaking at room temperature. The cells are transferred onto 96-well filter plates (Millipore, Billerica, Mass.) Pre-soaked in 0.3% polyethyleneimine and washed three times with 200 μL of binding buffer supplemented with 0.5 M NaCl. The plate is dried and radioactivity is measured after addition of scintillation fluid in a Wallac Microbeta scintillation counter (Perkin-Elmer Life Sciences, Boston, Mass.). The binding of CXCL9 can be evaluated similarly to CXCL10 and CXCL11.

In certain embodiments, the antibodies disclosed herein can prevent or reduce calcium flux into cells expressing CXCR3. In some embodiments, calcium flow can be detected in cells such as CXCR3 / 300-19 cells. Approximately 5 × 10 ⁶ cells are suspended in 2 mL of RPMI medium containing 1% bovine serum albumin (BSA). Add 15 micrograms of Fura-2 (Molecular Probes, Eugene, OR) and incubate the cells for 20 minutes at 37 ° C. Cells are washed twice in PBS and recreated in 2 mL of calcium flow buffer (145 mM NaCl, 4 mM KCl, 1 mM NaHPO ₄ , 1.8 mM CaCl ₂ , 25 mM HEPES, 0.8 mM MgCl ₂ , and 22 mM glucose). Muddy. Fluorescence readings are measured at 37 ° C. on a DeltaRAM fluorometer (Photon Technology International, Lawrenceville, NJ). Before and after the addition of chemokines (eg, CXCL9, CXCL10, or CXCL11), intracellular calcium concentrations are recorded as excitation fluorescence intensity emitted at 510 nm in response to sequential excitation at 340 nm and 380 nm and fluorescence at 340 nm. It is presented as the relative ratio of fluorescence at 380 nm.

In certain embodiments, CXCR3 neutralization can be assessed by measuring a decrease in receptor internalization. In some embodiments, the receptor internalization assay comprises 37 ° C. in RPMI medium containing about 2 × 10 ⁵ cells, eg, CXCR3 / 300-19 cells with 1% BSA with varying concentrations of CXCL10, CXCL11, or CXCL9. By 30 minutes incubation at. Cells can then be washed with ice cold flow cytometry staining buffer and then analyzed for surface expression of CXCR3 using PE-conjugated CXCR3 antibodies.

As assessed by any of the above assays, the neutralizing anti-CXCR3 antibody, in certain embodiments, is approximately 0.01, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7 , ND ₅₀ of 8, 9, 10, 15, 20, 40, 50, or 100 μg / mL. In certain embodiments, ND ₅₀ may be 0.5 to 12 μg / mL, and in more specific embodiments, 1 to 6 μg / mL.

Inhibition of migration, recruitment, or accumulation of cells by the antibodies or antigen binding fragments provided herein can be assessed by any method known to those of skill in the art. For example, such methods can be used in immunohistochemistry, flow cytometry, RT-PCR, etc., to assess the number of cells, such as CXCR3 ⁺ cells, in one or more populations of cells or in one or more locations in the body or organs. It may include the analysis of. Migration, recruitment, or accumulation of cells is at least or approximately 10%, 15%, 20%, 25%, 30%, 35 compared to migration, recruitment, or accumulation of cells in the absence of an antibody or antigen binding fragment provided herein. %, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more.

Nucleotide sequence

Also provided herein is a nucleotide sequence encoding an amino acid sequence disclosed herein. In some embodiments, the nucleotide sequence encodes an antibody or fragment capable of depleting CXCR3 ⁺ cells in vitro and / or in vivo. In certain embodiments, nucleotide sequences can be used in the preparation of expression vectors for expression of an anti-CXCR3 antibody or antigen binding fragment thereof in a cell (eg, expression in mammalian cells).

In certain embodiments, also disclosed herein are polynucleotides that are substantially identical to the polynucleotides encoding the amino acid sequences disclosed herein. Substantially identical sequences may be polymorphic sequences, ie alternative sequences or alleles (in a population). Substantially identical sequences may also include mutated sequences, including sequences comprising silent mutations. Mutations can include changes in one or more nucleotide residues, deletion of one or more nucleotide residues, or insertion of one or more additional nucleotide residues. Substantially identical sequences may also include various nucleotide sequences that encode the same amino acid at any given amino acid position in the amino acid sequences disclosed herein, due to the degeneracy of the nucleic acid code. Also included within substantially identical sequences are sequences encoding the chain (s) of the antibody that retains the ability to deplete CXCR3 ⁺ cells in vitro and / or in vivo.

In certain embodiments, a nucleic acid provided herein encodes an amino acid sequence of chain (s) in an antibody or fragment capable of depleting CXCR3-expressing cells provided herein, or wherein the nucleic acid is subjected to stringent conditions, such that the antibody or antigen thereof It may hybridize to a nucleic acid encoding the amino acid sequence of the chain (s) in the binding fragment.

In certain embodiments, the amino acid sequence of the VH domain of an anti-CXCR3 antibody or antigen-binding fragment thereof and at least about 80-100% (eg, about 80%, 85) of the nucleotide sequence encoding the heavy chain of this antibody %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) (or any percentage in between) containing the same nucleotide sequence A polynucleotide sequence is disclosed herein. In certain embodiments, the polynucleotide sequence comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (additional, deletion and conservative) relative to the nucleotide sequence encoding the heavy chain of the antibody. Nucleotide sequences, including substitutions such as substitutions).

In certain embodiments, the amino acid sequence of the VL domain of an anti-CXCR3 antibody or fragment and at least about 80-100% (eg, about 80%, 85%, 90) of the nucleotide sequence encoding the light chain of this antibody %, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) (or any percentage there between) polynucleotides comprising identical nucleotide sequences Sequences are disclosed herein. In certain embodiments, the polynucleotide sequence comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mutations (additional, deletion and conservative) relative to the nucleotide sequence encoding the light chain of the antibody. Nucleotide sequences, including substitutions such as substitutions).

In certain embodiments, at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with a VH amino acid sequence (Or any percentage therebetween) identical and at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, identical to the VL amino acid sequence, Disclosed herein are polynucleotide sequences comprising 99%, or 100% (or any percentage in between) identical nucleotide sequences, wherein the nucleotide sequences encode heavy and light chain amino acid sequences of any of the antibodies disclosed herein.

The disclosed polynucleotides can be obtained by any method known in the art. For example, if the nucleotide sequence of the antibody is known, polynucleotides encoding the antibody can be assembled from chemically synthesized oligonucleotides. This is, for example, by synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligation of such oligonucleotides, and then PCR of the ligated oligonucleotides. May involve amplification. The disclosed polynucleotides may also be generated from any other suitable nucleic acid source, such as an antibody cDNA library, or a cDNA library isolated from any tissue or cell expressing the antibody (eg, hybridoma cells selected to express the antibody). Can be.

term- CXCR3  Expression of Antibody or Antigen Binding Fragment thereof

After manipulation of the nucleic acid encoding a humanized anti-CXCR3 antibody or antigen binding fragment thereof provided herein, the coding nucleic acid is typically introduced into a host cell that can be used to generate the desired amount of the encoded antibody, or antigen binding fragment thereof. Is inserted into an expression vector for introduction. Suitable vectors for expression are known in the art. Suitable host cells include, for example, CHO, COS, Sf9 and / or other human or nonhuman cell lines. In some embodiments, the host cell provides a method for generating an antibody or fragment disclosed herein by transiently or stably expressing nucleic acid on a vector when cultured in a culture medium.

The term "vector" or "expression vector" is used herein to describe a vehicle for introducing and expressing a desired gene into a cell. As known to those skilled in the art, such vectors include, for example, plasmids, phages, viruses and retroviruses. In general, suitable vectors may include selection markers, restriction enzyme sites suitable to assist in cloning the desired gene and the ability to enter and / or replicate in eukaryotic or prokaryotic cells.

Many expression vector systems can be used for expression purposes of the anti-CXCR3 antibodies provided herein. For example, a class of vectors may contain DNA elements derived from animal viruses such as bovine papilloma virus, polyoma virus, adenovirus, vaccinia virus, baculovirus, retrovirus (RSV, MMTV or MOMLV) or SV40 virus. I use it. Others involve the use of polycistronic systems with internal ribosome binding sites. Additionally, cells in which DNA has been integrated into the cell's chromosome can be selected by introducing one or more markers that allow selection of transfected host cells. Such markers can provide autotrophs to the trophic host, or provide biocidal (eg antibiotic) resistance, or resistance to heavy metals such as copper. Selectable marker genes may be directly linked to the DNA sequence to be expressed or introduced into the same cell by cotransformation. Additional elements may also be required for optimal synthesis of mRNA. Such elements may include signal sequences, splicing signals, and transcriptional promoters, enhancers, and termination signals. In some embodiments, the cloned variable region genes are inserted into expression vectors along the heavy and light chain constant region genes as discussed above. In some embodiments, the heavy and light chain constant regions are human.

