US20040185040A1 - Modulating immune responses - Google Patents

Modulating immune responses Download PDF

Info

Publication number
US20040185040A1
US20040185040A1 US10/719,642 US71964203A US2004185040A1 US 20040185040 A1 US20040185040 A1 US 20040185040A1 US 71964203 A US71964203 A US 71964203A US 2004185040 A1 US2004185040 A1 US 2004185040A1
Authority
US
United States
Prior art keywords
seq
antibody
mammal
cells
antibodies
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US10/719,642
Inventor
Leon Garcia-Martinez
Yuching Chen
Dawn Andrews
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
UCB Celltech Ltd
Original Assignee
Celltech R&D Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from PCT/US2002/037738 external-priority patent/WO2003045318A2/en
Application filed by Celltech R&D Ltd filed Critical Celltech R&D Ltd
Priority to US10/719,642 priority Critical patent/US20040185040A1/en
Assigned to CELLTECH R & D LIMITED reassignment CELLTECH R & D LIMITED ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GARCIA-MARTINEZ, LEON FERNANDO, ANDREWS, DAWN, CHEN, YUCHING
Publication of US20040185040A1 publication Critical patent/US20040185040A1/en
Priority to US11/805,841 priority patent/US7700740B2/en
Priority to US12/699,636 priority patent/US7872103B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to multimerized antibodies directed against the CD83 gene product, and methods of modulating the immune response of an animal by using such multimerized antibodies.
  • CD83 is a 45 kilodalton glycoprotein that is predominantly expressed on the surface of dendritic cells and other cells of the immune system. Structural analysis of the predicted amino acid sequence of CD83 indicates that it is a member of the immunoglobulin superfamily. See, Zhou et al., J. Immunol. 149:735 (1992)). U.S. Pat. No. 5,316,920 and WO 95/29236 disclose further information about CD83. While such information suggests that CD83 plays a role in the immune system, that role is undefined, and the interrelationship of CD83 with cellular factors remains unclear.
  • the invention provides methods for modulating an immune response.
  • the invention relates to the surprising discovery that multimerized antibodies raised against the CD83 gene product can arrest cellular proliferation of immune cells.
  • the invention provides a method of modulating the immune response by modulating the activity or expression of the CD83 gene products, for example, by using such multimerized antibodies.
  • the production of a cytokine such as interleukin-2, interleukin-4, or interleukin-10 can be modulated by modulating the activity or expression of a CD83 polypeptide.
  • a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide.
  • the antibody can be administered to the mammal or the immune cell can be contacted with the antibody.
  • the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4 + T cells.
  • the invention also provides a method of modulating granulocyte macrophage colony stimulating factor production in a mammal or in an immune cell by modulating the activity or expression of CD83 polypeptides.
  • an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide.
  • the antibody can be administered to the mammal or the immune cell can be contacted with the antibody.
  • the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4 + T cells.
  • the invention also provides a method of modulating tumor necrosis factor production in a mammal or in a mammalian cell by modulating the activity or expression of CD83 polypeptides.
  • an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide.
  • the antibody can be administered to the mammal or the mammalian cell can be contacted with the antibody.
  • the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4 + T cells.
  • the invention further provides a method of inhibiting proliferation of a human peripheral blood mononuclear cell by modulating the activity or expression of CD83 polypeptides.
  • an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide.
  • the antibody can be administered to the mammal or the human peripheral blood mononuclear cell can be contacted with the antibody.
  • the invention also provides an antibody that can bind to a CD83 polypeptide comprising SEQ ID NO:4, SEQ ID NO:8 or SEQ ID NO:9, wherein activated CD4 + T-cells produce lower levels of interleukin-4 when the T-cells are contacted with the antibody.
  • the invention further provides an antibody that can bind to a CD83 polypeptide comprising SEQ ID NO:4, SEQ ID NO:8 or SEQ ID NO:9, wherein CD4 + T-cells proliferation is decreased when the T-cells are contacted with the antibody.
  • the antibody can be a multimerized antibody. Such multimerized antibodies can be bound to a solid support, covalently crosslinked or bound together by a second entity such as a secondary antibody.
  • antibodies of the invention include those that have an amino acid sequence that includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ
  • Nucleic acids encoding such an antibody can have, for example, a sequence that includes SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85 or SEQ ID NO:90.
  • the invention also provides a method for decreasing the activity of a CD83 gene product, comprising contacting the CD83 gene product with an antibody that comprises amino acid sequence includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO
  • the invention further provides a method for decreasing the translation of a CD83 gene product in a mammalian cell, comprising contacting the mammalian cell with a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • the invention provides a method for decreasing the translation of a CD83 gene product in a mammal, comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • the invention further provides a method for decreasing proliferation of CD4 + T-cells in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9.
  • the antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54
  • the invention also provides a method for decreasing interleukin-2 levels and increasing interleukin-4 levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9.
  • the antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54
  • the invention further provides a method for decreasing interleukin-2 levels and increasing interleukin-4 levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • the interleukin-2 levels are decreased and the interleukin-4 levels are increased to treat an autoimmune disease.
  • the interleukin-2 levels are decreased and the interleukin-4 levels are increased to stimulate production of Th2-associated cytokines in transplant recipients, for example, to prolong survival of transplanted tissues.
  • the invention also provides a method for increasing interleukin-10 levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9.
  • the antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54
  • the invention further provides a method for increasing interleukin-10 levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • the interleukin-10 levels are increased to treat neoplastic disease. In other embodiments, the interleukin-10 levels are increased to treat a tumor.
  • the invention also provides a method for increasing interleukin-2 levels in a mammal comprising administering to the mammal a functional CD83 polypeptide that comprises SEQ ID NO:9.
  • the invention further provides a method for increasing interleukin-2 levels in a mammal comprising: (a) transforming a T cell from the mammal with a nucleic acid encoding a functional CD83 polypeptide operably linked to a promoter functional in a mammalian cell, to generate a transformed T cell; (b) administering the transformed T cell to the mammal to provide increased levels of interleukin-2.
  • the CD83 polypeptide has a sequence that comprises SEQ ID NO:9 or the nucleic acid has a sequence that comprises SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • Such methods for increasing interleukin-2 levels can be used to treat an allergy or an infectious disease.
  • the invention also provides a method for increasing granulocyte macrophage colony stimulating factor levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9.
  • Such an antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:
  • the invention further provides a method for increasing granulocyte macrophage colony stimulating factor levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10.
  • the invention also provides a method for increasing tumor necrosis factor levels at a selected site in a mammal comprising administering to the site a functional CD83 polypeptide.
  • the invention provides a method for increasing tumor necrosis factor levels in a selected mammalian cell comprising transforming the cell with a nucleic acid encoding a functional CD83 polypeptide.
  • the CD83 polypeptide employed can, for example, have a sequence comprising SEQ ID NO:9.
  • Animals such as mammals and birds may be treated by the methods and compositions described herein.
  • Such mammals and birds include humans, dogs, cats, and livestock, for example, horses, cattle, sheep, goats, chickens, turkeys and the like.
  • the invention further provides a mutant mouse that can serve as an animal model of diminished T cell activation or altered cytokine levels.
  • the mutant mouse has an altered CD83 gene that produces a larger gene product, having SEQ ID NO:4 or containing SEQ ID NO:8.
  • methods of using the mutant mouse model to study the effects of cytokines on the immune system, inflammation, the function and regulation of CD83, T cell and dendritic cell activity, the immune response and conditions and treatments related thereto.
  • the invention further provides a mutant mouse whose somatic and germ cells comprise a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8, wherein expression of the mutant CD83 gene reduces CD4+ T cell activation.
  • the mutant CD83 gene can, for example, comprise SEQ ID NO:3.
  • the invention further provides a method of identifying a compound that can modulate CD4+ T cell activation comprising administering a test compound to a mouse having a mutant or wild type transgenic CD83 gene and observing whether CD4+ T cell activation is decreased or increased.
  • the somatic and/or germ cells of the mutant mouse can comprise a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8.
  • the somatic and/or germ cells of the mouse can contain a wild type CD83 gene, for example, SEQ ID NO:1 or SEQ ID NO:9.
  • the invention also provides a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8.
  • the invention further provides a mutant CD83 gene comprising nucleotide sequence SEQ ID NO:3.
  • FIG. 1 summarizes flow cytometry data for G3 animals. As shown, reduced numbers of CD4+ T cells are seen in two animals from Pedigree 9, mouse 9.4.1 and mouse 9.4.9. All other animals analyzed on that day exhibit normal numbers of CD4+ T cells.
  • FIG. 2 provides a graph of flow cytometry data for G3 animals that may have a mutant CD83 gene. Each diamond symbol represents an individual animal. As shown, multiple animals from the N2 generation exhibit a reduced percentage of CD4+ T cells.
  • FIG. 3 provides the nucleotide sequence of wild type mouse CD83 (SEQ ID NO:1). The ATG start codon and the TGA stop codon are underlined.
  • FIGS. 4 A-B provides the nucleotide sequence of the mutant CD83 gene (SEQ ID NO:3) of the invention derived from the mutant LCD4.1 animal.
  • the ATG start codon, the mutation and the TGA stop codon are underlined.
  • FIG. 5 provides the amino acid sequence for wild type (top, SEQ ID NO:2) and mutant (bottom, SEQ ID NO:4) CD83 coding regions. The additional C-terminal sequences arising because of the CD83 mutation are underlined.
  • FIG. 6A illustrates that dendritic cells from wild type (?, WT DC) and mutant ( ⁇ , mutant DC) mice are capable of the allogeneic activation of CD4+ T cells.
  • CD4+ T cells were stimulated with 10,000, 1000 or 100 dendritic cells for 5 days and proliferation was measured by incorporation of tritiated thymidine.
  • FIG. 6B illustrates that CD4 + T cells from mutant mice ( ⁇ , mutant CD4) fail to respond to allogeneic stimulation with BALBc dendritic cells, although wild type animals (?, WT CD4+) respond normally.
