Patent Publication Number: US-2003232387-A1

Title: Antibodies that bind alphaE integrin

Description:
BACKGROUND OF THE INVENTION  
       [0001] Integrin receptors are important for regulating both lymphocyte recirculation and recruitment to sites of inflammation (Carlos, T. M. and Harlan, J. M.,  Blood  84:2068-2101 (1994)). The αE integrin αEβ7 is expressed on mucosal homing lymphocytes such as intestinal intraepithelial lymphocytes (IEL) and binds E-cadherin, which is expressed on epithelial cells, as well as a ligand on intestinal microvascular endothelial cell lines (Cepek, K. L. et al.,  Nature  372:190-193 (1994); Stauch U. G. et al.,  J. Immunol . 166:3506-3514 (2001)). As such, the αEβ7 integrin acts as a homing receptor that mediates lymphocyte migration to mucosal epithelium, such as intestinal epithelium (Schon, M. P. et al.,  J. Immunol . 162:6641-6649 (1999)).  
       [0002] αE integrins, like other integrins, can assume an activated or inactive conformation. Activated integrins bind ligand (e.g. E-cadherin) with high affinity. αE integrins, such as αEβ7, can be activated by divalent cations and/or by inside out signalling upon cellular stimulation with mitogens, growth factors and/or specific antigen (e.g., peptide/MHC).  
       [0003] Antibodies which bind αEβ7 integrin can interfere with αEβ7 integrin binding to its ligands (e.g., E-cadherin) and inhibit leukocyte migration to mucosal inflammatory sites (see, e.g., Ludviksson, B. R. et al.,  J. Immunol . 162:4975-4982 (1999); WO 00/30681 (Ludviksson, B. R. et al.)). However, a problem with using murine antibodies or other non-human antibodies for in vivo applications (e.g., diagnostic methods, therapeutic methods) in humans is that they are highly immunogenic and quickly induce a human anti-foreign antibody response (e.g., a human anti-mouse antibody response, HAMA). Such a human anti-foreign antibody response can result in rapid clearance of the foreign antibody and severely limit diagnostic or therapeutic uses or abrogate any therapeutic benefits.  
       [0004] Thus, a need exists for improved antibodies and antigen-binding fragments that can be used to diagnose and/or treat subjects having mucosal inflammatory disorders.  
       SUMMARY OF THE INVENTION  
       [0005] The invention relates to antibodies and antigen-binding fragments of antibodies which bind an αE integrin (e.g., αEβ7 or other integrin comprising an αE chain). In one aspect, the invention is an antibody or antigen-binding fragment thereof that binds an activation-induced epitope on integrin αE chain, such as an epitope induced by exposure of an αE integrin to a divalent cation (e.g., Mn 2+ ). For example, the activation-induced epitope can comprise amino acid residues in the I domain of integrin αE chain. In one embodiment, the antibody or antigen-binding fragment thereof binds an activation-induced epitope on human integrin αE chain. In another embodiment, the antibody or antigen-binding fragment thereof can inhibit the binding of a ligand (e.g., E-cadherin) to an αE integrin (e.g., αEβ7). In other embodiments, the antibody or antigen-binding fragment can inhibit αE integrin-mediated adhesion of a first cell expressing an αE integrin to a second cell bearing a ligand of an αE integrin, such as epithelial cells (e.g., intestinal epithelial cells) or endothelial cells. In particular embodiments, the antibody or antigen-binding fragment competitively inhibits binding of mAb 3G6 to αEβ7 integrin, or has the epitopic specificity of mAb 3G6.  
       [0006] In other embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 3G6. For example, in a particular embodiment the antibody can comprise the heavy chain variable region of mAb 3G6 (SEQ ID NO: 4) and the light chain variable region of mAb 3G6 (SEQ ID NO: 9).  
       [0007] In other embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 5E4. For example, in a particular embodiment the antibody can comprise the heavy chain variable region of mAb 5E4 (SEQ ID NO: 14) and the light chain variable region of mAb 5E4 (SEQ ID NO: 19).  
       [0008] In additional embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 8D5. For example, in a particular embodiment the antibody can comprise the heavy chain variable region of mAb 8D5 (SEQ ID NO: 24) and the light chain variable region of mAb 8D5 (SEQ ID NO: 29).  
       [0009] Preferred antibodies that bind an αE integrin (e.g., selectively bind an activation-induced epitope on integrin αE chain) include chimeric antibodies, humanized antibodies and antigen-binding fragments of the foregoing. Particularly preferred antibodies are of human origin. In specific embodiments, the invention is mAb 3G6, mAb 5E4 or mAb 8D5 or an antigen-binding fragment of mAb 3G6, mAb 5E4 or mAb 8D5.  
       [0010] The invention also relates to the heavy chains, light chains and portions of the heavy chains and light chains of the antibodies described herein. The invention also relates to fusion proteins comprising an antibody or portion thereof (e.g., heavy chain, light chain, variable region) of the invention and a non-immunoglobulin moiety. The invention also relates to immuno-conjugates comprising an antibody or antigen-binding fragment of the invention and a second moiety, such as a toxin (e.g., cytotoxin, cytotoxic agent), a therapeutic agent (e.g., a chemotherapeutic agent, an antimetabolite, an alkylating agent, an anthracycline, an antibiotic, an anti-mitotic agent, a biological response modifier (e.g., a cytokine (e.g., an interleukin, an interferon, a tumor necrosis factor), a growth factor (e.g., a neurotrophic factor)), a plasminogen activator, a radionuclide (e.g, a radioactive ion) or enzyme, for example.  
       [0011] The invention also relates to isolated and/or recombinant nucleic acids encoding the antibodies, antigen-binding fragments, heavy chains, light chains and portions of the heavy chains and light chains of the antibodies described herein, and to expression constructs or vectors comprising same. The invention also relates to a host cell that comprises a nucleic acid of the invention. In specific embodiments, the invention is hybridoma 3G6, hybridoma 5E4 or hybridoma 8D5.  
       [0012] The invention also relates to a method of treating a subject having an inflammatory disease or disorder comprising administering to said subject an effective amount of an antibody or antigen-binding fragment of the invention. In particular embodiments, the subject is a human. In other particular embodiments, the subject has an inflammatory bowel disease, such as ulcerative colitis or Crohn&#39;s disease.  
       [0013] The invention also relates to a method for detecting an activated αE integrin (e.g., activated αEβ7) comprising contacting a composition comprising an αE integrin with an antibody or antigen-binding fragment thereof which binds an activation-induced epitope on integrin αE chain and detecting formation of a complex between said antibody or antigen-binding fragment and said activated αE integrin.  
       [0014] The invention further relates to an antibody, antigen-binding fragment of an antibody, fusion protein or immuno-conjugate as described herein for use in therapy (including prophylaxis) or diagnosis, and to the use of an antibody, antigen-binding fragment of an antibody, fusion protein or immuno-conjugate of the invention for the manufacture of a medicament for the treatment of a particular disease or condition as described herein (e.g., a mucosal inflammatory disease (e.g., inflammatory bowel disease (e.g., ulerative colitis, Crohn&#39;s disease)), cancer (e.g., leukemia, lymphoma)). 
     