In other embodiments, the anti-CXCR3 antibodies provided herein, or antigen binding fragments thereof, can be expressed using polycistronic constructs. In such expression systems, multiple gene products of interest, such as the heavy and light chains of an antibody, can be generated from a single polycistronic construct. Such systems advantageously use an internal ribosome entry site (IRES) to provide relatively high levels of polypeptides provided herein in eukaryotic host cells. Compatible IRES sequences are disclosed in US Pat. No. 6,193,980, which is incorporated herein by reference. Those skilled in the art will appreciate that such expression systems can be used to effectively produce the full range of polypeptides disclosed herein.

Once the vector or DNA sequence encoding the antibody, or antigen binding fragment thereof, is prepared, the expression vector can be introduced into a suitable host cell. That is, host cells can be transformed. Introduction of plasmids into host cells can be accomplished by various techniques well known to those skilled in the art. This includes, but is not limited to, transfection (including electrophoresis and electroporation), plasma fusion, calcium phosphate precipitation, fusion of cells and enveloped DNA, microinfusion, and infection with intact viruses. It doesn't work. Ridgway, A. A. G. "Mammalian Expression Vectors" Chapter 24.2, pp. 470-472 Vectors, Rodriguez and Denhardt, Eds. (Butterworths, Boston, Mass. 1988). In some embodiments, plasmid introduction into the host is via electroporation. Transformed cells are grown under conditions appropriate for the production of encoded amino acid sequences, such as antibody light and heavy chains, and analyzed for generation of encoded amino acid sequences. Exemplary assay techniques include enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), flow cytometry, immunohistochemistry, and the like.

"Host cell" refers to a cell constructed using recombinant nucleic acid technology and introduced with a vector encoding at least one heterologous protein. In descriptions of processes for isolation of polypeptides from recombinant hosts, the terms "cell" and "cell culture" refer to a source of encoded protein, eg, an antibody or antigen-binding fragment thereof, unless expressly specified otherwise. It is used interchangeably to indicate. In other words, recovery of a polypeptide from a "cell" can mean whole cell spin down, or recovery from a cell culture containing both medium and suspended cells.

In one embodiment, the host cell line used for antibody expression is of mammalian origin and one skilled in the art can determine the particular host cell line that is best suited for the desired gene product expressed in the host cell. Exemplary host cell lines DG44 and DUXB11 (Chinese hamster ovary cell line, DHFR minus), HELA (human cervical carcinoma), CVI (monkey kidney cell line), COS (derivatives of CVI with SV40 T antigen), R1610 (Chinese hamster fiber Parental cells), BALBC / 3T3 (mouse fibroblasts), SP2 / O (mouse myeloma), BFA-1c1BPT (bovine endothelial cells), RAJI (human lymphocytes), 293 (human kidneys). In one embodiment, NS0 cells can be used. In some embodiments, CHO cells are used. Host cell lines are typically available from the American Tissue Culture Collection or published literature, which is a commercial service.

In one embodiment, the cell line provides altered glycosylation of the antibody expressed in the cell line, eg, afucosylation (eg, PER.C6.RTM. (Crucell) or FUT8-knockout CHO). Cell line (Potelligent. RTM. Cells) (Biowa, Princeton, NJ). Alternatively, the cells may lack one or more glycosidase required for the initial processing of N-glycans and / or allow the culture conditions to inhibit the activity of one or more of these glycosidases. For example, the cell may lack one or more glycosidases such as alpha-glucosidase I, alpha-glucosidase II, and alpha-mannosidase I. In addition, or alternatively, the engineered cells can be contacted with inhibitors of one or more glycosidases such as alpha-glucosidase I, alpha-glucosidase II, and alpha-mannosidase I. In certain embodiments, the inhibitor is an inhibitor of alpha-mannosidase I, eg, an alpha-mannosidase I specific inhibitor, kifunensin. Exemplary methods of culturing host cells using kifunensine and other inhibitors are disclosed in US Pat. No. 8,071,336, which is incorporated herein by reference in its entirety. In certain embodiments, quipu nensin process is a major species at least _-9 Man ₅ (GlcNAc) and 50% to generate antibodies having ₂ N- glycan, wherein, with the N- glycans Man8-containing and Man9-.

In vitro production allows for scaling up to provide large amounts of desired polypeptides. Techniques for culturing mammalian cells under tissue culture conditions are known in the art, which are, for example, homogeneous suspension culture in an air flotation reactor or continuous stirred reactor, or, for example, hollow fibers, microcapsules Fixed or entrapped cell culture on internal, agarose microbeads or ceramic cartridges. If necessary and / or desired, the solution of the polypeptide may be subjected to conventional chromatography methods such as gel filtration, ion-exchange chromatography, chromatography on DEAE-cellulose and / or (immune-) affinity chromatography. Can be purified.

Nucleic acids encoding the anti-CXCR3 antibodies, or fragments thereof, provided herein can also be expressed in non-mammalian cells, such as bacteria or yeast or plant cells. In this regard, it will be appreciated that various single-cell non-mammalian microorganisms, such as bacteria, ie those that can be grown in culture or fermentation, may also be used for expression of the antibodies and antigen-binding fragments thereof provided herein. Suitable bacteria include members of the intestinal bacteria, such as Escherichia coli or Salmonella; Bacillus, for example Bacillus subtilis; Pneumococcal; Streptococcus, and strains of Haemophilus influenza. It will further be appreciated that when expressed in bacteria, the polypeptide may be part of an inclusion body. Polypeptides must be isolated, purified and then assembled into functional molecules.

In addition to prokaryotes, eukaryotic microorganisms may also be used. Saccharomyces cerevisiae, or common baker's yeast, is the most commonly used eukaryotic microorganism, although many other states are generally available. For expression in Saccharomyces, plasmid YRp7, for example, Stinchcomb et al., Nature, 282: 39 (1979); Kingsman et al., Gene, 7: 141 (1979); Plasmid YRp7 in Tschemper et al., Gene, 10: 157 (1980) is generally used. Such plasmids contain the TRP1 gene, which provides a selection marker for yeast mutants, for example ATCC number 44076 or PEP4-1, which already lack the ability to grow on tryptophan (Jones, Genetics, 85:12). (1977)]). The presence of Trpl damage as a feature of the yeast host cell genome provides an effective environment for detecting transformation by subsequent growth in the absence of tryptophan.

Pharmaceutical Compositions and Methods of Administration

Provided herein are pharmaceutical compositions comprising a humanized anti-human CXCR3 antibody disclosed herein, and a pharmaceutically acceptable carrier.

Methods of making the antibodies or antigen-binding fragments thereof provided herein and methods of administering them to a subject are well known to those skilled in the art or are readily determined by those skilled in the art. Routes of administration of the antibody or fragment thereof include oral, parenteral (eg, intravenous, intramuscular, intraperitoneal, or subcutaneous) administration, inhalation, or topical administration. In some embodiments, the antibodies provided herein are formulated for intravenous administration. In some embodiments, pharmaceutical compositions suitable for injection include buffers (eg, acetate, phosphate or citrate buffers), surfactants (eg, polysorbates), optionally stabilizers (eg, human albumin). And the like.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (if water soluble) or dispersions and sterile powders for the instant preparation of sterile injectable solutions or dispersions. The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyols (eg, glycerol, propylene glycol, liquid polyethylene glycols, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable composition may occur by including an agent in the composition that delays absorption, such as aluminum monostearate and gelatin.

In any case, the sterile injectable solution, in combination with one of the components listed herein or a combination of the above components, incorporates the active compound (eg, an antibody alone or in combination with other active agents) in a suitable amount in the required amount. And, if desired, can then be prepared by filtration sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation include vacuum drying and freeze-drying which produce a powder of the active ingredient + any further desired ingredient from the previously sterile filtered solution. Injectable preparations are processed under sterile conditions according to methods known in the art, filled and sealed into containers such as ampoules, bags, bottles, syringes or vials. The article of manufacture will preferably have a label or package insert indicating that the relevant composition is useful for the treatment of a subject suffering from or susceptible to an autoimmune or neoplastic disorder.