  • CD4+ T cells were stimulated with 10,000, 1000 or 100 dendritic cells for 5 days and proliferation measured by incorporation of tritiated thymidine.
  • FIG. 7 provides a bar graph illustrating IL-2, IL-4, IL-5, TNFa, and IFN? production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with 1 ⁇ g/ml of anti-CD3 antibodies and 0.2 ⁇ g/ml of anti-CD28 antibodies for 72 hours. As illustrated, IL-2 levels are lower, and IL-4 levels are higher in the CD83 mutant T cells.
  • FIG. 8 provides a bar graph illustrating IL-10 production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with 0.1 ⁇ g/ml of anti-CD28 antibodies and 1 to 10 ⁇ g/ml of anti-CD3 antibodies for 72 hours. As illustrated, IL-10 levels are higher in the CD83 mutant T cells.
  • FIG. 9 provides a bar graph illustrating GM-CSF production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, GM-CSF production is higher in the CD83 mutant cells than in wild type cells.
  • FIG. 10A provides a bar graph illustrating IL-4 mRNA levels from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, the IL-4 mRNA levels are higher in the CD83 mutant cells.
  • FIG. 10B provides a bar graph illustrating IL-10 mRNA levels from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, the IL-10 mRNA levels are higher in the CD83 mutant cells.
  • FIG. 11 provides a graph illustrating that various preparations of anti-CD83 antibodies inhibit IL-4 production in anti-CD3 and anti-CD28 antibody stimulated T cells.
  • the amount of IL-4 produced by T cells in pg/ml is plotted versus the concentration of different anti-CD83 antibody preparations, including the 20B08 (?) anti-CD83 preparation, the 20D04 ( ⁇ ) anti-CD83 preparation, the 14C12 (?) anti-CD83 preparation and the 11 G05 (X) anti-CD83 antibody preparation.
  • FIG. 12 provides a graph illustrating that various preparations of anti-CD83 antibodies inhibit T cell proliferation.
  • the graph plots the incorporation of radioactive thymidine in cpms, which was used as an indicator of the amount of T cell proliferation, versus the concentration of the different anti-CD83 antibody preparations, including the 20D04 (?) anti-CD83 preparation, the 11G05 ( ⁇ ) anti-CD83 antibody preparation, the 14C12 (?) anti-CD83 preparation and the 6G05 anti-CD83 preparation (X).
  • FIG. 13 provides a graph illustrating that transgenic mice that over-express wild type CD83 have increased T cell proliferation.
  • the graph plots the incorporation of radioactive thymidine in cpms, which was used as an indicator of the amount of T cell proliferation, versus the concentration of OVA peptide.
  • the transgenic mice utilized had a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide that can activate T-cells.
  • OVA ovalbumin
  • transgenic CD4+ T cells had increased T-cell proliferation.
  • transgenic dendritic cells could not substantially increase wild type CD4+ T cell proliferation.
  • Transgenic CD83 CD4+T cells mixed with wild type dendritic cells (?); transgenic CD83 CD4+ T cells mixed with transgenic dendritic cells ( ⁇ ); wild type CD4+ T cells mixed with transgenic dendritic cells (?); and wild type CD4+ T cells mixed with wild type dendritic cells (X).
  • FIG. 14 provides a schematic diagram of the structural elements included in the mouse CD83 protein used for generating antibodies.
  • FIG. 15 provides a graph of ELISA data illustrating the titer obtained for different isolates of polyclonal anti-CD83 anti-sera.
  • the first (?), second ( ⁇ ) and third (?) isolates had similar titers, though the titer of the second isolate ( ⁇ ) was somewhat higher.
  • FIG. 16 illustrates that proliferation of PHA-activated human PBMCs was inhibited by antibodies raised against the external region of the mouse CD83 protein (?). Pre-immune serum ( ⁇ ) had little effect on the proliferation of human PBMCs.
  • FIG. 17A provides a sequence alignment of anti-CD83 heavy chain variable regions isolated by the invention. Sequences for isolates 20B08H (SEQ ID NO:52), 6G05H (SEQ ID NO:53), 20D04H (SEQ ID NO:54), 11 G05 (SEQ ID NO:66) and 14C12 (SEQ ID NO:67) are provided. The CDR regions are highlighted in bold.
  • FIG. 17B provides a sequence alignment of anti-CD83 light chain variable regions isolated by the invention. Sequences for isolates 20B08L (SEQ ID NO:55), 6G05L (SEQ ID NO:56), 20D04L (SEQ ID NO:57), 11G05L (SEQ ID NO:68) and 14C12L (SEQ ID NO:69) are provided. The CDR regions are highlighted in bold.
  • FIG. 18 graphically illustrates that cells expressing CD83 can be detected and sorted using an anti-CD83 antibody preparation.
  • a Hodgkin's lymphoma cell line, KMH2, and a commercially available anti-CD83 antibody preparation was used for FACS sorting.
  • FIGS. 19 A-B shows that two antibody preparations of the invention can bind to endogenously produced human CD83, as detected by FACS sorting of KMH2 cells (see also FIG. 18). Note that “Beer” is another name used for CD83.
  • FIG. 20 illustrates that the 95F04 and 96G08 antibody preparations described herein can inhibit proliferation of human peripheral blood mononuclear cells as detected by [ 3 H] thymidine incorporation.
  • the 95F04 (?) antibody preparation when 30 ⁇ g/ml was present, incorporation of [ 3 H] thymidine dropped to about 2000 cpm.
  • 30 ⁇ g/ml 96G08 antibody preparation (?) was added to human peripheral blood mononuclear cells, [ 3 H] thymidine incorporation was reduced to about 300 cpm.
  • a third antibody preparation (98B 11, ⁇ ) provided slight inhibition of PBMC proliferation.
  • FIG. 21 provides nucleotide and amino acid sequences for the monoclonal antibody 96G08 light chain.
  • FIG. 22 provides nucleotide and amino acid sequences for the monoclonal antibody 96G08 heavy chain.
  • FIG. 23 provides nucleotide and amino acid sequences for the monoclonal antibody 95F04 light chain.
  • FIG. 24 provides nucleotide and amino acid sequences for the monoclonal antibody 95F04 heavy chain.
  • FIGS. 25 A-B provides the results of one screen of anti-CD83 antibody preparations that were multimerized by binding them to microtiter plates.
  • the plate-bound antibodies were screened for an ability to inhibit lymphocyte proliferation as measured by tritiated thymidine incorporation.
  • many plate-bound anti-CD83 antibody preparations inhibit proliferation of activated lymphocytes, including the 94c09, 98a02, 94d08, 98d11, 101b08, 6g05, 20d04, 14c12, 11g05, 12g04, 32f12 and 98b11 preparations.
  • FIG. 25B further illustrates that some antibody preparations are highly effective inhibitors (e.g. 117G12) but others are not (e.g. 824pb and 98g08).
  • FIG. 26 illustrates that the inhibitory activity of the multimerized (plate-bound) 6g05 antibody preparation is quenched by soluble mouse CD83 protein (mCD83rFc). Lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown, the multimerized 6g05 antibody preparation is strongly inhibitory of proliferation when no CD83 protein is added. However, when the mouse CD83 protein is added to assay, little or no inhibition of lymphocyte proliferation is observed. The 98g08 antibody preparation was used as a negative control because it exhibited little or no lymphocyte inhibition in previous tests (see FIG. 25B).
  • FIG. 27 is a bar graph showing that both plate-bound and cross-linked 6g05 antibodies are highly effective inhibitors of lymphocyte proliferation. Lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown on the left side of the graph above “plate-bound” the presence of plate-bound 6g05 antibodies in the lymphocyte proliferation assay cause little tritiated thymidine incorporation (about 1000 cpm). Similarly, as shown on the right side of the graph above “1 st Ab (1 ⁇ g/ml)” soluble cross-linked 6g05 antibodies also cause little tritiated thymidine incorporation (about 1800 cpm).
  • FIG. 28 is a bar graph showing that several preparations of soluble cross-linked anti-CD83 antibodies are highly effective inhibitors of lymphocyte proliferation.
  • Antibody preparations were cross-linked with the rabbit anti-mouse secondary antibody and lymphocyte proliferation was measured by tritiated thymidine incorporation.
  • soluble cross-linked antibody preparations including the 6g05, 11g04, 12g04, 14c12, 20d04, 32f12, 94c09, 94d08, 98a02, 98d11(3), 101B08(2.7) and 117g12 preparations caused little tritiated thymidine incorporation.
  • FIG. 29 shows that soluble, multimerized anti-CD83 antibodies exhibit inhibitory activity in mixed lymphocyte reaction assays.
  • a series of anti-CD83 antibody preparations that were cross-linked using a rabbit anti-mouse antibody and then screened for inhibition of CD4 + T cellular proliferation after activation of the CD4 + T cells with CD11 cells in a mixed lymphocyte reaction assay.
  • the 98a02, 98d11, 20d04, 14c12, 12g04, and 117g12 inhibit lymphocyte proliferation in this assay.
  • FIG. 30 shows that many anti-CD83 antibody preparations can inhibit the recall response of lymphocytes.
  • BALBc mice were first immunized with keyhole limpet hemocyanin (KLH) prior to spleen removal and CD 11 c and CD4+cell isolation.
  • CD11c and CD4+cells were mixed and added to microtiter wells coated with anti-CD83 antibodies. Lymphocyte proliferation was measured by tritiated thymidine incorporation.
  • the 94c09, 98a02, 6g05, 20d04, and 117104 antibody preparations inhibited proliferation of activated lymphocytes exposed to an antigen (KLH) to which they had been immunized.
  • FIGS. 31 A-B shows that soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations not only inhibit activated lymphocyte cell proliferation (FIG. 31B) but also have very low caspase activity (FIG. 31A). Caspase activity was determined using a fluorogenic substrate and plotted as relative fluorescent units (RFU) on the y axis.