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
     [0015] FIGS.  1 A- 1 H are fluorescent histograms showing binding of mAb 3G6 (IgG1) to transfected K562 cells that expressed an αEβ7 integrin under a variety of buffer conditions. The transfected cells were stained with isotype control antibody (human IgG1) in standard staining buffer (PBS/5% FBS) (FIG. 1A), with mAb 3G6 (IgG1) in standard staining buffer (FIG. 1B) or in buffer that contained EDTA (5 mM; FIG. 1C), in buffer that contained MnCl 2  (1 mM; FIG. 1D), in buffer that contained MgCl 2  (1 mM; FIG. 1E), in buffer that contained CaCl 2  (1 mM; FIG. 1F), in buffer that contained MgCl 2  and CaCl 2  (1 mM each; FIG. 1G) or in buffer that contained MgCl 2 , CaCl 2  and MnCl 2  (1 mM each; FIG. 1H), and bound antibody was detected using a fluorescein isothiocyanate (FITC) labeled anti-human IgG antibody. The results show that binding of mAb 3G6 (IgG1) to the transfected cells was enhanced in buffer that contained Mn 2+  (FIGS. 1D and 1H) and inhibited in buffer that contained EDTA (FIG. 1C) relative to binding in standard buffer.  
     [0016]FIG. 2A is an illustration of a nucleic acid sequence encoding the mature heavy chain variable region of mAb 3G6 (SEQ ID NO: 3) and the encoded amino acid sequence of the mature heavy chain variable region of mAb 3G6 (SEQ ID NO: 4). Complementarity determining region (CDR) 1 consists of amino acid residues 31-35 of SEQ ID NO: 4 (SEQ ID NO: 5), CDR 2 consists of amino acid residues 50-66 of SEQ ID NO: 4 (SEQ ID NO: 6), CDR 3 consists of amino acid residues 99-112 of SEQ ID NO: 4 (SEQ ID NO: 7).  
     [0017]FIG. 2B is an illustration of a nucleic acid sequence encoding the mature kappa light chain variable region of mAb 3G6 (SEQ ID NO: 8) and the encoded amino acid sequence of the mature light chain variable region of mAb 3G6 (SEQ ID NO: 9). Complementarity determining region (CDR) 1 consists of amino acid residues 24-34 of SEQ ID NO: 9 (SEQ ID NO: 10), CDR 2 consists of amino acid residues 50-56 of SEQ ID NO: 9 (SEQ ID NO: 11), CDR 3 consists of amino acid residues 89-98 of SEQ ID NO: 9 (SEQ ID NO: 12).  
     [0018]FIG. 3A is an illustration of a nucleic acid sequence encoding the mature heavy chain variable region of mAb 5E4 (SEQ ID NO:13) and the encoded amino acid sequence of the mature heavy chain variable region of mAb 5E4 (SEQ ID NO: 14). Complementarity determining region (CDR) 1 consists of amino acid residues 31-35 of SEQ ID NO: 14 (SEQ ID NO: 15), CDR 2 consists of amino acid residues 50-66 of SEQ ID NO: 14 (SEQ ID NO: 16), CDR 3 consists of amino acid residues 99-107 of SEQ ID NO: 14 (SEQ ID NO: 17).  
     [0019]FIG. 3B is an illustration of a nucleic acid sequence encoding the mature kappa light chain variable region of mAb 5E4 (SEQ ID NO: 18) and the encoded amino acid sequence of the mature light chain variable region of mAb 5E4 (SEQ ID NO: 19). Complementarity determining region (CDR) 1 consists of amino acid residues 24-34 of SEQ ID NO: 19 (SEQ ID NO: 20), CDR 2 consists of amino acid residues 50-56 of SEQ ID NO: 19 (SEQ ID NO: 21), CDR 3 consists of amino acid residues 89-98 of SEQ ID NO: 19 (SEQ ID NO: 22).  
     [0020]FIG. 4A is an illustration of a nucleic acid sequence encoding the mature heavy chain variable region of mAb 8D5 (SEQ ID NO: 23) and the encoded amino acid sequence of the mature heavy chain variable region of mAb 8D5 (SEQ ID NO: 24). Complementarity determining region (CDR) 1 consists of amino acid residues 31-35 of SEQ ID NO: 24 (SEQ ID NO: 25), CDR 2 consists of amino acid residues 50-65 of SEQ ID NO: 24 (SEQ ID NO: 26), CDR 3 consists of amino acid residues 98-117 of SEQ ID NO: 24 (SEQ ID NO: 27).  
     [0021]FIG. 4B is an illustration of a nucleic acid sequence encoding the mature kappa light chain variable region of mAb 8D5 (SEQ ID NO: 28) and the encoded amino acid sequence of the mature light chain variable region of mAb 8D5 (SEQ ID NO: 29). Complementarity determining region (CDR) 1 consists of amino acid residues 24-34 of SEQ ID NO: 29 (SEQ ID NO: 30), CDR 2 consists of amino acid residues 50-56 of SEQ ID NO: 29 (SEQ ID NO: 31), CDR 3 consists of amino acid residues 89-97 of SEQ ID NO: 29 (SEQ ID NO: 32). 
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
     [0022] As used herein, “activation-induced epitope” refers to an epitope that is present on an activated αE integrin (e.g., integrin αE chain (CD103), an αEβ7 integrin) but not on non-activated αE integrin. An activated αE integrin is an αE integrin that binds ligand (e.g., E-cadherin) with high affinity, while a non-activated αE integrin binds the ligand with low affinity. (See, Higgins, J. M. G. et al.,  J. Biol. Chem . 140:197-210 (1998).) An αE integrin can be activated, for example, by exposure to divalent cations (e.g., Mn 2+ ). When the αE integrin is expressed on the surface of a cell, it can be activated upon exposure of the cell to phorbol esters (e.g., Phorbol 12-myristate 13-acetate (PMA)), or to suitable growth factors and/or mitogens (e.g., concanavalin A). An αE integrin expressed on a T cell can be activated by signals transduced through the T cell receptor (TCR) complex (e.g., upon TCR binding to specific MHC-peptide complexes, crosslinking with anti-CD3 antibody).  
     [0023] An antibody that “binds an activation-induced epitope” on integrin αE chain binds integrin αE chain under activation conditions (e.g., in the presence of divalent cations (e.g., Mn 2+ )) but does not significantly bind in the absence of activation (e.g., when a suitable chelating agent (e.g., Ethylenediaminetetraacetic acid (EDTA)) is present).  
     [0024] As used herein, an antibody and antigen-binding fragment thereof that “binds” an αE integrin (e.g., an activated αE integrin, an αEβ7 integrin, an integrin αE chain (CD103)) has binding specificity for the αE integrin. The terms “binding specificity” or “specific” when referring to an antibody-antigen interaction indicate that the antibody can discriminate between one or more αE integrins (e.g., an activated αE integrin, an αEβ7 integrin, an integrin αE chain (CD103)) and other antigens, rather than to indicate that the antibody can bind only one antigen. For example, in certain embodiments, the antibody or antigen-binding fragments of the invention can “selectively bind” an αE integrin. Such selective antibodies or antigen-binding fragments may bind another antigen with low affinity, but bind said αE integrin with higher affinity. Under appropriate binding conditions (e.g., physiological conditions), an antibody or antigen-binding fragment thereof that selectively binds an αE integrin will bind the αE integrin but will not significantly bind other antigens. An antibody or antigen-binding fragment of an antibody does not “significantly bind” an antigen when the extent of binding is less than about 25%, preferably less than about 15%, more preferably less than about 10%, most preferably less than about 5% or less than about 2% or 1% of the level of binding to an antigen that is “selectively” bound under the same conditions (e.g., physiological conditions). The concentration of antibody and other conditions required to provide selectivity for an αE integrin (e.g., an antibody concentration and pH which reduces or eliminates non-selective binding) can be readily determined using any suitable method, such as titration.  
     [0025] As used herein, the term “functionally rearranged” refers to a segment of DNA from an immunoglobulin locus which has undergone V(D)J recombination, with or without insertion or deletion of nucleotide(s) (e.g., N nucleotides, P nucleotides) and/or somatic mutation, thereby producing an immunoglobulin gene which encodes an immunoglobulin variable region or immunoglobulin chain (e.g., heavy chain, light chain). A functionally rearranged immunoglobulin gene can be directly or indirectly identified using suitable methods, such as, for example, nucleotide sequencing, hybridization (e.g., Southern blotting, Northern blotting) using probes which can anneal to coding joints between gene segments (e.g., VH, VL, D, JH, JL) or enzymatic amplification of immunoglobulin genes (e.g., polymerase chain reaction) with primers which can anneal to coding joints between gene segments. Whether a cell produces an antibody comprising a particular variable region or a variable region comprising a particular sequence (e.g., a CDR sequence) can also be determined using suitable methods. In one example, mRNA can be isolated from an antibody producing cell (e.g., a hybridoma) and used to produce cDNA. The cDNA can be cloned and sequenced or can be amplified (e.g., by polymerase chain reaction) using a first primer which anneals specifically to a portion of the variable region of interest (e.g., CDR, coding joint) and a second primer which anneals specifically to non-variable region sequences (e.g., C H 1, C L ).  
     [0026] As used herein, the phrase “of human origin” refers to antibodies, antigen-binding fragments of antibodies and portions or regions of antibodies (e.g., variable regions, complementarity determining regions (CDRs), framework regions (FRs), constant regions) having amino acid sequences that are encoded by nucleotide sequences derived from human ( Homo sapiens ) germ line immunoglobulin genes. For example, an antibody of human origin can be encoded by human germ line immunoglobulin genes that have been functionally rearranged to produce a functional gene that can be expressed to produce an antibody. As described herein, functionally rearranged genes that encode an antibody chain can include sequences that are not found in the germ line, such as N nucleotides and P nucleotides, and mutations that can occur as part of the processes that produce high-affinity antibodies (e.g., somatic mutation, affinity maturation, clonal selection). Functionally rearranged immunoglobulin genes of human origin, including those that include non-germ line sequences, can be generated via natural processes in a suitable in vivo expression system (e.g., a human, a human-antibody transgenic animal), artificially using any suitable methods (e.g., recombinant DNA technology, phage display) or any combination of natural and artificial processes. Antibodies, antigen-binding fragments of antibodies and portions or regions of antibodies of human origin can be produced, for example, by expression of a nucleic acid of non-human origin (e.g., a synthetic nucleic acid) that has the requisite nucleotide sequence.  
     [0027] An antibody, antigen-binding fragment of an antibody or a portion of an antibody (e.g., a framework region) “of human origin” can have an amino acid sequence that is encoded by a nucleic acid that has a nucleotide sequence that is a consensus of the nucleotide sequences of a number of naturally occurring human antibody genes or human germ line sequences, or have an amino acid sequence that is a consensus of the amino acid sequences of a number of naturally occurring human antibodies or amino acid sequences encoded in the human germ line. A number of human antibody consensus sequences are available, including consensus sequences for the different subgroups of human variable regions (see, Kabat, E. A., et al.,  Sequences of Proteins of Immunological Interest , Fifth Edition, U.S. Department of Health and Human Services, U.S. Government Printing Office (1991). The Kabat database and its applications are freely available on line. (See, Johnson, G. and Wu, T. T.,  Nucleic Acids Research  29:205-206 (2001).)  
     [0028] As used herein, the phrase “human antibody” refers to antibodies or antigen-binding fragments of antibodies in which the variable and constant regions (if present) have amino acid sequences that are encoded by nucleotide sequences derived from human ( Homo sapiens ) germline immunoglobulin genes. A “human antibody” can include sequences that are not encoded in the germline (e.g., due to N nucleotides, P nucleotides, and mutations that can occur as part of the processes that produce high-affinity antibodies such as, somatic mutation, affinity maturation, clonal selection)) that occur as a result of biological processes in a suitable in vivo expression system (e.g., a human, a human-antibody transgenic animal). Antibodies, antigen-binding fragments of antibodies and portions or regions of human antibodies can be produced, for example, by expression of a nucleic acid of non-human origin (e.g., a synthetic nucleic acid) that has the requisite nucleotide sequence.  
     [0029] As used herein, the phrase “CDR-grafted” antibody refers to antibodies and antigen-binding fragments of antibodies that comprise a CDR that is not naturally associated with the framework regions of the antibody or antigen-binding fragment. Generally the CDR is from an antibody from a first species and the framework regions and constant regions (if present) are from an antibody from a different species. The CDR-grafted antibody can be a “humanized antibody.” 
     [0030] As used herein, “humanized antibody” refers to an antibody or antigen-binding fragment thereof comprising a CDR that is not of human origin and framework and/or constant regions that are of human origin. For example, a humanized antibody can comprise a CDR derived from an antibody of nonhuman origin (e.g., natural antibody such as a murine (e.g., mouse, rat) antibody, artificial antibody) that binds an αE integrin, preferably integrin αE chain (CD103), and framework and constant regions (if present) of human origin (e.g., a human framework region, a human consensus framework region, a human constant region (e.g., CL, CH1, hinge, CH2, CH3, CH4)). CDR-grafted single chain antibodies containing a CDR of non-human origin and framework and constant regions (if present) of human origin (e.g., CDR-grafted scFV) are also encompassed by the term humanized antibody.  
     [0031] As used herein, the term “chimeric antibody” refers to an antibody or antigen-binding fragment thereof comprising a variable region from an antibody from a first species and a constant region from an antibody from a different species. None of the portions which comprise a chimeric antibody need to be of human origin. For example, a chimeric antibody can comprise a variable region from a rodent (e.g., mouse) antibody and a constant region of a non-human primate antibody (e.g., a chimpanzee constant region).  
     [0032] The antibody of the invention can be a single chain antibody (e.g., a single chain Fv (scFv)) and can include a linker moiety (e.g., a linker peptide) not found in native antibodies. For example, an scFv can comprise a linker peptide, such as two to about twenty glycine residues or other suitable linker, which connects a heavy chain variable region to a light chain variable region. For the purposes of the invention, the presence of such a linker does not affect the status of the single chain antibody as being “of human origin” or “human.” For example, a human scFv can comprise a human heavy chain variable region and a human light chain variable region that are connected through a suitable peptide linker.  
     [0033] “Conservative amino acid substitution” refers to the replacement of a first amino acid by a second amino acid that has chemical and/or physical properties (e.g., charge, structure, polarity, hydrophobicity/hydrophilicity) which are similar to those of the first amino acid. For example, replacement of one amino acid by another within the following groups is a conservative amino acid substitution: Ala, Val, Leu, and Ile; Ser and Thr; Asp and Glu; Asn and Gln; Lys and Arg; Phe and Tyr.  
     [0034] A nucleotide sequence encoding a human ( Homo sapiens ) integrin αE chain (CD103), used in the studies described herein and deposited in GenBank under accession number L25851, is presented as SEQ ID NO: 1. (See also, Shaw et al.,  J. Biol. Chem . 269:6016-6025 (1994).) The nucleotide sequence has an open-reading frame beginning at position 126. The amino acid sequence of a human integrin αE chain encoded by SEQ ID NO: 1 is presented as SEQ ID NO: 2. The human integrin αE chain contains a signal peptide (amino acid residues −18 to −1 of SEQ ID NO: 2), an X-domain (amino acid residues 126-180 of SEQ ID NO: 2) and an I-Domain (residues 181-372 of SEQ ID NO: 2). The entire teachings of GenBank Accession No. L25851 are incorporated herein by reference.  
     [0035] A nucleotide sequence encoding a human ( Homo sapiens ) E-cadherin used in the studies described herein and deposited in GenBank under accession number L08599 is presented as SEQ ID NO:33. The nucleotide sequence has an open-reading frame beginning at position 109. The amino acid sequence of a human E-cadherin encoded by SEQ ID NO: 33 is presented as SEQ ID NO: 34. The entire teachings of GenBank Accession No. L08599 are incorporated herein by reference.  
     [0036] A nucleotide sequence encoding a human ( Homo sapiens ) integrin α4 chain used in the studies described herein and deposited in GenBank under accession number L12002 is presented as SEQ ID NO: 35. The nucleotide sequence has an open-reading frame beginning at position 411. The amino acid sequence of an integrin α4 chain encoded by SEQ ID NO: 35 is presented as SEQ ID NO:36. The entire teachings of GenBank Accession No. L12002 are incorporated herein by reference.  
     [0037] A nucleotide sequence encoding a human ( Homo sapiens ) integrin β7 chain used in the studies described herein and deposited in GenBank under accession number M62880 is presented as SEQ ID NO:37. The nucleotide sequence has an open-reading frame beginning at position 114. The amino acid sequence of an integrin β7 chain encoded by SEQ ID NO: 37 is presented as SEQ ID NO: 38. The entire teachings of GenBank Accession No. M62880 are incorporated herein by reference.  
     [0038] Antibodies and Antibody Producing Cells  
     [0039] The antibody of the invention can be polyclonal or monoclonal, and the term “antibody” is intended to encompass both polyclonal and monoclonal antibodies. The terms polyclonal and monoclonal refer to the degree of homogeneity of an antibody preparation, and are not intended to be limited to particular methods of production. The term “antibody” as used herein encompasses antigen-binding fragments of antibodies, including antigen-binding fragments of human, humanized, chimeric, CDR-grafted, veneered or single-chain antibodies.  
     [0040] Antibodies which bind an αE integrin can be selected from a suitable collection of natural or artificial antibodies or raised against an appropriate immunogen in a suitable host. For example, antibodies can be raised by immunizing a suitable host (e.g., mouse, human antibody-transgenic mouse) with a suitable immunogen, such as an isolated or purified αE integrin (e.g., αEβ7) or cells expressing a recombinant αE integrin (e.g., cell that expresses an exogenous nucleic acid encoding human integrin αE chain (CD103)). In addition, cells expressing a recombinant αE integrin, such as transfected cells, can be used in a screen for antibody which binds thereto (See e.g., Chuntharapai et al.,  J. Immunol ., 152: 1783-1789 (1994); Chuntharapai et al., U.S. Pat. No. 5,440,021).  
     [0041] Preparation of immunizing antigen, and polyclonal and monoclonal antibody production can be performed using any suitable technique. A variety of methods have been described. (See, e.g., Kohler et al.,  Nature , 256: 495-497 (1975) and  Eur. J. Immunol . 6: 511-519 (1976); Milstein et al.,  Nature  266: 550-552 (1977); Koprowski et al., U.S. Pat. No. 4,172,124; Harlow, E. and D. Lane, 1988 , Antibodies: A Laboratory Manual , (Cold Spring Harbor Laboratory: Cold Spring Harbor, N.Y.);  Current Protocols In Molecular Biology , Vol. 2 (Supplement 27, Summer &#39;94), Ausubel, F. M. et al., Eds., (John Wiley &amp; Sons: New York, N.Y.), Chapter 11, (1991).) Generally, where a monoclonal antibody is desired, a hybridoma is produced by fusing a suitable immortal cell line (e.g., a myeloma cell line such as SP2/0, P3X63Ag8.653 or a heteromyeloma) with antibody-producing cells. Antibody-producing cells can be obtained from the peripheral blood or, preferably the spleen or lymph nodes, of humans, human-antibody transgenic animals or other suitable animals immunized with the antigen of interest. Cells that produce antibodies of human origin (e.g., a human antibody) can be produced using suitable methods, for example, fusion of a human antibody-producing cell and a heteromyeloma or trioma, or immortalization of an activated human B cell via infection with Epstein Barr virus. (See, e.g., U.S. Pat. No. 6,197,582 (Trakht); Niedbala et al.,  Hybridoma , 17:299-304 (1998); Zanella et al.,  J Immunol Methods , 156:205-215 (1992); Gustafsson et al.,  Hum Antibodies Hybridomas , 2:26-32 (1991).) The fused or immortalized antibody-producing cells (hybridomas) can be isolated using selective culture conditions, and cloned by limiting dilution. Cells which produce antibodies with the desired specificity can be identified using a suitable assay (e.g., ELISA).  
     [0042] Other suitable methods of producing or isolating antibodies or antigen-binding fragments of the desired specificity can be used, including, for example, methods which select a recombinant antibody or antigen-binding fragment thereof from a library, such as a phage display library. Such libraries can contain antibodies or antigen-binding fragments of antibodies that contain natural or artificial amino acid sequences. For example, the library can contain Fab fragments which contain artificial CDRs (e.g., random amino acid sequences) and human framework regions. (See, for example, U.S. Pat. No. 6,300,064 (Knappik, et al.), the entire teachings of which are incorporated herein by reference.)  
     [0043] Human antibodies and nucleic acids encoding same can be obtained from a human or from human-antibody transgenic animals. Human-antibody transgenic animals (e.g., mice) are animals that are capable of producing a repertoire of human antibodies, such as XENOMOUSE (Abgenix, Fremont, Calif.), HUMAB-MOUSE, KIRIN TC MOUSE or KM-MOUSE (MEDAREX, Princeton, N.J.). Generally, the genome of human-antibody transgenic animals has been altered to include a transgene comprising DNA from a human immunoglobulin locus that can undergo functional rearrangement. An endogenous immunoglobulin locus in a human-antibody transgenic animal can be disrupted or deleted to eliminate the capacity of the animal to produce antibodies encoded by an endogenous gene. Suitable methods for producing human-antibody transgenic animals are well known in the art. (See, for example, U.S. Pat. Nos. 5,939,598 and 6,075,181 (Kucherlapati et al.), U.S. Pat. Nos. 5,569,825, 5,545,806, 5,625,126, 5,633,425, 5,661,016, and 5,789,650 (Lonberg et al.), Jakobovits et al.,  Proc. Natl. Acad. Sci. USA , 90: 2551-2555 (1993), Jakobovits et al.,  Nature , 362: 255-258 (1993), Jakobovits et al. WO 98/50433, Jakobovits et al. WO 98/24893, Lonberg et al. WO 98/24884, Lonberg et al. WO 97/13852, Lonberg et al. WO 94/25585, Lonberg et al. EP 0 814 259 A2, Lonberg et al. GB 2 272 440 A, Lonberg et al.,  Nature  368:856-859 (1994), Lonberg et al.,  Int Rev Immunol  13(1):65-93 (1995), Kucherlapati et al. WO 96/34096, Kucherlapati et al. EP 0 463 151 B1, Kucherlapati et al. EP 0 710 719 A1, Surani et al. U.S. Pat. No. 5,545,807, Bruggemann et al. WO 90/04036, Bruggemann et al. EP 0 438 474 B1, Taylor et al.,  Int. Immunol . 6(4)579-591 (1994), Taylor et al.,  Nucleic Acids Research  20(23):6287-6295 (1992), Green et al,  Nature Genetics  7:13-21 (1994), Mendez et al.,  Nature Genetics  15:146-156 (1997), Tuaillon et al.,  Proc Natl Acad Sci USA  90(8)3720-3724 (1993) and Fishwild et al.,  Nat Biotechnol  14(7):845-851 (1996), the teachings of each of the foregoing are incorporated herein by reference in their entirety.)  
     [0044] As described herein, human-antibody transgenic animals can be immunized with a suitable composition comprising an antigen of interest (e.g., a recombinant cell expressing an αEβ7 integrin). Antibody producing cells can be isolated and fused to form hybridomas using conventional methods. Hybridomas that produce human antibodies having the desired characteristics (e.g., specificity, affinity) can be identified using any suitable assay (e.g, ELISA) and, if desired, selected and subcloned using suitable culture techniques.  
     [0045] Human-antibody transgenic animals provide a source of nucleic acids that can be enriched in nucleic acids that encode antibodies having desired properties, such as specificity and affinity. For example, nucleic acids encoding antibodies or antibody variable regions can be isolated from human-antibody transgenic mice that have been immunized with an αE integrin. The isolated nucleic acids or portions thereof (e.g., portions encoding variable regions, CDRs, framework regions) can be expressed using any suitable method (e.g., phage display) to produce a library of antibodies or antigen-binding fragments of antibodies (e.g., single chain antigen-binding fragments, double chain antigen-binding fragments) that is enriched for antibodies or antigen-binding fragments that bind αE. Such a library can exhibit enhanced diversity (e.g., combinatorial diversity through pairing of heavy chain variable regions and light chain variable regions) relative to the repertoire of antibodies produced in the immunized human-antibody transgenic animal. The library can be screened using any suitable assay (e.g., an αE binding assay) to identify antibodies or antigen-binding fragments having desired properties (e.g., specificity, affinity). The nucleic acids encoding antibody or antigen-binding fragments having desired properties can be recovered using any suitable methods. (See, e.g., U.S. Pat. No. 5,871,907 (Winter et al.) and U.S. Pat. No. 6,057,098 (Buechler et al.), the entire teachings of each of the foregoing are incorporated herein by reference.)  
     [0046] The antibody of the invention can be a CDR-grafted (e.g., humanized) antibody or an antigen-binding fragment thereof. The CDRs of a CDR-grafted antibody can be derived from a suitable antibody which binds an αE integrin (referred to as a donor antibody). For example, suitable CDRs can be derived from mAb 3G6, mAb 5E4 or mAb 8D5 which, as described herein, bind integrin αE chain (CD103) or from any other suitable antibody. Other sources of suitable CDRs include natural and artificial αE integrin-specific antibodies obtained from nonhuman sources, such as rodent (e.g., mouse, rat), rabbit, pig, goat, non-human primate (e.g., monkey) or non-human library.  
     [0047] The framework regions of a CDR-grafted antibody are preferably of human origin, and can be derived from any human antibody variable region having sequence similarity to the analogous or equivalent region (e.g., light chain variable region) of the antigen binding region of the donor antibody. Other sources of framework regions of human origin include human variable region consensus sequences. (See, e.g., Kettleborough, C. A. et al.,  Protein Engineering  4:773-783 (1991); Carter et al., WO 94/04679; Kabat, E. A., et al.,  Sequences of Proteins of Immunological Interest , Fifth Edition, U.S. Department of Health and Human Services, U.S. Government Printing Office (1991)).  
     [0048] In one embodiment, the framework regions of a CDR-grafted (e.g., humanized) antibody chain can be derived from a variable region of human origin having at least about 65% overall amino acid sequence identity, and preferably at least about 70% overall amino acid sequence identity, with the amino acid sequence of the variable region of the donor antibody. A suitable framework region can also be derived from a antibody of human origin having at least about 65% amino acid sequence identity, and preferably at least about 70%, 80%, 90% or 95% amino acid sequence identity over the length of the framework region within the amino acid sequence of the equivalent portion (e.g., framework region) of the donor antibody. For example, a suitable framework region of human origin can be derived from an antibody of human origin (e.g., a human antibody) having at least about 65% amino acid sequence identity, and preferably at least about 70%, 80%, 90% or 95% amino acid sequence identity, over the length of the particular framework region being used, when compared to the amino acid sequence of the equivalent portion (e.g., framework region) of the donor antibody. Amino acid sequence identity can be determined using a suitable amino acid sequence alignent algorithm, such as CLUSTAL W, using the default parameters. (Thompson J. D. et al.,  Nucleic Acids Res . 22:4673-4680 (1994).)  
     [0049] Framework regions of human origin can include amino acid substitutions or replacements, such as “back mutations” which replace an amino acid residue in the framework region of human origin with a residue from the corresponding position of the donor antibody. One or more mutations in the framework region can be made, including deletions, insertions and substitutions of one or more amino acids. Preferably, the CDR-grafted (e.g., humanized) antibody binds αE integrin with an affinity similar to, substantially the same as, or better than that of the donor antibody. Variants can be produced by a variety of suitable methods, including mutagenesis of nonhuman donor or acceptor human chains. (See, e.g., U.S. Pat. Nos. 5,693,762 (Queen et al.) and 5,859,205 (Adair et al.), the entire teachings of which are incorporated herein by reference.)  
     [0050] Constant regions of antibodies, antibody chains (e.g, heavy chain, light chain) or fragments or portions thereof of the invention, if present, can be derived from any suitable source. For example, constant regions of human, humanized and certain chimeric antibodies, antibody chains (e.g, heavy chain, light chain) or fragments or portions thereof, if present can be of human origin and can be derived from any suitable human antibody or antibody chain. For example, a constant region of human origin or portion thereof can be derived from a human κ or λ light chain, and/or a human γ (e.g., γ1, γ2, γ3, γ4), μ, α (e.g., α1, α2), δ or ε heavy chain, including allelic variants. In certain embodiments, the antibody or antigen-binding fragment (e.g., antibody of human origin, human antibody) can include amino acid substitutions or replacements that alter or tailor function (e.g., effector function). For example, a constant region of human origin (e.g., γ1 constant region, γ2 constant region) can be designed to reduce complement activation and/or Fc receptor binding. (See, for example, U.S. Pat. Nos. 5,648,260 (Winter et al.), 5,624,821 (Winter et al.) and 5,834,597 (Tso et al.), the entire teachings of which are incorporated herein by reference.) Preferably, the amino acid sequence of a constant region of human origin that contains such amino acid substitutions or replacements is at least about 95% identical over the full length to the amino acid sequence of the unaltered constant region of human origin, more preferably at least about 99% identical over the full length to the amino acid sequence of the unaltered constant region of human origin.  
     [0051] Humanized antibodies or antigen-binding fragments of a humanized antibody can be prepared using any suitable method. Several such methods are well-known in the art. (See, e.g., U.S. Pat. No. 5,225,539 (Winter), U.S. Pat. No. 5,530,101 (Queen et al.).) The portions of a humanized antibody (e.g., CDRs, framework, constant region) can be obtained or derived directly from suitable antibodies (e.g., by de novo synthesis of a portion), or nucleic acids encoding an antibody or chain thereof having the desired property (e.g., binds αE integrin) can be produced and expressed. Humanized immunoglobulins comprising the desired portions (e.g., CDR, FR, constant region) of human and nonhuman origin can be produced using synthetic and/or recombinant nucleic acids to prepare a nucleic acid (e.g., cDNA) encoding the desired humanized chain. To prepare a portion of a chain, one or more stop codons can be introduced at the desired position. For example, nucleic acid (e.g., DNA) sequences coding for newly designed humanized variable regions can be constructed using PCR mutagenesis methods to alter existing DNA sequences. (See, e.g., Kamman, M., et al.,  Nucl. Acids Res . 17:5404 (1989).) PCR primers coding for the new CDRs can be hybridized to a DNA template of a previously humanized variable region which is based on the same, or a very similar, human variable region (Sato, K., et al.,  Cancer Research  53:851-856 (1993)). If a similar DNA sequence is not available for use as a template, a nucleic acid comprising a sequence encoding a variable region sequence can be constructed from synthetic oligonucleotides (see e.g., Kolbinger, F.,  Protein Engineering  8:971-980 (1993)). A sequence encoding a signal peptide can also be incorporated into the nucleic acid (e.g., on synthesis, upon insertion into a vector). The natural signal peptide sequence from the acceptor antibody, a signal peptide sequence from another antibody or other suitable sequence can be used (see, e.g., Kettleborough, C. A.,  Protein Engineering  4:773-783 (1991)). Using these methods, methods described herein or other suitable methods, variants can be readily produced. In one embodiment, cloned variable regions can be mutated, and sequences encoding variants with the desired specificity can be selected (e.g., from a phage library; see, e.g., U.S. Pat. No. 5,514,548 (Krebber et al.) and WO 93/06213 (Hoogenboom et al.)).  
     [0052] The antibody of the invention can be a chimeric antibody or an antigen-binding fragment of a chimeric antibody. Preferably, the chimeric antibody or antigen-binding fragment thereof comprises a variable region of non-human origin and a constant region of human origin (e.g., a human constant region).  
     [0053] Chimeric antibodies and antigen-binding fragments of chimeric antibodies that bind αE integrin can be prepared using any suitable method. Several suitable methods are well-known in the art. (See, e.g., U.S. Pat. No. 4,816,567 (Cabilly et al.), U.S. Pat. No. 5,116,946 (Capon et al.).) Generally, chimeric antibodies are produced by preparing, for each of the light and heavy chain components of the chimeric immunoglobulin, a recombinant nucleic acid comprising a first nucleotide sequence encoding at least the variable region of an antibody from a first species that binds αE integrin that is joined in frame to a second nucleotide sequence encoding at least a part of a constant region from an antibody of a different species. Generally, the recombinant nucleic acid encodes a chimeric heavy chain or a chimeric light chain. However, if desired, a single recombinant nucleic acid encoding a chimeric heavy chain and a chimeric light chain can be prepared. The recombinant nucleic acids can be assembled in or inserted into an expression vector. The recombinant nucleic acid(s) can be introduced into a suitable host cell that is capable of expressing the chimeric antibody or chimeric antibody chain using any suitable method (e.g., transfection, transformation, infection) to produce a recombinant host cell. The recombinant host cell can be maintained under conditions suitable for expression of the chimeric antibody or chimeric antibody chain and the antibody or chain can be recovered.  
     [0054] Nucleic acids encoding the variable region of antibody light and heavy chains can be obtained from cells (e.g., B cells, hybridoma cells) that produce an antibody that binds αE integrin. For example, nucleic acids that encode human heavy and light chain variable regions that can bind αE integrin can be obtained from hybridomas 3G6, 5E4 and 8D5, and from recombinant cell lines CHO 3G6 C1.2D6 and CHO 5E4 A1.2C12, described herein. Nucleic acids that encode constant regions can be obtained from suitable sources using any suitable technique, such a conventional techniques of recombinant DNA technology. The nucleotide sequences of nucleic acids encoding human κ or λ light chain constant regions, and γ (e.g., γ1, γ2, γ3, γ4), μ, α (e.g., α1, α2), δ or ε human heavy chain constant regions are readily available.  
     [0055] The invention also relates to a bispecific antibody or antigen-binding fragment thereof (e.g., F(ab′) 2 ), which binds an αE integrin and at least one other antigen. In a particular embodiment, the bispecific antibody, or antigen-binding fragment thereof binds an activation-induced epitope on an αE integrin (e.g., integrin αE chain (CD103)). In other embodiments, the bispecific antibody or antigen-binding fragment thereof has the epitopic specificity of mAb 3G6, mAb 5E4 or mAb 8D5 and at least one other antibody. Bispecific antibodies can be secreted by triomas and hybrid hybridomas. Generally, triomas are formed by fusion of a hybridoma and a lymphocyte (e.g., antibody secreting B cell) and hybrid hybridomas are formed by fusion of two hybridomas. Each of the cells that are fused to produce a trioma or hybrid hybridoma produces a monospecific antibody. However, triomas and hybrid hybridomas can produce an antibody containing antigen binding sites which recognize different antigens. The supernatants of triomas and hybrid hybridomas can be assayed for bispecific antibody using a suitable assay (e.g., ELISA), and bispecific antibodies can be purified using conventional methods. (See, e.g., U.S. Pat. No. 5,959,084 (Ring et al.) U.S. Pat. No. 5,141,736 (Iwasa et al.), U.S. Pat. Nos. 4,444,878, 5,292,668 and 5,523,210 (Paulus et al.) and U.S. Pat. No. 5,496,549 (Yamazaki et al.).)  
     [0056] The various portions of an antibody (e.g., mouse antibody, human antibody, humanized antibody, chimeric antibody and antigen-binding fragments of the foregoing) can be joined together chemically using conventional techniques, or can be prepared as a continuous polypeptide chain by expression (in vivo or in vitro) of a nucleic acid (one or more nucleic acids) encoding antibody. For example, nucleic acids encoding a human, humanized or chimeric chain can be expressed in vivo or in vitro to produce a continuous polypeptide chain. See, e.g., Cabilly et al., U.S. Pat. No. 4,816,567; Cabilly et al., European Patent No. 0,125,023 B1; Boss et al., U.S. Pat. No. 4,816,397; Boss et al., European Patent No. 0,120,694 B1; Neuberger, M. S. et al., WO 86/01533; Neuberger, M. S. et al., European Patent No. 0,194,276 B1; Winter, U.S. Pat. No. 5,225,539; Winter, European Patent No. 0,239,400 B1; Queen et al., European Patent No. 0 451 216 B1; and Padlan, E. A. et al., EP 0 519 596 A1. See also, Newman, R. et al.,  BioTechnology , 10: 1455-1460 (1992), regarding primatized antibody, and Ladner et al., U.S. Pat. No. 4,946,778 and Bird, R. E. et al.,  Science , 242: 423-426 (1988)) regarding single chain antibodies.  
     [0057] The invention also relates to antigen-binding fragments of antibodies that retain the capacity to bind antigen (e.g., an αE integrin, an activation-induced epitope on integrin αE chain). Such antigen-binding fragments of antibodies retain the antigen binding function of a corresponding full-length antibody (e.g., binding specificity for an αE integrin), and preferably inhibit binding of ligand (e.g., E-cadherin) to an αE integrin (e.g., αEβ7). Antigen-binding fragments of antibodies encompassed by the invention include, Fv fragments (e.g., single chain Fv fragments (scFv)), Fab fragments, Fab′ fragments and F(ab′) 2  fragments, for example. Such antigen-binding fragments can be produced using any suitable method, for example by enzymatic cleavage and/or using recombinant DNA technology. For example, an antibody can be cleaved with papain or pepsin to yield a Fab fragment or F(ab′) 2  fragment, respectively. Other proteases with the requisite substrate specificity can also be used to generate antigen-binding fragments of antibodies, such as Fab fragments or F(ab′) 2  fragments. Similarly, Fv fragments can be prepared by digesting an antibody with a suitable protease or using recombinant DNA technology. For example, a nucleic acid can be prepared that encodes a light chain variable region and heavy chain variable region that are connected by a suitable peptide linker, such as a chain of two to about twenty Glycyl residues. The nucleic acid can be introduced into a suitable host (e.g.,  E. coli ) using any suitable technique (e.g., transfection, transformation, infection), and the host can be maintained under conditions suitable for expression of a single chain Fv fragment. A variety of antigen-binding fragments of antibodies can be prepared using antibody genes in which one or more stop codons has been introduced upstream of the natural stop site. For example, an expression construct encoding a F(ab′) 2  portion of an immunoglobulin heavy chain can be designed by introducing a translation stop codon at the 3′ end of the sequence encoding the hinge region of the heavy chain.  
     [0058] The invention also relates to the individual heavy and light chains of the antibodies (e.g., mouse antibodies, human antibodies, humanized antibodies, chimeric antibodies) that bind an αE integrin and to antigen-binding portions thereof. The heavy chains or light chains (and antigen-binding portions thereof) of the invention can bind an αE integrin when paired with a complementary light or heavy chain, respectively. Complementary chains can be identified using any suitable method (e.g., phage display, transgenic animals). For example, a transgenic animal comprising a functionally rearranged nucleic acid encoding a desired heavy chain can be prepared. The heavy-chain transgenic animal can be immunized with the antigen of interest and hybridomas produced. Because of allelic exclusion at immunoglubulin loci, the heavy-chain transgenic mouse may not significantly express endogenous heavy chains and substantially all antibodies elicited by immunization can comprise the heavy chain of interest and a complementary light chain.  
     [0059] The antigen-binding properties (e.g., specificity, affinity) of antibodies and antigen-binding fragments of antibodies can be elucidated using any suitable method. For example, binding specificity can be determined using assays in which formation of a complex between antibody or antigen-binding fragment and an αE integrin, such as an αEβ7 integrin, is detected or measured. Compositions which comprise an αE integrin and which can be used to assess antigen-binding properties of the antibodies and antigen-binding fragments described herein include, a membrane fraction of a cell comprising an αEβ7 integrin, a cell bearing an αEβ7 integrin, such as a human lymphocyte, human lymphocyte cell line or recombinant host cell comprising a nucleic acid encoding αE and/or β7 which expresses an αEβ7 integrin, a recombinant soluble αEβ7, such as ts.αEβ7.coil described herein, and the like. Binding and/or adhesion assays or other suitable methods can also be used in procedures for the identification and/or isolation of antibodies (e.g., human and/or humanized antibodies) having the requisite specificity (e.g., an assay in which adhesion between a cell bearing an αEβ7 integrin and a ligand thereof (e.g., a second cell expressing E-cadherin, an immobilized E-cadherin fusion protein (e.g., E-cadherin-Fc fusion protein) is detected and/or measured), or other suitable methods.  
     [0060] The antibodies of the invention bind an αE integrin (e.g., αEβ7) and preferably bind integrin αE chain (CD103). In a preferred embodiment, the antibody or antigen-binding fragment selectively binds an activation-induced epitope on an integrin αE chain (CD103). The activation-induced epitope can be induced by activation with a divalent cation, such as Mn 2+ , Mg + , Ca 2+  or any combination of the foregoing. The activation-induced epitope on an integrin αE chain expressed on the surface of a cell (e.g., as integrin αEβ7) can also be induced by exposing the cell to phorbol esters (e.g., PMA), or suitable mitogens and/or growth factors. When the cell expressing an integrin αE chain is a T cell, the activation-induced epitope can be induced by signals transduced through the T cell receptor complex. Thus, antibodies that selectively bind an activation-induced epitope can be used to detect or identify activated T cells that express an αE integrin for diagnostic and/or therapeutic purposes.  
     [0061] In one embodiment, the antibody of the invention binds an activation-induced epitope that is induced by exposure of the αE integrin to a divalent cation. Such antibodies bind an integrin αE chain (CD103) in the presence of a divalent cation, such as Mn 2+ , but do not significantly bind an integrin αE chain in the absence of a divalent cation or in the presence of a suitable divalent cation chelating agent (e.g., EDTA).  
     [0062] In certain embodiments, the antibody selectively binds an activation-induced epitope on an integrin αE chain that comprises amino acid residues in the I domain (amino acids 199-390 of SEQ ID NO:2) of integrin αE chain.  
     [0063] In other embodiments, the antibody binds an αE integrin (e.g., selectively binds an activation-induced epitope on integrin αE chain) and inhibits binding of ligand, such as E-cadherin, to the αE integrin (e.g., αEβ7 integrin). For example, the antibody can inhibit αE integrin mediated adhesion of a cell expressing an αE integrin (e.g., αEβ7) to cells expressing a ligand for an αE integrin (e.g. E-cadherin), such as epithelial cells and/or endothelial cells. Preferably, the antibodies do not bind the X domain of integrin αE chain (amino acids 144-198 of SEQ ID NO: 2).  
     [0064] Preferred antibodies that bind an αE integrin (e.g., selectively bind an activation-induced epitope on an integrin αE chain) include chimeric antibodies, humanized antibodies and antigen-binding fragments of the foregoing. Particularly preferred antibodies are human antibodies and antigen-binding fragments of human antibodies.  
     [0065] As described herein, human antibodies designated mAb 3G6, mAb 5E4 and mAb 8D5 which bind integrin αE chain (CD103) have been produced. mAb 3G6 and mAb 5E4 were originally produced as IgM antibodies and mAb 8D5 was originally produced as an IgG2 antibody. As described herein, IgG1 forms of mAbs 3G6, 5E4 and 8D5 have also been produced.  
     [0066] mAb 3G6 (IgM) can be produced by hybridoma 3G6, also referred to as hybridoma 241 3G6.1.15, which was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4201. The invention relates to hybridoma 3G6, to the antibody it produces, antigen-binding fragments thereof, and to nucleic acids encoding the antibody and portions thereof (e.g., heavy chain, heavy chain variable region, light chain, light chain variable region).  
     [0067] An IgG1 form of mAb 3G6 can be produced by “3G6 CHO stable cell line,” also referred to as CHO 3G6 C1.2D6, which was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4204. The invention relates to cell line CHO 3G6 C1.2D6, to the antibody it produces, antigen-binding fragments thereof, and to nucleic acids encoding the antibody and portions thereof (e.g., heavy chain, heavy chain variable region, light chain, light chain variable region).  
     [0068] mAb 5E4 can be produced by hybridoma 5E4, also referred to as hybridoma 233 5E4.3.10, which was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4202. The invention relates to hybridoma 5E4, to the antibody it produces, antigen-binding fragments thereof, and to nucleic acids encoding the antibody and portions thereof (e.g., heavy chain, heavy chain variable region, light chain, light chain variable region).  
     [0069] An IgG1 form of mAb 5E4 can be produced by “5E4 CHO stable cell line,” also referred to as CHO 5G4 A1.2C12, which was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4205. The invention relates to cell line CHO 5G4 A1.2C12, to the antibody it produces, antigen-binding fragments thereof, and to nucleic acids encoding the antibody and portions thereof (e.g., heavy chain, heavy chain variable region, light chain, light chain variable region).  
     [0070] mAb 8D5 can be produced by hybridoma 8D5, also referred to as hybridoma 321 8D5.3.11.8, which was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4203. The invention relates to hybridoma 8D5, to the antibody it produces, antigen-binding fragments thereof, and to nucleic acids encoding the antibody and portions thereof (e.g., heavy chain, heavy chain variable region, light chain, light chain variable region). As described herein, hybridoma 8D5 produces an IgG2 antibody.  
     [0071] The antibodies and antigen-binding fragments of the invention can bind to the same or similar epitope as mAb 3G6, mAb 5E4 or mAb 8D5. Antibodies and antigen-binding fragments that bind the same or similar epitope as mAb 3G6, mAb 5E4 or mAb 8D5 be identified using any suitable method, such as a competitive binding assay. For example, as described herein, an antibody can be tested for the ability to competitively inhibit binding of mAb 3G6, mAb 5E4 or mAb 8D5 to a fusion protein comprising the I domain of integrin αE chain or to an αE integrin (e.g., αEβ7) expressed on the surface of a cell. Competitive inhibition of binding of mAb 3G6, mAb 5E4 or mAb 8D5 in this type of assay is indicative that the test antibody binds the same or similar epitope as mAb 3G6, mAb 5E4 or mAb 8D5.  
     [0072] In particular embodiments, the antibody can have the epitopic specificity of mAb 3G6, mAb 5E4 or mAb 8D5. The fine epitopic specificity of an antibody can be determined using any suitable method, such as mutational analysis. For example, as described herein, a series of integrin αE chain variants comprising amino acid replacements can be prepared and an antibody can be tested for the ability to bind each variant. Inhibited or abrogated binding to a variant comprising a particular amino acid substitution is indicative that the substituted amino acid is part of the epitope that the antibody binds. (See, Higgins et al.,  J. Biol. Chem . 275:25652-25664 (2000).) In one embodiment, the antibody of the invention has the epitopic specificity of mAb 8D5 and binds an epitope that comprises Phe298 of integrin αE chain (SEQ ID NO: 1).  
     [0073] In more particular embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 3G6. In more particular embodiments, the antibody comprises the heavy chain variable region of mAb 3G6 (SEQ ID NO: 4) and the light chain variable region of mAb 3G6 (SEQ ID NO: 9).  
     [0074] In other particular embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 5E4. In more particular embodiments, the antibody comprises the heavy chain variable region of mAb 5E4 (SEQ ID NO: 14) and the light chain variable region of mAb 5E4 (SEQ ID NO: 19).  
     [0075] In additional particular embodiments, the antibody comprises one, two or three heavy chain complementarity determining regions (HCDR1, HCDR2 and/or HCDR3) having the amino acid sequences of the heavy chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each heavy chain CDR can be conservatively substituted, and one, two or three light chain complementarity determining regions (LCDR1, LCDR2 and/or LCDR3) having the amino acid sequences of the light chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each light chain CDR can be conservatively substituted. Preferably, the antibody comprises the three heavy chain CDRs and the three light chain CDRs of mAb 8D5. In more particular embodiments, the antibody comprises the heavy chain variable region of mAb 8D5 (SEQ ID NO: 24) and the light chain variable region of mAb 8D5 (SEQ ID NO: 29).  
     [0076] In additional embodiments, the invention provides novel heavy chains and light chains of the antibodies and antigen-binding fragments described herein. In particular embodiments, the antibody heavy chains or antigen-binding portions thereof comprise at least two and preferably three CDRs having the amino acid sequences of the heavy chain CDRs of mAb 3G6, the heavy chain CDRs of mAb 5E4 or the heavy chain CDRs of mAb 8D5. Optionally, one or two amino acid residues in each heavy chain CDR can be conservatively substituted. In preferred embodiments, the antibody heavy chains or antigen-binding portions thereof comprise three CDRs that have the amino acid sequences of the three CDRs of the heavy chain of mAb 3G6, the three CDRs of the heavy chain of mAb 5E4 or the three CDRs of the heavy chain of mAb 8D5. In other embodiments, the antibody heavy chains or antigen-binding portions thereof comprise the heavy chain variable region of mAb 3G6, mAb 5E4 or mAb 8D5. For example, the antibody heavy chains can comprise an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 14 and SEQ ID NO: 24. The antibody heavy chains and portions thereof can comprise any suitable framework regions and/or constant regions (as described herein).  
     [0077] In certain embodiments, the antibody light chains or antigen-binding portions thereof comprise at least two and preferably three CDRs having the amino acid sequences of the light chain CDRs of mAb 3G6, or the light chain CDRs of mAb 5E4 or the light chain CDRs of mAb 8D5. Optionally, one or two amino acid residues in each light chain CDR can be conservatively substituted. In preferred embodiments, the antibody light chains or antigen-binding portions thereof comprise three CDRs that have the amino acid sequences of the three CDRs of the light chain of mAb 3G6, the three CDRs of the light chain of mAb 5E4 or the three CDRs of the light chain of mAb 8D5. In other embodiments, the antibody light chains or antigen-binding portions thereof comprise the light chain variable region of mAb 3G6, mAb 5E4 or mAb 8D5. For example, the antibody light chains can comprise an amino acid sequence selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 19 and SEQ ID NO: 29. The antibody light chains and portions thereof can comprise any suitable framework regions and/or constant regions (as described herein).  
     [0078] Fusion Proteins and Immuno-Conjugates  
     [0079] Fusion proteins and immunoconjugates can be produced in which an antibody moiety (e.g., antibody or antigen-binding fragment thereof, antibody chain or antigen-binding portion thereof) is linked directly or indirectly to a non-immunoglobulin moiety (i.e., a moiety which does not occur in immunoglobulins as found in nature). Fusion proteins comprise an antibody moiety and a non-immunoglobulin moiety that are components of a single continuous polypeptide chain. The non-immunoglobulin moiety can be located N-terminally, C-terminally or internally with respect to the antibody moiety. For example, some embodiments can be produced by the insertion of a nucleic acid encoding immunoglobulin sequences into a suitable expression vector, such as a pET vector (e.g., pET-15b, Novagen), a phage vector (e.g., pCANTAB 5 E, Pharmacia), or other vector (e.g., pRIT2T Protein A fusion vector, Pharmacia). The resulting construct can be expressed (e.g., in vivo by a suitable host cell, in vitro) to produce antibody chains that comprise a non-immunoglobulin moiety (e.g., Histidine tag, E tag, Protein A IgG binding domain). Fusion proteins can be isolated or recovered using any suitable technique, such as chromatography using a suitable affinity matrix (see e.g.,  Current Protocols in Molecular Biology  (Ausubel, F. M. et al., eds., Vol. 2, Suppl. 26, pp. 16.4.1-16.7.8 (1991)).  
     [0080] In other embodiments, the antibody moiety and non-immunoglobulin moiety may not be part of a continuous polypeptide chain, but can be connected or conjugated directly or indirectly through any suitable linker. Suitable methods for connecting or conjugating the moieties are well known in the art. (See, e.g., Ghetie et al.,  Pharmacol. Ther . 63:209-34 (1994)). A variety of suitable linkers (e.g., heterobifunctional reagents) and methods for preparing immuno-conjugates are well known in the art. (See, for example, Hermanson, G. T.,  Bioconjugate Techniques , Academic Press: San Diego, Calif. (1996).) Suitable non-immunoglobulin moieties for inclusion in an immuno-conjugate include a therapeutic moiety such as a toxin (e.g., cytotoxin, cytotoxic agent), a therapeutic agent (e.g., a chemotherapeutic agent, an antimetabolite, an alkylating agent, an anthracycline, an antibiotic, an anti-mitotic agent, a biological response modifier (e.g., a cytokine (e.g., an interleukin, an interferon, a tumor necrosis factor), a growth factor (e.g., a neurotrophic factor)), a plasminogen activator), a radionuclide (e.g, a radioactive ion), an enzyme and the like. Suitable cytotoxins or cytotoxic agents include any agent that is detrimental to cells. Examples of suitable cytotoxins or cytotoxic agents include TAXOL (paclitaxel, Bristol-Myers Squibb Company), cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin (e.g, mitomycin C), etoposide, tenoposide, vincristine, vinblastine, colchicine, doxorubicin, daunorubicin, dihydroxy anthracindione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, puromycin, maytansinoids (e.g., maytansinol (see U.S. Pat. No. 5,208,020), CC-1065 (see U.S. Pat. Nos. 5,475,092, 5,585,499, 5,846,545), DM1) and analogs or homologs of any of the forgoing agents. Suitable therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepachlorambucil, CC-1065, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine, vinblastine, TAXOL (paclitaxel, Bristol-Myers Squibb Company) and maytansinoids (e.g., maytansinol (see U.S. Pat. No. 5,208,020), CC-1065 (see U.S. Pat. Nos. 5,475,092, 5,585,499, 5,846,545), DM1)). Suitable radionuclides include, for example iodine (e.g., iodine-125, -126) yttrium (e.g., yttrium-90, -91) and praseodymium (e.g., praseodymium-144, -145).  
     [0081] In certain embodiments, the therapeutic agent can be a protein or polypeptide possessing a desired biological activity. Such proteins or polypeptides can include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as a tumor necrosis factor (e.g., TNFα, TNFβ), and interferon (e.g., α-interferon, β-interferon, γ-interferion), a neurotrophic factor (e.g., nerve growth factor), a growth factor (e.g., platelet derived growth factor), a plasminogen activator (e.g., tissue plasminogen activator); or biological response modifiers such as, for example, cytokines and lymphokines, (e.g., interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophase colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”)), or other growth factors. In other embodiments, the antibody or antigen-binding fragment of the invention can be conjugated to a second antibody or antigen-binding fragment to form an antibody heteroconjugate. (See, e.g., U.S. Pat. No. 4,676,980 (Segal).)  
     [0082] Nucleic Acids and Constructs  
     [0083] The present invention also relates to isolated and/or recombinant (including, e.g., essentially pure) nucleic acids comprising sequences which encode an antibody or antigen-binding fragment (e.g., a human, humanized, chimeric antibody or light or heavy chain of any of the foregoing) or fusion protein of the invention.  
     [0084] Nucleic acids referred to herein as “isolated” are nucleic acids which have been separated away from other material (e.g., other nucleic acids such as genomic DNA, cDNA and/or RNA) in its original environment (e.g., in cells or in a mixture of nucleic acids such as a library). An isolated nucleic acid can be isolated as part of a vector (e.g., a plasmid). Nucleic acids can be naturally occurring, produced by chemical synthesis, by combinations of biological and chemical methods (e.g., semisynthetic), and be isolated using any suitable methods.  
     [0085] Nucleic acids referred to herein as “recombinant” are nucleic acids which have been produced by recombinant DNA methodology, including methods which rely upon artificial recombination, such as cloning into a vector or chromosome using, for example, restriction enzymes, homologous recombination, viruses and the like, and nucleic acids prepared using the polymerase chain reaction (PCR). “Recombinant” nucleic acids are also those that result from recombination of endogenous or exogenous nucleic acids through the natural mechanisms of cells or cells modified to allow recombination (e.g., cells modified to express Cre or other suitable recombinase), but are selected for after the introduction to the cells of nucleic acids designed to allow and make recombination probable. For example, a functionally rearranged human-antibody transgene is a recombinant nucleic acid.  