The dose of an antibody or antigen-binding fragment thereof provided herein for treating a condition described above may be determined by the means of administration, the target site, the physiological condition of the patient, whether the patient is human or animal, other drugs administered, and the treatment. It depends on many other factors, including whether it is prophylactic or therapeutic. Usually, the patient is a human, but non-human mammals, including transgenic mammals, can also be treated.

In some embodiments, the dose may range from, for example, about 0.0001 to 100 mg, or 0.01 to 5 mg per kg of host body weight.

The subject may be administered the dose on a daily, every other day, weekly or any other schedule determined by experimental analysis. The antibodies provided herein, or antigen binding fragments thereof, can be administered multiple times. Intervals between single doses can be, for example, daily, weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of the polypeptide or target molecule in the patient.

Antibodies or antigen-binding fragments thereof provided herein may optionally be administered in combination with other agents used to treat a disorder or condition in need of (eg, prophylactic or therapeutic) treatment. Preferred additional agents are known in the art and are administered standardly for certain disorders.

CXCR3 Methods of treating associated diseases or disorders

CXCR3 antibodies, or antigen binding fragments thereof, provided herein are useful for antagonizing CXCR3 activity. In some embodiments, the antibody and antigen binding fragments inhibit CXCR3 binding to one or more ligands such as CXCL9, CXCL10, and / or CXCL11; Inhibit the migration, accumulation, recruitment, or infiltration of CXCR3 ⁺ cells (eg to inflammatory sites); It is used in a method for depleting CXCR3 ⁺ cells. In some embodiments, the antibody and antigen binding fragment are used in a method for depleting CXCR3 ⁺ cells in vivo. CXCR3 ⁺ cells include, but are not limited to, CXCR3 ⁺ / CD4 ⁺ T cells, CXCR3 ⁺ / CD8 ⁺ T cells, and CXCR3 ⁺ / CD19 ⁺ B cell subsets.

In certain embodiments, a method of treating a CXCR3-associated disease or disorder is provided, the method comprising administering to a subject in need thereof a pharmaceutical composition comprising one or more CXCR3 antibodies, or antigen binding fragments thereof. In one embodiment, a method of reducing or treating the progression of T-cell-mediated autoimmune disease is provided. The method includes the step of reducing or treating the progression of T-cell-mediated autoimmune disease by administering to a subject in need thereof a humanized anti-human CXCR3 antibody or antigen binding fragment thereof. In certain embodiments, the T-cell-mediated autoimmune disease is vitiligo. In some embodiments, a subject in need thereof comprises a subject diagnosed with a CXCR3-associated disease or a T-cell-mediated autoimmune disease or a well-developed CXCR3-associated or T-cell-mediated autoimmune disease, This is as described herein.

Subjects to be treated by the methods provided herein can include humans or other mammals. In one embodiment, the subject is a human. In various embodiments, the subject can be treated after the onset of any condition associated with prophylactic or deviant CXCR3 activity or any condition where disruption of CXCR3 signaling can be therapeutically beneficial.

In some embodiments, the subject can be treated prophylactically or after the onset of vitiligo.

A “subject with vitiligo” is any subject with a punctate reduced skin pigment, usually of autoimmune or unknown cause, consistent with the medical diagnosis of vitiligo. The generated area may include the face, hands, feet, arms, legs, lips, or any combination thereof, but may include other areas, such as the neck or torso. In some cases, there is a reduction in hair pigment.

Example

The antibodies, compositions, and methods are further illustrated by the following examples, which should not be construed as further limiting. The contents of the Sequence Listing, the drawings, and all references, the patents cited throughout the present application, and published patent applications are expressly incorporated herein by reference.

Example 1: Generation of Humanized Anti-CXCR3 Monoclonal Antibodies

Anti-CXCR3 monoclonal antibodies were generated as described in International Publication No. 2013/109974. Humanized clone 53 monoclonal antibodies were generated as described below.

A humanized version of clone 53 was prepared that can induce depletion of CXCR3-expressing cells. Humanized clone 53 monoclonal antibodies with VH and VK sequences as shown in Table 4 were generated.

The germline index scores of each humanized variant are shown in Table 4 . The heavy chain is compared with the IGHV3-23 * 03 / IGHJ4 * 03 germline sequence; The light chain is compared to the IGKV1-9 * 01 / IGKJ2 * 01 germline sequence.

TABLE 4

The binding properties of each humanized monoclonal antibody provided herein are illustrated in Table 5 .

TABLE 5

Table 6 shows the binding properties and wiring properties of clone 53 (53), chimeric clone 53 (Ch53) and humanized version of clone 53 (53Hu1-53Hu20).

TABLE 6

As the data indicate, the humanized antibodies provided herein have significantly improved binding properties while having an advantageous germline index.

Example 2 CDR Optimization

Many VH CDRs and / or VL CDR variants were prepared. Binding affinity to recombinant human CXCR3 was measured using Biacore. Mutants with as strong binding for at least 53Hu37 are shown in Table 3.

Example 3 : CXCR3-173 is a depleting antibody.

Hamster anti-mouse CXCR3 monoclonal (clone CXCR3-173) was used as surrogate antibody in preclinical experiments. CXCR3-173 was previously disclosed as a blocking antibody that does not deplete CD4 + T cells in vivo. (Uppaluri et al., Transplantation 86: 137-47 (2008)).

The following Fc variants of hamster CXCR3-173 mAb were prepared to test the effector function of the antibodies: unglycosylated N297G variant of mouse IgG1 (CXCR3 mIgG1 agly); Wild type mouse IgG2a chimera (CXCR3 mIgG2a WT); Mouse IgG2a chimera (CXCR3 mIgG2a Dab) modified to abolish the depletion capacity; And wild type mouse IgG3 (CXCR3 mIgG3).

Blood was collected after 24 h for 8-10 week old C57BL / 6 mice (Jackson Laboratories, n = 5 / group) with a single 5 mg / kg intravenous dose antibody followed for flow cytometry analysis using the following markers: TCRab, CD4 and CD8 T cells by CD4, and CD8; Memory CD4 and CD8 T cells by TCRab, CD4, CD8, CD62L and CD44. Cells were quantified using Count Bright bead (Invitrogen) according to the manufacturer's protocol to determine total cells per microliter of blood.

Gating results in total T lymphocytes and T lymphocyte subsets are shown in FIG. 1. As shown in the figure, it was demonstrated for the first time that hamster CXCR3-173 was depleted in CD4 + and CD8 + memory T cells when administered to mice.

Example 4 Effector Function of Fc Variants of CXCR3-173

Mouse Fcγ receptor binding of the Fc-engineered version of CXCR3-173 was assessed using an antibody capture approach using the Biacore 3000 instrument. Recombinant protein A / G (Pierce) was covalently immobilized on a CM5 sensor chip using amine chemistry. CXCR3-173 antibody was diluted to 5 μg / mL in HBS-EP buffer and injected into Protein A / G chips for 30 seconds at a flow rate of 10 μL / min. Recombinant mouse FcγRI (CD64), FcγRIIb (CD32), FcγRIII (CD16), and FcγIV (CD16-2) from R & D Systems were diluted three-fold from 300 nM to 3.7 nM in HBS-EP buffer and flowed at 30 μL / min. Injected in duplicate into the antibody captured. The surface was regenerated with glycine 2.0 (GE Healthcare). Binding reactions were normalized to the RU amount of protein A / G capture. The results are shown in Figs. 2A to 2D.

As shown in the figure, hamster CXCR3-173 modified to have wild-type murine IgG2a isotypes binds to all four recombinant mouse (rm) Fcγ receptors, but dAB mutations significantly reduce this binding. CXCR3-173 modified to have wild-type murine and IgG3 isotypes also bind to all four recombinant mouse Fcγ receptors. The original unmodified hamster CXCR3-173 binds better to rmFcγRIIb and rmFcγRIII than the IgG2a isotype variant, and the unglycosylated mIgG1 isotype variant does not bind any rmFcγR.

Example 5 Effector Function in Vitro

Humanized anti-CXCR3 mAb and its Fc-engineered version were studied in a series of ADCC assays. Fc-engineered versions of the humanized anti-CXCR3 mAb were prepared using standard methods. Defucosylated versions were prepared by culturing cells expressing humanized mAb in the presence of kifunensine.

ADCC analysis was performed using NK9.2 cell line (Conkwest) overexpressing primary human NK cells or valine polymorphism (Conkwest) as effector cells and CHO transfected cells (A isotype) overexpressing human CXCR3 as target cells. It was.