  • REU relative fluorescent units
  • FIG. 32 shows that the percentage of activated lymphocytes that express annexin V is reduced after treatment with soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations.
  • FIG. 33 shows that the activation marker CD69 is expressed on splenocytes that were activated with Concavalin A and exposed to anti-CD83 antibodies.
  • the anti-CD83 antibodies employed were the 6g05, 14c12, 98b11 and 112d08 anti-CD83 antibody preparations that were shown to inhibit activated splenocyte proliferation.
  • FIGS. 34 A-E shows that a population of activated splenocytes mixed with anti-CD83 antibody preparations have lost the blasting (dividing) cells as detected by FACS sorting.
  • the antibody preparations employed were the rabbit anti-mouse antibody, called the 2 nd Ab (FIG. 34A), the 6g05 antibody preparation (FIG. 34B), the 98b11 antibody preparation (FIG. 34C), the 14c12 antibody preparation (FIG. 34D), and the 112d08 antibody preparation (FIG. 34E).
  • FIG. 35A shows that the proportion of cells in the G1/G0 phase of the cell cycle is increased when a population of activated splenocytes is treated with anti-CD83 antibody preparations.
  • the antibody preparations employed were the control rabbit anti-mouse antibody, called the 2 nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation. Both of the 6g05 and 14c12 antibody preparations arrest the activated splenocytes in the G1/G0 phase of the cell cycle.
  • FIG. 35B shows the proportion of cells in the G2/M phase of the cell cycle after a population of activated splenocytes is treated with anti-CD83 antibody preparations.
  • the antibody preparations employed were the control rabbit anti-mouse antibody, called the 2 nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation.
  • FIG. 35C shows that the proportion of cells in the S phase of the cell cycle is decreased when a population of activated splenocytes is treated with anti-CD83 antibody preparations.
  • the antibody preparations employed were the control rabbit anti-mouse antibody, called the 2 nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation.
  • Activated splenocytes treated with either of the 6g05 or 14c12 antibody preparations have lesser numbers of cells in the S phase of the cell cycle.
  • the invention provides methods for modulating the immune system. For example, according to the invention, loss or reduction of CD83 activity in vivo results in decreased numbers of immune cells, for example, decreased numbers of T cells.
  • binding entities such as monoclonal antibodies that are capable of inhibiting the function of CD83 are provided for use in the invention. In other embodiments the binding entities or antibodies are multimerized.
  • the compositions and methods of the invention can be used for treating conditions involving an inappropriate immune response, for example, autoimmune diseases, inflammation, tissue rejection, arthritis, atherosclerosis and the like.
  • CD83 is a lymphocyte and dendritic cell activation antigen that is expressed by activated lymphocytes and dendritic cells.
  • CD83 is also a single-chain cell-surface glycoprotein with a molecular weight of about 45,000 that is believed to be a member of the Ig superfamily.
  • the structure predicted from the CD83 amino acid sequence indicates that CD83 is a membrane glycoprotein with a single extracellular Ig-like domain, a transmembrane domain and cytoplasmic domain of approximately forty amino acids.
  • the mature CD83 protein has about 186 amino acids and is composed of a single extracellular V type immunoglobulin (Ig)-like domain, a transmembrane domain and a thirty nine amino acid cytoplasmic domain.
  • Ig immunoglobulin
  • CD83 is translated from three mRNA transcripts of about 1.7, 2.0 and 2.5 kb that are expressed by lymphoblastoid cell lines. It is likely that CD83 undergoes extensive post-translational processing because CD83 is expressed as a single chain molecule, but the determined molecular weight is twice the predicted size of the core protein. See U.S. Pat. No. 5,766,570.
  • FIG. 9 An example of a human CD83 gene product that can be used in the invention is provided below (SEQ ID NO:9): 1 MSRGLQLLLL SCAYSLAPAT PEVKVACSED VDLPCTAPWD 41 PQVPYTVSWV KLLEGGEERM ETPQEDHLRG QHYHQKGQNG 81 SFDAPNERPY SLKIRNTTSC NSGTYRCTLQ DPDGQRNLSG 121 KVILRVTGCP AQRKEETFKK YRAEIVLLLA LVIFYLTLII 161 FTCKFARLQS IFPDFSKAGM ERAFLPVTSP NKHLGLVTPH 201 KTELV
  • Such a CD83 gene product can be encoded by a number of different nucleic acids.
  • a human CD83 nucleic acid is provided below (SEQ ID NO:10).
  • 1 CCTGGCGCAG CCGCAGCAGC GACGCGAGCG AACTCGGCCG 41 GGCCCGGGCG CGCGGGGGCG GGACGCGCAC GCGGCGAGGG 81 CGGCGGGTGA GCCGGGGGCG GGGACGGGGG CGGGACGGGG 121 GCGAAGGGGG CGGGGACGGGCGCCCGCC GGCCTAACGG 161 GATTAGGAGG GCGCGCCACC CGCTTCCGCT GCCCGCCGGG 201 GAATCCCCCG GGTGGCGCCC AGGGAAGTTC CCGAACGGGC 241 GGGCATAAAA GGGCAGCCGC GCCGGCCC CACAGCTCTG 281 CAGCTCGTGG CAGCGGCGCA GCTCCAGC CATGTCGCGC 321 GGCCTCCAGC TTCT
  • a sequence of a wild type mouse CD83 gene that can be used in the invention is provided herein as SEQ ID NO:1.
  • SEQ ID NO:1 is provided below with the ATG start codon and the TGA stop codon identified by underlining.
  • 1 GCGCTCCAGC CGC ATG TCGC AAGGCCTCCA GCTCCTGTTT 41 CTAGGCTGCG CCTGCAGCCT GGCACCCGCG ATGGCGATGC 81 GGGAGGTGAC GGTGGCTTGC TCCGAGACCG CCGACTTGCC 121 TTGCACAGCG CCCTGGGACC CGCAGCTCTC CTATGCAGTG 161 TCCTGGGCCA AGGTCTCCGA GAGTGGCACT GAGAGTGTGG 201 AGCTCCCGGA GAGCAAGCAA AACAGCTCCT TCGAGGCCCC 241 CAGGAGAAGG GCCTATTCCC TGACGATCCA AAACACTACC 281 ATCTGCAGCT CGGGCACCTA CAGGTGTGCC CTGCAGGAGC 321 TCGG
  • Nucleic acids having SEQ ID NO:1 encode a mouse polypeptide having SEQ ID NO:2, provided below.
  • CD83 activity in vivo results in a decreased immune response, for example, decreased numbers of T cells.
  • the effect of CD83 on the immune response was initially ascertained through use of a mutant mouse that encodes a mutant CD83.
  • a mutant mouse has a CD83 gene encoding SEQ ID NO:4, with added C-terminal sequences provided by SEQ ID NO:8.
  • the mutant CD83 gene of the invention has SEQ ID NO:3.
  • SEQ ID NO:3 is provided below with the ATG start codon, the mutation, and the TGA stop codon are identified by underlining.
  • mutant CD83 nucleic acids having SEQ ID NO:3 encode an elongated polypeptide having SEQ ID NO:4, provided below, where the extra amino acids are underlined.
  • the invention provides mutant CD83 nucleic acids that include SEQ ID NO:5. 1 ATG TCGCAAG GCCTCCAGCT CCTGTTTCTA GGCTGCGCCT 41 GCAGCCTGGC ACCCGCGATG GCGATGCGGG AGGTGACGGT 81 GGCTTGCTCC GAGACCGCCG ACTTGCCTTG CACAGCGCCC 121 TGGGACCCGC AGCTCTCCTA TGCAGTGTCC TGGGCCAAGG 161 TCTCCGAGAG TGGCACTGAG AGTGTGGAGC TCCCGGAGAG 201 CAAGCAAAAC AGCTCCTTCG AGGCCCCCAG GAGAAGGGCC 241 TATTCCCTGA CGATCCAAAA CACTACCATC TGCAGCTCGG 281 GCACCTACAG GTGTGCCCTG CAGGAGCTCG GAGGGCAGCG 321 CAACTTGAGC GGCACCGTGG TTCTGAAGGT GACAGGATGC 361 CCCAAGGAAG CTACAGAGTC AACT
  • Nucleic acids having SEQ ID NO:5 also encode a polypeptide having SEQ ID NO:4.
  • the invention provides mutant CD83 nucleic acids that include SEQ ID NO:7. 1 A GAGTAGGAT CTCCACTGGT TTTTACAAAG CCAAGGGCAC 41 ATCAGATCAG TGTGCCTGAA TGCCACCCGG ACAAGAGAAG 81 AATGAGCTCC ATCCTCAGAT GGCAACCTTT CTTTGAAGTC 121 CTTCACCTGA CAGTGGGCTC CACACTACTC CCTGACACAG 161 GGTCT TGA
  • the invention also provides a mutant CD83 containing SEQ ID NO:8, provided below.
  • SEQ ID NO:8 contains read through sequences that are not present in the wild type CD83 polypeptide but are present in the mutant CD83 gene product provided by the invention.
  • the CD83 gene product is used for generating antibodies. While any of the CD83 gene products described herein can be employed for immunization of animal, in some embodiments the extracellular Ig-like domain of the CD83 gene product is used for immunization, or antibodies are screened for reactivity with the extracellular Ig-like domain.
  • the extracellular Ig-like domain of the human CD83 gene product spans amino acids 21-127, and is provided below (SEQ ID NO:97): 21 PEVKVACSED VDLPCTAPWD 41 PQVPYTVSWV KLLEGGEERM ETPQEDHLRG QHYHQKGQNG 81 SFDAPNERPY SLKIRNTTSC NSGTYRCTLQ DPDGQRNLSG 121 KVILRVT
  • the invention provides antibody preparations directed against the mutant and wild type CD83 polypeptides of the invention, for example, against a polypeptide having SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8 or SEQ ID NO:9.