     [0086] The present invention also relates more specifically to nucleic acids that encode the heavy chains and/or light chains of the antibodies and antigen-binding portions described herein. For example, in one embodiment, the nucleic acid can encode a heavy chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the heavy chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In another embodiment, the nucleic acid can encode a heavy chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the heavy chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In another embodiment, the nucleic acid can encode a heavy chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the heavy chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In preferred embodiments, the nucleic acid encodes an antibody heavy chain or antigen-binding portion thereof that comprises three CDRs that have the amino acid sequences of the three CDRs of the heavy chain of mAb 3G6, the three CDRs of the heavy chain of mAb 5E4 or the three CDRs of the heavy chain of mAb 8D5. In other embodiments, the nucleic acid encodes an antibody heavy chain or antigen-binding portion thereof that comprises the heavy chain variable region of mAb 3G6, mAb 5E4 or mAb 8D5. For example, the nucleic acid can comprise a nucleotide sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 13 and SEQ ID NO: 23. The antibody heavy chains and portions thereof can comprise any suitable framework regions and/or constant regions (as described herein).  
     [0087] In another embodiment, the nucleic acid can encode a light chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the light chain CDRs of mAb 3G6 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In another embodiment, the nucleic acid can encode a light chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the light chain CDRs of mAb 5E4 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In another embodiment, the nucleic acid can encode a light chain or antigen-binding portion thereof that comprises at least one, two or preferably three CDRs having the amino acid sequences of the light chain CDRs of mAb 8D5 wherein, optionally, one or two amino acids in each CDR can be conservatively substituted. In preferred embodiments, the nucleic acid encodes an antibody light chain or antigen-binding portion thereof that comprises three CDRs that have the amino acid sequences of the three CDRs of the light chain of mAb 3G6, the three CDRs of the light chain of mAb 5E4 or the three CDRs of the light chain of mAb 8D5. In other embodiments, the nucleic acid encodes an antibody light chain or antigen-binding portion thereof that comprises the light chain variable region of mAb 3G6, mAb 5E4 or mAb 8D5. For example, the nucleic acid can comprise a nucleotide sequence selected from the group consisting of SEQ ID NO: 8, SEQ ID NO: 18 and SEQ ID NO: 28. The antibody light chains and portions thereof can comprise any suitable framework regions and/or constant regions (as described herein).  
     [0088] Nucleic acid molecules of the present invention can be used in the production of antibodies (e.g., human antibodies, humanized antibodies, chimeric antibodies and antigen-binding fragments of the foregoing) that bind an αE integrin or integrin αE chain (CD103). For example, a nucleic acid (e.g., DNA) encoding an antibody of the invention can be incorporated into a suitable construct (e.g., an expression vector) for further manipulation or for production of the encoded polypeptide in suitable host cells.  
     [0089] Expression constructs or expression vectors suitable for the expression of a antibody or antigen-binding fragment that binds an αE integrin are also provided. For example, a nucleic acid encoding all or part of a desired antibody can be inserted into a nucleic acid vector, such as a plasmid or virus, for expression. The vector can be capable of replication in a suitable biological system (e.g., a replicon). A variety of suitable vectors are known in the art, including vectors which are maintained in single copy or multiple copy, or which become integrated into the host cell chromosome.  
     [0090] Suitable expression vectors can contain a number of components, for example, an origin of replication, a selectable marker gene, one or more expression control elements, such as a transcription control element (e.g., promoter, enhancer, terminator) and/or one or more translation signals, a signal sequence or leader sequence, and the like. Expression control elements and a signal or leader sequence, if present, can be provided by the vector or other source. For example, the transcriptional and/or translational control sequences of a cloned nucleic acid encoding an antibody chain can be used to direct expression.  
     [0091] A promoter can be provided for expression in a desired host cell. Promoters can be constitutive or inducible. For example, a promoter can be operably linked to a nucleic acid encoding an antibody, antibody chain or portion thereof, such that it directs transcription of the nucleic acid. A variety of suitable promoters for procaryotic (e.g., lac, tac, T3, T7 promoters for  E. coli ) and eucaryotic (e.g., simian virus 40 early or late promoter, Rous sarcoma virus long terminal repeat promoter, cytomegalovirus promoter, adenovirus late promoter, EG-1a promoter) hosts are available.  
     [0092] In addition, expression vectors typically comprise a selectable marker for selection of host cells carrying the vector, and, in the case of a replicable expression vector, an origin or replication. Genes encoding products which confer antibiotic or drug resistance are common selectable markers and may be used in procaryotic (e.g., β-lactamase gene (ampicillin resistance), Tet gene for tetracycline resistance) and eucaryotic cells (e.g., neomycin (G418 or geneticin), gpt (mycophenolic acid), ampicillin, or hygromycin resistance genes). Dihydrofolate reductase marker genes permit selection with methotrexate in a variety of hosts. Genes encoding the gene product of auxotrophic markers of the host (e.g., LEU2, URA3, HIS3) are often used as selectable markers in yeast. Use of viral (e.g., baculovirus) or phage vectors, and vectors which are capable of integrating into the genome of the host cell, such as retroviral vectors, are also contemplated.  
     [0093] Suitable expression vectors for expression in mammalian cells include, for example, pCDM8, pCDNA1.1/amp, pcDNA3.1, pRc/RSV, pEF-1 (Invitrogen, Carlsbad, Calif.), pCMV-SCRIPT, pFB, pSG5, pXT1 (Stratagene, La Jolla, Calif.), pCDEF3 (Goldman, L. A., et al.,  Biotechniques , 21:1013-1015 (1996)), pSVSPORT (GibcoBRL, Rockville, Md.), pEF-Bos (Mizushima, S., et al.,  Nucleic Acids Res ., 18:5322 (1990)) and the like. Expression vectors which are suitable for use in various expression hosts, such as prokaryotic cells ( E. coli ), insect cells (Drosophila Schnieder S2 cells, Sf9) and yeast ( P. methanolica, P. pastoris, S. cerevisiae ) are also available.  
     [0094] Thus, the invention provides an expression vector comprising a nucleic acid encoding an antibody, antigen-binding fragment of an antibody (e.g., a human, humanized, chimeric antibody or antigen-binding fragment of any of the foregoing), antibody chain (e.g., heavy chain, light chain) or antigen-binding portion of an antibody chain that binds an αE integrin (e.g., an integrin αE chain (CD103)).  
     [0095] Recombinant Host Cells and Methods of Production  
     [0096] In another aspect, the invention relates to recombinant host cells and a method of preparing an antibody or antigen-binding fragment, antibody chain (e.g., heavy chain, light chain) or antigen-binding portion of an antibody chain, or fusion protein of the invention. The antibody or antigen-binding fragment can be obtained, for example, by the expression of one or more recombinant nucleic acids encoding an antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding portion of an antibody chain that binds an αE integrin in a suitable host cell, or using other suitable methods. For example, the expression constructs described herein can be introduced into a suitable host cell, and the resulting cell can be maintained (e.g., in culture, in an animal, in a plant) under conditions suitable for expression of the constructs. Suitable host cells can be prokaryotic, including bacterial cells such as  E. coli, B. subtilis  and/or other suitable bacteria; eucaryotic cells, such as fungal or yeast cells (e.g.,  Pichia pastoris , Aspergillus sp.,  Saccharomyces cerevisiae, Schizosaccharomyces pombe, Neurospora crassa ), or other lower eukaryotic cells, and cells of higher eucaryotes such as those from insects (e.g., Drosophila Schnieder S2 cells, Sf9 insect cells (WO 94/26087 (O&#39;Connor)), mammals (e.g., COS cells, such as COS-1 (ATCC Accession No. CRL-1650) and COS-7 (ATCC Accession No. CRL-1651), CHO (e.g., ATCC Accession No. CRL-9096), 293 (ATCC Accession No. CRL-1573), HeLa (ATCC Accession No. CCL-2), CV1 (ATCC Accession No. CCL-70), WOP (Dailey, L., et al.,  J. Virol ., 54:739-749 (1985), 3T3, 293T (Pear, W. S., et al.,  Proc. Natl. Acad. Sci. U.S.A ., 90:8392-8396 (1993)) NSO cells, SP2/0, HuT 78 cells and the like, or plants (e.g., tobacco). (See, for example, Ausubel, F. M. et al., eds.  Current Protocols in Molecular Biology , Greene Publishing Associates and John Wiley &amp; Sons Inc. (1993).)  
     [0097] The invention also relates to a recombinant host cell which comprises a (one or more) recombinant nucleic acid or expression construct comprising a nucleic acid encoding an antibody, antigen-binding fragment of an antibody (e.g., a human, humanized, chimeric antibody or antigen-binding fragment of any of the foregoing), antibody chain (e.g., heavy chain, light chain) or antigen-binding portion of an antibody chain that binds an αE integrin (e.g., an integrin αE chain (CD103)). In particular embodiments, the recombinant host cell is hybridoma 3G6, hybridoma 5E4, hybridoma 8D5, CHO 3G6 C1.2D6 or CHO 5G4 A1.2C12.  
     [0098] The invention also includes a method of preparing an antibody, antigen-binding fragment of an antibody (e.g., a human, humanized, chimeric antibody or antigen-binding fragment of any of the foregoing), antibody chain (e.g., heavy chain, light chain) or antigen-binding portion of an antibody chain that binds an αE integrin (e.g., an integrin αE chain (CD103)), comprising maintaining a recombinant host cell of the invention under conditions appropriate for expression of an antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding fragment of an antibody chain. The method can further comprise the step of isolating or recovering the antibody, antigen-binding fragment of an antibody, antibody chain or antigen-binding fragment of an antibody chain, if desired.  
     [0099] For example, a nucleic acid molecule (i.e., one or more nucleic acid molecules) encoding the heavy and light chains of a human antibody that binds an integrin αE chain, or an expression construct (i.e., one or more constructs) comprising such nucleic acid molecule(s), can be introduced into a suitable host cell to create a recombinant host cell using any method appropriate to the host cell selected (e.g., transformation, transfection, electroporation, infection), such that the nucleic acid molecule(s) are operably linked to one or more expression control elements (e.g., in a vector, in a construct created by processes in the cell, integrated into the host cell genome). The resulting recombinant host cell can be maintained under conditions suitable for expression (e.g., in the presence of an inducer, in a suitable animal, in suitable culture media supplemented with appropriate salts, growth factors, antibiotics, nutritional supplements, etc.), whereby the encoded polypeptide(s) are produced. If desired, the encoded protein can be isolated or recovered (e.g., from the animal, the host cell, medium, milk). This process encompasses expression in a host cell of a transgenic animal (see, e.g., WO 92/03918, GenPharm International).  
     [0100] The antibodies, antigen-binding fragments, antibody chains and antigen-binding portions thereof described herein can also be produced in a suitable in vitro expression system, by chemical synthesis or by any other suitable method.  
     [0101] Diagnostic and Therapeutic Methods  
     [0102] The antibodies (including fragments), fusion proteins and immuno-conjugates described herein can bind an αE integrin and can be used to detect, measure, select, isolate and/or purify an αE integrin (e.g., αEβ7 integrin) or variants thereof (e.g., by affinity purification or other suitable methods), and to study αE integrin structure (e.g., conformation) and function. The antibodies, fusion proteins and immuno-conjugates of the present invention can also be used in diagnostic applications (e.g., in vitro, ex vivo) and/or in therapeutic applications.  
     [0103] The antibodies, fusion proteins and immuno-conjugates can be used to detect and/or measure the level of an αE integrin (e.g., αEβ7 integrin) in a sample (e.g., tissues or body fluids, such as an inflammatory exudate, bronchial lavage, blood, serum, bowel fluid, biopsy). In one example, a sample (e.g., tissue and/or body fluid) can be obtained from an individual and a suitable immunological method can be used to detect and/or measure αE integrin expression. Suitable immunological methods for detecting or measuring αE integrin expression include enzyme-linked immunosorbent assays (ELISA), radioimmunoassay, immunohistology, flow cytometry, and the like.  
     [0104] In one embodiment, the invention is a method of detecting or measuring an activated αE integrin in a sample (e.g., a biological sample) comprising contacting a sample (e.g., a biological sample) with an antibody or antigen-binding fragment thereof that binds an activation-induced epitope on an αE integrin (e.g., on an integrin αE chain (CD103)) under conditions suitable for binding of the antibody or antigen-binding fragment to the αE integrin and detecting and/or measuring binding of the antibody or antigen-binding fragment to the αE integrin. Binding of the antibody or antigen-binding fragment thereof to the αE integrin indicates the presence of the αE integrin in the sample. In an application of the method, an antibody or antigen-binding fragment of the invention can be used to analyze normal versus inflamed tissues (e.g., from a human) for activated αE integrin reactivity and/or expression to detect associations between disease (e.g., inflammatory bowel disease, graft rejection) and increased expression of activated αE (e.g., in affected tissues). In embodiments where the antibody or antigen-binding fragment binds an activation-induced epitope, the antibodies, antigen-binding fragments, fusion proteins and immuno-conjugates of the invention can be used to detect, measure, select, isolate and/or purify activated αE integrin or cells expressing an activated αE integrin.  
     [0105] The antibodies, fusion proteins and/or immuno-conjugates of the present invention permit assessment of the presence of an αE integrin in normal versus inflamed tissues, through which the presence or severity of disease, disease progress and/or the efficacy of therapy can be assessed. For example, therapy can be monitored and efficacy assessed. In one example, an αE integrin can be detected and/or measured in a first sample obtained from a subject having an inflammatory disease and therapy can be initiated. Later, a second sample can be obtained from the subject and αE integrin in the sample can be detected and/or measured. A decrease in the quantity of αE integrin detected or measured in the second sample can be indicative of therapeutic efficacy.  
     [0106] The antibodies, fusion proteins and immuno-conjugates described herein can modulate an activity or function of an αE integrin (e.g., αEβ7 integrin), such as ligand binding (e.g., E-cadherin) and/or leukocyte infiltration function, including recruitment and/or accumulation of leukocytes (e.g., T cells) in tissues. Antibodies, fusion proteins and immuno-conjugates that bind an activation-induced epitope can be used to selectively target cells expressing activated αE integrin (e.g., αEβ7 integrin) for therapy. For example, an antibody that binds an activation-induced epitope on an αEβ7 integrin and is capable of activating complement (e.g., a human IgG1 antibody) can be administered to selectively deplete cells expressing activated αEβ7 through, for example, complement-mediated lysis.  
     [0107] Preferably the antibodies, fusion proteins and immuno-conjugates can selectively bind an αE integrin (e.g., αEβ7 integrin) and inhibit αE integrin-mediated interactions, such as αE integrin-mediated adhesion of a cell (e.g., T cell) to endothelial cells. In particularly preferred embodiments, the antibodies, fusion proteins and immuno-conjugates can inhibit the interaction of αEβ7 with E-cadherin.  
     [0108] The antibodies, fusion proteins and immuno-conjugates described herein can be administered to a subject to modulate an inflammatory response or to treat an inflammatory disease or disorder. For example, an antibody which inhibits the binding of an αE integrin to a ligand (i.e., one or more ligands) can be administered in the treatment of diseases associated with leukocyte (e.g., lymphocyte, monocyte) infiltration of tissues, particularly of mucosal tissues. An effective amount of an antibody, fusion protein and/or immuno-conjugate (i.e., one or more) can be administered to a subject (e.g., a mammal, such as a human or other primate) in order to treat such a disease. For example, inflammatory diseases, including diseases which are associated with leukocyte infiltration of the gastrointestinal tract (including gut-associated endothelium), other mucosal tissues, or tissues expressing the molecule E-cadherin (e.g., mucosal epithelial surfaces), can be treated according to the present method. Similarly, an individual having a disease associated with leukocyte infiltration of tissues as a result of binding of leukocytes to cells (e.g., epithelial cells) expressing E-cadherin can be treated according to the present invention.  
     [0109] Examples of inflammatory diseases associated with mucosal tissues which can be treated according to the present method include mastitis (mammary gland), cholecystitis, cholangitis or pericholangitis (bile duct and surrounding tissue of the liver), chronic bronchitis, chronic sinusitis, asthma, and graft versus host disease (e.g., in the gastrointestinal tract). As seen in Crohn&#39;s disease, mucosal inflammation often extends beyond the mucosal surface. Accordingly chronic inflammatory diseases of the lung which result in interstitial fibrosis, such as hypersensitivity pneumonitis, collagen diseases, sarcoidosis, and other idiopathic conditions can be amenable to treatment.  
     [0110] According to the method, the severity of symptoms associated with an inflammatory condition can be inhibited (reduced) in whole or in part. When the subject has a relapsing or chronic condition, an effective amount of an antibody, fusion protein and/or immuno-conjugate of the invention can be administered to treat the subject, and therapy can be continued (maintenance therapy) with the same or different dosing as indicated, to inhibit relapse or renewed onset of symptoms. Preferably, the antibodies, fusion proteins and/or immuno-conjugates are administered to treat a subject having a mucosal inflammatory diseases, such as an inflammatory disease of the respiratory tract (e.g., bronchus, lung), urogenital tract (e.g., kidney, urinary bladder) or alimentary canal and associated organs and tissues (e.g., mouth, salivary glands, esophagus, stomach, small intestine, colon, pancreas, liver, gall bladder).  
     [0111] In a particularly preferred embodiment, the subject to be treated has an inflammatory bowel disease (IBD), such as ulcerative colitis, Crohn&#39;s disease, ileitis, Celiac disease, nontropical Sprue, enteropathy associated with seronegative arthropathies, colitis (e.g., microscopic or collagenous colitis), gastroenteritis (e.g., eosinophilic gastroenteritis), or pouchitis resulting after proctocolectomy and ileoanal anastomosis. Subjects having pancreatitis or insulin-dependent diabetes mellitus can also be treated using the present method. In another embodiment, the subject to be treated has an has on oral inflammatory disease, Sjogren&#39;s syndrome or Behcet&#39;s syndrome.  
     [0112] In another embodiment, the subject to be treated has a pulmonary inflammatory disease, such as a chronic obstructive lung disease (e.g., chronic bronchitis, asthma, silicosis, chronic obstructive pulmonary disease), hypersensitivity pneumonitis, pulmonary fibrosis (e.g., idiopathic pulmonary fibrosis) or sarcoidosis. In another embodiment, the subject to be treated has a cutaneous inflammatory disease, such as psoriasis or inflammatory dermatoses.  
     [0113] In another embodiment, the invention is a method of inhibiting graft rejection (e.g., allograft rejection, xenograft rejection) or graft versus host disease, comprising administering to a subject in need thereof an effective amount of an antibody, fusion protein and/or immuno-conjugate of the invention. In particular embodiments, the transplanted graft is a mucosa-associated organ or tissue, such as kidney, liver, lung and the like.  
     [0114] The invention also relates to a method of inhibiting αE integrin (e.g. αEβ7 integrin) mediated homing of leukocytes in a subject, comprising to a subject in need thereof an effective amount of an antibody, fusion protein and/or immuno-conjugate of the invention. For example, the homing of leukocytes to mucosal sites (e.g., gut, lung) can be inhibited.  
     [0115] As used herein, “subject” refers to humans and animals such as mammals, including, primates, cows, sheep, goats, horses, dogs, cats, rabbits, guinea pigs, rats, mice or other bovine, ovine, equine, canine, feline, rodent or murine species.  
     [0116] Diseases and conditions associated with inflammation, infection, and cancer can be treated using the method. In a preferred embodiment, the disease or condition is one in which the actions of cells bearing an αE integrin (e.g., αEβ7), such as lymphocytes (e.g., activated or stimulated T lymphocytes), are to be inhibited or promoted for therapeutic or prophylactic purposes.  
     [0117] Diseases or conditions, including chronic diseases, of humans or other species which can be treated with the antibodies, fusion proteins and/or immuno-conjugates of the invention, include, but are not limited to:  
     [0118] inflammatory or allergic diseases and conditions, including systemic anaphylaxis or hypersensitivity responses, drug allergies (e.g., to penicillin, cephalosporins), insect sting allergies; inflammatory bowel diseases, such as Crohn&#39;s disease, ulcerative colitis, celiac disease, ileitis and enteritis; sarcoidosis; vaginitis; psoriasis and inflammatory dermatoses such as dermatitis, eczema, atopic dermatitis, allergic contact dermatitis, urticaria; vasculitis (e.g., necrotizing, cutaneous, and hypersensitivity vasculitis); spondyloarthropathies; scleroderma; respiratory allergic diseases such as asthma, allergic rhinitis, hypersensitivity lung diseases, hypersensitivity pneumonitis, interstitial lung diseases (ILD) (e.g., idiopathic pulmonary fibrosis, or ILD associated with rheumatoid arthritis, or other autoimmune conditions);  
     [0119] autoimmune diseases, such as arthritis (e.g., rheumatoid arthritis, psoriatic arthritis), multiple sclerosis, systemic lupus erythematosus, myasthenia gravis, diabetes, including diabetes mellitus and juvenile onset diabetes, glomerulonephritis and other nephritides, autoimmune thyroiditis, Behcet&#39;s syndrome;  
     [0120] graft rejection (e.g., in transplantation), including allograft rejection or graft-versus-host disease;  
     [0121] viral infection, e.g., infection by hepatitis C virus (HCV), human papilloma virus (HPV), respiratory syncytial virus, influenza virus, simian immunodeficiency virus (SIV) or human immunodeficiency virus (HIV);  
     [0122] cancers and/or neoplastic diseases, such as leukemias and lymphomas;  
     [0123] other diseases or conditions in which undesirable inflammatory responses are to be inhibited can be treated, including, but not limited to, atherosclerosis (e.g., transplant accelerated atherosclerosis), restenosis, cytokine-induced toxicity, myositis (including polymyositis, dermatomyositis).  
     [0124] Modes of Administration  
     [0125] According to the method, an (i.e., one or more) antibody, antigen-binding fragment thereof, fusion protein and/or immuno-conjugate can be administered to the subject by an appropriate route, either alone or in combination with another drug. An “effective amount” of antibody, fusion protein and/or immuno-conjugate is administered. An “effective amount” is an amount sufficient to achieve the desired therapeutic or prophylactic effect, under the conditions of administration, such as an amount sufficient to inhibit binding of αE integrin (αEβ7 integrin) to E-cadherin expressed on epithelial cells, and thereby, inhibit αE integrin-mediated function, such as leukocyte binding, extravasation and/or retention (e.g., as intra-epithelial lymphocytes (IEL)). The antibody, fusion protein and/or immuno-conjugate can be administered in a single dose or multiple doses. The antibody or antigen-binding fragment can be administered as a bolus and/or infusion (e.g., continuous infusion). The dosage can be determined by methods known in the art and is dependent, for example, upon the antibody, antigen-binding fragment, fusion protein and/or immuno-conjugate chosen, the subject&#39;s age, sensitivity and tolerance to drugs, and overall well-being. Typically, an effective amount can range from about 0.01 mg per day to about 100 mg per day for an adult. Preferably, the dosage ranges from about 1 mg per day to about 100 mg per day or from about 1 mg per day to about 10 mg per day. Human, humanized and chimeric antibodies can often be administered with less frequency than other types of therapeutics. For example, an effective amount of a human, humanized or chimeric antibody (or antigen-binding fragment of any of the foregoing) can range from about 0.01 mg/kg to about 5 or 10 mg/kg administered daily, weekly, biweekly or monthly.  
     [0126] A variety of routes of administration are possible including, for example, oral, dietary, topical, transdermal, rectal, parenteral (e.g., intravenous, intraarterial, intramuscular, subcutaneous, intradermal, intraperatoneal injection), and inhalation (e.g., intrabronchial, intranasal or oral inhalation, intranasal drops) routes of administration, depending on the agent and disease or condition to be treated. Administration can be local or systemic as indicated. The preferred mode of administration can vary depending upon the agent chosen, and the condition (e.g., disease) being treated, however, oral or parenteral administration is generally preferred.  
     [0127] The antibody, fusion protein and/or immuno-conjugate and any other therapeutic agent to be administered can be administered as a neutral compound or as a salt. Salts of compounds (e.g., an antibody) containing an amine or other basic group can be obtained, for example, by reacting with a suitable organic or inorganic acid, such as hydrogen chloride, hydrogen bromide, acetic acid, perchloric acid and the like. Compounds with a quaternary ammonium group also contain a counteranion such as chloride, bromide, iodide, acetate, perchlorate and the like. Salts of compounds containing a carboxylic acid or other acidic functional group can be prepared by reacting with a suitable base, for example, a hydroxide base. Salts of acidic functional groups contain a countercation such as sodium, potassium and the like.  
     [0128] The antibody, fusion protein and/or immuno-conjugate can be administered to the individual as part of a pharmaceutical composition for modulation (e.g., inhibition) of αE integrin function (e.g., ligand binding and/or leukocyte infiltration), or treating a subject having a disease described herein. The pharmaceutical composition can comprise an antibody, antigen-binding fragment, fusion protein and/or immuno-conjugate of the invention and a pharmaceutically or physiologically acceptable carrier. Formulation will vary according to the route of administration selected (e.g., solution, emulsion, capsule). Suitable pharmaceutical and physiological carriers can contain inert ingredients which do not interact with the antibody, fusion protein and/or immuno-conjugate. Standard pharmaceutical formulation techniques can be employed, such as those described in Remington&#39;s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. Suitable pharmaceutical carriers for parenteral administration include, for example, sterile water, physiological saline, bacteriostatic saline (saline containing about 0.9% benzyl alcohol), phosphate-buffered saline, Hank&#39;s solution, Ringer&#39;s-lactate and the like. Methods for encapsulating compositions (such as in a coating of hard gelatin or cyclodextran) are known in the art (Baker, et al, “Controlled Release of Biological Active Agents”, John Wiley and Sons, 1986). For inhalation, the agent can be solubilized and loaded into a suitable dispenser for administration (e.g., an atomizer, nebulizer or pressurized aerosol dispenser).  
     [0129] Furthermore, the antibody or fusion protein of the invention and other therapeutic agents that are proteins can be administered via in vivo expression of the recombinant protein. In vivo expression can be accomplished via somatic cell expression according to suitable methods (see, e.g. U.S. Pat. No. 5,399,346). In this embodiment, a nucleic acid encoding the protein can be incorporated into a retroviral, adenoviral or other suitable vector (preferably, a replication deficient infectious vector) for delivery, or can be introduced into a transfected or transformed host cell capable of expressing the protein for delivery. In the latter embodiment, the cells can be implanted (alone or in a barrier device), injected or otherwise introduced in an amount effective to express the protein in a therapeutically effective amount.  
     [0130] The present invention will now be illustrated by the following Examples, which are not intended to be limiting in any way.  
     EXAMPLES  
     Methods and Materials  
     [0131] E-cadherin-IgG Fusion Protein  
     [0132] A DNA fragment encoding human E-cadherin extracellular domain (residues 1-695 of SEQ ID NO: 34) was isolated by PCR using full-length E-cadherin cDNA as template. Synthetic primers Spe-ECAD(5) (gcactagtccaccatgggcccttggagccgc; SEQ ID NO: 42) and ECAD-XHO(3) (ccctcgagaggctgtgccttcctaca; SEQ ID NO: 43) were designed so that SpeI and XhoI restriction sites were incorporated at the 5′ and 3′ of the PCR product, respectively. The PCR product was digested with SpeI and XhoI.  
     [0133] A DNA fragment coding for an human IgG Fc fragment (including the hinge, CH2 and CH3) was isolated by PCR using a fusion construct that encodes a fusion protein that contains a human IgG1 constant region that has been mutated to inhibit binding to Fc receptor as template and synthetic primers Xho-IgG(5) (atctcgagcccaaatcttgtgac; SEQ ID NO: 44) and IgGNot(3) (tagcggccgctcatttacccggagacag; SEQ ID NO: 45) which introduced XhoI and NotI sites at the 5′ and 3′ ends of the product, respectively. The product was cut with XhoI and NotI.  
     [0134] The PCR products (E-cadherin and IgG Fc) were ligated into vector pCDEF3 (Goldman, L. A., et al.,  Biotechniques , 21:1013-1015 (1996)) that had been linearized with SpeI and NotI. Vector pCDEF3 is a derivative of pcDNA (Invitrogen, Carlsbad, Calif.) and contains the EF-1 promoter. The sequence of the resulting E-cadherin-IgG fusion construct in pCDEF3 was confirmed by DNA sequencing.  
     [0135] The fusion construct encoded a fusion protein that contained a Leucine residue between the E-cadherin portion and the IgG1 Fc portion, and the IgG1 Fc portion contained mutations to reduce binding to Fc receptor.  
     [0136] Expression and Purification  
     [0137] The E-cadherin-IgG fusion construct was transiently transfected in 293T cells using calcium phosphate transfection method. 10 μg of the expression vector was used to transfect one 10 cm plate of 293T cells (Pear, W. S., et al.,  Proc. Natl. Acad. Sci. U.S.A ., 90:8392-8396 (1993)). For large scale purification of the fusion protein, 30-35 plates of cells were typically transfected. 7-11 hours post-transfection, the culture medium was changed to media supplemented with 10% ultra low IgG fetal bovine serum (Gibco). The transfected cells were cultured and the culture supernatant (10 mL) was collected daily for three days. The human IgG was isolated from the collected supernatant by chromatography using a protein A column at 4° C. The column was washed with TBS/Ca (20 mM Tris, pH 7.5, 140 mM NaCl, supplemented with 1 mM CaCl 2 ), and eluted with 100 mM Glycine-HCl (pH 2.3), 1 mM CaCl 2 . The eluate was immediately neutralized with 1M Tris pH 9.0 (1/15, v/v). Protein fractions were pooled and dialyzed in TBS/Ca overnight at 4° C. Protein concentration was determined by the Bradford method (Bio-Rad, Hercules, Calif.) using bovine IgG as standard, and protein purity was evaluated by SDS-PAGE.  
     [0138] Biotinylation  
     [0139] E-cadherin-IgG fusion protein was dialyzed in 10 mM Na-borate (pH 8.4), 0.5 mM CaCl 2  for overnight at 4° C., and labeled with aminohexanoyl-biotin-N-hydroxysuccinimide (AH-BNHS, Zymed, South San Francisco, Calif.) at a ratio of 1:10 (AH-BNHS/protein, w/w) for 1 hour at room temperature. The labeled protein was dialyzed in TBS (20 mM Tris-HCL, pH 7.5, 150 mM NaCl) supplemented with 1 mM CaCl 2  at 4° C. Protein concentration was determined using the Bradford method (Bio-Rad).  
     [0140] Soluble Recombinant αEβ7 Protein (ts αEβ7.coil)  
     [0141] A nucleic acid encoding the extracellular domain of integrin αE chain (amino acid residues 1-1105 of SEQ ID NO: 2) was fused with a nucleic acid encoding a 30 amino acid acidic peptide (AQLEKELQALEKENAQLEWELQALEKELAQ, SEQ ID NO: 39) to create a construct designated αE-acid. A nucleic acid encoding the extracellular domain of the β7 subunit (amino acid residues 1-707 of SEQ ID NO: 38) was fused with a nucleic acid encoding a 30 amino acid basic peptide (AQLKKKLQALKKKNAQLKWKLQALKKKLAQ, SEQ ID NO: 40) to create a construct designated β7-base. When expressed, the acidic and basic peptides form a heterodimeric coiled coil. (See, Lu et al,  J. Biol. Chem . 276:14642-14648 (2001); O&#39;Shea et al.,  Curr. Biol . 3:658-667 (1993).) A nucleic acid encoding a linker of six amino acid residues (GGSTGG, SEQ ID NO: 41) was inserted into both constructs (between the αE sequence and the acidic peptide, as well as between the β7 and the basic peptide). The αE and β7 fusion constructs were separately cloned into the expression vector AprM8 (see, Lu and Springer,  J. Immunol . 159:268-278 (1997)), and sequences were confirmed by DNA sequencing.  
     [0142] Expression and Purification  
     [0143] The αE-acid and β7-base constructs were transiently transfected into 293T cells and co-expressed to produce a soluble αEβ7. The secretion of soluble αEβ7 heterodimer (ts αEβ7.coil) by the transfected cells was confirmed by ELISA and immunoprecipitation using several antibodies that bound the αE or β7 subunit. For large-scale purification, 30-35 10 cm-plates of 293T cells were co-transfected with αE-acid and β7-base constructs, and culture supernatant was collected as described above.  
     [0144] ts αEP7.coil was purified by column chromatography using an anti-β7 antibody (mAb 6F7; Millennium Pharmaceuticals Inc., Cambridge, Mass.) affinity column. mAb 6F7 was covalently coupled to CNBr-activated SEPHAROSE 4B beads (beaded agarose, Pharmacia). Culture supernatant containing ts αEβP7.coil was applied to the column at 4° C. The column was washed with TBS (20 mM Tris-base, pH 7.5, 150 mM NaCl), 1 mM CaCl 2  and 1 mM MgCl 2  in cold, and eluted in 50 mM triethylamine (TEA), pH 11.5, 150 mM NaCl, 1 mM CaCl 2  and 1 mM MgCl 2 . The eluate was immediately neutralized with 1 mM Tris-HCl, pH 6.8, 5 mM CaCl 2  and 5 mM MgCl 2  (1/10, v/v). Protein fractions were pooled, concentrated using CENTRICON-30 membrane concentrator (Millipore, Bedford, Mass.), and buffer was changed to TBS, pH 7.5 containing 1 mM CaCl 2  and 1 mM MgCl 2 . The protein concentration was determined, and the purified sample was aliquoted and stored at −70° C. The purity of ts αEβ7.coil protein was about 90% as judged by SDS-PAGE and silver staining.  
     [0145] Transfectants  
     [0146] L1.2 cells (murine B lymphoma cell line) were cultured in RPMI/10% FetalClone (Hyclone). K562 cells were maintained in RPMI/10%FBS (Gibco). For stable expression of αEβ7, 20 μg of αE full length cDNA (SEQ ID NO: 1) in AprM8 and 20 μg of β7 cDNA (SEQ ID NO: 37) in AprM8 were linearized and cotransfected with 1 μg linear PEFpuro (see, Lu and Springer,  J. Immunol . 159:268-278 (1997)), which contains puromycin selection marker, by electroporation at 250 V, 960 μF using 0.4 cm cuvette. 48 hours post transfection, cells were collected, and resuspended in culture medium supplemented with 2 μg/ml or 4 μg/ml puromycin for L1.2 transfectants and K562 transfectants, respectively. Cells were subsquently subcloned in 96-well plates. Clones of transfectants were tested for αEβ7 cell surface expression by staining with mouse anti-αE and anti-β7 antibodies. Selected clones were subcloned again.  
     [0147] Mouse anti-αE mAb αE7.1 was described previously. (Russel, G. J. et al.,  Eur. J. Immunol . 24:2832-2841 (1994).) 293T cells (human embryonic kidney epithelial cell line) were maintained in Dulbecco&#39;s Modified Eagles Medium/10% FBS (Gibco), supplemented with essential amino acids and sodium pyruvate.  
     [0148] Generation of αEβ7-Specific Human Antibodies  
     [0149] Human monoclonal antibodies, mAb 3G6, mAb 5E4 and mAb 8D5, were generated using human-antibody transgenic mice that express human immunoglobulin genes. mAb 5E4 and mAb 3G6 were produced using HUMAB mice (MEDAREX, Princeton, N.J.), and mAb 8D5 was produced using XENOMOUSE mice (Abgenix, Fremont, Calif.). The same immunization, fusion and antibody screening protocols were used to produce human monoclonal antibody 3G6, human monoclonal antibody 5E4 and human monoclonal antibody 8D5.  
     [0150] Immunization  
     [0151] L1.2 transfectants that express human αEβ7 were treated with mitomycin C at 100 μg/ml for 30 minutes at 37° C. Cells were washed twice with phosphate buffered saline (PBS), and resuspended at 2×10 7  cells/ml in PBS. Mice were injected with about 0.5 ml of the resulting cell suspension (intraperitonial injection (IP), 10 7  cells/mouse/injection) at about two week intervals. After 4 IP injections, mice were boosted with purified recombinant αEβ7 protein (ts αEβ7.coil)(15 μg/mouse, intravenous (IV) injection). 4 days after the IV boost, mice were tested for αEβ7-specific human IgG response in the serum. Spleens from positive mice were used for fusion.  
     [0152] Titration of αEβ7-Specific Human IgG  
     [0153] A sandwich ELISA was used to titrate mouse sera containing human IgG antibodies that bind αEβ7 integrin. ELISA plates were coated with 15 μg/ml mouse anti-β7 mAb 6F7 (50 μl/well) at 37° C. for 2 hours. The plates were then washed with PBS and incubated with 50 μl culture supernatant containing recombinant αEβ7 protein overnight at 4° C. The plate was washed twice with PBS, and incubated with mouse anti-serum at various dilutions in PBS at 37° C., for 1 hour. Then, the plates were washed twice, and the plate was incubated with HRP-conjugated goat anti-human IgG at 37° C. for 1 hour. The plates were then washed again and human antibodies that bound αEβ7 were detected by addition of peroxidase substrate, and absorbance was read on an ELISA reader at 410 nM wavelength.  
     [0154] Hybridomas that Produce Antibodies that Bind αEβ7  
     [0155] Spleens were removed from mice that produced anti-αEβ7 antibodies and splenocytes were fused with myeloma cells (SP2/0) to produce hybridomas.  
     [0156] Hybridomas were Screened for Production of Anti-αEβ7 Antibodies using a Flow Cytometry Assay and an ELISA.  
     [0157] L1.2 αEβ7 transfectants or untransfected cells (negative control) were collected by centrifugation, and resuspended to 10 7  cells/ml in PBS/5% FBS. 50 μl of cell suspension (5×10 5  cells) was incubated with 50 μl hybridoma supernatant in a 96-well plate for 30 minutes on ice. The cells were washed once with PBS/5% FBS, and incubated with FITC-conjugated anti-human IgG or IgM for 30 minutes on ice. The cells were washed again, resuspended in PBS, and antibody binding was measured by flow cytometry using a FACS instrument. Hybridoma supernatants that stained L1.2 αEβ7 transfectants but not the untransfected parental L1.2 cells were saved and screened further by αEβ7-specific ELISA. The protocol for the ELISA was identical to the ELISA described above except that 50 μl hybridoma supernatant was used instead of diluted serum. Positive hybridomas were further tested for αE specificity.  
     [0158] Screen for αE-Specific Antibodies  
     [0159] FACS staining of K562 transfectants that express either αEβ7 or α4β7 integrin was used. FACs staining protocol was the same as described above. Hybridomas that stained αEβ7 transfectants but not α4β7 transfectants were selected as producing αE-specific antibody. αE-specific hybridomas were further subcloned at least twice by limiting dilution.  
     [0160] Assays for Selecting Antibodies that Inhibit Binding of αEβP7 to E-cadherin  
     [0161] Cell Adhesion Assay.  
     [0162] ELISA plates were coated with 100 ng/well E-cadherin-IgG fusion protein in TBS (20 mM Tris, 140 mM NaCl, pH 9)/1 mM CaCl 2  overnight at 4° C. Plates were washed with wash buffer (HBSS/1 mM CaCl 2 ), and blocked with HBSS/1 mM CaCl 2 /2% BSA for one hour at 37° C. After blocking, plates were washed twice with wash buffer. K562 transfectants at log growth stage were collected, washed once in HBSS/0.2% BSA/1 mM CaCl 2 /1 mM MgCl 2 , and resuspended to 4×10 6  cells/mL in the same buffer. Cells were labeled with the fluorecent dye BCECF-AM (Molecular Probes, 4 μg/ml final concentration) for 15 minutes at 3° C. Labeled cells were washed twice, and resuspended in assay buffer (HBSS/0.2% BSA/1 mM CaCl 2 /1 mM MgCl 2 /1 mM MnCl 2 ) to 8×10 5  cells/mL. 50 μl of the cell suspension was added to the E-cadherin-IgG coated well (4×10 4  cells/well), and mixed with 50 μl assay buffer containing antibodies with desired concentration, or isotype-matched control antibody. The plate was then incubated at room temperature for 1 hour. The fluorescence content in each well was read on a Fluorescent Concentration Analyser (IDEXX, Westbrook, Me.) before and after three washes with HBSS/0.5 mM CaCl 2 /0.5 mM MgCl 2 /0.5 mM MnCl 2  using a Microplate Autowasher (Bio-Tek instruments, Winooski, Vt.). The Microplate Autowasher was programmed with parameters: 250 μl wash volume, 1× wash cycle, 0 soak time, and aspiration tube depth of 70. The bound cells (after washes) were expressed as a percentage of total input cells (before washes) in each well. Each sample was set up in triplicate wells.  
     [0163] The effect of activation of αEβ7 integrin by divalent cations was evaluated in cell adhesion assays using transfected K562 cells that expressed αEβ7 integrin. The transfected K562 cells were fluorescently labeled and added to assay wells that were coated with E-cadherin-IgG fusion protein (100 ng/well). The assay media contained CaCl 2  and MgCl 2  (1 mM each; Ca+Mg); CaCl 2 , MgCl 2  and MnCl 2  (1 mM each; Ca+Mg+Mn); or the divalent cation chelating agent EDTA (5 mM). The fluorescently labeled cells were allowed to adhere to the plate-bound E-cadherin-IgG fusion protein, unbound cells were washed away and bound cells are detected by measuring fluorescence. Cell binding was enhanced in media that contained MnCl 2  and inhibited in media that contained EDTA (relative to media that contained media contained CaCl 2  and MgCl 2 ).  
     [0164] Cell-Free αEβ7/E-Cadherin Binding Assay.  
     [0165] Purified recombinant αEβ7 (ts αEβ7.coil) was diluted to 5 μg/ml in TBS, pH 8/Ca+Mg (20 mM Tris, pH 8, 140 mM NaCl, 1 mM CaCl 2  and 1 mM MgCl 2 ), and 50 μl was used to coat each well of 96-well ELISA plate overnight at 4° C. The plate was washed in wash buffer (20 mM Tris, pH 7.5, 140 mM NaCl, 1 mM CaCl 2  and 1 mM MgCl 2 ), and blocked with 300 μl/well blocking buffer (20 mM Tris, pH 7.5, 140 mM NaCl, 1 mM CaCl 2  and 1 mM MgCl 2 , 2% BSA) for 2 hours at 37° C. 25 μl of biotin-labeled E-cadherin-IgG fusion protein diluted to 20 μg/ml in assay buffer (20 mM Tris, pH 7.5, 140 mM NaCl, 1 mM CaCl 2 , 1 mM MgCl 2 , 1 mM MnCl 2 , and 1%BSA) was added to each αEβ7-coated wells, and mixed with 25 μl assay buffer containing test antibodies at desired concentration, or isotype-matched control antibody. The plate was then incubated for 90 minutes at 37° C. The plate was then washed twice with wash buffer, and 50 μl HRP-streptavidin (1:1000 dilution in assay buffer) was added to each well, and the plate was incubated for 1 hour at 37° C. Color was developed by adding substrate buffer (ABTS substrate for HRP, Zymed), and absorbance was read on an ELISA plate reader (410 nm).  
     [0166] The effect of activation of αEβ7 integrin by divalent cations was evaluated in this cell-free adhesion assays using assay buffer that contained CaCl 2  and MgCl 2  (1 mM each; Ca+Mg); CaCl 2 , MgCl 2  and MnCl 2  (1 mM each; Ca+Mg+Mn); or the divalent cation chelating agent EDTA (5 mM).  
     [0167] Conversion of 5E4 (IgM), 3G6 (IgM) and 8D5 (IgG2) to Human IgG1-FcRmut Isotype  
     [0168] RNA was prepared from 1×10 7  hybridoma cells using QIAGEN RNEASY RNA isolation kit (QIAGEN, Valencia, Calif.) according to manufacturer&#39;s instruction. cDNA was synthesized, and variable regions of light and heavy chains were cloned out by PCR. VL(kappa) regions were cloned using human IG-PRIMER oligonucleotide primers (Novagen, Madison, Wis.), and VH regions were made using synthetic primers AB85-89 (SEQ ID NOS: 46-50) and AB90 (MEDAREX, Princeton, N.J.; SEQ ID NO: 51) for hybridomas 5E4 and 3G6 or synthetic primers pHuVH1-7 (SEQ ID NOS: 52-58) and NHuIgG2p3 (SEQ ID NO: 59) for hybridoma 8D5.  
     [0169] PCR fragments were cloned into PCR2.1-TOPO vector using a TOPO cloning kit (Invitrogen, Carlsbad, Calif.), and 6-8 clones from each PCR reaction were sequenced to determine consensus of variable region sequences. The variable regions were subsequently isolated from PCR2.1-TOPO vectors by PCR using primers with restriction enzyme sites incorporated at both ends for subcloning (MfeI and BlpI sites for VH; EcoRI and BsiWI for 5E4 VL and 3G6 VL; or PpuMI and BsiWI for 8D5 VL). Primers p3G6VH5 (SEQ ID NO: 60) and pAEB7VH3 (SEQ ID NO: 62) were used for the 3G6 VH, primers pAEB7VH5 (SEQ ID NO: 61) and pAEB7VH3 (SEQ ID NO: 62) were used for the 5E4 VH, primers pAEB7VK5 (SEQ ID NO: 63) and pAEB7VK3 (SEQ ID NO: 64) were used for the 3G6 and 5E4 VLs. The primers for the VL of 5E4 and 3G6 include the VL leader sequence whereas all other primers allow cloning into antibody expression vectors that contain VH and VL leaders.  
     [0170] The PCR products encoding the VH of either 5E4 or 3G6 were separately subcloned into the MfeI and BlpI sites of pLKTOK30. pLKTOK30 is based on the pCDNA3 vector with the CMV promoter replaced with the EF-1a promoter. pLKTOK30 contains sequences encoding a VH leader and a human IgG1 constant region that are separated by the desired cloning sites. The human IgG1 constant region encoded by this vector contains the Leu 235 to Ala 235 and Gly 237 to Ala 237 mutations that interfere with the antibody binding to Fc receptors (human IgG1-FcR mut region). The MfeI site is within the bases VH3-4 and the BlpI site is at the junction of VH and CH.  
     [0171] The PCR products encoding the VL of either 5E4 or 3G6 were separately subcloned into the EcoRI and BsiWI sites of pLKTOK25. pLKTOK25 has a similar structure to pLKTOK30 with the exception that it contains a sequence that encodes a human kappa constant region instead of a human IgG1 constant region and does not contain a sequence encoding a leader. In this vector, the Kozak sequence and sequence encoding a VL leader are included with the adapted VL gene fragments.  
     [0172] The heavy and light chain containing vectors for each antibody (5E4 or 3G6) were cotransfected in 293T cells to evaluate IgG1 production. When production of functional antibody was confirmed, the heavy chain including the promoter region was excised from TOK30 vector with HindIII and XbaI and ligated into the same sites (HindIII and XbaI) of the light chain containing TOK25 vector to generate a single IgG1 expression vector. The single IgG1 expression vectors were used to make stable CHO cells expressing either 5E4 or 3G6 antibody as described below.  
     [0173] The VH and VL of 8D5 were adapted and cloned into the antibody expression vector pLKTOK59 using PCR. Vector pLKTOK59, like pLKTOK30, is based on the pCDNA3 vector. However, pLKTOK59 contains two EF-1a promoters, one of which drives expression of the heavy chain while the other drives expression of the light chain. The 8D5 VH gene was adapted by PCR using synthetic primers p8D5VH5 (SEQ ID NO: 65) and p8D5VH3 (SEQ ID NO: 66) to add the cloning sites MfeI and BlpI and cloned between the VH leader and Human IgG1-FcRmut region of pLKTOK59D. The 8D5 VL gene was adapted by PCR using synthetic primers p8D5VK5 (SEQ ID NO: 67) and p8D5VK3 (SEQ ID NO: 68) to add the cloning sites PpuMI and BsiWI and cloned between the VL leader and human kappa constant region of pLKTOK59D-8D5-VH to create pLKTOK59D-8D5-VHVK.  
     [0174] Expression of Converted IgG1 Antibodies and Preparation of Stable CHO Cells  
     [0175] Medium scale production of 3G6 (IgG1) and 5E4 (IgG1) was done in 293T cells using calcium phosphate transfection. 10 μg of each heavy and light chain expression vector were used to transfect one 10 cm plate of 293T cells. 7-11 hour post-transfection, the culture medium was changed to media supplemented with 10% ultra low IgG FBS (Gibco). The transfected cells were cultured and the culture supernatant (10 mL) was collected daily for three days. A total of about 900 mL supernatant for each antibody was collected.  
     [0176] Stable CHO cell lines were generated using the single IgG1 expression vectors described above that contain both heavy and light chains of the converted IgG1 antibodies. CHO (DG44) stable transfection was performed using FUGENE non-liposomal lipid transfection (Boehringer Mannheim) according to manufacturer&#39;s instruction. 2 days after transfection, CHO cells were collected, resuspended in selection medium (alphaMEM, 10% Hyclone serum, 800 mg/L G418), and subcloned into 96 well plates. Several stable CHO clones that secreted IgG1 antibodies were selected and the high producers were subcloned again. The yield of IgG1 production by the stable CHO lines was determined by ELISA assay using human IgG1 as standard, and the functional activity of the IgG1 antibodies was determined by binding to αEβ7 transfectants and blocking αEβ7 interaction with E-cadherin.  
     [0177] The “3G6 CHO stable cell line,” also referred to as CHO 3G6 C1.2D6, which produces an IgG1 form of mAb 3G6 was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4204.  
     [0178] The “5E4 CHO stable cell line,” also referred to as CHO 5G4 A1.2C12, which produces an IgG1 form of mAb 5E4 was deposited on Apr. 3, 2002, on behalf of Millennium Pharmaceuticals Inc., 75 Sidney Street, Cambridge, Mass., 02139, USA, at the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110, U.S.A., under Accession No. PTA-4205.  
     [0179] Purification of IgG Antibodies  
     [0180] The converted 3G6 (IgG1) and 5E4 (IgG1) antibodies were purified from culture supernatant of transiently transfected 293T cells, and the 8D6 IgG2 antibody was purified from hybridoma supernatant. Protein A agarose (Gibco) columns were used to purify both IgG1 and IgG2 antibodies. Briefly, antibody-containing supernatants were run through the Protein A column overnight at 4° C. at a slow flow rate. Then, the column was washed with TBS (20 mM Tris-HCl pH 7.5, 140 mM NaCl) at 4° C. and eluted with 100 mM Glycine-HCl pH 2.3. The eluate was immediately neutralized with 1M Tris-HCl pH 9.0 (1/15 v/v). Fractions were pooled and dialyzed in PBS at 4° C. Antibody concentration was determined by the Bradford method (Bio-Rad) using bovine IgG as standard. Antibody purity was analyzed by SDS-PAGE.  
     [0181] Determination of Half Saturation Concentration of mAb Binding to αEβ7 transfctants  
     [0182] Purified antibodies were serially diluted in PBS/5% FBS. FACS staining using the diluted antibodies and K562 transfectants that express αEβ7 on the cell surface was performed as described above. FITC-conjugated anti-human IgG1 or FITC-conjugated anti-human IgG2 was used as secondary antibody. The degree of staining (mean fluorescence intensity) detected was plotted against the concentration of antibody used (μg/ml), and half saturation concentrations were determined using the plot. To determine Mn 2+  effect on antibody binding, 1 mM MnCl 2  (final concentration) was included in the staining buffer in some studies.  
     [0183] Antibody IC50 Determination  
     [0184] IC50 was determined using the cell adhesion assay and cell-free αEβ7/E-cadherin binding assays described above. The percentage of input cells that bound αEβ7 (cell adhesion assay) or amount of E-cadherin-IgG fusion that bound αEβ7 (measured by absorbance in the cell-free binding assay) was plotted against the concentration of antibody used (μg/ml), an inhibition curve was drawn, and IC50 values were determined using the curve.  
     [0185] Culture of Human Peripheral Blood Lymphocytes  
     [0186] Human PBL were purified from fresh whole blood using standard Histopaque gradient centrifugation, and cultured at 1×10 6  cells per mL in RPMI 1640 with 10% FBS, supplemented with TGF-β1 and IL-2 to increase αEβ7 expression on the surface. After culture for 10-15 days, cells were collected for FACS staining as described above.  
     [0187] Epitope Mapping  
     [0188] Construction and Expression of αE I-domain-Fc Fusion Protein.  
     [0189] A nucleic acid encoding the αE chain I-domain (amino acids 161-379 of SEQ ID NO: 2) was isolated from full-length αE cDNA (SEQ ID NO: 1) by PCR using synthetic primers aEXID(5) (tcggatccgctctggagaaggaggag, SEQ ID NO: 69) and aEIDS(3) (gcgaattcaagggcgtctccaaccgt, SEQ ID NO: 70). This nucleic acid was joined in-frame with a nucleic acids encoding the αE secretion signal sequence (amino acids −18 to −1 of SEQ ID NO: 2) and a human IgG1 Fc region that contained mutations to reduce binding to Fc receptor, to produce a construct encoding the αE I-domain Fc fusion protein.  
     [0190] The fusion construct encoded a fusion protein that contains GlySer at the amino-terminus of the I domain and GluPhe between the I domain and the Fc region sequences. The I domain encoded by the fusion construct also includes a portion of the X domain. These X domain sequences ensure proper folding and secretion of the I domain fusion protein.  
     [0191] The αE I-domain-Fc fusion construct was cloned in vector pCDEF3, which was transiently expressed in 293T cells. Culture supernatant that contained the fusion protein was collected as described above.  
     [0192] Binding of mAb to αE I-domain-Fc Fusion Protein  
     [0193] An ELISA assay was established to evaluate secretion and folding of the I-domain fusion protein using mouse antibodies previously mapped to the αE I-domain. (Higgins, J. M. G. et al.,  J. Biol. Chem . 275:25652-25664 (2000).) In this assay, anti-human IgG was immobilized to capture the αE I-domain-Fc fusion protein from the supernatant of 293T transfectants, and binding of two mouse antibodies, αE7.1 and HML-1, was tested. (Higgins, J. M. G. et al.,  J. Biol. Chem . 275:25652-25664 (2000).) The two mouse mAb bound to the αE I-domain-Fc fusion protein. Binding of human antibodies was determined using similar assay conditions. Goat anti-human IgG (15 μg/ml in 20 mM Tris, pH9, 140 mM NaCl) was used to coat 96 well ELISA plate overnight at 4° C. The plate was blocked with 2%BSA and incubated with 50 μl culture supernatant of 293T transfected with the αE I-domain-Fc fusion construct or with vector alone (control) for 1 hour at 37° C. After washing, the plate was incubated with 50 μl of hybridoma supernatant of 5E4 IgM, 3G6 IgM or 8D5 IgG2. Binding of IgM antibody to the captured αE I-domain-Fc fusion protein was detected by HRP-conjugated anti-human IgM or anti-human IgG2.  
     [0194] Antibody Competition Assay (Cytometry Assay)  
     [0195] 5×10 5  K562 transfectants expressing αEβ7 were incubated with mouse mAb αE7.1 (15 μg/ml)(Russel, G. J. et al.,  Eur. J. Immunol . 24:2832-2841 (1994)), human mAb 5E4 (IgM hybridoma supernatant), human 3G6 (IgM hybridoma supernatant), or medium control on ice for 30 minutes. Then, the cells were washed and incubated with human mAb 8D5 (IgG2, 15 μg/ml) for 30 minutes on ice. Cells were then washed twice and incubated with FITC-anti-human IgG, and analyzed by fluorescence flow cytometry.  
     [0196] Fine Specificity  
     [0197] The fine specificity of mAb 5E4 was determined using a panel of transfected K562 cells that expressed various mutant αEβ7 integrins and detecting antibody binding to the transfectants by flow cytometry. The mutants proteins and methods used have been previously described in Higgins, J. M. G. et al.,  J. Biol. Chem . 275:25652-25664 (2000). The mutant αEβ7 integrins used contained the following mutations in the αE chain: R159S/R160S; ΔE163-E180; ΔE176; D190A; G193A; D199A; R202A/D205A; G230A/V231A; D240A; F298A; P311H/E345A/T346A; E325A; and Y354W. (See, Higgins, J. M. G. et al.,  J. Biol. Chem . 275:25652-25664 (2000).)  
     Results  
     [0198] Hybridomas that produce human antibodies which bind αEβ7 integrin were produced, and the antibodies produced by three of the hybridomas were characterized. The supernatants of hybridomas 3G6 (which produces an IgM), 5E4 (which produces an IgM) and 8D5 (which produces an IgG2) were tested for αEβ7 binding specificity by flow cytometry. mAb 3G6 (IgG1), mAb 5E4 (IgG1) and mAb 8D5 each bound transfected L1.2 cells and transfected K562 cells that expressed αEβ7 integrin, but none of these antibodies bound transfected K562 cells that expressed α4β7 integrin, indicating that the mAbs have binding specificity for integrin αE chain. Each mAb (mAb 3G6 (IgM), mAb 3G6 (IgG1), mAb 5E4 (IgM), mAb 5E4 (IgG1) and mAb 8D5) inhibited binding of αEβ7 integrin to its ligand E-cadherin using an in vitro cell adhesion assay and also inhibited binding of soluble E-cadherin-Fc to immobilized αEβ7 integrin in a cell free adhesion assay.  
     [0199] The variable regions of mAb 3G6, mAb 5E4 and mAb 8D5 were cloned and constructs encoding these antibodies with a human IgG1 constant region were produced. The IgG1 versions of mAb 3G6 (IgG1) and mAb 5E4 (IgG1) were used in some of the studies described herein.  
     [0200] The concentration of mAb 3G6 (IgG1), mAb 5E4 (IgG1) and mAb 8D5 that resulted in half saturation of antibody binding sites on transfected K562 cells that expressed αEβ7 was determined using flow cytometry. mAb 3G6 (IgG1) and mAb 5E4 (IgG1) both had a half saturation concentration of 1 μg/mL, while mAb 3G6 had a half saturation concentration of 2.5 μg/mL. The concentrations of antibody that inhibited binding in the cell adhesion assay and the cell free binding assay (IC50) were also determined for mAb 3G6 (IgG1), mAb 5E4 (IgG1) and mAb 8D5. The IC50 for mAb 3G6 (IgG1) was about 2.04 μg/mL (13.4 nM) in the cell adhesion assay, and about 0.089 μg/mL (0.59 nM) in the cell free assay. The IC50 for mAb 5E4 (IgG1) was about 1.29 μg/mL (8.5 nM) in the cell adhesion assay, and about 1.02 μg/mL (6.7 nM) in the cell free assay. The IC50 for mAb 8D5 (IgG1) was about 0.715 μg/mL (4.7 nM) in the cell adhesion assay, and about 0.197 μg/mL (1.30 nM) in the cell free assay.  
     [0201] Integrin molecules, such as αEβ7, bind their ligands with high affinity when activated by, for example, divalent cations (e.g., Mn 2+ ). The results of cellular binding studies revealed that recombinant αEβ7 expressed on transfected K562 cells is activated by divalent cation ions, particularly Mn 2+ , and binding to immobilized E-cadherin is enhanced under conditions where Mn 2+  is present. Similar results were obtained in studies in which transfected K562 cells were stained with Biotin-E-cadherin-IgG. The result of the cellular binding assay are presented in Table 1, and the results of the staining assay are presented in Table 2.  
                   TABLE 1                       Assay Buffer   % cells bound to immobilized E-cadherin                  Ca 2+  and Mg 2+  (1 mM each)   ˜40%       Ca 2+  and Mg 2+  and Mn 2+     ˜60%       (1 mM each)       EDTA (5 mM)   &lt;10%                  
 