For assays where primary NK cells were used as effectors, NK cells were purified from leucopak of normal donors and incubated for 24 h in IL-2, followed by CHO-human CXCR3 target cells labeled overnight with chromium 5: Plated at E: T ratio of 1. Cultures were incubated in a tissue culture incubator for 3 hours and then washed, dissolved in 1% Triton-X and the supernatants read on a gamma counter.

For assays in which NK9.2 cells were used as effectors, NK9.2 cells were grown for two weeks in IL-2 according to the manufacturer's recommendations. On the day of analysis, NK9.2 cells (70,000 cells) were labeled with calcein AM (Invitrogen) and incubated with appropriately diluted antibody for 30 minutes to bind antibodies to CXCR3 on target cells. NK cells were plated with an effector to target cell ratio of 3: 1 and the cultures were incubated for 1 hour in a tissue culture incubator. Cells were lysed with Triton X-100 at the end of the culture period and plates were read using an M5 plate reader (492 nm excitation and 515 nm emission).

Human IgG1 (Sigma) was used as a negative control and lysis of CD52-overexpressing CHO cells treated with alemtuzumab (monoclonal anti-CD52 antibody) served as a positive control for lysis. The signal is expressed in arbitrary fluorescence units (AFU). Percent toxicity is expressed by (experimental dissolution-spontaneous dissolution) / (maximum dissolution-spontaneous dissolution) x 100%.

Humanized anti-CXCR3 mAb 53Hu37 and Fc-engineered versions with human IgG1 Fc were tested in ADCC assays. Fc-engineered versions M1 (S239D / D332E (EU notation), M2 (G236A / S267E / H268F / S324T / I332E (EU notation), "AEFTE"), and M3 (S239D / H268F / S324T / I332E (EU notation) , “DFTE”) contain amino acid changes that allow for enhanced ADCC or CDC activity A fourth Fc-engineered version was generated by kifunensin treatment of cell lines making wild type antibodies.

Anti-human CXCR3 clones Clone 4 (CXCR3 CL4), Clone 12 (CXCR3 CL12), Clone 82 (CXCR3 CL82), Clone 135 (CXCR3 CL135) were also tested. In independent experiments, the assay was performed using different effector cells, effector: target (E: T) ratios and concentrations of antibody. Representative results are shown in FIGS. 3B and 3C.

3A summarizes the effector function of the M1, M2, M3, and afufusylated versions of 53H37. 3B shows the results of the assay using primary NK cells as effectors. 3C shows the results of the assay using NK9.2 cells as effectors.

Example 6 Fcγ Receptor Binding

Human and mouse Fcγ receptor binding affinities of Fc-engineered versions of humanized anti-CXCR3 mAb 53Hu37 and 53Hu37 were assessed using Biacore T200 equipment. Protein A from Sigma was immobilized on CM5 series S chips using amine chemistry. Antibodies were injected into the Protein A chip and various concentrations of recombinant human and mouse Fcγ receptors (R & D Systems) were injected into the captured antibody. A wide range of receptors were used to span the low and high affinity binders (1.2 nM to 5 μM). Each sample was injected in duplicate. Binding sensorgrams were fitted to a 1: 1 kinetic binding model. Quantitative results are summarized in FIG. 4.

As shown in FIG. 4, the M1 and M3 Fc-engineered versions had improved affinity for both hFcγRIII and mFcγRIV, M2 had increased binding to hFcγRIIa, and kifunensine-treated 53H37 was Fc -Moderate increases in hFcγRIII and mFcγRIV compared to the engineered version.

Example 7 :

Cynomolgus monkeys were given a single intravenous infusion of 2 mg 53Hu37, M1 variant, or kifunensine-treated version (n = 8 / antibody treated group), or vehicle control (n = 6) per kg body weight. . Blood samples were collected and analyzed by flow cytometry for total T cells and T-cell subsets prior to and 1 day after infusion, 3 days, 7 days, and 14 days. For flow cytometry, cells were stained with antibodies against the following markers to lyse red blood cells and identify total and memory CD4 and CD8 T cells: CD3 (clone SP34-2), CD4 (clone OKT4), CD8a (clone RPA). -T8), CD45RA (clone 5H9), CCR7 (clone G043H7), and CXCR3 (clone 1C6). Cells / microliters (cells / μL) were determined by quantifying cells using count bright beads (Invitrogen). The data is expressed as the average percentage of the value of pretreatment blood in each cell subset of the group over time. Histological characteristics of spleen samples obtained from a subset of each treatment group at 14 days post-injection were studied by staining fixed sample sections for CXCR3 using anti-human CXCR3 clone 4 antibody and appropriate secondary antibody. The results are shown in Figs. 5A to 5C. Serum from blood samples was also analyzed for pharmacokinetic properties by measuring circulating concentrations of the administered antibodies using peptide ELISA.

As shown in the figure, M1 version of 53Hu37, 53Hu37, and Kif-treated 53Hu37 reduced effector memory CD4 T cells, and 53Hu37, M1 version of Hu37, and Kif-treated 53Hu37 reduced effector memory CD8 T cells I was. In addition, the M1 version of 53Hu37, and Kif-treated 53Hu37, dramatically reduced CXCR3 staining in the spleen 14 days after a single intravenous infusion of antibody.

Example 8 Biochemical Analysis

Thermal stability, stability to shear stress and virus inactivation, stability to freeze-thaw stress, stability to agitation stress and pH stability were measured for 53Hu37, M1 version and kifunensine-treated 53Hu37.

Differential Scanning Calorimetry (DSC)

Samples were analyzed using high treatment VP-DSC (Microcal). Samples were diluted to approximately 0.5 mg / mL with corresponding buffers and loaded onto 96 well plates. Scan parameters consisted of a start temperature of 25 ° C. and a final temperature of 100 ° C. A scan rate of 200 ° C./h was used.

Turbidity

Sample absorbance at 340-360 nm in 5 nm increments was measured on Spectramax Plus (Molecular Devices) and averaged to obtain final haze measurements. 96-well UV plate plates were used with 150-200 μL of material. The plate was read before the sample addition and a path check was applied to the sample values. To classify the degree of turbidity, see "A Turbidimetric Method to Determine Visual Appearances of Protein Solutions" by Brigitte Eckhardt, Technology Applications Vol. 48, No. 2 Mar.-Apr. 1994 was compared with values obtained from the UV absorbance of the turbidity standard.

Size Exclusion Chromatography (SEC)

HP1100 or 1200 series systems were equipped with a TSK SW _XL size exclusion column combined with a SW _XL guard column. Samples were run for 35 minutes using a mobile phase of 20 mM sodium phosphate, 500 mM NaCl, pH 6.0. A flow rate of 0.5 mL / min was used. 50 μg was injected and the UV signal was monitored at 280 nm.

Heat-induced relative cohesion trend (TI-RAP)

0.2 mg / mL anti-human CXCR3 antibody (individual or antibody mixture) was 10 minutes in PBS buffer and 10 mM histidine and 9% sucrose buffer at 5 ° C (control), 64 ° C, 67 ° C, 70 ° C, or 73 ° C Incubation during the production of temperature-induced aggregation. After heat incubation, the samples were centrifuged at 7000 × g for 2 minutes at 5 ° C. to remove insoluble protein precipitates and the supernatants were analyzed by cation exchange chromatography (CEX). The percentage of soluble monomer (and relative aggregation trend) was calculated by normalizing the chromatographic peak area of the thermally stressed sample using the peak area of the control (5 ° C.) sample.

Agitation-induced relative cohesion trend (AI-RAP)

For agitation-induced relative aggregation trend, 9% water with 0 and 0.01% polysorbate 80 and a solution containing 0.2 mg / mL final protein concentration of anti-human CXCR3 antibody in PBS buffer and 10 mM histidine at 5 ° C. The cross buffer was subjected to vigorous stirring stress at 5 ° C. A solution of anti-human CXCR3 antibody was stirred at maximum speed from VX-2500 Multi-Tube Vortexer (VWR, West Chester, Pa.) For a total of 24 h.

Various time points were analyzed throughout the study by taking out a small sample aliquot for CEX analysis. Sample aliquots were centrifuged at 7000 × g (5 ° C.) for 2 minutes to remove insoluble precipitate and the supernatants were analyzed by CEX. The percentage of soluble monomers (and relative aggregation trend) was calculated by normalizing the chromatographic peak area of the stirred sample using the peak area of the control (0 h) sample.