  • Other antibodies of interest can bind to the cytoplasmic tail of CD83.
  • the anti-CD83 antibodies are multimerized antibodies.
  • multimerized anti-CD83 antibodies are surprisingly effective inhibitors of lymphocyte cell proliferation.
  • an “multimerized” anti-CD83 antibody is a collection of anti-CD83 antibodies that are in close proximity. While such multimerized antibodies can be covalently linked, no such covalent linkage is necessary. Instead, multimerization of anti-CD83 antibodies can simply involve bringing the antibodies into close proximity, for example, by attachment to a solid support such as a plate or a bead. Alternatively, the antibodies can be non-covalently linked together through another entity, for example, any convenient non-covalent binding entity or secondary antibody. Hence, any available means for bringing the anti-CD83 antibodies into proximity can be used to generate the multimerized antibodies of the invention.
  • the anti-CD83 binding proteins or antibodies can be chemically cross-linked or genetically fused with any available crosslinking reagent.
  • Crosslinking can be achieved using one or a combination of a wide variety of multifunctional reagents.
  • Such crosslinking agents include glutaraldehyde, succinaldehyde, octanedialdehyde and glyoxal.
  • Additional multifunctional crosslinking agents include halo-triazines, e.g., cyanuric chloride; halo-pyrimidines, e.g., 2,4,6-trichloro/bromo-pyrimidine; anhydrides or halides of aliphatic or aromatic mono- or di-carboxylic acids, e.g., maleic anhydride, (meth)acryloyl chloride, chloroacetyl chloride; N-methylol compounds, e.g., N-methylol-chloro acetamide; di-isocyanates or di-isothiocyanates, e.g., phenylene-1,4-di-isocyanate and aziridines.
  • halo-triazines e.g., cyanuric chloride
  • halo-pyrimidines e.g., 2,4,6-trichloro/bromo-pyrimidine
  • crosslinking agents include epoxides, such as, for example, di-epoxides, tri-epoxides and tetra-epoxides.
  • Other crosslinking agents include, for example, dimethyl 3,3′-dithiobispropionimidate-HCl (DTBP); dithiobis (succinimidylpropionate) (DSP); bismaleimidohexane (BMH); bis[Sulfosuccinimidyl]suberate (BS); 1,5-difluoro-2,4-dinitrobenzene (DFDNB); dimethylsuberimidate-2HCl (DMS); disuccinimidyl glutarate (DSG); disulfosuccinimidyl tartarate (Sulfo-DST); 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC); ethylene glycolbis [sulfo-succinimidy
  • crosslinkers useful with various preparations of anti-CD83 antibodies of this invention include (1) those which create covalent links from one cysteine side chain of a protein to another cysteine side chain, (2) those which create covalent links from one lysine side chain of a protein to another, or (3) those which create covalent links from one cysteine side chain of a protein to a lysine side chain.
  • the anti-CD83 antibodies are reversibly crosslinked.
  • Such reversibly crosslinked antibodies are useful for short term use, for example, for short term control of the immune response either in vivo or in vitro, or for controlled dissipation of the anti-CD83 antibodies at a localized site after administration for short term therapeutic purposes.
  • reversible crosslinkers are described in T. W. Green, Protective Groups in Organic Synthesis, John Wiley & Sons (Eds.) (1981).
  • Other types of reversible crosslinkers are disulfide bond-containing crosslinkers. The crosslinks formed by such crosslinkers can be broken by the addition of reducing agent, such as cysteine, to the environment of the crosslinked anti-CD83 antibodies.
  • Disulfide crosslinkers are described in the Pierce Catalog and Handbook (1994-1995).
  • crosslinkers examples include: Homobifunctional (Symmetric); DSP—Dithiobis(succinimidylpropionate), also know as Lomant,'s Reagent; DTSSP—3-3′-Dithiobis (sulfosuccinimidyl-propionate), water soluble version of DSP; DTBP—Dimethyl 3,3′-dithiobispropionimidate-HCl; BASED—Bis-(13-[4-azidosalicylamido] ethyl)disulfide; DPDPB—1,4-Di-(3′-[2′-pyridyldithio]-propionamido)butane; Heterobifunctional (Asymmetric); SPDP—N-Succinimidyl-3-(2-pyridyldithio)propionate; LC-SPDP—Succinimidyl-6-(3-[2-pyrid
  • a fusion protein can be made with a selected anti-CD83 antibody to allow a domain to be attached to one or both of the polypeptides comprising the anti-CD83 antibody to be bound to a solid substrate.
  • glutathione-S-transferase/anti-CD83 fusion proteins can be linked to another anti-CD83 preparation having glutathione attached thereto or the glutathione-S-transferase/anti-CD83 fusion proteins can be adsorbed onto a solid support having glutathione attached thereto, such as glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtiter plate.
  • DSP-crosslinked antibodies can be linked to protein G agarose beads.
  • Other techniques for immobilizing polypeptides on solid support materials can also be used.
  • an anti-CD83 antibody can be immobilized utilizing conjugation of biotin and streptavidin.
  • Biotinylated anti-CD83 polypeptides can be prepared from biotin-NHS(N-hydroxy-succinimide) using techniques known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized with a streptavidin-linked antiCD83 antibody preparation, streptavidin-coated beads or another solid support material.
  • the invention provides antibodies capable of reducing CD83 activity and decreasing an immune response in a mammal.
  • Such antibodies can be multimerized antibodies. These antibodies may be used as CD83 inhibitory agents in the methods of the invention as described herein.
  • the antibodies of the invention can activate CD83 activity. Such activating antibodies may be used as CD83 stimulatory agents.
  • All antibody molecules belong to a family of plasma proteins called immunoglobulins, whose basic building block, the immunoglobulin fold or domain, is used in various forms in many molecules of the immune system and other biological recognition systems.
  • a typical immunoglobulin has four polypeptide chains, containing an antigen binding region known as a variable region and a non-varying region known as the constant region.
  • Native antibodies and immunoglobulins are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end.
  • VH variable domain
  • VL variable domain at one end
  • the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain.
  • Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains (Clothia et al., J. Mol. Biol. 186, 651-66, 1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA 82, 4592-4596(1985).
  • immunoglobulins can be assigned to different classes. There are at least five (5) major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g. IgG-1, IgG-2, IgG-3 and IgG-4; IgA-1 and IgA-2.
  • the heavy chains constant domains that correspond to the different classes of immunoglobulins are called alpha (a), delta (d), epsilon (e), gamma (?) and mu ( ⁇ ), respectively.
  • the light chains of antibodies can be assigned to one of two clearly distinct types, called kappa (?) and lambda (?), based on the amino sequences of their constant domain.
  • kappa ?
  • lambda ?
  • variable in the context of variable domain of antibodies, refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies.
  • the variable domains are for binding and determine the specificity of each particular antibody for its particular antigen.
  • variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) also known as hypervariable regions both in the light chain and the heavy chain variable domains.
  • CDRs complementarity determining regions
  • variable domains The more highly conserved portions of variable domains are called the framework (FR).
  • the variable domains of native heavy and light chains each comprise four FR regions, largely adopting a ⁇ -sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the ⁇ -sheet structure.
  • the CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies.
  • the constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector function, such as participation of the antibody in antibody-dependent cellular toxicity.
  • an antibody that is contemplated for use in the present invention thus can be in any of a variety of forms, including a whole immunoglobulin, an antibody fragment such as Fv, Fab, and similar fragments, a single chain antibody that includes the variable domain complementarity determining regions (CDR), and the like forms, all of which fall under the broad term “antibody,” as used herein.
  • the multimerized antibodies of the invention can be an aggregation or multimerization of whole immunoglobulins.
  • the multimerized antibodies of the invention can be an aggregation or multimerization of antibody fragments such as Fv, Fab, single chain antibodies that include the variable domain complementarity determining regions (CDR), CDRs and the like.
  • Such intact antibodies or antibody fragments can be multimerized by any convenient means, including covalent linkage or non-covalent association.
  • the present invention contemplates the use of any specificity of an antibody, polyclonal or monoclonal, and is not limited to antibodies that recognize and immunoreact with a specific epitope.
  • an antibody or fragment thereof is used that is immunospecific for an extracellular portion of the CD83 protein.
  • antibody fragment refers to a portion of a full-length antibody, generally the antigen binding or variable region.
  • antibody fragments include Fab, Fab′, F(ab′) 2 and Fv fragments.
  • Papain digestion of antibodies produces two identical antigen binding fragments, called the Fab fragment, each with a single antigen binding site, and a residual “Fc” fragment, so-called for its ability to crystallize readily.
  • Pepsin treatment yields an F(ab′) 2 fragment that has two antigen binding fragments, which are capable of cross-linking antigen, and a residual other fragment (which is termed pFc′).
  • Additional fragments can include diabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments.
  • “functional fragment” with respect to antibodies refers to Fv, F(ab) and F(ab′) 2 fragments.
  • Antibody fragments retain some ability to selectively bind with its antigen or receptor and are defined as follows:
  • Fab is the fragment that contains a monovalent antigen-binding fragment of an antibody molecule.
  • a Fab fragment can be produced by digestion of whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain.
  • Fab′ is the fragment of an antibody molecule can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain. Two Fab′ fragments are obtained per antibody molecule. Fab′ fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH 1 domain including one or more cysteines from the antibody hinge region.
  • (Fab′) 2 is the fragment of an antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction.
  • F(ab′) 2 is a dimer of two Fab′ fragments held together by two disulfide bonds.
  • Fv is the minimum antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy and one light chain variable domain in a tight, non-covalent association (VH-V L dimer). It is in this configuration that the three CDRs of each variable domain interact to define an antigen binding site on the surface of the VH-V L dimer. Collectively, the six CDRs confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.
  • Single chain antibody defined as a genetically engineered molecule containing the variable region of the light chain, the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule.