     [0202]                       TABLE 2                       Biotin-E-       Binding       cadhering-IgG       (mean fluorescence       used (μg/ml)   Staining Buffer   intensity)                                            2.5   Ca 2+  and Mg 2+  (1 mM each)   &lt;25            Ca 2+  and Mg 2+  and Mn 2+     50-75           (1 mM each)       5   Ca 2+  and Mg 2+  (1 mM each)   25-50           Ca 2+  and Mg 2+  and Mn 2+     ˜100           (1 mM each)       10   Ca 2+  and Mg 2+  (1 mM each)   ˜75            Ca 2+  and Mg 2+  and Mn 2+     150-175           (1 mM each)                    
     [0203] To determine whether mAbs 3G6, 5E4 or 8D5 bound an activation-induced epitope, αEβ7 expressing K562 transfectants were stained with antibodies using a buffer that contained Mn 2+  and using a buffer that did not contain MW 2+ , and antibody binding was detected by fluorescence flow cytometry. The results of these studies demonstrated that binding of mAb 3G6 (IgG1) was enhanced in the presence of Mn 2+ , but that the binding of mAb 5E4 (IgG1), mAb 8D5 and mAb αE7.1 to αEβ7 integrin on the K562 transfectants was about equivalent in buffers that contained or did not contain Mn 2+ . The results show that mAb 3G6 (IgG1) preferentially bound Mn 2+  activated αEβ7 integrin on transfected K562 cells.  
     [0204] These results were confirmed in antibody binding studies using primary human peripheral blood mononuclear cells. The human PBMC were cultured in IL-2 and TGF-β for 10-15 days, which increased the percentage of CD3+αE+cells to about 30-40%. The cells were then stained with mAb 3G6 (IgG1), mAb 5E4 (IgG1), mAb 8D5 or mAb αE7.1 using a buffer that contained Mn 2+  and using a buffer that did not contain Mn 2+ , and antibody binding to CD3+ cells was detected by fluorescence flow cytometry. As with transfected K563 cells, binding of mAb 3G6 (IgG1) was enhanced in the presence of Mn 2+  (positive cells in buffer without Mn 2+ , &lt;5%; positive cells in buffer with Mn 2+ , ˜20%), whereas binding of mAb 5E4 (IgG1), mAb 8D5 and mAb αE7.1 was about equivalent in buffers that contained or did not contain Mn 2+ . These results demonstrate that integrin αE chain can adopt an activated conformation and that mAb 3G6 preferentially binds an activation-induced epitope on integrin αE chain.  
     [0205] Epitopic specificity of the mAbs was studied further using an αE I-domain-Fc fusion protein, a panel of transfected K562 cells that expressed various mutant αEβ7 integrins (see, Higgins, J. M. et al.,  J. Biol. Chem . 275:25652-25664 (2000)), and antibody blocking studies using transfected K562 cells that expressed αEβ7. Mabs 3G6 (IgM), 5E4 (IgM) and 8D5 (hybridoma culture supernatants) each bound αE I-domain-Fc fusion protein coated wells in the ELISA, but binding above control levels was not detected in wells coated with supernatants from mock transfected 293T cells that did not produce the αE I-domain-Fc fusion protein, demonstrating that each antibody binds an epitope that includes amino acids in the I domain of E-cadherin.  
     [0206] The fine specificity of mAb 5E4 was examined using transfected K562 cells that expressed αEβ7 integrin or mutated version of αEβ7 integrins, and antibody binding was detected by flow cytometry. The results are shown in Table 3. Antibody binding was inhibited by deletion of amino acid residues 163-180 (ΔE163-E180; amino acid residues 163-180 of SEQ ID NO: 2), which are in the X domain of integrin αE chain, and was essentially abrogated by mutation of amino acid residue 298 (amino acid residue 298 of SEQ ID NO:2), which is in the I-domain, from Phenylalanine to Alanine (F298A).  
                           TABLE 3                           % Control       % Control       αE Mutation   Staining   αE Mutation   Staining                  none   100%   R202A/D205A   75-100%       R159S/R160S   75-100%   G230A/V231A   75-100%       ΔE163-E180   ˜25%   D240A   75-100%       ΔE176   75-100%   F298A   no binding detected       D190A   50-75%    P311H/E345A/T346A   50-75%        G193A   75-100%   E325A   75-100%       D199A   75-100%   Y354W   ˜75%                  
 