Cation Exchange Chromatography (CEX)

CEX analysis was performed on an Agilent 1290 infinity HPLC system at 25 ° C. using a ProPac WCX-10 analytical column (weak cation exchange, 4 × 250 mm, Thermo Scientific). 20 microgram protein samples were loaded on the column and analyzed at a flow rate of 0.8 mL / min. Equilibrate the column with Buffer A (20 mM Sodium Acetate, 0.0025% Sodium Azide, pH 5.2) and protein in linear gradient from 0 to 100% Buffer B (20 mM Sodium Acetate, 1 M Sodium Chloride, 0.0025 over 40 minutes). % Sodium azide, pH 5.2). The absorbance at 280 nm was measured and the protein peak area was determined by integrating the 280 nm absorbance peak.

Experimental Design

pH / temperature stress

Some of the material from each clone was dialyzed into 20 mM sodium phosphate at pH 5.0 and sodium phosphate at pH 7.0 using Slide-A-Lyzers (Thermo Scientific PN 66810). This material was diluted with the corresponding buffer and filtered at approximately 2 mg / mL using Millex GV filter (Millipore PN SLGV033RB). Samples in sodium phosphate at pH 5 and pH 7 were stored at 37 ° C. and tested at 3 and 5 weeks.

Freeze-thaw stress

The indicated anti-human CXCR3 antibodies were dialyzed into 20 mM sodium phosphate at pH 6.0 using Slide-A-Lyzers (Thermo Scientific PN 66810). This material was diluted with the corresponding buffer and filtered at approximately 1 mg / mL using Millex GV filter (Millipore PN SLGV033RB). Freeze-thaw Stress Samples were frozen five times at −80 ° C. and thawed at room temperature. After one and three freeze-thaw cycles, material was removed for testing by selection assay. Complete assay tests were performed after the final freeze-thaw.

Shear stress

Antibody samples were dialyzed into 20 mM sodium phosphate at pH 6.0 using Slide-A-Lyzers (Thermo Scientific PN 66810). This material was diluted with the corresponding buffer and filtered at approximately 1 mg / mL using Millex GV filter (Millipore PN SLGV033RB). Shear stress samples were repeatedly pipetted into 200 μL pipettes a total of 50 times.

Mock virus inactivation

Antibody samples were dialyzed into 20 mM sodium phosphate at pH 6.0 using Slide-A-Lyzers (Thermo Scientific PN 66810). This material was diluted with the corresponding buffer and filtered at approximately 1 mg / mL using Millex GV filter (Millipore PN SLGV033RB). The virus inactivation sample was brought to pH 3.5 with 1 N HCl and kept at that pH for 100 minutes at room temperature. Following the maintenance period, the sample was brought back to pH 7.2 with 1 N NaOH and held for 100 minutes. The sample was then brought to pH 6.0 with 1 N HCl and tested.

result

Differential scanning calorimetry

The M1 version (D / E mutant) was found to be slightly less stable than the Kif version, and the Kif version was slightly less stable than 53Hu37.

Shear stress:

The M1 version (D / E mutant) was found to be slightly less stable than the 53H37 and Kif versions, with the latter two being approximately equally stable.

Virus Inactivation Stress:

The M1 version (D / E mutant) was found to be more stable than the Kif version, both less stable than 53Hu37.

Freeze-thaw stress:

The M1 version (D / E mutant), Kif version, and 53Hu37 were found to be approximately equivalent.

Agitation-induced relative cohesion trend:

Approximately 80% of all versions of the antibody precipitated within 2 hours of stirring, but at pH 5.5, the M1 version (D / E mutant) was relatively more stable than the other two versions.

Aggregate Formation:

Under control (non-accelerated) conditions at pH 5.0, the M1 version (D / E mutant) and 53 Hu37 were essentially stable over a period of 5 weeks, but the Kif version became milky; This same pattern was found under accelerated conditions. Under control (non-accelerated) conditions at pH 7.0, the three antibody versions were approximately identical over a five week period. Under accelerated conditions, DE mutants were slightly more stable than 53H37 and 53Hu37 were more stable than the Kif version.

Example 9: Depleted anti-CXCR3 is protective and therapeutic in mouse models of vitiligo, but non-depleted anti-CXCR3 is not.

mouse. KRT14-Kitl * 4XTG2Bjl (Krt14-Kitl *) mice were donations from BJ Longley of the University of Wisconsin. PMEL TCR transgenic mice are Thy1.1 ⁺ and were obtained from The Jackson Laboratory (Stock No. 005023, B6.Cg Thy1 ^a / CyTg (TcraTcrb) 8Rest / J). All mice were set against C57BL / 6J and maintained in a pathogen free facility and procedures were performed according to the NIH Guide for the Care and Use of Laboratory Animals.

Antibodies. deltaAB CXCR3 is an antibody that neutralizes CXCR3 activity (also referred to herein as IgG2a-dAB, dAB, or ΔAB). WT mouse CXCR3 is an antibody with wild type mouse IgG2a Fc with depleting activity.

Induction of vitiligo and antibody treatment. Vitiligo was induced through adoptive transfer of pre-melanosome specific (PMEL) CD8 ⁺ T cells as described previously. Harris et al., J. Invest. Dermatol. 132: 1869-76 (2012). Briefly, PMEL CD8 ⁺ T cells were isolated from the spleen of PMEL TCR transgenic mice (Jackson Laboratory, Stock No. 005023) via negative selection on microbeads (Miltenyi Biotech) according to the manufacturer's instructions. Purified CD8 ⁺ T cells (1 × 10 ⁶ ) were injected intravenously into a lethal dose (500 rad one day before delivery) Krt14-Kitl * host (Jackson Laboratory Stock No. 009687, 8-12 weeks old). Receptive mice also received intraperitoneal injections of 1 × 10 ⁶ pfu of rVV-hPMEL (N Restifo, NCI, NIH) on the same delivery day.

Treatment was performed by intraperitoneal injection of 100 μg of antibody or isotype three times weekly for the duration of the observation period.

For prophylactic studies, mice were treated from weeks 2 to 6 after induction of vitiligo. This time is significant because it occurs after the removal of the virus used to induce the disease and before the onset of autoimmunity. Vitiligo scores were quantified using a point scale based on the degree of depigmentation at four easily visible locations including ear, nose, hind paw, and tail as previously described. Harris et al., J. Invest. Dermatol. 132: 1869-76 (2012). Investigate each site and estimate the degree of depigmentation as a percentage of the dissection site; Both the left and right ears and the left and right hind paws were estimated together and thus evaluated as a sweetwill site. Points were awarded as follows: No evidence of depigmentation (0%) received a score of 0,> 0-10% = 1 point,> 10-25% = 2 points,> 25-75% = 3 Points,> 75 to <100% = 4 points, and 100% = 5 points. The "vitiligo score" was the sum of scores at all four sites with a maximum score of 20 points.

In the pigmentation study, mice with greater than 75% depigmentation in the tail were treated with WT mouse CXCR3 depleted Ab or isotype controls from weeks 12-20 after induction of vitiligo if the disease was normally stable. Pigmentation assays were performed using ImageJ as previously described. Agarwal et al., J. Invest. Dermatol. 135: 1080-8 (2015).

Effect of CXCR3 Antibody on Cell Counts in Blood and Spleen. Mice were injected with 100 μg of each antibody and counted memory CD8 ⁺ T cells from blood (cells per microliter determined using Countbright beads) and spleen (total cell count using Via-Cell XR instrument from Becton Coulter). Was evaluated at 24 hours. The dosed antibody is essentially the same antibody (same CDRs) as the antibody used for CXCR3 staining, and memory CD8 ⁺ T cells (as defined by CD44 and CD62L staining) because they can block binding of the staining antibody to CXCR3. Depletion was assessed using the CXCR3-independent scale by evaluating the number of (because more than 50% of these cells express CXCR3).