  • Such single chain antibodies are also referred to as “single-chain Fv” or “sFv” antibody fragments.
  • the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for antigen binding.
  • diabodies refers to a small antibody fragments with two antigen-binding sites, which fragments comprise a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (VH-VL).
  • VH heavy chain variable domain
  • VL light chain variable domain
  • VH-VL polypeptide chain
  • polyclonal antibodies are well-known to those skilled in the art. See, for example, Green, et al., Production of Polyclonal Antisera, in: Immunochemical Protocols (Manson, ed.), pages 1-5 (Humana Press); Coligan, et al., Production of Polyclonal Antisera in Rabbits, Rats Mice and Hamsters, in: Current Protocols in Immunology , section 2.4.1 (1992), which are hereby incorporated by reference.
  • the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler and Milstein, Nature 256, 495 (1975), or they may be made by recombinant methods, for example, as described in U.S. Pat. No. 4,816,567.
  • the monoclonal antibodies for use with the present invention may also be isolated from antibody libraries using the techniques described in Clackson et al. Nature 352: 624-628 (1991), as well as in Marks et al., J. Mol. Biol. 222: 581-597 (1991).
  • Monoclonal antibodies can be isolated and purified from hybridoma cultures by a variety of well-established techniques. Such isolation techniques include affinity chromatography with Protein-A Sepharose, size-exclusion chromatography, and ion-exchange chromatography. See, e.g., Coligan, et al., sections 2.7.1-2.7.12 and sections 2.9.1-2.9.3; Bames, et al., Purification of Immunoglobulin G (IgG), in: Methods in Molecular Biology , Vol. 10, pages 79-104 (Humana Press (1992).
  • SLAM Selected Lymphocyte Antibody Method
  • an animal is immunized with a source of specific antigen.
  • the animal can be a rabbit, mouse, rat, or any other convenient animal.
  • This immunization may consist of purified protein, in either native or recombinant form, peptides, DNA encoding the protein of interest or cells expressing the protein of interest.
  • blood, spleen or other tissues are harvested from the animal. Lymphocytes are isolated from the blood and cultured under specific conditions to generate antibody-forming cells, with antibody being secreted into the culture medium.
  • Another method involves humanizing a monoclonal antibody by recombinant means to generate antibodies containing human specific and recognizable sequences. See, for review, Holmes, et al., J. Immunol., 158:2192-2201 (1997) and Vaswani, et al., Annals Allergy, Asthma & Immunol., 81:105-115 (1998).
  • the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site.
  • each monoclonal antibody is directed against a single determinant on the antigen.
  • the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other immunoglobulins.
  • the modifier “monoclonal” indicates the antibody is obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • the monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567); Morrison et al. Proc. Natl. Acad. Sci. 81, 6851-6855 (1984).
  • chimeric antibodies immunoglobulins in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies
  • Antibody fragments of the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. Coli of DNA encoding the fragment.
  • Antibody fragments can be obtained by pepsin or papain digestion of whole antibodies conventional methods.
  • antibody fragments can be produced by enzymatic cleavage of antibodies with pepsin to provide a 5S fragment denoted F(ab′) 2 .
  • a thiol reducing agent optionally a blocking group for the sulfhydryl groups resulting from cleavage of disulfide linkages
  • an enzymatic cleavage using pepsin produces two monovalent Fab′ fragments and an Fc fragment directly.
  • Fv fragments comprise an association of VH and VL chains. This association may be noncovalent or the variable chains can be linked by an intermolecular disulfide bond or cross-linked by chemicals such as glutaraldehyde.
  • the Fv fragments comprise VH and VL chains connected by a peptide linker.
  • sFv single-chain antigen binding proteins
  • CDR peptides (“minimal recognition units”) can be obtained by constructing genes encoding the CDR of an antibody of interest. Such genes are prepared, for example, by using the polymerase chain reaction to synthesize the variable region from RNA of antibody-producing cells. See, for example, Larrick, et al., Methods: a Companion to Methods in Enzymology , Vol. 2, page 106 (1991).
  • the invention further contemplates human and humanized forms of non-human (e.g. murine) antibodies.
  • humanized antibodies can be chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′) 2 or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a nonhuman species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity.
  • CDR complementary determining region
  • humanized antibodies may comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance.
  • humanized antibodies can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the Fv regions are those of a human immunoglobulin consensus sequence.
  • the humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • mutant antibody refers to an amino acid sequence variant of an antibody.
  • one or more of the amino acid residues in the mutant antibody is different from what is present in the reference antibody.
  • Such mutant antibodies necessarily have less than 100% sequence identity or similarity with the reference amino acid sequence.
  • mutant antibodies have at least 75% amino acid sequence identity or similarity with the amino acid sequence of either the heavy or light chain variable domain of the reference antibody.
  • mutant antibodies have at least 80%, more preferably at least 85%, even more preferably at least 90%, and most preferably at least 95% amino acid sequence identity or similarity with the amino acid sequence of either the heavy or light chain variable domain of the reference antibody.
  • the antibodies of the invention are isolated antibodies.
  • An isolated antibody is one that has been identified and separated and/or recovered from a component of the environment in which it was produced. Contaminant components of its production environment are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.
  • isolated antibody also includes antibodies within recombinant cells because at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step.
  • the antibodies of the invention can be purified by any available procedure.
  • the antibodies can be affinity purified by binding an antibody preparation to a solid support to which the antigen used to raise the antibodies is bound. After washing off contaminants, the antibody can be eluted by known procedures.
  • Those of skill in the art will know of various techniques common in the immunology arts for purification and/or concentration of polyclonal antibodies, as well as monoclonal antibodies (see for example, Coligan, et al., Unit 9 , Current Protocols in Immunology , Wiley Interscience, 1991, incorporated by reference).
  • the antibody will be purified as measurable by at least three different methods: 1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight; 2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator; or 3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomasie blue or, preferably, silver stain.
  • the invention also provides antibodies that can bind to CD83 polypeptides. Sequences of complementarity determining regions (CDRs) or hypervariable regions from light and heavy chains of these anti-CD83 antibodies are provided. For example, a heavy chain variable region having a CDR1 sequence of SYDMT (SEQ ID NO:23), SYDMS (SEQ ID NO:24), DYDLS (SEQ ID NO:25) or SYDMS (SEQ ID NO:26) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products and/or modulate the immune response.
  • CDRs complementarity determining regions
  • SEQ ID NO:24 SYDMS
  • DYDLS SEQ ID NO:25
  • SYDMS SEQ ID NO:26
  • a heavy chain variable region having a CDR2 sequence of YASGSTYY (SEQ ID NO:27), SSSGTTYY (SEQ ID NO:28), YASGSTYY (SEQ ID NO:29), AIDGNPYY (SEQ ID NO:30) or STAYNSHY (SEQ ID NO:31) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system.
  • a heavy chain variable region having a CDR3 sequence of EHAGYSGDTGH (SEQ ID NO:32), EGAGVSMT (SEQ ID NO:33), EDAGFSNA (SEQ ID NO:34), GAGD (SEQ ID NO:35) or GGSWLD (SEQ ID NO:36) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system.
  • a light chain variable region having a CDR1 sequence of RCAYD (SEQ ID NO:37), RCADVV (SEQ ID NO:38), or RCALV (SEQ ID NO:39) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system.
  • a light chain variable region having a CDR2 sequence of QSISTY (SEQ ID NO:40), QSVSSY (SEQ ID NO:41), ESISNY (SEQ ID NO:42), KNVYNNNW (SEQ ID NO:43), or QSVYDNDE (SEQ ID NO:98) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system.
  • a light chain variable region having a CDR3 sequence of QQGYTHSNVDNV (SEQ ID NO:44), QQGYSISDIDNA (SEQ ID NO:45), QCTSGGKFISDGAA (SEQ ID NO:46), AGDYSSSSDNG (SEQ ID NO:47), or QATHYSSDWLTY (SEQ ID NO:48) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products.
  • Light and heavy chains that can bind CD83 polypeptides are also provided by the invention.
  • the invention provides a 20D04 light chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 20D04 light chain is provided below (SEQ ID NO:11).
  • a nucleic acid sequence for this 20D04 anti-CD83 light chain is provided below (SEQ ID NO:12).
  • 1 ATGGACATGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC 41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCG ATGTCGTGAT 81 GACCCAGACT CCAGCCTCCG TGTCTGCAGC TGTGGGAGGC 121 ACAGTCACCA TCAATTGCCA GGCCAGTGAA AGCATTAGCA 161 ACTACTTATC CTGGTATCAG CAGAAACCAG GGCAGCCTCC 201 CAAGCTCCTG ATCTACAGGA CATCCACTCT GGCATCTGGG 241 GTCTCATCGC GGTTCAAAGG CAGTGGATCT GGGACAGAGT 281 ACACTCTCAC CATCAGCGGC GTGCAGTGTG ACGATGTTGC 321 CACTTACTAC TGTCAATGCA CTTCTGGTGG GAAGTTCATT 361 AGTGATGGTG CTGCTTTCGG CGG C
  • the invention provides a 20D04 heavy chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 20D04 heavy chain is provided below (SEQ ID NO:13).
  • 1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC 41
  • a nucleic acid sequence for this 20D04 anti-CD83 heavy chain is provided below (SEQ ID NO:14). 1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC 41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG 81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC 121 ACCGTCTCTG GATTCTCCCT CAGTAACAAT GCAATAAACT 161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTAG AGTGGATCGG 201 ATACATTTGG AGTGGTGGGC TTACATACTA CGCGAACTGG 241 GCGGAAGGCC GATTCACCAT CTCCAAAACC TCGACTACGG 281 TGGATCTGAA GATGACCAGT CCGACAATCG AGGACACGGC 321 CACCTATTTC TGTGCCAGAG GGATTAATAA CTCCGCTTTG 361 TGGGGCCCAG GCACCCT
  • the invention provides a 111 G05 light chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 11 G05 light chain is provided below (SEQ ID NO:15).