     [0207] The fine specificity of several anti-αE antibodies have been evaluated using this method and the mutant αEβ7 integrins. For most antibodies tested, binding to the ΔE163-E180 mutant is inhibited relative to binding to un-mutated αEβ7. Thus, this inhibition appears to be nonspecific and may be the result of instability of the mutant and/or proteolytic degradation. In contrast, antibody binding was essentially abrogated by mutation of amino acid 298, which is in the I-domain, from Phenylalanine to Alanine (F298A), indicating that epitope bound by mAb 5E4 includes Phe298.  
     [0208] The results of flow cytometry based antibody blocking studies revealed that pre-incubating transfected K562 cells that expressed αEβ7 integrin with mAb αE7.1 partially inhibited binding of mAb 8D5, indicating that these antibodies may bind adjacent or overlapping epitopes. However, the inhibition could be the result of steric interference. Binding of mAb 8D5 was not significantly inhibited when the transfected cells were pre-incubated with mAb 5E4 (IgM) or mAb 3G6 (IgM), demonstrating that mAbs 3G6, 5E4 and 8D5 bind distinct epitopes.  
     [0209] While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.  
    
     
       
         1 
         
           
             70  
           
           
             1  
             3927  
             DNA  
             Homo sapiens  
             
               CDS  
               (126)...(3665)  
               Sig-Peptide (126-179)  
             
           
            1 

gaattccggc ccccgtgtct gggcgtccgc ctcctggcct cctggctgag gggaagctga     60 

gtgggccacg gcccatgtgt cgcactcgcc tcggctccca cacagccgcc tctgctccag    120 

caagg atg tgg ctc ttc cac act ctg ctc tgc ata gcc agc ctg gcc ctg    170 
      Met Trp Leu Phe His Thr Leu Leu Cys Ile Ala Ser Leu Ala Leu 
       1               5                   10                  15 

ctg gcc gct ttc aat gtg gat gtg gcc cgg ccc tgg ctc acg ccc aag      218 
Leu Ala Ala Phe Asn Val Asp Val Ala Arg Pro Trp Leu Thr Pro Lys 
                 20                  25                  30 

gga ggt gcc cct ttc gtg ctc agc tcc ctt ctg cac caa gac ccc agc      266 
Gly Gly Ala Pro Phe Val Leu Ser Ser Leu Leu His Gln Asp Pro Ser 
             35                  40                  45 

acc aac cag acc tgg ctc ctg gtc acc agc ccc aga acc aag agg aca      314 
Thr Asn Gln Thr Trp Leu Leu Val Thr Ser Pro Arg Thr Lys Arg Thr 
         50                  55                  60 

cca ggg ccc ctc cat cga tgt tcc ctt gtc cag gat gaa atc ctt tgc      362 
Pro Gly Pro Leu His Arg Cys Ser Leu Val Gln Asp Glu Ile Leu Cys 
     65                  70                  75 

cat cct gta gag cat gtc ccc atc ccc aag ggg agg cac cgg gga gtg      410 
His Pro Val Glu His Val Pro Ile Pro Lys Gly Arg His Arg Gly Val 
 80                  85                  90                  95 

acc gtt gtc cgg agc cac cac ggt gtt ttg ata tgc att caa gtg ctg      458 
Thr Val Val Arg Ser His His Gly Val Leu Ile Cys Ile Gln Val Leu 
                100                 105                 110 

gtc cgg cgg cct cac agc ctc agc tca gaa ctc aca ggc acc tgt agc      506 
Val Arg Arg Pro His Ser Leu Ser Ser Glu Leu Thr Gly Thr Cys Ser 
            115                 120                 125 

ctc ctg ggc cct gac ctc cgt ccc cag gct cag gcc aac ttc ttc gac      554 
Leu Leu Gly Pro Asp Leu Arg Pro Gln Ala Gln Ala Asn Phe Phe Asp 
        130                 135                 140 

ctt gaa aat ctc ctg gat cca gat gca cgt gtg gac act gga gac tgc      602 
Leu Glu Asn Leu Leu Asp Pro Asp Ala Arg Val Asp Thr Gly Asp Cys 
    145                 150                 155 

tac agc aac aaa gaa ggc ggt gga gaa gac gat gtg aac aca gcc agg      650 
Tyr Ser Asn Lys Glu Gly Gly Gly Glu Asp Asp Val Asn Thr Ala Arg 
160                 165                 170                 175 

cag cgc cgg gct ctg gag aag gag gag gag gaa gac aag gag gag gag      698 
Gln Arg Arg Ala Leu Glu Lys Glu Glu Glu Glu Asp Lys Glu Glu Glu 
                180                 185                 190 

gaa gac gag gag gag gag gaa gct ggc acc gag att gcc atc atc ctg      746 
Glu Asp Glu Glu Glu Glu Glu Ala Gly Thr Glu Ile Ala Ile Ile Leu 
            195                 200                 205 

gat ggc tca gga agc att gat ccc cca gac ttt cag aga gcc aaa gac      794 
Asp Gly Ser Gly Ser Ile Asp Pro Pro Asp Phe Gln Arg Ala Lys Asp 
        210                 215                 220 

ttc atc tcc aac atg atg agg aac ttc tat gaa aag tgt ttt gag tgc      842 
Phe Ile Ser Asn Met Met Arg Asn Phe Tyr Glu Lys Cys Phe Glu Cys 
    225                 230                 235 

aac ttt gcc ttg gtg cag tat gga gga gtg atc cag act gag ttt gac      890 
Asn Phe Ala Leu Val Gln Tyr Gly Gly Val Ile Gln Thr Glu Phe Asp 
240                 245                 250                 255 

ctt cgg gac agc cag gat gtg atg gcc tcc ctc gcc aga gtc cag aac      938 
Leu Arg Asp Ser Gln Asp Val Met Ala Ser Leu Ala Arg Val Gln Asn 
                260                 265                 270 

atc act caa gtg ggg agt gtc acc aag act gcc tca gcc atg caa cac      986 
Ile Thr Gln Val Gly Ser Val Thr Lys Thr Ala Ser Ala Met Gln His 
            275                 280                 285 

gtc tta gac agc atc ttc acc tca agc cac ggc tcc agg aga aag gca     1034 
Val Leu Asp Ser Ile Phe Thr Ser Ser His Gly Ser Arg Arg Lys Ala 
        290                 295                 300 

tcc aag gtc atg gtg gtg ctc acc gat ggt ggc ata ttc gag gac ccc     1082 
Ser Lys Val Met Val Val Leu Thr Asp Gly Gly Ile Phe Glu Asp Pro 
    305                 310                 315 

ctc aac ctt acg aca gtc atc aac tcc ccc aaa atg cag ggt gtt gag     1130 
Leu Asn Leu Thr Thr Val Ile Asn Ser Pro Lys Met Gln Gly Val Glu 
320                 325                 330                 335 

cgc ttt gcc att ggg gtg gga gaa gaa ttt aag agt gct agg act gcg     1178 
Arg Phe Ala Ile Gly Val Gly Glu Glu Phe Lys Ser Ala Arg Thr Ala 
                340                 345                 350 

agg gaa ctg aac ctg atc gcc tca gac ccg gat gag acc cat gct ttc     1226 
Arg Glu Leu Asn Leu Ile Ala Ser Asp Pro Asp Glu Thr His Ala Phe 
            355                 360                 365 

aag gtg acc aac tac atg gcg ctg gat ggg ctg ctg agc aaa ctg cgg     1274 
Lys Val Thr Asn Tyr Met Ala Leu Asp Gly Leu Leu Ser Lys Leu Arg 
        370                 375                 380 

tac aac atc atc agc atg gaa ggc acg gtt gga gac gcc ctt cac tac     1322 
Tyr Asn Ile Ile Ser Met Glu Gly Thr Val Gly Asp Ala Leu His Tyr 
    385                 390                 395 

cag ctg gca cag att ggc ttc agt gct cag atc ctg gat gag cgg cag     1370 
Gln Leu Ala Gln Ile Gly Phe Ser Ala Gln Ile Leu Asp Glu Arg Gln 
400                 405                 410                 415 

gtg ctg ctc ggc gcc gtc ggg gcc ttt gac tgg tcc gga ggg gcg ttg     1418 
Val Leu Leu Gly Ala Val Gly Ala Phe Asp Trp Ser Gly Gly Ala Leu 
                420                 425                 430 

ctc tac gac aca cgc agc cgc cgg ggc cgc ttc ctg aac cag aca gcg     1466 
Leu Tyr Asp Thr Arg Ser Arg Arg Gly Arg Phe Leu Asn Gln Thr Ala 
            435                 440                 445 

gcg gcg gcg gca gac gcg gag gct gcg cag tac agc tac ctg ggt tac     1514 
Ala Ala Ala Ala Asp Ala Glu Ala Ala Gln Tyr Ser Tyr Leu Gly Tyr 
        450                 455                 460 

gct gtg gcc gtg ctg cac aag acc tgc agc ctc tcc tac gtc gcg ggg     1562 
Ala Val Ala Val Leu His Lys Thr Cys Ser Leu Ser Tyr Val Ala Gly 
    465                 470                 475 

gct cca cag tac aaa cat cat ggg gcc gtg ttt gag ctc cag aag gag     1610 
Ala Pro Gln Tyr Lys His His Gly Ala Val Phe Glu Leu Gln Lys Glu 
480                 485                 490                 495 

ggc aga gag gcc agc ttc ctg cca gtg ctg gag gga gag cag atg ggg     1658 
Gly Arg Glu Ala Ser Phe Leu Pro Val Leu Glu Gly Glu Gln Met Gly 
                500                 505                 510 

tcc tat ttt ggc tct gag ctg tgc cct gtg gac att gac atg gat gga     1706 
Ser Tyr Phe Gly Ser Glu Leu Cys Pro Val Asp Ile Asp Met Asp Gly 
            515                 520                 525 

agc acg gac ttc ttg ctg gtg gct gct cca ttt tac cac gtt cat gga     1754 
Ser Thr Asp Phe Leu Leu Val Ala Ala Pro Phe Tyr His Val His Gly 
        530                 535                 540 

gaa gaa ggc aga gtc tac gtg tac cgt ctc agc gag cag gat ggt tct     1802 
Glu Glu Gly Arg Val Tyr Val Tyr Arg Leu Ser Glu Gln Asp Gly Ser 
    545                 550                 555 

ttc tcc ttg gca cgc ata ctg agt ggg cac ccc ggg ttc acc aat gcc     1850 
Phe Ser Leu Ala Arg Ile Leu Ser Gly His Pro Gly Phe Thr Asn Ala 
560                 565                 570                 575 

cgc ttt ggc ttt gcc atg gcg gct atg ggg gat ctc agt cag gat aag     1898 
Arg Phe Gly Phe Ala Met Ala Ala Met Gly Asp Leu Ser Gln Asp Lys 
                580                 585                 590 

ctc aca gat gtg gcc atc ggg gcc ccc ctg gaa ggt ttt ggg gca gat     1946 
Leu Thr Asp Val Ala Ile Gly Ala Pro Leu Glu Gly Phe Gly Ala Asp 
            595                 600                 605 

gat ggt gcc agc ttc ggc agt gtg tat atc tac aat gga cac tgg gac     1994 
Asp Gly Ala Ser Phe Gly Ser Val Tyr Ile Tyr Asn Gly His Trp Asp 
        610                 615                 620 

ggc ctc tcc gcc agc ccc tcg cag cgg atc aga gcc tcc acg gtg gcc     2042 
Gly Leu Ser Ala Ser Pro Ser Gln Arg Ile Arg Ala Ser Thr Val Ala 
    625                 630                 635 

cca gga ctc cag tac ttc ggc atg tcc atg gct ggt ggc ttt gat att     2090 
Pro Gly Leu Gln Tyr Phe Gly Met Ser Met Ala Gly Gly Phe Asp Ile 
640                 645                 650                 655 

agt ggc gac ggc ctt gcc gac atc acc gtg ggc act ctg ggc cag gcg     2138 
Ser Gly Asp Gly Leu Ala Asp Ile Thr Val Gly Thr Leu Gly Gln Ala 
                660                 665                 670 

gtt gtg ttc cgc tcc cgg cct gtg gtt cgc ctg aag gtc tcc atg gcc     2186 
Val Val Phe Arg Ser Arg Pro Val Val Arg Leu Lys Val Ser Met Ala 
            675                 680                 685 

ttc acc ccc agc gca ctg ccc atc ggc ttc aac ggc gtc gtg aat gtc     2234 
Phe Thr Pro Ser Ala Leu Pro Ile Gly Phe Asn Gly Val Val Asn Val 
        690                 695                 700 

cgt tta tgt ttt gaa atc agc tct gta acc aca gcc tct gag tca ggc     2282 
Arg Leu Cys Phe Glu Ile Ser Ser Val Thr Thr Ala Ser Glu Ser Gly 
    705                 710                 715 

ctc cgt gag gca ctt ctc aac ttc acg ctg gat gtg gat gtg ggg aag     2330 
Leu Arg Glu Ala Leu Leu Asn Phe Thr Leu Asp Val Asp Val Gly Lys 
720                 725                 730                 735 

cag agg aga cgg ctg cag tgt tca gac gta aga agc tgt ctg ggc tgc     2378 
Gln Arg Arg Arg Leu Gln Cys Ser Asp Val Arg Ser Cys Leu Gly Cys 
                740                 745                 750 

ctg agg gag tgg agc agc gga tcc cag ctt tgt gag gac ctc ctg ctc     2426 
Leu Arg Glu Trp Ser Ser Gly Ser Gln Leu Cys Glu Asp Leu Leu Leu 
            755                 760                 765 

atg ccc aca gag gga gag ctc tgt gag gag gac tgc ttc tcc aat gcc     2474 
Met Pro Thr Glu Gly Glu Leu Cys Glu Glu Asp Cys Phe Ser Asn Ala 
        770                 775                 780 

agt gtc aaa gtc agc tac cag ctc cag acc cct gag gga cag acg gac     2522 
Ser Val Lys Val Ser Tyr Gln Leu Gln Thr Pro Glu Gly Gln Thr Asp 
    785                 790                 795 

cat ccc cag ccc atc ctg gac cgc tac act gag ccc ttt gcc atc ttc     2570 
His Pro Gln Pro Ile Leu Asp Arg Tyr Thr Glu Pro Phe Ala Ile Phe 
800                 805                 810                 815 

cag ctg ccc tat gag aag gcc tgc aag aat aag ctg ttt tgt gtc gca     2618 
Gln Leu Pro Tyr Glu Lys Ala Cys Lys Asn Lys Leu Phe Cys Val Ala 
                820                 825                 830 

gaa tta cag ttg gcc acc acc gtc tct cag cag gag ttg gtg gtg ggt     2666 
Glu Leu Gln Leu Ala Thr Thr Val Ser Gln Gln Glu Leu Val Val Gly 
            835                 840                 845 

ctc aca aag gag ctg acc ctg aac att aac cta act aac tcc ggg gaa     2714 
Leu Thr Lys Glu Leu Thr Leu Asn Ile Asn Leu Thr Asn Ser Gly Glu 
        850                 855                 860 

gat tcc tac atg aca agc atg gcc ttg aat tac ccc aga aac ctg cag     2762 
Asp Ser Tyr Met Thr Ser Met Ala Leu Asn Tyr Pro Arg Asn Leu Gln 
    865                 870                 875 

ttg aag agg atg caa aag cct ccc tct cca aac att cag tgt gat gac     2810 
Leu Lys Arg Met Gln Lys Pro Pro Ser Pro Asn Ile Gln Cys Asp Asp 
880                 885                 890                 895 

cct cag ccg gtt gct tct gtc ctg atc atg aac tgc agg att ggt cac     2858 
Pro Gln Pro Val Ala Ser Val Leu Ile Met Asn Cys Arg Ile Gly His 
                900                 905                 910 

ccc gtc ctc aag agg tca tct gct cat gtt tca gtc gtt tgg cag cta     2906 
Pro Val Leu Lys Arg Ser Ser Ala His Val Ser Val Val Trp Gln Leu 
            915                 920                 925 

gag gag aat gcc ttt cca aac agg aca gca gac atc act gtg act gtc     2954 
Glu Glu Asn Ala Phe Pro Asn Arg Thr Ala Asp Ile Thr Val Thr Val 
        930                 935                 940 

acc aat tcc aat gaa aga cgg tct ttg gcc aac gag acc cac acc ctt     3002 
Thr Asn Ser Asn Glu Arg Arg Ser Leu Ala Asn Glu Thr His Thr Leu 
    945                 950                 955 

caa ttc agg cat ggc ttc gtt gca gtt ctg tcc aaa cca tcc ata atg     3050 
Gln Phe Arg His Gly Phe Val Ala Val Leu Ser Lys Pro Ser Ile Met 
960                 965                 970                 975 

tac gtg aac aca ggc cag ggg ctt tct cac cac aaa gaa ttc ctc ttc     3098 
Tyr Val Asn Thr Gly Gln Gly Leu Ser His His Lys Glu Phe Leu Phe 
                980                 985                 990 

cat gta cat ggg gag aac ctc ttt gga gca gaa tac cag ttg caa att     3146 
His Val His Gly Glu Asn Leu Phe Gly Ala Glu Tyr Gln Leu Gln Ile 
             995                1000                1005 

tgc gtc cca acc aaa tta cga ggt ctc cag gtt gca gca gtg aag aag     3194 
Cys Val Pro Thr Lys Leu Arg Gly Leu Gln Val Ala Ala Val Lys Lys 
        1010                1015                1020 

ctg acg agg act cag gcc tcc acg gtg tgc acc tgg agt cag gag cgc     3242 
Leu Thr Arg Thr Gln Ala Ser Thr Val Cys Thr Trp Ser Gln Glu Arg 
    1025                1030                1035 

gct tgt gcg tac agt tcg gtt cag cat gtg gaa gaa tgg cat tca gtg     3290 
Ala Cys Ala Tyr Ser Ser Val Gln His Val Glu Glu Trp His Ser Val 
1040                1045                1050                1055 

agc tgt gtc atc gct tca gat aaa gaa aat gtc acc gtg gct gca gag     3338 
Ser Cys Val Ile Ala Ser Asp Lys Glu Asn Val Thr Val Ala Ala Glu 
                1060                1065                1070 

atc tcc tgg gat cac tct gag gag tta cta aaa gat gta act gaa ctg     3386 
Ile Ser Trp Asp His Ser Glu Glu Leu Leu Lys Asp Val Thr Glu Leu 
            1075                1080                1085 

cag atc ctt ggt gaa ata tct ttc aac aaa tct cta tat gag gga ctg     3434 
Gln Ile Leu Gly Glu Ile Ser Phe Asn Lys Ser Leu Tyr Glu Gly Leu 
        1090                1095                1100 

aat gca gag aac cac aga act aag atc act gtc gtc ttc ctg aaa gat     3482 
Asn Ala Glu Asn His Arg Thr Lys Ile Thr Val Val Phe Leu Lys Asp 
    1105                1110                1115 

gag aag tac cat tct ttg cct atc atc att aaa ggc agc gtt ggt gga     3530 
Glu Lys Tyr His Ser Leu Pro Ile Ile Ile Lys Gly Ser Val Gly Gly 
1120                1125                1130                1135 

ctt ctg gtg ttg atc gtg att ctg gtc atc ctg ttc aag tgt ggc ttt     3578 
Leu Leu Val Leu Ile Val Ile Leu Val Ile Leu Phe Lys Cys Gly Phe 
                1140                1145                1150 

ttt aaa aga aaa tat caa caa ctg aac ttg gag agc atc agg aag gcc     3626 
Phe Lys Arg Lys Tyr Gln Gln Leu Asn Leu Glu Ser Ile Arg Lys Ala 
            1155                1160                1165 

cag ctg aaa tca gag aat ctg ctc gaa gaa gag aat tag gacctgctat      3675 
Gln Leu Lys Ser Glu Asn Leu Leu Glu Glu Glu Asn  * 
        1170                1175 

ccactgggag aggctatcag ccagtcctgg gacttggaga cccagcatcc tttgcattac  3735 

tttttccttc aggatgatct agagcagcat ggagctgttg gtagaatatt agtttttaac  3795 

catacattgt cccaaaagtg tctgtgcatt gtgcaaaaag taaacttagg aaacatttgg  3855 

tattaaataa atttacactt ttctttgcaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa  3915 

aaaaaaaaaa aa                                                      3927 

 
           
             2  
             1179  
             PRT  
             Homo sapiens  
             
               SIGNAL  
               (1)...(18)  
             
           
            2 

Met Trp Leu Phe His Thr Leu Leu Cys Ile Ala Ser Leu Ala Leu Leu 
            -15                 -10                 -5 

Ala Ala Phe Asn Val Asp Val Ala Arg Pro Trp Leu Thr Pro Lys Gly 
         1               5                  10 

Gly Ala Pro Phe Val Leu Ser Ser Leu Leu His Gln Asp Pro Ser Thr 
15                  20                  25                  30 

Asn Gln Thr Trp Leu Leu Val Thr Ser Pro Arg Thr Lys Arg Thr Pro 
                35                  40                  45 

Gly Pro Leu His Arg Cys Ser Leu Val Gln Asp Glu Ile Leu Cys His 
            50                  55                  60 

Pro Val Glu His Val Pro Ile Pro Lys Gly Arg His Arg Gly Val Thr 
        65                  70                  75 

Val Val Arg Ser His His Gly Val Leu Ile Cys Ile Gln Val Leu Val 
    80                  85                  90 

Arg Arg Pro His Ser Leu Ser Ser Glu Leu Thr Gly Thr Cys Ser Leu 
95                  100                 105                 110 

Leu Gly Pro Asp Leu Arg Pro Gln Ala Gln Ala Asn Phe Phe Asp Leu 
                115                 120                 125 

Glu Asn Leu Leu Asp Pro Asp Ala Arg Val Asp Thr Gly Asp Cys Tyr 
            130                 135                 140 

Ser Asn Lys Glu Gly Gly Gly Glu Asp Asp Val Asn Thr Ala Arg Gln 
        145                 150                 155 

Arg Arg Ala Leu Glu Lys Glu Glu Glu Glu Asp Lys Glu Glu Glu Glu 
    160                 165                 170 

Asp Glu Glu Glu Glu Glu Ala Gly Thr Glu Ile Ala Ile Ile Leu Asp 
175                 180                 185                 190 

Gly Ser Gly Ser Ile Asp Pro Pro Asp Phe Gln Arg Ala Lys Asp Phe 
                195                 200                 205 

Ile Ser Asn Met Met Arg Asn Phe Tyr Glu Lys Cys Phe Glu Cys Asn 
            210                 215                 220 

Phe Ala Leu Val Gln Tyr Gly Gly Val Ile Gln Thr Glu Phe Asp Leu 
        225                 230                 235 

Arg Asp Ser Gln Asp Val Met Ala Ser Leu Ala Arg Val Gln Asn Ile 
    240                 245                 250 

Thr Gln Val Gly Ser Val Thr Lys Thr Ala Ser Ala Met Gln His Val 
255                 260                 265                 270 

Leu Asp Ser Ile Phe Thr Ser Ser His Gly Ser Arg Arg Lys Ala Ser 
                275                 280                 285 

Lys Val Met Val Val Leu Thr Asp Gly Gly Ile Phe Glu Asp Pro Leu 
            290                 295                 300 

Asn Leu Thr Thr Val Ile Asn Ser Pro Lys Met Gln Gly Val Glu Arg 
        305                 310                 315 

Phe Ala Ile Gly Val Gly Glu Glu Phe Lys Ser Ala Arg Thr Ala Arg 
    320                 325                 330 