Flow cytometry. Blood, tail, spleen, and skin-drained lymph nodes were harvested and analyzed via flow cytometry. The spleens were destroyed and erythrocyte cells were lysed using RBC lysis buffer (Sigma Aldrich). Blood samples were stained and treated with Biolegend lysis / fix buffer (according to manufacturer's protocol). For separation of dermis and epidermis, tail skin samples were incubated with 5 U mL ^-1 of Dispase II (Roche) at 37 ° C. for 1 hour. 1 mg mL to remove the epidermis, destroyed mechanically with a 70 μm cell strainer (strainer), and including a DNAse I (Sigma-Aldrich) in 1 ~ 2 mg mL ^-1 for 1 hour dermal sample in 37 ℃ ^- Incubation with ¹ collagenase IV. All cells were filtered through a 70 μm mesh and analyzed. To identify PMEL and its phenotypes, samples were stained with 1: 200 of the following antibodies in FACS buffer (1% BSA in PBS): CD45 AF700, CD8b PerCP-Cy5.5, Thy1.1 Pacific Blue, ( Biolegend). Live / Dead Blue (Invitrogen) was used as 1: 1000 in FACS buffer to distinguish live cells. Data was collected and analyzed using a BD LSR II flow cytometer (BD Biosciences) and FlowJo software version 10 (Tree Star, Inc.).

result. Effect of CXCR3 Antibody on Cell Counts in Blood and Spleen. In blood and spleen, the total number of memory CD8 ⁺ T cells was significantly reduced in WT mice or mice treated with hamster CXCR3 antibodies, while the number of cells did not change in mice treated with deltaAB CXCR3 (untreated mice and In comparison).

prevention. Mice were treated with hamster anti-mouse CXCR3 (depleted), WT mouse CXCR3 (excellent depleted), deltaAB CXCR3 (neutralized; non-depleted), or isotype control antibodies. Of these, the depleted antibody WT mouse CXCR3 worked best in the prevention of clinical disease, as evidenced by a significantly reduced score after 5 weeks of treatment (one-way ANOVA p = 0.0066, Dunnett's post-test for isotype ns; Tukey's post-test is significant for ΔAB for Wt and ΔAB for hamsters. This observation is consistent with data indicating that WT mouse CXCR3 Ab has better depletion activity than hamster CXCR3 Ab. All Abs tested in vitiligo prevention in mice led to less PMEL in the skin. PMEL numbers were significantly reduced in epidermis after treatment with any CXCR3 Ab (two-way ANOVA p <0.0001 by Bonferroni's post-test compared to isotype control) In the dermis, there was a decreasing trend. However, neutralizing antibodies, despite their ability to reduce the number of PMELs in the skin, were less effective than depleted antibodies, perhaps due to the fact that the low threshold number of PMELs was sufficient for the overall clinical disease. The effect of CXCR3 depleted antibodies on host T cells was assessed by measuring the total number of CD45 ⁺ cells and host CD8 ⁺ T cells in lymph nodes (LN), spleen, blood, epidermis, and dermis. The number of bypass host CD8 ⁺ T cells was significantly reduced in the spleen and lymph nodes (LN) by hamster or WT mouse CXCR3 Ab treatment, but not in the skin (binary dispersion by Bonferroni's post-test compared to isotype control). Analysis p <0.0001). However, the total number of CD45 ⁺ cells did not change after treatment with any CXCR3 Ab.

Pigment redeposition. Mouse WT CXCR3 Ab treatment significantly reversed clinical disease and reduced the number of PMEL in the epidermis. The number of host CD8 ⁺ T cells decreased slightly in treated mice, but the total CD45 ⁺ cell number did not change. Taken together, these data indicate that CXCR3 depleted Abs can reverse disease and reduce autoreactive T cell counts with minimal effect on other compartments of the immune system.

The foregoing embodiments are intended to be illustrative and do not limit the invention in any way. Other embodiments of the disclosed compounds and methods will become apparent to those skilled in the art upon consideration of the specification and the practice of the compounds and methods disclosed herein.