  • a nucleic acid sequence for this 11G05 anti-CD83 light chain is provided below (SEQ ID NO:16). 1 ATGGACACCA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC 41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCG ACGTCGTGAT 81 GACCCAGACT CCAGCCTCCG TGTCTGCAGC TGTGGGAGGC 121 ACAGTCACCA TCAATTGCCA GTCCAGTAAG AATGTTTATA 161 ATAACAACTG GTTATCCTGG TTTCAGCAGA AACCAGGGCA 201 GCCTCCCAAG CTCCTGATCT ATTATGCATC CACTCTGGCA 241 TCTGGGGTCC CATCGCGGTT CAGAGGCAGT GGATCTGGGA 281 CACAGTTCAC TCACCATT AGCGACGTGC AGTGTGACGA 321 TGCTGCCACT TACTACTGTG CAGGCGATTA TAGTAGTAGT 361 AGTGATAATG GTTTCGGCGG AGGG
  • the invention provides a 111 G05 heavy chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 11G05 heavy chain is provided below (SEQ ID NO:17).
  • 1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC 41
  • TVSGFTISDY DLSWVRQAPG EGLKYIGFIA IDGNPYYATW 81 AKGRFTISKT STTVDLKITA PTTEDTATYF
  • CARGAGDLWG 121 PGTLVTVSSG QPKAPSVFPL APCCGDTPSS TVTLGCLVKG 161 YLPEPVTVTW NSGTLTNGVR TFPSVRQSSG LYSLSSVVSV 201 TSSSQPVTCN VAHPATNTKV DKTVAPSTCS KPTCPPPELL 241 GGPSVFIFPP KPKDTLMISR TPEVTCVVVD VSQDDPEVQF 281 TWYINNEQVR TARPPLREQQ FNSTI
  • a nucleic acid sequence for this 11 G05 anti-CD83 heavy chain is provided below (SEQ ID NO:18). 1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC 41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG 81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC 121 ACAGTCTCTG GATTCACCAT CAGTGACTAC GACTTGAGCT 161 GGGTCCGCCA GGCTCCAGGG GAGGGGCTGA AATACATCGG 201 ATTCATTGCT ATTGATGGTA ACCCATACTA CGCGACCTGG 241 GCAAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG 281 TGGATCTGAA AATCACCGCT CCGACAACCG AAGACACGGC 321 CACGTATTTC TGCCAGAG GGGCAGGGGA CCTCTGGGGC 361 CCAGGGACCC TCG
  • the invention provides a 14C 12 light chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 14C12 light chain is provided below (SEQ ID NO:19).
  • a nucleic acid sequence for this 14C12 anti-CD83 light chain is provided below (SEQ ID NO:20).
  • the invention provides a 14C 12 heavy chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this 14C 12 heavy chain is provided below (SEQ ID NO:21).
  • 1 METGLRWLLL VAVLKGVHCQ SVEESGGRLV TPGTPLTLTC 41 TASGFSRSSY DMSWVRQAPG KGLEWVGVIS TAYNSHYASW 81 AKGRFTISRT STTVDLKMTS LTTEDTATYF CARGGSWLDL 121 WGQGTLVTVS SGQPKAPSVF PLAPCCGDTP SSTVTLGCLV 161 KGYLPEPVTV TWNSGTLTNG VRTFPSVRQS SGLYSLSSVV 201 SVTSSSQPVT CNVAHPATNT KVDKTVAPST CSKPTCPPPE 241 LLGGPSVFIF PPKPKDTLMI SRTPEVTCVV VDVSQDDPEV 281 QFTWYINNEQ VRTARPPLRE QQFNSTIRV
  • a nucleic acid sequence for this 14C12 anti-CD83 heavy chain is provided below (SEQ ID NO:22). 1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC 41 TCAAAGGTGT CCACTGTCAG TCGGTGGAGG AGTCCGGGGG 81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC 121 ACAGCCTCTG GATTCTCCCG CAGCAGCTAC GACATGAGCT 161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGGTCGG 201 AGTCATTAGT ACTGCTTATA ACTCACACTA CGCGAGCTGG 241 GCAAAAGGCC GATTCACCAT CTCCAGAACC TCGACCACGG 281 TGGATCTGAA AATGACCAGT CTGACAACCG AAGACACGGC 321 CACCTATTTC TGTGCCAGAG GGGGTAGTTG GTTGGATCTC 361 TGGGGCCAGG
  • the invention provides a M83 020B08L light chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this M83 020B08L light chain is provided below (SEQ ID NO:58).
  • a nucleic acid sequence for this M83 020B08L anti-CD83 light chain is provided below (SEQ ID NO:59).
  • 1 ATGGACATGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC 41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCT ATGATATGAC 81 CCAGACTCCA GCCTCTGTGG AGGTAGCTGT GGGAGGCACA 121 GTCACCATCA AGTGCCAGGC CAGTCAGAGC ATTAGTACCT 161 ACTTAGACTG GTATCAGCAG AAACCAGGGC AGCCTCCCAA 201 GCTCCTGATC TATGATGCAT CCGATCTGGC ATCTGGGGTC 241 CCATCGCGGT TCAAAGGCAG TGGATCTGGG ACACAGTTCA 281 CTCTCACCAT CAGCGACCTG GAGTGTGCCG ATGCTGCCAC 321 TTACTACTGT CAACAGGGTT ATACACATAG TAATGTTGAT 361 AATGTTTTCG GCGG
  • the invention provides a M83 020B08H heavy chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this M83 020B08H heavy chain is provided below (SEQ ID NO:60).
  • 1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC 41
  • QSSGLYSLSS 201 VVSVTSSSQP VTCNVAHPAT NTKVDKTVAP
  • STCSKPTCPP 241 PELLGGPSVF IFPPKPKDTL MISRTPEVTC VVVDVSQDDP 281 EVQFTWYINN
  • a nucleic acid sequence for this M83 020B08H anti-CD83 heavy chain is provided below (SEQ ID NO:61).
  • the invention provides a M83 006G05L light chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this M83 006G05L light chain is provided below (SEQ ID NO:62).
  • a nucleic acid sequence for M83 006G05L anti-CD83 light chain is provided below (SEQ ID NO:63).
  • 1 ATGGACATGA GGGCCCCCAC TCAACTGCTG GGGCTCCTGC 41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCT ATGATATGAC 81 CCAGACTCCA GCCTCTGTGG AGGTAGCTGT GGGAGGCACA 121 GTCGCCATCA AGTGCCAGGC CAGTCAGAGC GTTAGTAGTT 161 ACTTAGCCTG GTATCAGCAG AAACCAGGGC AGCCTCCCAA 201 GCCCCTGATC TACGAAGCAT CCATGCTGGC GGCTGGTC 241 TCATCGCGGT TCAAAGGCAG TGGATCTGGG ACAGACTTCA 281 CTCTCACCAT CAGCGACCTG GAGTGTGACG ATGCTGCCAC 321 TTACTATTGT CAACAGGGTT ATTCTATCAG TGATATTGAT 361 AATGCTTTCG GCGG
  • the invention provides a M83 006G05L heavy chain that can bind to CD83 polypeptides.
  • the amino acid sequence for this M83 006G05L heavy chain is provided below (SEQ ID NO:64).
  • 1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV SPGTPLTLTC 41 TASGFSLSSY DMSWVRQAPG KGLEYIGIIS SSGSTYYASW 81 AKGRFTISKT STTVDLEVTS LTTEDTATYF CSREHAGYSG 121 DTGHLWGPGT LVTVSSGQPK APSVFPLAPC CGDTPSSTVT 161 LGCLVKGYLP EPVTVTWNSG TLTNGVRTFP SVRQSSGLYS 201 LSSVVSVTSS SQPVTCNVAH PATNTKVDKT VAPSTCSKPT 241 CPPPELLGGP SVFIFPPKPK DTLMISRTPE VTCVVVDVSQ 281 DDPEVQFTWY INN
  • a nucleic acid sequence for this M83 006G05L anti-CD83 heavy chain is provided below (SEQ ID NO:65).
  • the invention provides a 96G08 light chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • the CDR regions in the 96G08 light chain include amino acid sequences QSSQSVYNNDFLS (SEQ ID NO:71), YASTLAS (SEQ ID NO:72), and TGTYGNSAWYEDA (SEQ ID NO:73).
  • a nucleic acid sequence for this 96G08 anti-CD83 light chain is provided below (SEQ ID NO:74).
  • the CDR regions in the 96G08 light chain include nucleic acid sequences CAGTCCAGTCAGAGTGTTTATAATA (SEQ ID NO:75), ATGCATCCACTCTGGCATCT (SEQ ID NO:76), and ACAGGCACTTATGGT AATAGTGCTT (SEQ ID NO:77).
  • the invention provides a 96G08 heavy chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • the CDR regions in the 96G08 heavy chain include amino acid sequences SDGIS (SEQ ID NO:79), IISSGGNTYYASWAKG (SEQ ID NO:80) and VVGGTYSI (SEQ ID NO:81).
  • a nucleic acid sequence for the 96G08 anti-CD83 heavy chain is provided below (SEQ ID NO:82). 1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC 41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG 81 TCGCCTGGTC ACACCTGGGA CACCCCTGAC ACTCACCTGC 121 ACAGTGTCTG GAATCGACCT CAGTAGCGAT GGAATAAGCT 161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGATCGG 201 AATCATTAGT AGTGGTGGTA ACACATACTA CGCGAGCTGG 241 GCAAAAGGCC GATTCACCAT CTCCAGAACC TCGACCACGG 281 TGGATCTGAA GATGACCAGT CTGACAACCG AGGACACGGC 321 CACCTATTTC TGTGCCAGAG TTGTTGGTGG TACTTATAGC 361 ATCTGGGGCC AGGGC
  • the CDR regions in the 96G08 anti-CD83 heavy chain include AGCGATGGAATAAGC (SEQ ID NO:83), ATCATTAGTAGTGGTGGTA ACACATACTACGCGAGCTGGGCAAAAGGC (SEQ ID NO:84), and G TTGTTGGTGG TACTTATAGC ATC (SEQ ID NO:85).