Glu Leu Asn Leu Ile Ala Ser Asp Pro Asp Glu Thr His Ala Phe Lys 
335                 340                 345                 350 

Val Thr Asn Tyr Met Ala Leu Asp Gly Leu Leu Ser Lys Leu Arg Tyr 
                355                 360                 365 

Asn Ile Ile Ser Met Glu Gly Thr Val Gly Asp Ala Leu His Tyr Gln 
            370                 375                 380 

Leu Ala Gln Ile Gly Phe Ser Ala Gln Ile Leu Asp Glu Arg Gln Val 
        385                 390                 395 

Leu Leu Gly Ala Val Gly Ala Phe Asp Trp Ser Gly Gly Ala Leu Leu 
    400                 405                 410 

Tyr Asp Thr Arg Ser Arg Arg Gly Arg Phe Leu Asn Gln Thr Ala Ala 
415                 420                 425                 430 

Ala Ala Ala Asp Ala Glu Ala Ala Gln Tyr Ser Tyr Leu Gly Tyr Ala 
                435                 440                 445 

Val Ala Val Leu His Lys Thr Cys Ser Leu Ser Tyr Val Ala Gly Ala 
            450                 455                 460 

Pro Gln Tyr Lys His His Gly Ala Val Phe Glu Leu Gln Lys Glu Gly 
        465                 470                 475 

Arg Glu Ala Ser Phe Leu Pro Val Leu Glu Gly Glu Gln Met Gly Ser 
    480                 485                 490 

Tyr Phe Gly Ser Glu Leu Cys Pro Val Asp Ile Asp Met Asp Gly Ser 
495                 500                 505                 510 

Thr Asp Phe Leu Leu Val Ala Ala Pro Phe Tyr His Val His Gly Glu 
                515                 520                 525 

Glu Gly Arg Val Tyr Val Tyr Arg Leu Ser Glu Gln Asp Gly Ser Phe 
            530                 535                 540 

Ser Leu Ala Arg Ile Leu Ser Gly His Pro Gly Phe Thr Asn Ala Arg 
        545                 550                 555 

Phe Gly Phe Ala Met Ala Ala Met Gly Asp Leu Ser Gln Asp Lys Leu 
    560                 565                 570 

Thr Asp Val Ala Ile Gly Ala Pro Leu Glu Gly Phe Gly Ala Asp Asp 
575                 580                 585                 590 

Gly Ala Ser Phe Gly Ser Val Tyr Ile Tyr Asn Gly His Trp Asp Gly 
                595                 600                 605 

Leu Ser Ala Ser Pro Ser Gln Arg Ile Arg Ala Ser Thr Val Ala Pro 
            610                 615                 620 

Gly Leu Gln Tyr Phe Gly Met Ser Met Ala Gly Gly Phe Asp Ile Ser 
        625                 630                 635 

Gly Asp Gly Leu Ala Asp Ile Thr Val Gly Thr Leu Gly Gln Ala Val 
    640                 645                 650 

Val Phe Arg Ser Arg Pro Val Val Arg Leu Lys Val Ser Met Ala Phe 
655                 660                 665                 670 

Thr Pro Ser Ala Leu Pro Ile Gly Phe Asn Gly Val Val Asn Val Arg 
                675                 680                 685 

Leu Cys Phe Glu Ile Ser Ser Val Thr Thr Ala Ser Glu Ser Gly Leu 
            690                 695                 700 

Arg Glu Ala Leu Leu Asn Phe Thr Leu Asp Val Asp Val Gly Lys Gln 
        705                 710                 715 

Arg Arg Arg Leu Gln Cys Ser Asp Val Arg Ser Cys Leu Gly Cys Leu 
    720                 725                 730 

Arg Glu Trp Ser Ser Gly Ser Gln Leu Cys Glu Asp Leu Leu Leu Met 
735                 740                 745                 750 

Pro Thr Glu Gly Glu Leu Cys Glu Glu Asp Cys Phe Ser Asn Ala Ser 
                755                 760                 765 

Val Lys Val Ser Tyr Gln Leu Gln Thr Pro Glu Gly Gln Thr Asp His 
            770                 775                 780 

Pro Gln Pro Ile Leu Asp Arg Tyr Thr Glu Pro Phe Ala Ile Phe Gln 
        785                 790                 795 

Leu Pro Tyr Glu Lys Ala Cys Lys Asn Lys Leu Phe Cys Val Ala Glu 
    800                 805                 810 

Leu Gln Leu Ala Thr Thr Val Ser Gln Gln Glu Leu Val Val Gly Leu 
815                 820                 825                 830 

Thr Lys Glu Leu Thr Leu Asn Ile Asn Leu Thr Asn Ser Gly Glu Asp 
                835                 840                 845 

Ser Tyr Met Thr Ser Met Ala Leu Asn Tyr Pro Arg Asn Leu Gln Leu 
            850                 855                 860 

Lys Arg Met Gln Lys Pro Pro Ser Pro Asn Ile Gln Cys Asp Asp Pro 
        865                 870                 875 

Gln Pro Val Ala Ser Val Leu Ile Met Asn Cys Arg Ile Gly His Pro 
    880                 885                 890 

Val Leu Lys Arg Ser Ser Ala His Val Ser Val Val Trp Gln Leu Glu 
895                 900                 905                 910 

Glu Asn Ala Phe Pro Asn Arg Thr Ala Asp Ile Thr Val Thr Val Thr 
                915                 920                 925 

Asn Ser Asn Glu Arg Arg Ser Leu Ala Asn Glu Thr His Thr Leu Gln 
            930                 935                 940 

Phe Arg His Gly Phe Val Ala Val Leu Ser Lys Pro Ser Ile Met Tyr 
        945                 950                 955 

Val Asn Thr Gly Gln Gly Leu Ser His His Lys Glu Phe Leu Phe His 
    960                 965                 970 

Val His Gly Glu Asn Leu Phe Gly Ala Glu Tyr Gln Leu Gln Ile Cys 
975                 980                 985                 990 

Val Pro Thr Lys Leu Arg Gly Leu Gln Val Ala Ala Val Lys Lys Leu 
                995                 1000                1005 

Thr Arg Thr Gln Ala Ser Thr Val Cys Thr Trp Ser Gln Glu Arg Ala 
            1010                1015                1020 

Cys Ala Tyr Ser Ser Val Gln His Val Glu Glu Trp His Ser Val Ser 
        1025                1030                1035 

Cys Val Ile Ala Ser Asp Lys Glu Asn Val Thr Val Ala Ala Glu Ile 
    1040                1045                1050 

Ser Trp Asp His Ser Glu Glu Leu Leu Lys Asp Val Thr Glu Leu Gln 
1055                1060                1065                1070 

Ile Leu Gly Glu Ile Ser Phe Asn Lys Ser Leu Tyr Glu Gly Leu Asn 
                1075                1080                1085 

Ala Glu Asn His Arg Thr Lys Ile Thr Val Val Phe Leu Lys Asp Glu 
            1090                1095                1100 

Lys Tyr His Ser Leu Pro Ile Ile Ile Lys Gly Ser Val Gly Gly Leu 
        1105                1110                1115 

Leu Val Leu Ile Val Ile Leu Val Ile Leu Phe Lys Cys Gly Phe Phe 
    1120                1125                1130 

Lys Arg Lys Tyr Gln Gln Leu Asn Leu Glu Ser Ile Arg Lys Ala Gln 
1135                1140                1145                1150 

Leu Lys Ser Glu Asn Leu Leu Glu Glu Glu Asn 
                1155                1160 

 
           
             3  
             369  
             DNA  
             Homo sapiens  
           
            3 

gaggtgcagc tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaagatc     60 

tcctgtaagg gttctggata cagctttacc agctattgga tcggctgggt gcgccagatg    120 

cccgggaaag gcctggagtg gatggggatc atctatcctg gtgactctgg tcccagatac    180 

agcccgtcct tccaaggcca ggtcaccatc tcagccgaca agtccatcag caccgcctac    240 

ctgcagtgga gcagcctgaa ggcctcggac accgccatgt attactgtgc gcgactgtcg    300 

tataccagca cctggtaccc gtactacttt gactactggg gccagggaac cctggtcacc    360 

gtctcctca                                                            369 

 
           
             4  
             123  
             PRT  
             Homo sapiens  
             
               SITE  
               (31)...(35)  
               CDR1  
             
           
            4 

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 
 1               5                  10                  15 

Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 
            20                  25                  30 

Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 
        35                  40                  45 

Gly Ile Ile Tyr Pro Gly Asp Ser Gly Pro Arg Tyr Ser Pro Ser Phe 
    50                  55                  60 

Gln Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr 
65                  70                  75                  80 

Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 
                85                  90                  95 

Ala Arg Leu Ser Tyr Thr Ser Thr Trp Tyr Pro Tyr Tyr Phe Asp Tyr 
            100                 105                 110 

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 
        115                 120 

 
           
             5  
             5  
             PRT  
             Homo sapiens  
           
            5 

Ser Tyr Trp Ile Gly 
 1               5 

 
           
             6  
             17  
             PRT  
             Homo sapiens  
           
            6 

Ile Ile Tyr Pro Gly Asp Ser Gly Pro Arg Tyr Ser Pro Ser Phe Gln 
 1               5                  10                  15 

Gly 

 
           
             7  
             14  
             PRT  
             Homo sapiens  
           
            7 

Leu Ser Tyr Thr Ser Thr Trp Tyr Pro Tyr Tyr Phe Asp Tyr 
 1               5                  10 

 
           
             8  
             324  
             DNA  
             Homo sapiens  
           
            8 

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc     60 

ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct    120 

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc    180 

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct    240 

gaagattttg cagtttatta ctgtcagcag cgtagcaact ggcctccggg gacgttcggc    300 

caagggacca aggtggaaat caaa                                           324 

 
           
             9  
             108  
             PRT  
             Homo sapiens  
             
               SITE  
               (24)...(34)  
               CDR1  
             
           
            9 

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 
 1               5                  10                  15 

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 
            20                  25                  30 

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
        35                  40                  45 

Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60 

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 
65                  70                  75                  80 

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro 
                85                  90                  95 

Gly Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
            100                 105 

 
           
             10  
             11  
             PRT  
             Homo sapiens  
           
            10 

Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala 
 1               5                  10 

 
           
             11  
             7  
             PRT  
             Homo sapiens  
           
            11 

Asp Ala Ser Asn Arg Ala Thr 
 1               5 

 
           
             12  
             10  
             PRT  
             Homo sapiens  
           
            12 

Gln Gln Arg Ser Asn Trp Pro Pro Gly Thr 
 1               5                  10 

 
           
             13  
             354  
             DNA  
             Homo sapiens  
           
            13 

gaggtgcagt tggtggagtc tgggggaggc ttggtccagc ctggagggtc cctgagactc     60 

tcctgtgcag cctctggatt cacctttagt aacttttgga tgagctgggt ccgccaggct    120 

ccagggaaag ggctggagtg gatggccaac ataaagcaag atggaagtga gaaatactat    180 

gtggactctg tgaagggccg attcaccatc tccagagaca acgccaagag ctcactgttt    240 

ctgcaaatga acagcctgag agtcgacgac acggctgtat atttctgtgc gggggattac    300 

tatgattcgg ggagtttcta ctggggccag ggaaccctgg tcaccgtctc ctca          354 

 
           
             14  
             118  
             PRT  
             Homo sapiens  
             
               SITE  
               (31)...(35)  
               CDR1  
             
           
            14 

Glu Val Gln Leu Val 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 Ser Asn Phe 
            20                  25                  30 

Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 
        35                  40                  45 

Ala Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val 
    50                  55                  60 

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Ser Ser Leu Phe 
65                  70                  75                  80 

Leu Gln Met Asn Ser Leu Arg Val Asp Asp Thr Ala Val Tyr Phe Cys 
                85                  90                  95 

Ala Gly Asp Tyr Tyr Asp Ser Gly Ser Phe Tyr Trp Gly Gln Gly Thr 
            100                 105                 110 

Leu Val Thr Val Ser Ser 
        115 

 
           
             15  
             5  
             PRT  
             Homo sapiens  
           
            15 

Asn Phe Trp Met Ser 
 1               5 

 
           
             16  
             17  
             PRT  
             Homo sapiens  
           
            16 

Asn Ile Lys Gln Asp Gly Ser Glu Lys Tyr Tyr Val Asp Ser Val Lys 
 1               5                  10                  15 

Gly 

 
           
             17  
             9  
             PRT  
             Homo sapiens  
           
            17 

Asp Tyr Tyr Asp Ser Gly Ser Phe Tyr 
 1               5 

 
           
             18  
             324  
             DNA  
             Homo sapiens  
           
            18 

gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc     60 

ctctcctgca gggccagtca gagtgttagg agcaacttag cctggtacca acagaaacct    120 

ggccaggctc ccaggctcct catctatgat gcatccaaca gggccattgg catcccagcc    180 

aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag tttagagcct    240 

gaagattttg tagtttatta ctgtcagcag cgtagcaact ggcctccgtg gacgttcggc    300 

caagggacca aggtggaaat caaa                                           324 

 
           
             19  
             108  
             PRT  
             Homo sapiens  
             
               SITE  
               (24)...(34)  
               CDR1  
             
           
            19 

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 
 1               5                  10                  15 

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Arg Ser Asn 
            20                  25                  30 

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
        35                  40                  45 

Tyr Asp Ala Ser Asn Arg Ala Ile Gly Ile Pro Ala Arg Phe Ser Gly 
    50                  55                  60 

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 
65                  70                  75                  80 

Glu Asp Phe Val Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Pro 
                85                  90                  95 

Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
            100                 105 

 
           
             20  
             11  
             PRT  
             Homo sapiens  
           
            20 

Arg Ala Ser Gln Ser Val Arg Ser Asn Leu Ala 
 1               5                  10 

 
           
             21  
             7  
             PRT  
             Homo sapiens  
           
            21 

Asp Ala Ser Asn Arg Ala Ile 
 1               5 

 
           
             22  
             10  
             PRT  
             Homo sapiens  
           
            22 

Gln Gln Arg Ser Asn Trp Pro Pro Trp Thr 
 1               5                  10 

 
           
             23  
             384  
             DNA  
             Homo sapiens  
           
            23 

caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc     60 

acctgcactg tctctggtgg ctccgtcagt agttactatt ggagctggat ccggcagccc    120 

ccagggaagg gactggagtg gattggccat atctattaca gtgggaatac caactacaac    180 

ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaatca gttctccctg    240 

aaactgagct ctgtgaccgc tgcggacacg gccgtgtatt tttgtgcgag agatagatgg    300 

aattattatg atagtagtcc cggctattat tattactacg gtatggacgt ctggggccaa    360 

gggaccacgg tcaccgtcag ctca                                           384 

 
           
             24  
             128  
             PRT  
             Homo sapiens  
             
               SITE  
               (31)...(35)  
               CDR1  
             
           
            24 

Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 
 1               5                  10                  15 

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Val Ser Ser Tyr 
            20                  25                  30 

Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 
        35                  40                  45 

Gly His Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 
    50                  55                  60 

Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu 
65                  70                  75                  80 

Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Phe Cys Ala 
                85                  90                  95 

Arg Asp Arg Trp Asn Tyr Tyr Asp Ser Ser Pro Gly Tyr Tyr Tyr Tyr 
            100                 105                 110 

Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 
        115                 120                 125 

 
           
             25  
             5  
             PRT  
             Homo sapiens  
           
            25 

Ser Tyr Tyr Trp Ser 
 1               5 

 
           
             26  
             16  
             PRT  
             Homo sapiens  
           
            26 

His Ile Tyr Tyr Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser 
 1               5                  10                  15 

 
           
             27  
             20  
             PRT  
             Homo sapiens  
           
            27 

Asp Arg Trp Asn Tyr Tyr Asp Ser Ser Pro Gly Tyr Tyr Tyr Tyr Tyr 
 1               5                  10                  15 

Gly Met Asp Val 
            20 

 
           
             28  
             321  
             DNA  
             Homo sapiens  
           
            28 

gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc     60 

atcacttgcc gggcgagtca gggcattaga aatgatttag gctggtatca gcaaaaacca    120 

gggaaagccc ctaagcgcct aatctttgct gcatcccatt tgcaaagtgg agtcccttca    180 

aggttcagcg gcagtggatc tgggacagag ttcactctca caatcagcag cctgcagcct    240 

gaagattttg caacttatta ctgtcaacag cataatagtt ccccattcac tttcggccct    300 

gggaccagag tggatatcaa a                                              321 

 
           
             29  
             107  
             PRT  
             Homo sapiens  
             
               SITE  
               (24)...(34)  
               CDR1  
             
           
            29 

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
 1               5                  10                  15 

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 
            20                  25                  30 

Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 
        35                  40                  45 

Phe Ala Ala Ser His Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 
    50                  55                  60 

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
65                  70                  75                  80 

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Asn Ser Ser Pro Phe 
                85                  90                  95 

Thr Phe Gly Pro Gly Thr Arg Val Asp Ile Lys 
            100                 105 

 
           
             30  
             11  
             PRT  
             Homo sapiens  
           
            30 

Arg Ala Ser Gln Gly Ile Arg Asn Asp Leu Gly 
 1               5                  10 

 
           
             31  
             7  
             PRT  
             Homo sapiens  
           
            31 

Ala Ala Ser His Leu Gln Ser 
 1               5 

 
           
             32  
             9  
             PRT  
             Homo sapiens  
           
            32 

Gln Gln His Asn Ser Ser Pro Phe Thr 
 1               5 

 
           
             33  
             2808  
             DNA  
             Homo sapiens  
             
               CDS  
               (109)...(2745)  
             
           
            33 

ggaaagcacc tgtgagcttg gcaagtcagt tcagagctcc agcccgctcc agcccggccc     60 

gacccgaccg cacccggcgc ctgcctcgct cgggctcccc ggccagcc atg ggc cct     117 
                                                     Met Gly Pro 
                                                      1 

tgg agc cgc agc ctc tcg ggc ctg ctg ctg ctg ctg agg tct cct ctt      165 
Trp Ser Arg Ser Leu Ser Gly Leu Leu Leu Leu Leu Arg Ser Pro Leu 
     5                   10                  15 

ggc tct cag gag cgg agc cct cct ccc tgt ttg acg cga gag cta cac      213 
Gly Ser Gln Glu Arg Ser Pro Pro Pro Cys Leu Thr Arg Glu Leu His 
 20                  25                  30                  35 

gtt cac ggt gcc ccg gcg cca cct gag aag agg ccg cgt ctg ggc aga      261 
Val His Gly Ala Pro Ala Pro Pro Glu Lys Arg Pro Arg Leu Gly Arg 
                 40                  45                  50 

gtg aat ttt gaa gat tgc acc ggt cga caa agg aca gct att ttc ctg      309 
Val Asn Phe Glu Asp Cys Thr Gly Arg Gln Arg Thr Ala Ile Phe Leu 
             55                  60                  65 

aca ccg att ccg aaa gtg ggc aca gat ggt gtg att aca gtc aaa agg      357 
Thr Pro Ile Pro Lys Val Gly Thr Asp Gly Val Ile Thr Val Lys Arg 
         70                  75                  80 

cct cta cgg ttt cat aac cca aca gat cca ttt ctt ggt cta cgc tgg      405 
Pro Leu Arg Phe His Asn Pro Thr Asp Pro Phe Leu Gly Leu Arg Trp 
     85                  90                  95 

gac tcc acc tac aga aag ttt tcc acc aaa gtc acg ctg aat aca gtg      453 
Asp Ser Thr Tyr Arg Lys Phe Ser Thr Lys Val Thr Leu Asn Thr Val 
100                 105                 110                 115 

ggg cac cac cac cgc ccc ccg ccc cat cag gcc tcc gtt tct gga atc      501 
Gly His His His Arg Pro Pro Pro His Gln Ala Ser Val Ser Gly Ile 
                120                 125                 130 

caa gca gaa ttg ctc aca ttt ccc aac tcc tct cct ggc ctc aga aga      549 
Gln Ala Glu Leu Leu Thr Phe Pro Asn Ser Ser Pro Gly Leu Arg Arg 
            135                 140                 145 

cag aag aga gac tgg gtt att cct ccc atc agc tgc cca gaa aat gaa      597 
Gln Lys Arg Asp Trp Val Ile Pro Pro Ile Ser Cys Pro Glu Asn Glu 
        150                 155                 160 

aaa ggc cca ttt cct aaa aac ctg gtt cag atc aaa tcc aac aaa gac      645 
Lys Gly Pro Phe Pro Lys Asn Leu Val Gln Ile Lys Ser Asn Lys Asp 
    165                 170                 175 

aaa gaa ggc aag gtt ttc tac agc atc act ggc caa gga gct gac aca      693 
Lys Glu Gly Lys Val Phe Tyr Ser Ile Thr Gly Gln Gly Ala Asp Thr 
180                 185                 190                 195 

ccc cct gtt ggt gtc ttt att att gaa aga gaa aca gga tgg ctg aag      741 
Pro Pro Val Gly Val Phe Ile Ile Glu Arg Glu Thr Gly Trp Leu Lys 
                200                 205                 210 

gtg aca gag cct ctg gat aga gaa cgc att gcc aca tac act ctc ttc      789 
Val Thr Glu Pro Leu Asp Arg Glu Arg Ile Ala Thr Tyr Thr Leu Phe 
            215                 220                 225 

tct cac gct gtg tca tcc aac ggg aat gca gtt gag gat cca atg gag      837 
Ser His Ala Val Ser Ser Asn Gly Asn Ala Val Glu Asp Pro Met Glu 
        230                 235                 240 

att ttg atc acg gta acc gat cag aat gac aac aag ccc gaa ttc acc      885 
Ile Leu Ile Thr Val Thr Asp Gln Asn Asp Asn Lys Pro Glu Phe Thr 
    245                 250                 255 

cag gag gtc ttt aag ggg tct gtc atg gaa ggt gct ctt cca gga acc      933 
Gln Glu Val Phe Lys Gly Ser Val Met Glu Gly Ala Leu Pro Gly Thr 
260                 265                 270                 275 

tct gtg atg gag gtc aca gcc aca gac gcg gac gat gat gtg aac acc      981 
Ser Val Met Glu Val Thr Ala Thr Asp Ala Asp Asp Asp Val Asn Thr 
                280                 285                 290 

tac aat gcc gcc atc gct tac acc atc ctc agc caa gat cct gag ctc     1029 
Tyr Asn Ala Ala Ile Ala Tyr Thr Ile Leu Ser Gln Asp Pro Glu Leu 
            295                 300                 305 

cct gac aaa aat atg ttc acc att aac agg aac aca gga gtc atc agt     1077 
Pro Asp Lys Asn Met Phe Thr Ile Asn Arg Asn Thr Gly Val Ile Ser 
        310                 315                 320 

gtg gtc acc act ggg ctg gac cga gag agt ttc cct acg tat acc ctg     1125 
Val Val Thr Thr Gly Leu Asp Arg Glu Ser Phe Pro Thr Tyr Thr Leu 
    325                 330                 335 

gtg gtt caa gct gct gac ctt caa ggt gag ggg tta agc aca aca gca     1173 
Val Val Gln Ala Ala Asp Leu Gln Gly Glu Gly Leu Ser Thr Thr Ala 
340                 345                 350                 355 

aca gct gtg atc aca gtc act gac acc aac gat aat cct ccg atc ttc     1221 
Thr Ala Val Ile Thr Val Thr Asp Thr Asn Asp Asn Pro Pro Ile Phe 
                360                 365                 370 

aat ccc acc acg tac aag ggt cag gtg cct gag aac gag gct aac gtc     1269 
Asn Pro Thr Thr Tyr Lys Gly Gln Val Pro Glu Asn Glu Ala Asn Val 
            375                 380                 385 

gta atc acc aca ctg aaa gtg act gat gct gat gcc ccc aat acc cca     1317 
Val Ile Thr Thr Leu Lys Val Thr Asp Ala Asp Ala Pro Asn Thr Pro 
        390                 395                 400 

gcg tgg gag gct gta tac acc ata ttg aat gat gat ggt gga caa ttt     1365 
Ala Trp Glu Ala Val Tyr Thr Ile Leu Asn Asp Asp Gly Gly Gln Phe 
    405                 410                 415 

gtc gtc acc aca aat cca gtg aac aac gat ggc att ttg aaa aca gca     1413 
Val Val Thr Thr Asn Pro Val Asn Asn Asp Gly Ile Leu Lys Thr Ala 
420                 425                 430                 435 

aag ggc ttg gat ttt gag gcc aag cag cag tac att cta cac gta gca     1461 
Lys Gly Leu Asp Phe Glu Ala Lys Gln Gln Tyr Ile Leu His Val Ala 
                440                 445                 450 

gtg acg aat gtg gta cct ttt gag gtc tct ctc acc acc tcc aca gcc     1509 
Val Thr Asn Val Val Pro Phe Glu Val Ser Leu Thr Thr Ser Thr Ala 
            455                 460                 465 

acc gtc acc gtg gat gtg ctg gat gtg aat gaa ggc ccc atc ttt gtg     1557 
Thr Val Thr Val Asp Val Leu Asp Val Asn Glu Gly Pro Ile Phe Val 
        470                 475                 480 

cct cct gaa aag aga gtg gaa gtg tcc gag gac ttt ggc gtg ggc cag     1605 
Pro Pro Glu Lys Arg Val Glu Val Ser Glu Asp Phe Gly Val Gly Gln 
    485                 490                 495 

gaa atc aca tcc tac act gcc cag gag cca gac aca ttt atg gaa cag     1653 
Glu Ile Thr Ser Tyr Thr Ala Gln Glu Pro Asp Thr Phe Met Glu Gln 
500                 505                 510                 515 

aaa ata aca tat cgg att tgg aga gac act cgc aac tgg ctg gag att     1701 
Lys Ile Thr Tyr Arg Ile Trp Arg Asp Thr Arg Asn Trp Leu Glu Ile 
                520                 525                 530 

aat ccg gac act ggt gcc att tcc act cgg gct gag ctg gac agg gag     1749 
Asn Pro Asp Thr Gly Ala Ile Ser Thr Arg Ala Glu Leu Asp Arg Glu 
            535                 540                 545 

gat ttt gag cac gtg aag aac agc acg tac aca gcc cta atc ata gct     1797 
Asp Phe Glu His Val Lys Asn Ser Thr Tyr Thr Ala Leu Ile Ile Ala 
        550                 555                 560 

aca gac aat ggt tct cca gtt gct act gga aca ggg aca ctt ctg ctg     1845 
Thr Asp Asn Gly Ser Pro Val Ala Thr Gly Thr Gly Thr Leu Leu Leu 
    565                 570                 575 

atc ctg tct gat gtg aat gac aac gcc ccc ata cca gaa cct cga act     1893 
Ile Leu Ser Asp Val Asn Asp Asn Ala Pro Ile Pro Glu Pro Arg Thr 
580                 585                 590                 595 

ata ttc ttc tgt gag agg aat cca aag cct cag gtc ata aac att cat     1941 
Ile Phe Phe Cys Glu Arg Asn Pro Lys Pro Gln Val Ile Asn Ile His 
                600                 605                 610 

gat gca gac ctt cct ccc aat aca tct ccc ttc aca gca gaa cta aca     1989 
Asp Ala Asp Leu Pro Pro Asn Thr Ser Pro Phe Thr Ala Glu Leu Thr 
            615                 620                 625 

cac ggg cga gtg ccc aac tgg acc att cag tac aac gac cca acc caa     2037 
His Gly Arg Val Pro Asn Trp Thr Ile Gln Tyr Asn Asp Pro Thr Gln 
        630                 635                 640 

gaa tct atc att ttg aag cca aag atg gcc tta gag gtg ggt gac tac     2085 
Glu Ser Ile Ile Leu Lys Pro Lys Met Ala Leu Glu Val Gly Asp Tyr 
    645                 650                 655 

aaa atc aat ctc aag ctc atg gat aac cag aat aaa gac caa gtg acc     2133 
Lys Ile Asn Leu Lys Leu Met Asp Asn Gln Asn Lys Asp Gln Val Thr 
660                 665                 670                 675 

acc tta gag gtc agc gtg tgt gac tgt gaa ggg gcc gcc ggc gtc tgt     2181 
Thr Leu Glu Val Ser Val Cys Asp Cys Glu Gly Ala Ala Gly Val Cys 
                680                 685                 690 

agg aag gca cag cct gtc gaa gca gga ttg caa att cct gcc att ctg     2229 
Arg Lys Ala Gln Pro Val Glu Ala Gly Leu Gln Ile Pro Ala Ile Leu 
            695                 700                 705 

ggg att ctt gga gga att ctt gct ttg cta att ctg att ctg ctg ctc     2277 
Gly Ile Leu Gly Gly Ile Leu Ala Leu Leu Ile Leu Ile Leu Leu Leu 
        710                 715                 720 

ttg ctg ttt ctt cgg agg aga gcg gtg gtc aaa gag ccc tta ctg ccc     2325 
Leu Leu Phe Leu Arg Arg Arg Ala Val Val Lys Glu Pro Leu Leu Pro 
    725                 730                 735 

cca gag gat gac acc cgg gac aac gtt tat tac tat gat gaa gaa gga     2373 
Pro Glu Asp Asp Thr Arg Asp Asn Val Tyr Tyr Tyr Asp Glu Glu Gly 
740                 745                 750                 755 

ggc gga gaa gag gac cag gac ttt gac ttg agc cag ctg cac agg ggc     2421 
Gly Gly Glu Glu Asp Gln Asp Phe Asp Leu Ser Gln Leu His Arg Gly 
                760                 765                 770 

ctg gac gct cgg cct gaa gtg act cgt aac gac gtt gca cca acc ctc     2469 
Leu Asp Ala Arg Pro Glu Val Thr Arg Asn Asp Val Ala Pro Thr Leu 
            775                 780                 785 

atg agt gtc ccc cgg tat ctt ccc cgc cct gcc aat ccc gat gaa att     2517 
Met Ser Val Pro Arg Tyr Leu Pro Arg Pro Ala Asn Pro Asp Glu Ile 
        790                 795                 800 

gga aat ttt att gat gaa aat ctg aaa gcg gct gat act gac ccc aca     2565 
Gly Asn Phe Ile Asp Glu Asn Leu Lys Ala Ala Asp Thr Asp Pro Thr 
    805                 810                 815 

gcc ccg cct tat gat tct ctg ctc gtg ttt gac tat gaa gga agc ggt     2613 
Ala Pro Pro Tyr Asp Ser Leu Leu Val Phe Asp Tyr Glu Gly Ser Gly 
820                 825                 830                 835 

tcc gaa gct gct agt ctg agc tcc ctg aac tcc tca gag tca gac aaa     2661 
Ser Glu Ala Ala Ser Leu Ser Ser Leu Asn Ser Ser Glu Ser Asp Lys 
                840                 845                 850 

gac cag gac tat gac tac ttg aac gaa tgg ggc aat ccg ttc aag aag     2709 
Asp Gln Asp Tyr Asp Tyr Leu Asn Glu Trp Gly Asn Pro Phe Lys Lys 
            855                 860                 865 

ctg gct gac atg tac gga ggc ggc gag gac cac tag gggactcgag          2755 
Leu Ala Asp Met Tyr Gly Gly Gly Glu Asp His  * 
        870                 875 

agaggcggcc cagaccatgt gcagaaatgc agaaatcagc gttctggtgt ttt          2808 

 
           