                         SEQUENCE LISTING <110> SANOFI   <120> ANTI-HUMAN CXCR3 ANTIBODIES FOR TREATMENT OF VITILIGO <130> 597513: SA9-197PC <150> 62 / 437,889 <151> 2016-12-22 <150> EP 17306770.3 <151> 2017-12-14 <160> 141 <170> PatentIn version 3.5 <210> 1 <211> 368 <212> PRT <213> Homo sapiens <400> 1 Met Val Leu Glu Val Ser Asp His Gln Val Leu Asn Asp Ala Glu Val 1 5 10 15 Ala Ala Leu Leu Glu Asn Phe Ser Ser Ser Tyr Asp Tyr Gly Glu Asn             20 25 30 Glu Ser Asp Ser Cys Cys Thr Ser Pro Pro Cys Pro Gln Asp Phe Ser         35 40 45 Leu Asn Phe Asp Arg Ala Phe Leu Pro Ala Leu Tyr Ser Leu Leu Phe     50 55 60 Leu Leu Gly Leu Leu Gly Asn Gly Ala Val Ala Ala Val Leu Leu Ser 65 70 75 80 Arg Arg Thr Ala Leu Ser Ser Thr Asp Thr Phe Leu Leu His Leu Ala                 85 90 95 Val Ala Asp Thr Leu Leu Val Leu Thr Leu Pro Leu Trp Ala Val Asp             100 105 110 Ala Ala Val Gln Trp Val Phe Gly Ser Gly Leu Cys Lys Val Ala Gly         115 120 125 Ala Leu Phe Asn Ile Asn Phe Tyr Ala Gly Ala Leu Leu Leu Ala Cys     130 135 140 Ile Ser Phe Asp Arg Tyr Leu Asn Ile Val His Ala Thr Gln Leu Tyr 145 150 155 160 Arg Arg Gly Pro Pro Ala Arg Val Thr Leu Thr Cys Leu Ala Val Trp                 165 170 175 Gly Leu Cys Leu Leu Phe Ala Leu Pro Asp Phe Ile Phe Leu Ser Ala             180 185 190 His His Asp Glu Arg Leu Asn Ala Thr His Cys Gln Tyr Asn Phe Pro         195 200 205 Gln Val Gly Arg Thr Ala Leu Arg Val Leu Gln Leu Val Ala Gly Phe     210 215 220 Leu Leu Pro Leu Leu Val Met Ala Tyr Cys Tyr Ala His Ile Leu Ala 225 230 235 240 Val Leu Leu Val Ser Arg Gly Gln Arg Arg Leu Arg Ala Met Arg Leu                 245 250 255 Val Val Val Val Val Val Ala Phe Ala Leu Cys Trp Thr Pro Tyr His             260 265 270 Leu Val Val Leu Val Asp Ile Leu Met Asp Leu Gly Ala Leu Ala Arg         275 280 285 Asn Cys Gly Arg Glu Ser Arg Val Asp Val Ala Lys Ser Val Thr Ser     290 295 300 Gly Leu Gly Tyr Met His Cys Cys Leu Asn Pro Leu Leu Tyr Ala Phe 305 310 315 320 Val Gly Val Lys Phe Arg Glu Arg Met Trp Met Leu Leu Leu Arg Leu                 325 330 335 Gly Cys Pro Asn Gln Arg Gly Leu Gln Arg Gln Pro Ser Ser Ser Arg             340 345 350 Arg Asp Ser Ser Trp Ser Glu Thr Ser Glu Ala Ser Tyr Ser Gly Leu         355 360 365 <210> 2 <211> 255 <212> PRT <213> Homo sapiens <400> 2 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 3 <211> 255 <212> PRT <213> Synthetic Peptide <400> 3 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Ala Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 4 <211> 255 <212> PRT <213> Synthetic Peptide <400> 4 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Asp         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 5 <211> 255 <212> PRT <213> Synthetic Peptide <400> 5 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Glu His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 6 <211> 255 <212> PRT <213> Synthetic Peptide <400> 6 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser Phe Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 7 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 7 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 8 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 8 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 9 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 9 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Asp         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser His Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 10 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 10 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Ala Gly Pro Ser         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Glu Phe Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 11 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 11 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 1 5 10 15 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr             20 25 30 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Asp         35 40 45 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg     50 55 60 Thr Pro Glu Val Thr Cys Val Val Asp Val Ser Phe Glu Asp Pro 65 70 75 80 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala                 85 90 95 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val             100 105 110 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr         115 120 125 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Glu Glu Lys Thr     130 135 140 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 145 150 155 160 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys                 165 170 175 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser             180 185 190 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp         195 200 205 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser     210 215 220 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 225 230 235 240 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly                 245 250 255 <210> 12 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 12 Gly Phe Thr Phe Thr Ser Tyr Ala 1 5 <210> 13 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 13 Ile Ser His Gly Gly Thr Tyr Thr 1 5 <210> 14 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 14 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 15 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 15 Ser Gly Val Asn Tyr 1 5 <210> 16 <211> 3 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 16 Phe Thr Ser One <210> 17 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 17 Gln Gln Phe Thr Ser Ser Pro Tyr Thr 1 5 <210> 18 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 18 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Val Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 19 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 19 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 20 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 20 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 21 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 21 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Pro Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 22 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 22 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Pro Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 23 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 23 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 24 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 24 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 25 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 25 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 26 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 26 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Glu Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 27 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 27 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 28 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 28 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 29 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 29 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Glu Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 30 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 30 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 31 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 31 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Val Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 32 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 32 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Val Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Glu Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 33 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 33 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Val Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Asp Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser Phe Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Thr Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Glu Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 34 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 34 Gly Phe Thr Phe Arg Ser Tyr Ala 1 5 <210> 35 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 35 Ile Ser His Lys Gly Thr Tyr Thr 1 5 <210> 36 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 36 Ile Ser His Lys Gly Lys Tyr Thr 1 5 <210> 37 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 37 Ile Ser His Arg Gly Thr Tyr Thr 1 5 <210> 38 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 38 Ile Ser His Arg Gly Arg Tyr Thr 1 5 <210> 39 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 39 Ile Ser Arg Gly Gly Thr Tyr Thr 1 5 <210> 40 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 40 Ile Ser Ser Gly Gly Thr Tyr Thr 1 5 <210> 41 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 41 Ile Ser His Gly Gly Lys Tyr Thr 1 5 <210> 42 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 42 Ile Ser His Gly Gly Arg Tyr Thr 1 5 <210> 43 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 43 Ile Ser His Gly Gly Trp Tyr Thr 1 5 <210> 44 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 44 Ile Ser His Gly Gly Tyr Tyr Thr 1 5 <210> 45 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 45 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 46 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 46 Ala Arg His Pro Ile Tyr Ser Gly Trp Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 47 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 47 Ala Arg His Pro Ile Tyr Ser Gly Gln Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 48 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 48 Ala Arg His Pro Ile Tyr Ala Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 49 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 49 Ala Arg His Pro Ile Tyr Cys Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 50 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 50 Ala Arg His Pro Ile Tyr Asp Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 51 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 51 Ala Arg His Pro Ile Tyr Glu Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 52 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 52 Ala Arg His Pro Ile Tyr Phe Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 53 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 53 Ala Arg His Pro Ile Tyr Gly Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 54 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 54 Ala Arg His Pro Ile Tyr His Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 55 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 55 Ala Arg His Pro Ile Tyr Ile Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 56 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 56 Ala Arg His Pro Ile Tyr Lys Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 57 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 57 Ala Arg His Pro Ile Tyr Leu Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 58 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 58 Ala Arg His Pro Ile Tyr Met Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 59 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 59 Ala Arg His Pro Ile Tyr Asn Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 60 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 60 Ala Arg His Pro Ile Tyr Pro Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 61 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 61 Ala Arg His Pro Ile Tyr Gln Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 62 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 62 Ala Arg His Pro Ile Tyr Arg Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 63 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 63 Ala Arg His Pro Ile Tyr Thr Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 64 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 64 Ala Arg His Pro Ile Tyr Val Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 65 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 65 Ala Arg His Pro Ile Tyr Trp Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 66 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 66 Ala Arg His Pro Ile Tyr Tyr Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 67 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 67 Ile Ser His Gly Gly Arg Tyr Thr 1 5 <210> 68 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 68 Ala Arg His Pro Ile His Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr 1 5 10 15 <210> 69 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 69 Ser Gly Val Ile Tyr 1 5 <210> 70 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 70 Ser Gly Val Lys Tyr 1 5 <210> 71 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 71 Ser Gly Val Arg Tyr 1 5 <210> 72 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 72 Ser Gly Val Ser Tyr 1 5 <210> 73 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 73 Ser Gly Val Trp Tyr 1 5 <210> 74 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 74 Ser Gly Val Tyr Tyr 1 5 <210> 75 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 75 Gln Gln Phe Thr Arg Ser Pro Tyr Thr 1 5 <210> 76 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 76 Gln Gln Phe Thr Ser Lys Pro Tyr Thr 1 5 <210> 77 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 77 Gln Gln Phe Lys Ser Ser Pro Tyr Thr 1 5 <210> 78 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 78 Gln Gln Phe Arg Ser Ser Pro Tyr Thr 1 5 <210> 79 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 79 Gln Gln Phe Tyr Ser Ser Pro Tyr Thr 1 5 <210> 80 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 80 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Lys Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 81 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 81 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Lys Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 82 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 82 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Lys Gly Lys Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 83 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 83 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Arg Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 84 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 84 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Arg Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 85 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 85 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Arg Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Trp Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 86 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 86 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Arg Gly Arg Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 87 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 87 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Arg Gly Arg Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 88 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 88 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser Arg Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 89 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 89 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser Arg Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Gln Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 90 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 90 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser Ser Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 91 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 91 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 92 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 92 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Trp Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 93 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 93 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ala Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 94 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 94 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Cys Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 95 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 95 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Asp Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 96 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 96 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Glu Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 97 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 97 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Phe Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 98 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 98 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Gly Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 99 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 99 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr His Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 100 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 100 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ile Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 101 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 101 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Lys Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 102 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 102 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Leu Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 103 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 103 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Met Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 104 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 104 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Asn Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 105 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 105 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Pro Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 106 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 106 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Gln Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 107 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 107 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Arg Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 108 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 108 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Thr Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 109 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 109 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Val Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 110 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 110 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Trp Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 111 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 111 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Tyr Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 112 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 112 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 113 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 113 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Lys Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 114 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 114 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Lys Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Trp Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 115 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 115 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Arg Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 116 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 116 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Arg Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Phe Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 117 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 117 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Trp Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 118 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 118 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Tyr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 119 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 119 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Lys Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 120 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 120 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile His Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 121 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 121 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Arg Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 122 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 122 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Trp Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser         115 120 <210> 123 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 123 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Ile Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 124 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 124 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Lys Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 125 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 125 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Arg Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 126 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 126 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Arg Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Arg Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 127 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 127 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Ser Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 128 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 128 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Trp Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 129 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 129 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Tyr Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 130 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 130 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Arg Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 131 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 131 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Lys Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 132 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 132 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Lys Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 133 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <133> 133 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Arg Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 134 <211> 106 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 134 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Tyr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys             100 105 <210> 135 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 135 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro             100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr         115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys     130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser                 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala             180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe         195 200 205 Asn Arg Gly Glu Cys     210 <210> 136 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 136 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Ser Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro             100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr         115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys     130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser                 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala             180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe         195 200 205 Asn Arg Gly Glu Cys     210 <210> 137 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 137 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Pro Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro             100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr         115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys     130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser                 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala             180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe         195 200 205 Asn Arg Gly Glu Cys     210 <210> 138 <211> 213 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 138 Asp Ile Gln Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Ser Gly Val Asn Tyr Leu             20 25 30 Tyr Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Trp Ile Tyr         35 40 45 Phe Thr Ser Thr Leu Ala Pro Gly Val Pro Ser Arg Phe Ser Gly Ser     50 55 60 Gly Ser Gly Asn Glu Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Thr Ser Ser Pro Tyr Thr                 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro             100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr         115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys     130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser                 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala             180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe         195 200 205 Asn Arg Gly Glu Cys     210 <139> <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 139 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 140 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 140 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Val Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Ala Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450 <210> 141 <211> 452 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Peptide <400> 141 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Thr Ser Tyr             20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val         35 40 45 Ala Thr Ile Ser His Gly Gly Thr Tyr Thr Tyr Tyr Pro Asp Ser Val     50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys                 85 90 95 Ala Arg His Pro Ile Tyr Ser Gly Asn Tyr Gln Gly Tyr Phe Asp Tyr             100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly         115 120 125 Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly     130 135 140 Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 145 150 155 160 Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe                 165 170 175 Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val             180 185 190 Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val         195 200 205 Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys     210 215 220 Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 225 230 235 240 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr                 245 250 255 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Asp Val             260 265 270 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val         275 280 285 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser     290 295 300 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 305 310 315 320 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala                 325 330 335 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro             340 345 350 Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln         355 360 365 Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala     370 375 380 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 385 390 395 400 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu                 405 410 415 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser             420 425 430 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser         435 440 445 Leu Ser Pro Gly     450

Claims (32)