  • the invention provides a 95F04 light chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • the CDR regions in the 95F04 anti-CD83 light chain include amino acid sequences QSSQSVYGNNELS (SEQ ID NO:87), QASSLAS (SEQ ID NO:88) and LGEYSISADNH (SEQ ID NO:89).
  • a nucleic acid sequence for this 95F04 anti-CD83 light chain is provided below (SEQ ID NO:90).
  • the invention provides a 95F04 heavy chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs).
  • PBMCs peripheral blood mononuclear cells
  • the CDR regions in the 95F04 anti-CD83 heavy chain include amino acid sequences SNAMI (SEQ ID NO:92), AMDSNSRTYYATWAKG (SEQ ID NO:93), and GDGGSSDYTEM (SEQ ID NO:94).
  • a nucleic acid sequence for this 95F04 anti-CD83 heavy chain is provided below (SEQ ID NO:95).
  • a related nucleic acid sequence for the 95F04 anti-CD83 light chain is provided below (SEQ ID NO:96).
  • the invention also provides compositions and methods for decreasing inappropriate immune responses in animals, including humans.
  • the CD83 gene has a profound effect upon T cell activity.
  • CD4+ T cells require CD83-related functions.
  • CD83 CD4+ T cell activation and/or proliferation is impaired.
  • the therapeutic manipulation of CD83 may thus represent a mechanism for the specific regulation of T cell function in the treatment of T cell mediated diseases, including autoimmune disorders.
  • antibodies capable of blocking CD83 function can be used as therapeutics in the treatment of immune diseases.
  • the CD83-related compositions and methods of the invention can be used in the treatment of autoimmune diseases.
  • Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against “self tissues” and that promote the production of cytokines and auto-antibodies involved in the pathology of the diseases.
  • Modulation of T cell activity by modulating CD83 can have an effect on the course of the autoimmune disease.
  • Non-limiting examples of autoimmune diseases and disorders having an autoimmune component that may be treated according to the invention include diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiple sclerosis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjogren's Syndrome, including keratoconjunctivitis sicca secondary to Sjogren's Syndrome, alopecia areata, allergic responses due to arthropod bite reactions, Crohn's disease, aphthous ulcer, ulceris, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis,
  • anti-CD83 antibodies can inhibit T cell proliferation.
  • the efficacy of anti-CD83-related compositions for treating autoimmune diseases can be tested in the animal models provided herein or other models of human diseases (e.g., EAE as a model of multiple sclerosis and the NOD mice as a model for diabetes).
  • animal models include the mrl/lpr/lpr mouse as a model for lupus erythematosus, murine collagen-induced arthritis as a model for rheumatoid arthritis, and murine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 840-856).
  • a CD83-modulatory (e.g., inhibitory) agent of the invention is administered to test animals and the course of the disease in the test animals is then monitored by the standard methods for the particular model being used. Effectiveness of the modulatory agent is evidenced by amelioration of the disease condition in animals treated with the agent as compared to untreated animals (or animals treated with a control agent).
  • compositions and methods of the invention that involve decreasing CD83 function can be used to decrease transplant rejection and prolong survival of the tissue graft.
  • These methods can be used both in solid organ transplantation and in bone marrow transplantation (e.g., to inhibit graft-versus-host disease).
  • These methods can involve either direct administration of a CD83 inhibitory agent to the transplant recipient or ex vivo treatment of cells obtained from the subject (e.g., T cells, Th1 cells, B cells, non-lymphoid cells) with an inhibitory agent followed by re-administration of the cells to the subject.
  • any agent that can modulate CD83 or to further decrease T cell levels can also be used in the compositions and methods of the invention.
  • anti-CD83 antibodies of the invention are used to either activate or inhibit CD83 activity.
  • any agent that can inhibit CD83 from performing its natural functions can be used in the compositions and methods of the invention as a CD83 inhibitory agent.
  • Indicators that CD83 activity is inhibited include decreased T cell counts, increased IL-4 cytokine levels, increased IL-10 levels, decreased IL-2 production, and decreased TNF levels relative to uninhibited levels in wild type CD83 cells.
  • CD83 inhibitors include anti-CD83 antibodies, CD83 anti-sense nucleic acids (e.g. nucleic acids that can hybridize to CD83 nucleic acids), organic compounds, peptides and agents that can mutate an endogenous CD83 gene.
  • CD83 anti-sense nucleic acids e.g. nucleic acids that can hybridize to CD83 nucleic acids
  • organic compounds e.g. peptides and agents that can mutate an endogenous CD83 gene.
  • the CD83 stimulatory or inhibitory agents are proteins, for example, CD83 gene products, anti-CD83 antibody preparations, CD83 inhibitors, peptides and protein factors that can promote CD83 transcription or translation.
  • the CD83 stimulatory or inhibitory agents are peptides or organic molecules. Such proteins, organic molecules and organic molecules can be prepared and/or purified as described herein or by methods available in the art, and administered as provided herein.
  • the CD83 inhibitory agents can be nucleic acids including recombinant expression vectors or expression cassettes encoding CD83 anti-sense nucleic acid, intracellular antibodies capable of binding to CD83 or dominant negative CD83 inhibitors.
  • nucleic acids can be operably linked to a promoter that is functional in a mammalian cell, and then introduced into cells of the subject mammal using methods known in the art for introducing nucleic acid (e.g., DNA) into cells.
  • the “promoter functional in a mammalian cell” or “mammalian promoter” is capable of directing transcription of a polypeptide coding sequence operably linked to the promoter.
  • the promoter should generally be active in T cells and antigen presenting cells and may be obtained from a gene that is expressed in T cells or antigen presenting cells. However, it need not be a T cell-specific or an antigen presenting cell specific-promoter. Instead, the promoter may be selected from any mammalian or viral promoter that can function in a T cell.
  • the promoter may be an actin promoter, an immunoglobulin promoter, a heat-shock promoter, or a viral promoter obtained from the genome of viruses such as adenoviruses, retroviruses, lentiviruses, herpes viruses, including but not limited to, polyoma virus, fowlpox virus, adenovirus 2, bovine papilloma virus, avian sarcoma virus, cytomegalovirus (CMV), hepatitis-B virus, Simian Virus 40 (SV40), Epstein Barr virus (EBV), feline immunodeficiency virus (FIV), and Sra, or are respiratory synsitial viral promoters (RSV) or long terminal repeats (LTRs) of a retrovirus, i.e., a Moloney Murine Leukemia Virus (MoMuLv) (Cepko et al. (1984) Cell 37:1053-1062).
  • the promoter functional in
  • any cloning procedure used by one of skill in the art can be employed to make the expression vectors or expression that comprise a promoter operably linked to a CD83 nucleic acid, CD83 transcription factor or a nucleic acid encoding an anti-CD83 antibody. See, e.g., Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y., 1989; Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y., 2001.
  • mammalian cells After constructing an expression vector or an expression cassette encoding CD83 transcription factors, CD83 anti-sense nucleic acid, intracellular antibodies capable of binding to CD83 or dominant negative CD83 inhibitors, mammalian cells can be transformed with the vector or cassette. Examples of such methods include:
  • Naked DNA can be introduced into cells in vivo by directly injecting the DNA into the cells (see e.g., Acsadi et al. (1991) Nature 332:815-818; Wolff et al. (1990) Science 247:1465-1468).
  • a delivery apparatus e.g., a “gene gun” for injecting DNA into cells in vivo can be used.
  • Such an apparatus is commercially available (e.g., from BioRad).
  • Naked DNA can also be introduced into cells in vivo by complexing the DNA to a cation, such as polylysine, which is coupled to a ligand for a cell-surface receptor (see for example Wu, G. and Wu, C. H. (1988) J. Biol. Chem. 263:14621; Wilson et al. (1992) J. Biol. Chem. 267:963-967; and U.S. Pat. No. 5,166,320). Binding of the DNA-ligand complex to the receptor facilitates uptake of the DNA by receptor-mediated endocytosis.
  • a cation such as polylysine
  • a DNA-ligand complex linked to adenovirus capsids that naturally disrupt endosomes, thereby releasing material into the cytoplasm can be used to avoid degradation of the complex by intracellular lysosomes (see for example Curiel et al. (1991) Proc. Natl. Acad. Sci. USA 88:8850; Cristiano et al. (1993) Proc. Natl. Acad. Sci. USA 90:2122-2126).
  • Retroviruses Defective retroviruses are well characterized for use in gene transfer for gene therapy purposes (for a review see Miller, A. D. (1990) Blood 76:271).
  • a recombinant retrovirus can be constructed having nucleotide sequences of interest incorporated into the retroviral genome. Additionally, portions of the retroviral genome can be removed to render the retrovirus replication defective. The replication defective retrovirus is then packaged into virions that can be used to infect a target cell through the use of a helper virus by standard techniques. Protocols for producing recombinant retroviruses and for infecting cells in vitro or in vivo with such viruses can be found in Current Protocols in Molecular Biology, Ausubel, F. M. et al.
  • retroviruses include pLJ, pZIP, pWE and pEM which are available to those skilled in the art.
  • suitable packaging virus lines include ? Crip, ? Cre, ? 2 and ? Am. Retroviruses have been used to introduce a variety of genes into many different cell types, including epithelial cells, endothelial cells, lymphocytes, myoblasts, hepatocytes, bone marrow cells, in vitro and/or in vivo (see for example Eglitis, et al. (1985) Science 230:1395-1398; Danos and Mulligan (1988) Proc. Natl.
  • Retroviral vectors require target cell division in order for the retroviral genome (and foreign nucleic acid inserted into it) to be integrated into the host genome to stably introduce nucleic acid into the cell. Thus, it may be necessary to stimulate replication of the target cell.