             34  
             878  
             PRT  
             Homo sapiens  
           
            34 

Met Gly Pro Trp Ser Arg Ser Leu Ser Gly Leu Leu Leu Leu Leu Arg 
 1               5                  10                  15 

Ser Pro Leu Gly Ser Gln Glu Arg Ser Pro Pro Pro Cys Leu Thr Arg 
            20                  25                  30 

Glu Leu His Val His Gly Ala Pro Ala Pro Pro Glu Lys Arg Pro Arg 
        35                  40                  45 

Leu Gly Arg Val Asn Phe Glu Asp Cys Thr Gly Arg Gln Arg Thr Ala 
    50                  55                  60 

Ile Phe Leu Thr Pro Ile Pro Lys Val Gly Thr Asp Gly Val Ile Thr 
65                  70                  75                  80 

Val Lys Arg Pro Leu Arg Phe His Asn Pro Thr Asp Pro Phe Leu Gly 
                85                  90                  95 

Leu Arg Trp Asp Ser Thr Tyr Arg Lys Phe Ser Thr Lys Val Thr Leu 
            100                 105                 110 

Asn Thr Val Gly His His His Arg Pro Pro Pro His Gln Ala Ser Val 
        115                 120                 125 

Ser Gly Ile Gln Ala Glu Leu Leu Thr Phe Pro Asn Ser Ser Pro Gly 
    130                 135                 140 

Leu Arg Arg Gln Lys Arg Asp Trp Val Ile Pro Pro Ile Ser Cys Pro 
145                 150                 155                 160 

Glu Asn Glu Lys Gly Pro Phe Pro Lys Asn Leu Val Gln Ile Lys Ser 
                165                 170                 175 

Asn Lys Asp Lys Glu Gly Lys Val Phe Tyr Ser Ile Thr Gly Gln Gly 
            180                 185                 190 

Ala Asp Thr Pro Pro Val Gly Val Phe Ile Ile Glu Arg Glu Thr Gly 
        195                 200                 205 

Trp Leu Lys Val Thr Glu Pro Leu Asp Arg Glu Arg Ile Ala Thr Tyr 
    210                 215                 220 

Thr Leu Phe Ser His Ala Val Ser Ser Asn Gly Asn Ala Val Glu Asp 
225                 230                 235                 240 

Pro Met Glu Ile Leu Ile Thr Val Thr Asp Gln Asn Asp Asn Lys Pro 
                245                 250                 255 

Glu Phe Thr Gln Glu Val Phe Lys Gly Ser Val Met Glu Gly Ala Leu 
            260                 265                 270 

Pro Gly Thr Ser Val Met Glu Val Thr Ala Thr Asp Ala Asp Asp Asp 
        275                 280                 285 

Val Asn Thr Tyr Asn Ala Ala Ile Ala Tyr Thr Ile Leu Ser Gln Asp 
    290                 295                 300 

Pro Glu Leu Pro Asp Lys Asn Met Phe Thr Ile Asn Arg Asn Thr Gly 
305                 310                 315                 320 

Val Ile Ser Val Val Thr Thr Gly Leu Asp Arg Glu Ser Phe Pro Thr 
                325                 330                 335 

Tyr Thr Leu Val Val Gln Ala Ala Asp Leu Gln Gly Glu Gly Leu Ser 
            340                 345                 350 

Thr Thr Ala Thr Ala Val Ile Thr Val Thr Asp Thr Asn Asp Asn Pro 
        355                 360                 365 

Pro Ile Phe Asn Pro Thr Thr Tyr Lys Gly Gln Val Pro Glu Asn Glu 
    370                 375                 380 

Ala Asn Val Val Ile Thr Thr Leu Lys Val Thr Asp Ala Asp Ala Pro 
385                 390                 395                 400 

Asn Thr Pro Ala Trp Glu Ala Val Tyr Thr Ile Leu Asn Asp Asp Gly 
                405                 410                 415 

Gly Gln Phe Val Val Thr Thr Asn Pro Val Asn Asn Asp Gly Ile Leu 
            420                 425                 430 

Lys Thr Ala Lys Gly Leu Asp Phe Glu Ala Lys Gln Gln Tyr Ile Leu 
        435                 440                 445 

His Val Ala Val Thr Asn Val Val Pro Phe Glu Val Ser Leu Thr Thr 
    450                 455                 460 

Ser Thr Ala Thr Val Thr Val Asp Val Leu Asp Val Asn Glu Gly Pro 
465                 470                 475                 480 

Ile Phe Val Pro Pro Glu Lys Arg Val Glu Val Ser Glu Asp Phe Gly 
                485                 490                 495 

Val Gly Gln Glu Ile Thr Ser Tyr Thr Ala Gln Glu Pro Asp Thr Phe 
            500                 505                 510 

Met Glu Gln Lys Ile Thr Tyr Arg Ile Trp Arg Asp Thr Arg Asn Trp 
        515                 520                 525 

Leu Glu Ile Asn Pro Asp Thr Gly Ala Ile Ser Thr Arg Ala Glu Leu 
    530                 535                 540 

Asp Arg Glu Asp Phe Glu His Val Lys Asn Ser Thr Tyr Thr Ala Leu 
545                 550                 555                 560 

Ile Ile Ala Thr Asp Asn Gly Ser Pro Val Ala Thr Gly Thr Gly Thr 
                565                 570                 575 

Leu Leu Leu Ile Leu Ser Asp Val Asn Asp Asn Ala Pro Ile Pro Glu 
            580                 585                 590 

Pro Arg Thr Ile Phe Phe Cys Glu Arg Asn Pro Lys Pro Gln Val Ile 
        595                 600                 605 

Asn Ile His Asp Ala Asp Leu Pro Pro Asn Thr Ser Pro Phe Thr Ala 
    610                 615                 620 

Glu Leu Thr His Gly Arg Val Pro Asn Trp Thr Ile Gln Tyr Asn Asp 
625                 630                 635                 640 

Pro Thr Gln Glu Ser Ile Ile Leu Lys Pro Lys Met Ala Leu Glu Val 
                645                 650                 655 

Gly Asp Tyr Lys Ile Asn Leu Lys Leu Met Asp Asn Gln Asn Lys Asp 
            660                 665                 670 

Gln Val Thr Thr Leu Glu Val Ser Val Cys Asp Cys Glu Gly Ala Ala 
        675                 680                 685 

Gly Val Cys Arg Lys Ala Gln Pro Val Glu Ala Gly Leu Gln Ile Pro 
    690                 695                 700 

Ala Ile Leu Gly Ile Leu Gly Gly Ile Leu Ala Leu Leu Ile Leu Ile 
705                 710                 715                 720 

Leu Leu Leu Leu Leu Phe Leu Arg Arg Arg Ala Val Val Lys Glu Pro 
                725                 730                 735 

Leu Leu Pro Pro Glu Asp Asp Thr Arg Asp Asn Val Tyr Tyr Tyr Asp 
            740                 745                 750 

Glu Glu Gly Gly Gly Glu Glu Asp Gln Asp Phe Asp Leu Ser Gln Leu 
        755                 760                 765 

His Arg Gly Leu Asp Ala Arg Pro Glu Val Thr Arg Asn Asp Val Ala 
    770                 775                 780 

Pro Thr Leu Met Ser Val Pro Arg Tyr Leu Pro Arg Pro Ala Asn Pro 
785                 790                 795                 800 

Asp Glu Ile Gly Asn Phe Ile Asp Glu Asn Leu Lys Ala Ala Asp Thr 
                805                 810                 815 

Asp Pro Thr Ala Pro Pro Tyr Asp Ser Leu Leu Val Phe Asp Tyr Glu 
            820                 825                 830 

Gly Ser Gly Ser Glu Ala Ala Ser Leu Ser Ser Leu Asn Ser Ser Glu 
        835                 840                 845 

Ser Asp Lys Asp Gln Asp Tyr Asp Tyr Leu Asn Glu Trp Gly Asn Pro 
    850                 855                 860 

Phe Lys Lys Leu Ala Asp Met Tyr Gly Gly Gly Glu Asp His 
865                 870                 875 

 
           
             35  
             3567  
             DNA  
             Homo sapiens  
             
               CDS  
               (411)...(3527)  
             
           
            35 

cgccatcccg cgctctgcgg actgggaggc ccgggccagg acgcgagtct gcgcagccga     60 

ggttccccag cgccccctgc agccgcgcgt aggcagagac ggagcccggc cctgcgcctc    120 

cgcaccacgc ccgggacccc acccagcggc ccgtacccgg agaagcagcg cgagcacccg    180 

aagctcccgg ctcggcggca gaaaccggga gtggggccgg gcgagtgcgc ggcatcccag    240 

gccggcccga acgtccgccc gcggtgggcc gacttcccct cctcttccct ctctccttcc    300 

tttagcccgc tggcgccgga cacgctgcgc ctcatctctt ggggcgttct tccccgttgg    360 

ccaaccgtcg catcccgtgc aactttgggg tagtggccgc ttagtgttga atg ttc       416 
                                                        Met Phe 
                                                         1 

ccc acc gag agc gca tgg ctt ggg aag cga ggc gcg aac ccg ggc ccc      464 
Pro Thr Glu Ser Ala Trp Leu Gly Lys Arg Gly Ala Asn Pro Gly Pro 
         5                   10                  15 

gaa gcc gcc gtc cgg gag acg gtg atg ctg ttg ctg tgc ctg ggg gtc      512 
Glu Ala Ala Val Arg Glu Thr Val Met Leu Leu Leu Cys Leu Gly Val 
     20                  25                  30 

ccg acc ggc cgc ccc tac aac gtg gac act gag agc gcg ctg ctt tac      560 
Pro Thr Gly Arg Pro Tyr Asn Val Asp Thr Glu Ser Ala Leu Leu Tyr 
 35                  40                  45                  50 

cag ggc ccc cac aac acg ctg ttc ggc tac tcg gtc gtg ctg cac agc      608 
Gln Gly Pro His Asn Thr Leu Phe Gly Tyr Ser Val Val Leu His Ser 
                 55                  60                  65 

cac ggg gcg aac cga tgg ctc cta gtg ggt gcg ccc act gcc aac tgg      656 
His Gly Ala Asn Arg Trp Leu Leu Val Gly Ala Pro Thr Ala Asn Trp 
             70                  75                  80 

ctc gcc aac gct tca gtg atc aat ccc ggg gcg att tac aga tgc agg      704 
Leu Ala Asn Ala Ser Val Ile Asn Pro Gly Ala Ile Tyr Arg Cys Arg 
         85                  90                  95 

atc gga aag aat ccc ggc cag acg tgc gaa cag ctc cag ctg ggt agc      752 
Ile Gly Lys Asn Pro Gly Gln Thr Cys Glu Gln Leu Gln Leu Gly Ser 
    100                 105                 110 

cct aat gga gaa cct tgt gga aag act tgt ttg gaa gag aga gac aat      800 
Pro Asn Gly Glu Pro Cys Gly Lys Thr Cys Leu Glu Glu Arg Asp Asn 
115                 120                 125                 130 

cag tgg ttg ggg gtc aca ctt tcc aga cag cca gga gaa aat gga tcc      848 
Gln Trp Leu Gly Val Thr Leu Ser Arg Gln Pro Gly Glu Asn Gly Ser 
                135                 140                 145 

atc gtg act tgt ggg cat aga tgg aaa aat ata ttt tac ata aag aat      896 
Ile Val Thr Cys Gly His Arg Trp Lys Asn Ile Phe Tyr Ile Lys Asn 
            150                 155                 160 

gaa aat aag ctc ccc act ggt ggt tgc tat gga gtg ccc cct gat tta      944 
Glu Asn Lys Leu Pro Thr Gly Gly Cys Tyr Gly Val Pro Pro Asp Leu 
        165                 170                 175 

cga aca gaa ctg agt aaa aga ata gct ccg tgt tat caa gat tat gtg      992 
Arg Thr Glu Leu Ser Lys Arg Ile Ala Pro Cys Tyr Gln Asp Tyr Val 
    180                 185                 190 

aaa aaa ttt gga gaa aat ttt gca tca tgt caa gct gga ata tcc agt     1040 
Lys Lys Phe Gly Glu Asn Phe Ala Ser Cys Gln Ala Gly Ile Ser Ser 
195                 200                 205                 210 

ttt tac aca aag gat tta att gtg atg ggg gcc cca gga tca tct tac     1088 
Phe Tyr Thr Lys Asp Leu Ile Val Met Gly Ala Pro Gly Ser Ser Tyr 
                215                 220                 225 

tgg act ggc tct ctt ttt gtc tac aat ata act aca aat aaa tac aag     1136 
Trp Thr Gly Ser Leu Phe Val Tyr Asn Ile Thr Thr Asn Lys Tyr Lys 
            230                 235                 240 

gct ttt tta gac aaa caa aat caa gta aaa ttt gga agt tat tta gga     1184 
Ala Phe Leu Asp Lys Gln Asn Gln Val Lys Phe Gly Ser Tyr Leu Gly 
        245                 250                 255 

tat tca gtc gga gct ggt cat ttt cgg agc cag cat act acc gaa gta     1232 
Tyr Ser Val Gly Ala Gly His Phe Arg Ser Gln His Thr Thr Glu Val 
    260                 265                 270 

gtc gga gga gct cct caa cat gag cag att ggt aag gca tat ata ttc     1280 
Val Gly Gly Ala Pro Gln His Glu Gln Ile Gly Lys Ala Tyr Ile Phe 
275                 280                 285                 290 

agc att gat gaa aaa gaa cta aat atc tta cat gaa atg aaa ggt aaa     1328 
Ser Ile Asp Glu Lys Glu Leu Asn Ile Leu His Glu Met Lys Gly Lys 
                295                 300                 305 

aag ctt gga tcg tac ttt gga gct tct gtc tgt gct gtg gac ctc aat     1376 
Lys Leu Gly Ser Tyr Phe Gly Ala Ser Val Cys Ala Val Asp Leu Asn 
            310                 315                 320 

gca gat ggc ttc tca gat ctg ctc gtg gga gca ccc atg cag agc acc     1424 
Ala Asp Gly Phe Ser Asp Leu Leu Val Gly Ala Pro Met Gln Ser Thr 
        325                 330                 335 

atc aga gag gaa gga aga gtg ttt gtg tac atc aac tct ggc tcg gga     1472 
Ile Arg Glu Glu Gly Arg Val Phe Val Tyr Ile Asn Ser Gly Ser Gly 
    340                 345                 350 

gca gta atg aat gca atg gaa aca aac ctc gtt gga agt gac aaa tat     1520 
Ala Val Met Asn Ala Met Glu Thr Asn Leu Val Gly Ser Asp Lys Tyr 
355                 360                 365                 370 

gct gca aga ttt ggg gaa tct ata gtt aat ctt ggc gac att gac aat     1568 
Ala Ala Arg Phe Gly Glu Ser Ile Val Asn Leu Gly Asp Ile Asp Asn 
                375                 380                 385 

gat ggc ttt gaa gat gtt gct atc gga gct cca caa gaa gat gac ttg     1616 
Asp Gly Phe Glu Asp Val Ala Ile Gly Ala Pro Gln Glu Asp Asp Leu 
            390                 395                 400 

caa ggt gct att tat att tac aat ggc cgt gca gat ggg atc tcg tca     1664 
Gln Gly Ala Ile Tyr Ile Tyr Asn Gly Arg Ala Asp Gly Ile Ser Ser 
        405                 410                 415 

acc ttc tca cag aga att gaa gga ctt cag atc agc aaa tcg tta agt     1712 
Thr Phe Ser Gln Arg Ile Glu Gly Leu Gln Ile Ser Lys Ser Leu Ser 
    420                 425                 430 

atg ttt gga cag tct ata tca gga caa att gat gca gat aat aat ggc     1760 
Met Phe Gly Gln Ser Ile Ser Gly Gln Ile Asp Ala Asp Asn Asn Gly 
435                 440                 445                 450 

tat gta gat gta gca gtt ggt gct ttt cgg tct gat tct gct gtc ttg     1808 
Tyr Val Asp Val Ala Val Gly Ala Phe Arg Ser Asp Ser Ala Val Leu 
                455                 460                 465 

cta agg aca aga cct gta gta att gtt gac gct tct tta agc cac cct     1856 
Leu Arg Thr Arg Pro Val Val Ile Val Asp Ala Ser Leu Ser His Pro 
            470                 475                 480 

gag tca gta aat aga acg aaa ttt gac tgt gtt gaa aat gga tgg cct     1904 
Glu Ser Val Asn Arg Thr Lys Phe Asp Cys Val Glu Asn Gly Trp Pro 
        485                 490                 495 

tct gtg tgc ata gat cta aca ctt tgt ttc tca tat aag ggc aag gaa     1952 
Ser Val Cys Ile Asp Leu Thr Leu Cys Phe Ser Tyr Lys Gly Lys Glu 
    500                 505                 510 

gtt cca ggt tac att gtt ttg ttt tat aac atg agt ttg gat gtg aac     2000 
Val Pro Gly Tyr Ile Val Leu Phe Tyr Asn Met Ser Leu Asp Val Asn 
515                 520                 525                 530 

aga aag gca gag tct cca cca aga ttc tat ttc tct tct aat gga act     2048 
Arg Lys Ala Glu Ser Pro Pro Arg Phe Tyr Phe Ser Ser Asn Gly Thr 
                535                 540                 545 

tct gac gtg att aca gga agc ata cag gtg tcc agc aga gaa gct aac     2096 
Ser Asp Val Ile Thr Gly Ser Ile Gln Val Ser Ser Arg Glu Ala Asn 
            550                 555                 560 

tgt aga aca cat caa gca ttt atg cgg aaa gat gtg cgg gac atc ctc     2144 
Cys Arg Thr His Gln Ala Phe Met Arg Lys Asp Val Arg Asp Ile Leu 
        565                 570                 575 

acc cca att cag att gaa gct gct tac cac ctt ggt cct cat gtc atc     2192 
Thr Pro Ile Gln Ile Glu Ala Ala Tyr His Leu Gly Pro His Val Ile 
    580                 585                 590 

agt aaa cga agt aca gag gaa ttc cca cca ctt cag cca att ctt cag     2240 
Ser Lys Arg Ser Thr Glu Glu Phe Pro Pro Leu Gln Pro Ile Leu Gln 
595                 600                 605                 610 

cag aag aaa gaa aaa gac ata atg aaa aaa aca ata aac ttt gca agg     2288 
Gln Lys Lys Glu Lys Asp Ile Met Lys Lys Thr Ile Asn Phe Ala Arg 
                615                 620                 625 

ttt tgt gcc cat gaa aat tgt tct gct gat tta cag gtt tct gca aag     2336 
Phe Cys Ala His Glu Asn Cys Ser Ala Asp Leu Gln Val Ser Ala Lys 
            630                 635                 640 

att ggg ttt ttg aag ccc cat gaa aat aaa aca tat ctt gct gtt ggg     2384 
Ile Gly Phe Leu Lys Pro His Glu Asn Lys Thr Tyr Leu Ala Val Gly 
        645                 650                 655 

agt atg aag aca ttg atg ttg aat gtg tcc ttg ttt aat gct gga gat     2432 
Ser Met Lys Thr Leu Met Leu Asn Val Ser Leu Phe Asn Ala Gly Asp 
    660                 665                 670 

gat gca tat gaa acg act cta cat gtc aaa cta ccc gtg ggt ctt tat     2480 
Asp Ala Tyr Glu Thr Thr Leu His Val Lys Leu Pro Val Gly Leu Tyr 
675                 680                 685                 690 

ttc att aag att tta gag ctg gaa gag aag caa ata aac tgt gaa gtc     2528 
Phe Ile Lys Ile Leu Glu Leu Glu Glu Lys Gln Ile Asn Cys Glu Val 
                695                 700                 705 

aca gat aac tct ggc gtg gta caa ctt gac tgc agt att ggc tat ata     2576 
Thr Asp Asn Ser Gly Val Val Gln Leu Asp Cys Ser Ile Gly Tyr Ile 
            710                 715                 720 

tat gta gat cat ctc tca agg ata gat att agc ttt ctc ctg gat gtg     2624 
Tyr Val Asp His Leu Ser Arg Ile Asp Ile Ser Phe Leu Leu Asp Val 
        725                 730                 735 

agc tca ctc agc aga gcg gaa gag gac ctc agt atc aca gtg cat gct     2672 
Ser Ser Leu Ser Arg Ala Glu Glu Asp Leu Ser Ile Thr Val His Ala 
    740                 745                 750 

acc tgt gaa aat gaa gag gaa atg gac aat cta aag cac agc aga gtg     2720 
Thr Cys Glu Asn Glu Glu Glu Met Asp Asn Leu Lys His Ser Arg Val 
755                 760                 765                 770 

act gta gca ata cct tta aaa tat gag gtt aag ctg act gtt cat ggg     2768 
Thr Val Ala Ile Pro Leu Lys Tyr Glu Val Lys Leu Thr Val His Gly 
                775                 780                 785 

ttt gta aac cca act tca ttt gtg tat gga tca aat gat gaa aat gag     2816 
Phe Val Asn Pro Thr Ser Phe Val Tyr Gly Ser Asn Asp Glu Asn Glu 
            790                 795                 800 

cct gaa acg tgc atg gtg gag aaa atg aac tta act ttc cat gtt atc     2864 
Pro Glu Thr Cys Met Val Glu Lys Met Asn Leu Thr Phe His Val Ile 
        805                 810                 815 

aac act ggc aat agt atg gct ccc aat gtt agt gtg gaa ata atg gta     2912 
Asn Thr Gly Asn Ser Met Ala Pro Asn Val Ser Val Glu Ile Met Val 
    820                 825                 830 

cca aat tct ttt agc ccc caa act gat aag ctg ttc aac att ttg gat     2960 
Pro Asn Ser Phe Ser Pro Gln Thr Asp Lys Leu Phe Asn Ile Leu Asp 
835                 840                 845                 850 

gtc cag act act act gga gaa tgc cac ttt gaa aat tat caa aga gtg     3008 
Val Gln Thr Thr Thr Gly Glu Cys His Phe Glu Asn Tyr Gln Arg Val 
                855                 860                 865 

tgt gca tta gag cag caa aag agt gca atg cag acc ttg aaa ggc ata     3056 
Cys Ala Leu Glu Gln Gln Lys Ser Ala Met Gln Thr Leu Lys Gly Ile 
            870                 875                 880 

gtc cag ttc ttg tcc aag act gat aag agg cta ttg tac tgc ata aaa     3104 
Val Gln Phe Leu Ser Lys Thr Asp Lys Arg Leu Leu Tyr Cys Ile Lys 
        885                 890                 895 

gct gat cca cat tgt tta aat ttc ttg tgt aat ttt ggg aaa atg gaa     3152 
Ala Asp Pro His Cys Leu Asn Phe Leu Cys Asn Phe Gly Lys Met Glu 
    900                 905                 910 

agt gga aaa gaa gcc agt gtt cat atc caa ctg gaa ggc cgg cca tcc     3200 
Ser Gly Lys Glu Ala Ser Val His Ile Gln Leu Glu Gly Arg Pro Ser 
915                 920                 925                 930 

att tta gaa atg gat gag act tca gca ctc aag ttt gaa ata aga gca     3248 
Ile Leu Glu Met Asp Glu Thr Ser Ala Leu Lys Phe Glu Ile Arg Ala 
                935                 940                 945 

aca ggt ttt cca gag cca aat cca aga gta att gaa cta aac aag gat     3296 
Thr Gly Phe Pro Glu Pro Asn Pro Arg Val Ile Glu Leu Asn Lys Asp 
            950                 955                 960 

gag aat gtt gcg cat gtt cta ctg gaa gga cta cat cat caa aga ccc     3344 
Glu Asn Val Ala His Val Leu Leu Glu Gly Leu His His Gln Arg Pro 
        965                 970                 975 

aaa cgt tat ttc acc ata gtg att att tca agt agc ttg cta ctt gga     3392 
Lys Arg Tyr Phe Thr Ile Val Ile Ile Ser Ser Ser Leu Leu Leu Gly 
    980                 985                 990 

ctt att gta ctt ctg ttg atc tca tat gtt atg tgg aag gct ggc ttc     3440 
Leu Ile Val Leu Leu Leu Ile Ser Tyr Val Met Trp Lys Ala Gly Phe 
 995                1000                1005                1010 

ttt aaa aga caa tac aaa tct atc cta caa gaa gaa aac aga aga gac     3488 
Phe Lys Arg Gln Tyr Lys Ser Ile Leu Gln Glu Glu Asn Arg Arg Asp 
                1015                1020                1025 

agt tgg agt tat atc aac agt aaa agc aat gat gat taa ggacttcttt      3537 
Ser Trp Ser Tyr Ile Asn Ser Lys Ser Asn Asp Asp  * 
            1030                1035 

caaattgaga gaatggaaaa cagcccgccc                                    3567 

 
           
             36  
             1038  
             PRT  
             Homo sapiens  
           
            36 

Met Phe Pro Thr Glu Ser Ala Trp Leu Gly Lys Arg Gly Ala Asn Pro 
 1               5                  10                  15 

Gly Pro Glu Ala Ala Val Arg Glu Thr Val Met Leu Leu Leu Cys Leu 
            20                  25                  30 

Gly Val Pro Thr Gly Arg Pro Tyr Asn Val Asp Thr Glu Ser Ala Leu 
        35                  40                  45 

Leu Tyr Gln Gly Pro His Asn Thr Leu Phe Gly Tyr Ser Val Val Leu 
    50                  55                  60 

His Ser His Gly Ala Asn Arg Trp Leu Leu Val Gly Ala Pro Thr Ala 
65                  70                  75                  80 

Asn Trp Leu Ala Asn Ala Ser Val Ile Asn Pro Gly Ala Ile Tyr Arg 
                85                  90                  95 

Cys Arg Ile Gly Lys Asn Pro Gly Gln Thr Cys Glu Gln Leu Gln Leu 
            100                 105                 110 

Gly Ser Pro Asn Gly Glu Pro Cys Gly Lys Thr Cys Leu Glu Glu Arg 
        115                 120                 125 

Asp Asn Gln Trp Leu Gly Val Thr Leu Ser Arg Gln Pro Gly Glu Asn 
    130                 135                 140 

Gly Ser Ile Val Thr Cys Gly His Arg Trp Lys Asn Ile Phe Tyr Ile 
145                 150                 155                 160 

Lys Asn Glu Asn Lys Leu Pro Thr Gly Gly Cys Tyr Gly Val Pro Pro 
                165                 170                 175 

Asp Leu Arg Thr Glu Leu Ser Lys Arg Ile Ala Pro Cys Tyr Gln Asp 
            180                 185                 190 

Tyr Val Lys Lys Phe Gly Glu Asn Phe Ala Ser Cys Gln Ala Gly Ile 
        195                 200                 205 

Ser Ser Phe Tyr Thr Lys Asp Leu Ile Val Met Gly Ala Pro Gly Ser 
    210                 215                 220 

Ser Tyr Trp Thr Gly Ser Leu Phe Val Tyr Asn Ile Thr Thr Asn Lys 
225                 230                 235                 240 

Tyr Lys Ala Phe Leu Asp Lys Gln Asn Gln Val Lys Phe Gly Ser Tyr 
                245                 250                 255 

Leu Gly Tyr Ser Val Gly Ala Gly His Phe Arg Ser Gln His Thr Thr 
            260                 265                 270 

Glu Val Val Gly Gly Ala Pro Gln His Glu Gln Ile Gly Lys Ala Tyr 
        275                 280                 285 

Ile Phe Ser Ile Asp Glu Lys Glu Leu Asn Ile Leu His Glu Met Lys 
    290                 295                 300 

Gly Lys Lys Leu Gly Ser Tyr Phe Gly Ala Ser Val Cys Ala Val Asp 
305                 310                 315                 320 

Leu Asn Ala Asp Gly Phe Ser Asp Leu Leu Val Gly Ala Pro Met Gln 
                325                 330                 335 

Ser Thr Ile Arg Glu Glu Gly Arg Val Phe Val Tyr Ile Asn Ser Gly 
            340                 345                 350 

Ser Gly Ala Val Met Asn Ala Met Glu Thr Asn Leu Val Gly Ser Asp 
        355                 360                 365 

Lys Tyr Ala Ala Arg Phe Gly Glu Ser Ile Val Asn Leu Gly Asp Ile 
    370                 375                 380 

Asp Asn Asp Gly Phe Glu Asp Val Ala Ile Gly Ala Pro Gln Glu Asp 
385                 390                 395                 400 

Asp Leu Gln Gly Ala Ile Tyr Ile Tyr Asn Gly Arg Ala Asp Gly Ile 
                405                 410                 415 