Humanized anti-human CXC motif chemokine receptor 3 (CXCR3) antibody comprises a heavy chain (HC) having a heavy chain variable region (VH) and a light chain (LC) having a light chain variable region (VL),
a) VH and VL are SEQ ID NOs: 24/20, 22/18, 80/130, 81/24, 82/130, 82/24, 83/126, 83/130, 83/24, 84/126, 84 / 24, 85/24, 86/126, 87/126, 87/133, 87/24, 88/24, 89/24, 90/126, 91/126, 91/130, 92/130, 92/24 , 93/126, 94/126, 95/126, 96/126, 97/126, 98/126, 99/126, 100/126, 101/126, 102/126, 103/126, 104/126, 105 / 126, 106/126, 107/126, 108/126, 109/126, 110/126, 111/126, 121/24, 113/130, 113/24, 114/130, 115/126, 115/130 , 115/24, 116/126, 116/130, 116/24, 117/126, 118/126, 20/123, 20/124, 20/125, 20/126, 20/127, 20/128, 20 Amino acid sequences of sequence pairs selected from the group consisting of / 129, 20/130, 20/131, 20/132, 20/133, 20/134, 119/126, 122/126, and 120/130,
b) the heavy chain is used in a method of treating vitiligo comprising a human IgG1 Fc region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 Humanized anti-human CXC motif chemokine receptor 3 (CXCR3) antibody or pharmaceutical composition thereof for The humanized anti-human CXCR3 antibody of claim 1, wherein the VH comprises the amino acid sequence of SEQ ID NO: 20 and the VL comprises the amino acid sequence of SEQ ID NO: 24. 3. The humanized anti-human CXCR3 antibody of claim 2, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2. 4. The humanized anti-human CXCR3 antibody of claim 2, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 9. 4. The humanized anti-human CXCR3 antibody of claim 2, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 10. 4. The humanized anti-human CXCR3 antibody of claim 2, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 11. 4. The humanized anti-human CXCR3 antibody of claim 1, wherein the VH comprises the amino acid sequence of SEQ ID NO: 18 and the VL comprises the amino acid sequence of SEQ ID NO: 22. 3. The humanized anti-human CXCR3 antibody of claim 7 wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2. 9. The humanized anti-human CXCR3 antibody of claim 8, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 9. 10. The humanized anti-human CXCR3 antibody of claim 8, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 10. 10. The humanized anti-human CXCR3 antibody of claim 8, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 11. 10. 36. The method of claim 35 wherein
a) HC comprises the amino acid sequence of SEQ ID NO: 25 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
b) HC comprises the amino acid sequence of SEQ ID NO: 26 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
c) HC comprises the amino acid sequence of SEQ ID NO: 27 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
d) HC comprises the amino acid sequence of SEQ ID NO: 139 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
e) HC comprises the amino acid sequence of SEQ ID NO: 31 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
f) HC comprises the amino acid sequence of SEQ ID NO: 32 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
g) HC comprises the amino acid sequence of SEQ ID NO: 32 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
h) HC comprises the amino acid sequence of SEQ ID NO: 140 and HC comprises the amino acid sequence of SEQ ID NO: 137 or a humanized anti-human CXCR3 antibody or pharmaceutical composition thereof. The humanized anti-human CXCR3 antibody or pharmaceutical composition thereof according to any one of claims 1 to 12, wherein the IgG1 Fc region has a reduced fucose content. The humanized anti-human CXCR3 antibody or pharmaceutical composition thereof according to any one of claims 1 to 12 produced in a host cell cultured in the presence of a glycation inhibitor. The humanized anti-human CXCR3 antibody or pharmaceutical composition thereof of claim 14, wherein the glycosylation inhibitor is kifunensine. A method of treating vitiligo, comprising administering to a subject in need thereof a humanized anti-human CXCR3 antibody comprising a heavy chain (HC) having a heavy chain variable region (VH) and a light chain (LC) having a light chain variable region (VL) Step, where:
a) VH and VL are SEQ ID NOs: 24/20, 22/18, 80/130, 81/24, 82/130, 82/24, 83/126, 83/130, 83/24, 84/126, 84 / 24, 85/24, 86/126, 87/126, 87/133, 87/24, 88/24, 89/24, 90/126, 91/126, 91/130, 92/130, 92/24 , 93/126, 94/126, 95/126, 96/126, 97/126, 98/126, 99/126, 100/126, 101/126, 102/126, 103/126, 104/126, 105 / 126, 106/126, 107/126, 108/126, 109/126, 110/126, 111/126, 121/24, 113/130, 113/24, 114/130, 115/126, 115/130 , 115/24, 116/126, 116/130, 116/24, 117/126, 118/126, 20/123, 20/124, 20/125, 20/126, 20/127, 20/128, 20 Amino acid sequences of sequence pairs selected from the group consisting of / 129, 20/130, 20/131, 20/132, 20/133, 20/134, 119/126, 122/126, and 120/130,
b) the heavy chain comprises a human IgG1 Fc region selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11. The method of claim 16, wherein the VH comprises the amino acid sequence of SEQ ID NO: 20 and the VL comprises the amino acid sequence of SEQ ID NO: 24. The humanized anti-human CXCR3 antibody of claim 17, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2. 18. The humanized anti-human CXCR3 antibody of claim 17, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 9. 18. The humanized anti-human CXCR3 antibody of claim 17, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 10. 18. The humanized anti-human CXCR3 antibody of claim 17, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 11. 18. The method of claim 16, wherein the VH comprises the amino acid sequence of SEQ ID NO: 18 and the VL comprises the amino acid sequence of SEQ ID NO: 22. The humanized anti-human CXCR3 antibody of claim 22, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 2. 23. The humanized anti-human CXCR3 antibody of claim 22, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 9. 23. The humanized anti-human CXCR3 antibody of claim 22, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 10. The humanized anti-human CXCR3 antibody of claim 22, wherein the human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 11. 23. The method of claim 16,
a) HC comprises the amino acid sequence of SEQ ID NO: 25 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
b) HC comprises the amino acid sequence of SEQ ID NO: 26 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
c) HC comprises the amino acid sequence of SEQ ID NO: 27 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
d) HC comprises the amino acid sequence of SEQ ID NO: 139 and LC comprises the amino acid sequence of SEQ ID NO: 135, or
e) HC comprises the amino acid sequence of SEQ ID NO: 31 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
f) HC comprises the amino acid sequence of SEQ ID NO: 32 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
g) HC comprises the amino acid sequence of SEQ ID NO: 32 and LC comprises the amino acid sequence of SEQ ID NO: 137, or
h) HC comprises the amino acid sequence of SEQ ID NO: 140 and HC comprises the amino acid sequence of SEQ ID NO: 137. 28. The method of any one of claims 16-27, wherein the HC comprises a human IgG1 Fc region with a reduced fucose content. The method of any one of claims 16-27, wherein the antibody is produced in a host cell cultured in the presence of a glycosylation inhibitor. The method of claim 29, wherein the glycosylation inhibitor is kifunensin. A method of treating vitiligo, comprising administering to a subject in need a humanized anti-human CXCR3 antibody comprising a heavy chain (HC) having a heavy chain variable region (VH) and a light chain (LC) having a light chain variable region (VL). Providing instructions for, where:
a) VH and VL are SEQ ID NOs: 24/20, 22/18, 80/130, 81/24, 82/130, 82/24, 83/126, 83/130, 83/24, 84/126, 84 / 24, 85/24, 86/126, 87/126, 87/133, 87/24, 88/24, 89/24, 90/126, 91/126, 91/130, 92/130, 92/24 , 93/126, 94/126, 95/126, 96/126, 97/126, 98/126, 99/126, 100/126, 101/126, 102/126, 103/126, 104/126, 105 / 126, 106/126, 107/126, 108/126, 109/126, 110/126, 111/126, 121/24, 113/130, 113/24, 114/130, 115/126, 115/130 , 115/24, 116/126, 116/130, 116/24, 117/126, 118/126, 20/123, 20/124, 20/125, 20/126, 20/127, 20/128, 20 Amino acid sequences of sequence pairs selected from the group consisting of / 129, 20/130, 20/131, 20/132, 20/133, 20/134, 119/126, 122/126, and 120/130,
b) the heavy chain comprises a human IgG1 Fc region having an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11. Kit for treating vitiligo, including:
a) Humanized anti-human CXCR3 antibody comprising a heavy chain (HC) having a heavy chain variable region (VH) and a light chain (LC) having a light chain variable region (VL), wherein
i) VH and VL are SEQ ID NOs: 24/20, 22/18, 80/130, 81/24, 82/130, 82/24, 83/126, 83/130, 83/24, 84/126, 84 / 24, 85/24, 86/126, 87/126, 87/133, 87/24, 88/24, 89/24, 90/126, 91/126, 91/130, 92/130, 92/24 , 93/126, 94/126, 95/126, 96/126, 97/126, 98/126, 99/126, 100/126, 101/126, 102/126, 103/126, 104/126, 105 / 126, 106/126, 107/126, 108/126, 109/126, 110/126, 111/126, 121/24, 113/130, 113/24, 114/130, 115/126, 115/130 , 115/24, 116/126, 116/130, 116/24, 117/126, 118/126, 20/123, 20/124, 20/125, 20/126, 20/127, 20/128, 20 Amino acid sequences of sequence pairs selected from the group consisting of / 129, 20/130, 20/131, 20/132, 20/133, 20/134, 119/126, 122/126, and 120/130,
ii) HC further comprises a human IgG1 Fc region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11); And
b) instructions for administering humanized anti-human CXCR3 to a human subject in need thereof.

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