Ser Ser Thr Phe Ser Gln Arg Ile Glu Gly Leu Gln Ile Ser Lys Ser 
            420                 425                 430 

Leu Ser Met Phe Gly Gln Ser Ile Ser Gly Gln Ile Asp Ala Asp Asn 
        435                 440                 445 

Asn Gly Tyr Val Asp Val Ala Val Gly Ala Phe Arg Ser Asp Ser Ala 
    450                 455                 460 

Val Leu Leu Arg Thr Arg Pro Val Val Ile Val Asp Ala Ser Leu Ser 
465                 470                 475                 480 

His Pro Glu Ser Val Asn Arg Thr Lys Phe Asp Cys Val Glu Asn Gly 
                485                 490                 495 

Trp Pro Ser Val Cys Ile Asp Leu Thr Leu Cys Phe Ser Tyr Lys Gly 
            500                 505                 510 

Lys Glu Val Pro Gly Tyr Ile Val Leu Phe Tyr Asn Met Ser Leu Asp 
        515                 520                 525 

Val Asn Arg Lys Ala Glu Ser Pro Pro Arg Phe Tyr Phe Ser Ser Asn 
    530                 535                 540 

Gly Thr Ser Asp Val Ile Thr Gly Ser Ile Gln Val Ser Ser Arg Glu 
545                 550                 555                 560 

Ala Asn Cys Arg Thr His Gln Ala Phe Met Arg Lys Asp Val Arg Asp 
                565                 570                 575 

Ile Leu Thr Pro Ile Gln Ile Glu Ala Ala Tyr His Leu Gly Pro His 
            580                 585                 590 

Val Ile Ser Lys Arg Ser Thr Glu Glu Phe Pro Pro Leu Gln Pro Ile 
        595                 600                 605 

Leu Gln Gln Lys Lys Glu Lys Asp Ile Met Lys Lys Thr Ile Asn Phe 
    610                 615                 620 

Ala Arg Phe Cys Ala His Glu Asn Cys Ser Ala Asp Leu Gln Val Ser 
625                 630                 635                 640 

Ala Lys Ile Gly Phe Leu Lys Pro His Glu Asn Lys Thr Tyr Leu Ala 
                645                 650                 655 

Val Gly Ser Met Lys Thr Leu Met Leu Asn Val Ser Leu Phe Asn Ala 
            660                 665                 670 

Gly Asp Asp Ala Tyr Glu Thr Thr Leu His Val Lys Leu Pro Val Gly 
        675                 680                 685 

Leu Tyr Phe Ile Lys Ile Leu Glu Leu Glu Glu Lys Gln Ile Asn Cys 
    690                 695                 700 

Glu Val Thr Asp Asn Ser Gly Val Val Gln Leu Asp Cys Ser Ile Gly 
705                 710                 715                 720 

Tyr Ile Tyr Val Asp His Leu Ser Arg Ile Asp Ile Ser Phe Leu Leu 
                725                 730                 735 

Asp Val Ser Ser Leu Ser Arg Ala Glu Glu Asp Leu Ser Ile Thr Val 
            740                 745                 750 

His Ala Thr Cys Glu Asn Glu Glu Glu Met Asp Asn Leu Lys His Ser 
        755                 760                 765 

Arg Val Thr Val Ala Ile Pro Leu Lys Tyr Glu Val Lys Leu Thr Val 
    770                 775                 780 

His Gly Phe Val Asn Pro Thr Ser Phe Val Tyr Gly Ser Asn Asp Glu 
785                 790                 795                 800 

Asn Glu Pro Glu Thr Cys Met Val Glu Lys Met Asn Leu Thr Phe His 
                805                 810                 815 

Val Ile Asn Thr Gly Asn Ser Met Ala Pro Asn Val Ser Val Glu Ile 
            820                 825                 830 

Met Val Pro Asn Ser Phe Ser Pro Gln Thr Asp Lys Leu Phe Asn Ile 
        835                 840                 845 

Leu Asp Val Gln Thr Thr Thr Gly Glu Cys His Phe Glu Asn Tyr Gln 
    850                 855                 860 

Arg Val Cys Ala Leu Glu Gln Gln Lys Ser Ala Met Gln Thr Leu Lys 
865                 870                 875                 880 

Gly Ile Val Gln Phe Leu Ser Lys Thr Asp Lys Arg Leu Leu Tyr Cys 
                885                 890                 895 

Ile Lys Ala Asp Pro His Cys Leu Asn Phe Leu Cys Asn Phe Gly Lys 
            900                 905                 910 

Met Glu Ser Gly Lys Glu Ala Ser Val His Ile Gln Leu Glu Gly Arg 
        915                 920                 925 

Pro Ser Ile Leu Glu Met Asp Glu Thr Ser Ala Leu Lys Phe Glu Ile 
    930                 935                 940 

Arg Ala Thr Gly Phe Pro Glu Pro Asn Pro Arg Val Ile Glu Leu Asn 
945                 950                 955                 960 

Lys Asp Glu Asn Val Ala His Val Leu Leu Glu Gly Leu His His Gln 
                965                 970                 975 

Arg Pro Lys Arg Tyr Phe Thr Ile Val Ile Ile Ser Ser Ser Leu Leu 
            980                 985                 990 

Leu Gly Leu Ile Val Leu Leu Leu Ile Ser Tyr Val Met Trp Lys Ala 
        995                 1000                1005 

Gly Phe Phe Lys Arg Gln Tyr Lys Ser Ile Leu Gln Glu Glu Asn Arg 
    1010                1015                1020 

Arg Asp Ser Trp Ser Tyr Ile Asn Ser Lys Ser Asn Asp Asp 
1025                1030                1035 

 
           
             37  
             2742  
             DNA  
             Homo sapiens  
             
               CDS  
               (114)...(2510)  
             
           
            37 

agcccagaga gaaagtctga cttgccccac agccagtgag tgactgcagc agcaccagaa     60 

tctggtctgt ttcctgtttg gctcttctac cactacggct tgggatctcg ggc atg       116 
                                                           Met 
                                                            1 

gtg gct ttg cca atg gtc ctt gtt ttg ctg ctg gtc ctg agc aga ggt      164 
Val Ala Leu Pro Met Val Leu Val Leu Leu Leu Val Leu Ser Arg Gly 
             5                   10                  15 

gag agt gaa ttg gac gcc aag atc cca tcc aca ggg gat gcc aca gaa      212 
Glu Ser Glu Leu Asp Ala Lys Ile Pro Ser Thr Gly Asp Ala Thr Glu 
         20                  25                  30 

tgg cgg aat cct cac ctg tcc atg ctg ggg tcc tgc cag cca gcc ccc      260 
Trp Arg Asn Pro His Leu Ser Met Leu Gly Ser Cys Gln Pro Ala Pro 
     35                  40                  45 

tcc tgc cag aag tgc atc ctc tca cac ccc agc tgt gca tgg tgc aag      308 
Ser Cys Gln Lys Cys Ile Leu Ser His Pro Ser Cys Ala Trp Cys Lys 
 50                  55                  60                  65 

caa ctg aac ttc acc gcg tcg gga gag gcg gag gcg cgg cgc tgc gcc      356 
Gln Leu Asn Phe Thr Ala Ser Gly Glu Ala Glu Ala Arg Arg Cys Ala 
                 70                  75                  80 

cga cga gag gag ctg ctg gct cga ggc tgc ccg ctg gag gag ctg gag      404 
Arg Arg Glu Glu Leu Leu Ala Arg Gly Cys Pro Leu Glu Glu Leu Glu 
             85                  90                  95 

gag ccc cgc ggc cag cag gag gtg ctg cag gac cag ccg ctc agc cag      452 
Glu Pro Arg Gly Gln Gln Glu Val Leu Gln Asp Gln Pro Leu Ser Gln 
        100                 105                 110 

ggc gcc cgc gga gag ggt gcc acc cag ctg gcg ccg cag cgg gtc cgg      500 
Gly Ala Arg Gly Glu Gly Ala Thr Gln Leu Ala Pro Gln Arg Val Arg 
    115                 120                 125 

gtc acg ctg cgg cct ggg gag ccc cag cag ctc cag gtc cgc ttc ctt      548 
Val Thr Leu Arg Pro Gly Glu Pro Gln Gln Leu Gln Val Arg Phe Leu 
130                 135                 140                 145 

cgt gct gag gga tac ccg gtg gac ctg tac tac ctt atg gac ctg agc      596 
Arg Ala Glu Gly Tyr Pro Val Asp Leu Tyr Tyr Leu Met Asp Leu Ser 
                150                 155                 160 

tac tcc atg aag gac gac ctg gaa cgc gtg cgc cag ctc ggg cac gct      644 
Tyr Ser Met Lys Asp Asp Leu Glu Arg Val Arg Gln Leu Gly His Ala 
            165                 170                 175 

ctg ctg gtc cgg ctg cag gaa gtc acc cat tct gtg cgc att ggt ttt      692 
Leu Leu Val Arg Leu Gln Glu Val Thr His Ser Val Arg Ile Gly Phe 
        180                 185                 190 

ggt tcc ttt gtg gac aaa acg gtg ctg ccc ttt gtg agc aca gta ccc      740 
Gly Ser Phe Val Asp Lys Thr Val Leu Pro Phe Val Ser Thr Val Pro 
    195                 200                 205 

tcc aaa ctg cgc cac ccc tgc ccc acc cgg ctg gag cgc tgc cag tca      788 
Ser Lys Leu Arg His Pro Cys Pro Thr Arg Leu Glu Arg Cys Gln Ser 
210                 215                 220                 225 

cca ttc agc ttt cac cat gtg ctg tcc ctg acg ggg gac gca caa gcc      836 
Pro Phe Ser Phe His His Val Leu Ser Leu Thr Gly Asp Ala Gln Ala 
                230                 235                 240 

ttc gag cgg gag gtg ggg cgc cag agt gtg tcc ggc aat ctg gac tcg      884 
Phe Glu Arg Glu Val Gly Arg Gln Ser Val Ser Gly Asn Leu Asp Ser 
            245                 250                 255 

cct gaa ggt ggc ttc gat gcc att ctg cag gct gca ctc tgc cag gag      932 
Pro Glu Gly Gly Phe Asp Ala Ile Leu Gln Ala Ala Leu Cys Gln Glu 
        260                 265                 270 

cag att ggc tgg aga aat gtg tcc cgg ctg ctg gtg ttc act tca gac      980 
Gln Ile Gly Trp Arg Asn Val Ser Arg Leu Leu Val Phe Thr Ser Asp 
    275                 280                 285 

gac aca ttc cat aca gct ggg gac ggg aag ttg ggc ggc att ttc atg     1028 
Asp Thr Phe His Thr Ala Gly Asp Gly Lys Leu Gly Gly Ile Phe Met 
290                 295                 300                 305 

ccc agt gat ggg cac tgc cac ttg gac agc aat ggc ctc tac agt cgc     1076 
Pro Ser Asp Gly His Cys His Leu Asp Ser Asn Gly Leu Tyr Ser Arg 
                310                 315                 320 

agc aca gag ttt gac tac cct tct gtg ggt cag gta gcc cag gcc ctc     1124 
Ser Thr Glu Phe Asp Tyr Pro Ser Val Gly Gln Val Ala Gln Ala Leu 
            325                 330                 335 

tct gca gca aat atc cag ccc atc ttt gct gtc acc agt gcc gca ctg     1172 
Ser Ala Ala Asn Ile Gln Pro Ile Phe Ala Val Thr Ser Ala Ala Leu 
        340                 345                 350 

cct gtc tac cag gag ctg agt aaa ctg att cct aag tct gca gtt ggg     1220 
Pro Val Tyr Gln Glu Leu Ser Lys Leu Ile Pro Lys Ser Ala Val Gly 
    355                 360                 365 

gag ctg agt gag gac tcc agc aac gtg gta cag ctc atc atg gat gct     1268 
Glu Leu Ser Glu Asp Ser Ser Asn Val Val Gln Leu Ile Met Asp Ala 
370                 375                 380                 385 

tat aat agc ctg tct tcc act gtg acc ctt gaa cac tct tca ctc cct     1316 
Tyr Asn Ser Leu Ser Ser Thr Val Thr Leu Glu His Ser Ser Leu Pro 
                390                 395                 400 

cct ggg gtc cac att tct tac gaa tcc cag tgt gag ggt cct gag aag     1364 
Pro Gly Val His Ile Ser Tyr Glu Ser Gln Cys Glu Gly Pro Glu Lys 
            405                 410                 415 

agg gag ggt aag gct gag gat cga gga cag tgc aac cac gtc cga atc     1412 
Arg Glu Gly Lys Ala Glu Asp Arg Gly Gln Cys Asn His Val Arg Ile 
        420                 425                 430 

aac cag acg gtg act ttc tgg gtt tct ctc caa gcc acc cac tgc ctc     1460 
Asn Gln Thr Val Thr Phe Trp Val Ser Leu Gln Ala Thr His Cys Leu 
    435                 440                 445 

cca gag ccc cat ctc ctg agg ctc cgg gcc ctt ggc ttc tca gag gag     1508 
Pro Glu Pro His Leu Leu Arg Leu Arg Ala Leu Gly Phe Ser Glu Glu 
450                 455                 460                 465 

ctg att gtg gag ttg cac acg ctg tgt gac tgt aat tgc agt gac acc     1556 
Leu Ile Val Glu Leu His Thr Leu Cys Asp Cys Asn Cys Ser Asp Thr 
                470                 475                 480 

cag ccc cag gct ccc cac tgc agt gat ggc cag gga cac cta caa tgt     1604 
Gln Pro Gln Ala Pro His Cys Ser Asp Gly Gln Gly His Leu Gln Cys 
            485                 490                 495 

ggt gta tgc agc tgt gcc cct ggc cgc cta ggt cgg ctc tgt gag tgc     1652 
Gly Val Cys Ser Cys Ala Pro Gly Arg Leu Gly Arg Leu Cys Glu Cys 
        500                 505                 510 

tct gtg gca gag ctg tcc tcc cca gac ctg gaa tct ggg tgc cgg gct     1700 
Ser Val Ala Glu Leu Ser Ser Pro Asp Leu Glu Ser Gly Cys Arg Ala 
    515                 520                 525 

ccc aat ggc aca ggg ccc ctg tgc agt gga aag ggt cac tgt caa tgt     1748 
Pro Asn Gly Thr Gly Pro Leu Cys Ser Gly Lys Gly His Cys Gln Cys 
530                 535                 540                 545 

gga cgc tgc agc tgc agt gga cag agc tct ggg cat ctg tgc gag tgt     1796 
Gly Arg Cys Ser Cys Ser Gly Gln Ser Ser Gly His Leu Cys Glu Cys 
                550                 555                 560 

gac gat gcc agc tgt gag cga cat gag ggc atc ctc tgc gga ggc ttt     1844 
Asp Asp Ala Ser Cys Glu Arg His Glu Gly Ile Leu Cys Gly Gly Phe 
            565                 570                 575 

ggt cgc tgc caa tgt gga gta tgt cac tgt cat gcc aac cgc acg ggc     1892 
Gly Arg Cys Gln Cys Gly Val Cys His Cys His Ala Asn Arg Thr Gly 
        580                 585                 590 

aga gca tgc gaa tgc agt ggg gac atg gac agt tgc atc agt ccc gag     1940 
Arg Ala Cys Glu Cys Ser Gly Asp Met Asp Ser Cys Ile Ser Pro Glu 
    595                 600                 605 

gga ggg ctc tgc agt ggg cat gga cgc tgc aaa tgc aac cgc tgc cag     1988 
Gly Gly Leu Cys Ser Gly His Gly Arg Cys Lys Cys Asn Arg Cys Gln 
610                 615                 620                 625 

tgc ttg gac ggc tac tat ggt gct cta tgc gac caa tgc cca ggc tgc     2036 
Cys Leu Asp Gly Tyr Tyr Gly Ala Leu Cys Asp Gln Cys Pro Gly Cys 
                630                 635                 640 

aag aca cca tgc gag aga cac cgg gac tgt gca gag tgt ggg gcc ttc     2084 
Lys Thr Pro Cys Glu Arg His Arg Asp Cys Ala Glu Cys Gly Ala Phe 
            645                 650                 655 

agg act ggc cca ctg gcc acc aac tgc agt aca gct tgt gcc cat acc     2132 
Arg Thr Gly Pro Leu Ala Thr Asn Cys Ser Thr Ala Cys Ala His Thr 
        660                 665                 670 

aat gtg acc ctg gcc ttg gcc cct atc ttg gat gat ggc tgg tgc aaa     2180 
Asn Val Thr Leu Ala Leu Ala Pro Ile Leu Asp Asp Gly Trp Cys Lys 
    675                 680                 685 

gag cgg acc ctg gac aac cag ctg ttc ttc ttc ttg gtg gag gat gac     2228 
Glu Arg Thr Leu Asp Asn Gln Leu Phe Phe Phe Leu Val Glu Asp Asp 
690                 695                 700                 705 

gcc aga ggc acg gtc gtg ctc aga gtg aga ccc caa gaa aag gga gca     2276 
Ala Arg Gly Thr Val Val Leu Arg Val Arg Pro Gln Glu Lys Gly Ala 
                710                 715                 720 

gac cac acg cag gcc att gtg ctg ggc tgc gta ggg ggc atc gtg gca     2324 
Asp His Thr Gln Ala Ile Val Leu Gly Cys Val Gly Gly Ile Val Ala 
            725                 730                 735 

gtg ggg ctg ggg ctg gtc ctg gct tac cgg ctc tcg gtg gaa atc tat     2372 
Val Gly Leu Gly Leu Val Leu Ala Tyr Arg Leu Ser Val Glu Ile Tyr 
        740                 745                 750 

gac cgc cgg gaa tac agt cgc ttt gag aag gag cag caa caa ctc aac     2420 
Asp Arg Arg Glu Tyr Ser Arg Phe Glu Lys Glu Gln Gln Gln Leu Asn 
    755                 760                 765 

tgg aag cag gac agt aat cct ctc tac aaa agt gcc atc acg acc acc     2468 
Trp Lys Gln Asp Ser Asn Pro Leu Tyr Lys Ser Ala Ile Thr Thr Thr 
770                 775                 780                 785 

atc aat cct cgc ttt caa gag gca gac agt ccc act ctc tga             2510 
Ile Asn Pro Arg Phe Gln Glu Ala Asp Ser Pro Thr Leu  * 
                790                 795 

aggagggagg gacacttacc caaggctctt ctccttggag gacagtggga actggagggt   2570 

gagaggaagg gtgggtctgt aagaccttgg taggggacta attcactggc gaggtgcggc   2630 

caccacccta cttcattttc agagtgacac ccaagagggc tgcttcccat gcctgcaacc   2690 

ttgcatccat ctgggctacc ccacccaagt atacaataaa gtcttacctc ag           2742 

 
           
             38  
             798  
             PRT  
             Homo sapiens  
           
            38 

Met Val Ala Leu Pro Met Val Leu Val Leu Leu Leu Val Leu Ser Arg 
 1               5                  10                  15 

Gly Glu Ser Glu Leu Asp Ala Lys Ile Pro Ser Thr Gly Asp Ala Thr 
            20                  25                  30 

Glu Trp Arg Asn Pro His Leu Ser Met Leu Gly Ser Cys Gln Pro Ala 
        35                  40                  45 

Pro Ser Cys Gln Lys Cys Ile Leu Ser His Pro Ser Cys Ala Trp Cys 
    50                  55                  60 

Lys Gln Leu Asn Phe Thr Ala Ser Gly Glu Ala Glu Ala Arg Arg Cys 
65                  70                  75                  80 

Ala Arg Arg Glu Glu Leu Leu Ala Arg Gly Cys Pro Leu Glu Glu Leu 
                85                  90                  95 

Glu Glu Pro Arg Gly Gln Gln Glu Val Leu Gln Asp Gln Pro Leu Ser 
            100                 105                 110 

Gln Gly Ala Arg Gly Glu Gly Ala Thr Gln Leu Ala Pro Gln Arg Val 
        115                 120                 125 

Arg Val Thr Leu Arg Pro Gly Glu Pro Gln Gln Leu Gln Val Arg Phe 
    130                 135                 140 

Leu Arg Ala Glu Gly Tyr Pro Val Asp Leu Tyr Tyr Leu Met Asp Leu 
145                 150                 155                 160 

Ser Tyr Ser Met Lys Asp Asp Leu Glu Arg Val Arg Gln Leu Gly His 
                165                 170                 175 

Ala Leu Leu Val Arg Leu Gln Glu Val Thr His Ser Val Arg Ile Gly 
            180                 185                 190 

Phe Gly Ser Phe Val Asp Lys Thr Val Leu Pro Phe Val Ser Thr Val 
        195                 200                 205 

Pro Ser Lys Leu Arg His Pro Cys Pro Thr Arg Leu Glu Arg Cys Gln 
    210                 215                 220 

Ser Pro Phe Ser Phe His His Val Leu Ser Leu Thr Gly Asp Ala Gln 
225                 230                 235                 240 

Ala Phe Glu Arg Glu Val Gly Arg Gln Ser Val Ser Gly Asn Leu Asp 
                245                 250                 255 

Ser Pro Glu Gly Gly Phe Asp Ala Ile Leu Gln Ala Ala Leu Cys Gln 
            260                 265                 270 

Glu Gln Ile Gly Trp Arg Asn Val Ser Arg Leu Leu Val Phe Thr Ser 
        275                 280                 285 

Asp Asp Thr Phe His Thr Ala Gly Asp Gly Lys Leu Gly Gly Ile Phe 
    290                 295                 300 

Met Pro Ser Asp Gly His Cys His Leu Asp Ser Asn Gly Leu Tyr Ser 
305                 310                 315                 320 

Arg Ser Thr Glu Phe Asp Tyr Pro Ser Val Gly Gln Val Ala Gln Ala 
                325                 330                 335 

Leu Ser Ala Ala Asn Ile Gln Pro Ile Phe Ala Val Thr Ser Ala Ala 
            340                 345                 350 

Leu Pro Val Tyr Gln Glu Leu Ser Lys Leu Ile Pro Lys Ser Ala Val 
        355                 360                 365 

Gly Glu Leu Ser Glu Asp Ser Ser Asn Val Val Gln Leu Ile Met Asp 
    370                 375                 380 

Ala Tyr Asn Ser Leu Ser Ser Thr Val Thr Leu Glu His Ser Ser Leu 
385                 390                 395                 400 

Pro Pro Gly Val His Ile Ser Tyr Glu Ser Gln Cys Glu Gly Pro Glu 
                405                 410                 415 

Lys Arg Glu Gly Lys Ala Glu Asp Arg Gly Gln Cys Asn His Val Arg 
            420                 425                 430 

Ile Asn Gln Thr Val Thr Phe Trp Val Ser Leu Gln Ala Thr His Cys 
        435                 440                 445 

Leu Pro Glu Pro His Leu Leu Arg Leu Arg Ala Leu Gly Phe Ser Glu 
    450                 455                 460 

Glu Leu Ile Val Glu Leu His Thr Leu Cys Asp Cys Asn Cys Ser Asp 
465                 470                 475                 480 

Thr Gln Pro Gln Ala Pro His Cys Ser Asp Gly Gln Gly His Leu Gln 
                485                 490                 495 

Cys Gly Val Cys Ser Cys Ala Pro Gly Arg Leu Gly Arg Leu Cys Glu 
            500                 505                 510 

Cys Ser Val Ala Glu Leu Ser Ser Pro Asp Leu Glu Ser Gly Cys Arg 
        515                 520                 525 

Ala Pro Asn Gly Thr Gly Pro Leu Cys Ser Gly Lys Gly His Cys Gln 
    530                 535                 540 

Cys Gly Arg Cys Ser Cys Ser Gly Gln Ser Ser Gly His Leu Cys Glu 
545                 550                 555                 560 

Cys Asp Asp Ala Ser Cys Glu Arg His Glu Gly Ile Leu Cys Gly Gly 
                565                 570                 575 

Phe Gly Arg Cys Gln Cys Gly Val Cys His Cys His Ala Asn Arg Thr 
            580                 585                 590 

Gly Arg Ala Cys Glu Cys Ser Gly Asp Met Asp Ser Cys Ile Ser Pro 
        595                 600                 605 

Glu Gly Gly Leu Cys Ser Gly His Gly Arg Cys Lys Cys Asn Arg Cys 
    610                 615                 620 

Gln Cys Leu Asp Gly Tyr Tyr Gly Ala Leu Cys Asp Gln Cys Pro Gly 
625                 630                 635                 640 

Cys Lys Thr Pro Cys Glu Arg His Arg Asp Cys Ala Glu Cys Gly Ala 
                645                 650                 655 

Phe Arg Thr Gly Pro Leu Ala Thr Asn Cys Ser Thr Ala Cys Ala His 
            660                 665                 670 

Thr Asn Val Thr Leu Ala Leu Ala Pro Ile Leu Asp Asp Gly Trp Cys 
        675                 680                 685 

Lys Glu Arg Thr Leu Asp Asn Gln Leu Phe Phe Phe Leu Val Glu Asp 
    690                 695                 700 

Asp Ala Arg Gly Thr Val Val Leu Arg Val Arg Pro Gln Glu Lys Gly 
705                 710                 715                 720 

Ala Asp His Thr Gln Ala Ile Val Leu Gly Cys Val Gly Gly Ile Val 
                725                 730                 735 

Ala Val Gly Leu Gly Leu Val Leu Ala Tyr Arg Leu Ser Val Glu Ile 
            740                 745                 750 

Tyr Asp Arg Arg Glu Tyr Ser Arg Phe Glu Lys Glu Gln Gln Gln Leu 
        755                 760                 765 

Asn Trp Lys Gln Asp Ser Asn Pro Leu Tyr Lys Ser Ala Ile Thr Thr 
    770                 775                 780 

Thr Ile Asn Pro Arg Phe Gln Glu Ala Asp Ser Pro Thr Leu 
785                 790                 795 

 
           
             39  
             30  
             PRT  
             Unknown  
             
               acidic peptide  
             
           
            39 

Ala Gln Leu Glu Lys Glu Leu Gln Ala Leu Glu Lys Glu Asn Ala Gln 
 1               5                  10                  15 

Leu Glu Trp Glu Leu Gln Ala Leu Glu Lys Glu Leu Ala Gln 
            20                  25                  30 

 
           
             40  
             30  
             PRT  
             Unknown  
             
               basic peptide  
             
           
            40 

Ala Gln Leu Lys Lys Lys Leu Gln Ala Leu Lys Lys Lys Asn Ala Gln 
 1               5                  10                  15 

Leu Lys Trp Lys Leu Gln Ala Leu Lys Lys Lys Leu Ala Gln 
            20                  25                  30 

 
           
             41  
             6  
             PRT  
             Artificial Sequence  
             
               amino acid linker sequence  
             
           
            41 

Gly Gly Ser Thr Gly Gly 
 1               5 

 
           
             42  
             31  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            42 

gcactagtcc accatgggcc cttggagccg c                                    31 

 
           
             43  
             26  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            43 

ccctcgagag gctgtgcctt cctaca                                          26 

 
           
             44  
             23  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            44 

atctcgagcc caaatcttgt gac                                             23 

 
           
             45  
             28  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            45 

tagcggccgc tcatttaccc ggagacag                                        28 

 
           
             46  
             21  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            46 

atggactgga cctggagcat c                                               21 

 
           
             47  
             20  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            47 

atggaattgg ggctgagctg                                                 20 

 
           
             48  
             20  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            48 

atggagtttg grctgagctg                                                 20 

 
           
             49  
             21  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            49 

atgaaacacc tgtggttctt c                                               21 

 
           
             50  
             20  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            50 

atggggtcaa ccgccatcct                                                 20 

 
           
             51  
             21  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            51 

tgccaggggg aagaccgatg g                                               21 

 
           
             52  
             33  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            52 

ttcttggtgg cagcagccac aggtgcccac tcc                                  33 

 
           
             53  
             30  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            53 

ctgctgacca tcccttcatg ggtcttgtcc                                      30 

 
           
             54  
             33  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            54 

ttccttgttg ctattttaaa aggtgtccag tgt                                  33 

 
           
             55  
             27  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            55 

gtggcagctc ccagatgggt cctgtcc                                         27 

 
           
             56  
             33  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            56 

ctcctcctga ctgttctcca aggagtctgt tcc                                  33 

 
           
             57  
             30  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            57 

gtgctgggcc tcccatgggg tgtcctgtca                                      30 

 
           
             58  
             30  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            58 

ttggtggcag cagcaacagg tgcccactcc                                      30 

 
           
             59  
             26  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            59 

gaaggtgtgc acgccgctgg tcagag                                          26 

 
           
             60  
             66  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            60 

gtgcaattgg tgcagtctgg agcagaggtg aaaaagcccg gggagtctct gaaaatctcc     60 

tgtaag                                                                66 

 
           
             61  
             30  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            61 

gtgcaattgg tggagtctgg gggaggcttg                                      30 

 
           
             62  
             32  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            62 

ctcgctgagc tgacggtgac cagggttccc tg                                   32 

 
           
             63  
             57  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            63 

ccggaattcc tcaccatgga aaccccagcg cagcttctct tcctcctgct actctgg        57 

 
           
             64  
             28  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            64 

acccgtacgt ttgatctcca ccttggtc                                        28 

 
           
             65  
             45  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            65 

ttacccaatt gtgtcctgtc ccaggtgcag ctgcaggagt cgggc                     45 

 
           
             66  
             40  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            66 

tggaggctga gctgacggtg accgtggtcc cttggcccca                           40 

 
           
             67  
             38  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            67 

ttcccagggt cccgttccga catccagatg acccagtc                             38 

 
           
             68  
             31  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            68 

agccaccgta cgtttgatat ccactctggt c                                    31 

 
           
             69  
             26  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            69 

tcggatccgc tctggagaag gaggag                                          26 

 
           
             70  
             26  
             DNA  
             Artificial Sequence  
             
               Synthetic oligonucleotide  
             
           
            70 

gcgaattcaa gggcgtctcc aaccgt                                          26