  • Adenoviruses The genome of an adenovirus can be manipulated such that it encodes and expresses a gene product of interest but is inactivated in terms of its ability to replicate in a normal lytic viral life cycle. See, for example, Berkner et al. (1988) BioTechniques 6:616; Rosenfeld et al. (1991) Science 252:431-434; and Rosenfeld et al. (1992) Cell 68:143-155.
  • Suitable adenoviral vectors derived from the adenovirus strain Ad type 5 dl 324 or other strains of adenovirus are available to those skilled in the art.
  • Recombinant adenoviruses are advantageous in that they do not require dividing cells to be effective gene delivery vehicles and can be used to infect a wide variety of cell types, including airway epithelium (Rosenfeld et al. (1992) cited supra), endothelial cells (Lemarchand et al. (1992) Proc. Natl. Acad. Sci. USA 89:6482-6486), hepatocytes (Herz and Gerard (1993) Proc. Natl. Acad. Sci. USA 90:2812-2816) and muscle cells (Quantin et al. (1992) Proc. Natl. Acad. Sci. USA 89:2581-2584).
  • introduced adenoviral DNA (and foreign DNA contained therein) is not integrated into the genome of a host cell but remains episomal, thereby avoiding potential problems that can occur as a result of insertional mutagenesis in situations where introduced DNA becomes integrated into the host genome (e.g., retroviral DNA).
  • the carrying capacity of the adenoviral genome for foreign DNA is large (up to 8 kilobases) relative to other gene delivery vectors (Berkner et al. cited supra; Haj-Ahmand and Graham (1986) J. Virol. 57:267).
  • Most replication-defective adenoviral vectors currently in use are deleted for all or parts of the viral E1 and E3 genes but retain as much as 80% of the adenoviral genetic material.
  • Adeno-associated virus is a naturally occurring defective virus that requires another virus, such as an adenovirus or a herpes virus, as a helper virus for efficient replication and a productive life cycle.
  • AAV Adeno-associated virus
  • AAV vector such as that described in Tratschin et al. (1985) Mol. Cell. Biol. 5:3251-3260 can be used to introduce DNA into cells.
  • a variety of nucleic acids have been introduced into different cell types using AAV vectors (see for example Hermonat et al. (1984) Proc. Natl. Acad. Sci. USA 81:6466-6470; Tratschin et al. (1985) Mol. Cell. Biol.
  • Transformed mammalian cells can then be identified and administered to the mammal from whence they came to permit expression of a CD83 transcription factor, CD83 anti-sense nucleic acid, intracellular antibody capable of binding to CD83 proteins, or dominant negative CD83 inhibitors.
  • the efficacy of a particular expression vector system and method of introducing nucleic acid into a cell can be assessed by standard approaches routinely used in the art.
  • DNA introduced into a cell can be detected by a filter hybridization technique (e.g., Southern blotting).
  • RNA produced by transcription of an introduced DNA can be detected, for example, by Northern blotting, RNase protection or reverse transcriptase-polymerase chain reaction (RT-PCR).
  • the CD83 gene product can be detected by an appropriate assay, for example, by immunological detection of a produced CD83 protein, such as with a CD83-specific antibody.
  • Anti-sense nucleic acids can be used to inhibit the function of CD83.
  • the function of CD83 RNA is inhibited, for example, by administering to a mammal a nucleic acid that can inhibit the functioning of CD83 RNA.
  • Nucleic acids that can inhibit the function of a CD83 RNA can be generated from coding and non-coding regions of the CD83 gene.
  • nucleic acids that can inhibit the function of a CD83 RNA are often selected to be complementary to CD83 nucleic acids that are naturally expressed in the mammalian cell to be treated with the methods of the invention.
  • the nucleic acids that can inhibit CD83 RNA functions are complementary to CD83 sequences found near the 5′ end of the CD83 coding region.
  • nucleic acids that can inhibit the function of a CD83 RNA can be complementary to the 5′ region of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10.
  • a nucleic acid that can inhibit the functioning of a CD83 RNA need not be 100% complementary to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. Instead, some variability the sequence of the nucleic acid that can inhibit the functioning of a CD83 RNA is permitted.
  • a nucleic acid that can inhibit the functioning of a CD83 RNA from a human can be complementary to a nucleic acid encoding either a human or a mouse CD83 gene product.
  • nucleic acids that can hybridize under moderately or highly stringent hybridization conditions to a nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10 are sufficiently complementary to inhibit the functioning of a CD83 RNA and can be utilized in the methods of the invention.
  • “Stringent hybridization conditions” and “stringent hybridization wash conditions” in the context of nucleic acid hybridization are somewhat sequence dependent, and may differ depending upon the environmental conditions of the nucleic acid. For example, longer sequences tend to hybridize specifically at higher temperatures.
  • An extensive guide to the hybridization of nucleic acids is found in Tijssen, Laboratory Techniques in Biochemistry and Molecular biology-Hybridization with Nucleic Acid Probes, page 1, chapter 2 “Overview of principles of hybridization and the strategy of nucleic acid probe assays” Elsevier, New York (1993). See also, J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, N.Y., pp 9.31-9.58 (1989); J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, N.Y. (3rd ed. 2001).
  • highly stringent hybridization and wash conditions are selected to be about 5° C. lower than the thermal melting point (T m ) for the specific double-stranded sequence at a defined ionic strength and pH.
  • T m thermal melting point
  • highly stringent hybridization and wash conditions are selected to be about 5° C. lower than the thermal melting point (T m ) for the specific double-stranded sequence at a defined ionic strength and pH.
  • T m thermal melting point
  • T m can be Approximated from the Equation of Meinkoth and Wahl Anal. Biochem. 138:267-284 (1984):
  • M is the molarity of monovalent cations
  • % GC is the percentage of guanosine and cytosine nucleotides in the DNA
  • % form is the percentage of formamide in the hybridization solution
  • L is the length of the hybrid in base pairs.
  • T m is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe.
  • Very stringent conditions are selected to be equal to the T m for a particular probe.
  • stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity can hybridize.
  • stringent conditions will be those in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60° C. for long probes (e.g., greater than 50 nucleotides).
  • Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide.
  • Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., and a wash in 0.5X to 1X SSC at 55 to 60° C.
  • Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1X SSC at 60 to 65° C.
  • the degree of complementarity or sequence identity of hybrids obtained during hybridization is typically a function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution.
  • the type and length of hybridizing nucleic acids also affects whether hybridization will occur and whether any hybrids formed will be stable under a given set of hybridization and wash conditions.
  • An example of stringent hybridization conditions for hybridization of complementary nucleic acids that have more than 100 complementary residues on a filter in a Southern or Northern blot is 50% formamide with 1 mg of heparin at 42° C., with the hybridization being carried out overnight.
  • An example of highly stringent conditions is 0.1 5 M NaCl at 72° C. for about 15 minutes.
  • An example of stringent wash conditions is a 0.2x SSC wash at 65° C. for 15 minutes (see also, Sambrook, infra). Often, a high stringency wash is preceded by a low stringency wash to remove background probe signal.
  • An example of medium stringency for a duplex of, e.g., more than 100 nucleotides, is 1x SSC at 45° C. for 15 minutes.
  • An example low stringency wash for a duplex of, e.g., more than 100 nucleotides, is 4-6x SSC at 40° C. for 15 minutes.
  • stringent conditions typically involve salt concentrations of less than about 1.0M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3, and the temperature is typically at least about 30° C.
  • Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.
  • destabilizing agents such as formamide.
  • a signal to noise ratio of 2x (or higher) than that observed for an unrelated probe in the particular hybridization assay indicates detection of a specific hybridization.
  • Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the proteins that they encode are substantially identical. This occurs, e.g., when a copy of a nucleic acid is created using the maximum codon degeneracy permitted by the genetic code.
  • a reference nucleotide sequence preferably hybridizes to the reference nucleotide sequence in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO 4 , 1 mM EDTA at 50° C. with washing in 2X SSC, 0.1% SDS at 50° C., more desirably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO 4 , 1 mM EDTA at 50° C.
  • SDS sodium dodecyl sulfate
  • T m is reduced by about 1° C. for each 1% of mismatching.
  • T m , hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired sequence identity. For example, if sequences with >90% identity are sought, the T m can be decreased 10° C.
  • stringent conditions are selected to be about 5° C. lower than the thermal melting point (T m ) for the specific sequence and its complement at a defined ionic strength and pH.
  • severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermal melting point (T m );
  • moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than the thermal melting point (T m ); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower than the thermal melting point (T m ).
  • Precise complementarity is therefore not required for successful duplex formation between a nucleic acid that can inhibit a CD83 RNA and the complementary coding sequence of a CD83 RNA.
  • Inhibitory nucleic acid molecules that comprise, for example, 2, 3, 4, or 5 or more stretches of contiguous nucleotides that are precisely complementary to a CD83 coding sequence, each separated by a stretch of contiguous nucleotides that are not complementary to adjacent CD83 coding sequences, can inhibit the function of CD83 RNA.
  • each stretch of contiguous nucleotides is at least 4, 5, 6, 7, or 8 or more nucleotides in length.
  • Non-complementary intervening sequences are preferably 1, 2, 3, or 4 nucleotides in length.
  • One skilled in the art can easily use the calculated melting point of an anti-sense nucleic acid hybridized to a sense nucleic acid to determine the degree of mismatching that will be tolerated between a particular anti-sense nucleic acid and a particular CD83 RNA.
  • nucleic acids that complementary a CD83 RNA can be administered to a mammal or to directly to the site of the inappropriate immune system activity.
  • nucleic acids that are complementary to a CD83 RNA can be generated by transcription from an expression cassette that has been administered to a mammal.
  • a complementary RNA can be transcribed from a CD83 nucleic acid that has been inserted into an expression cassette in the 3′ to 5′ orientation, that is, opposite to the usual orientation employed to generate sense RNA transcripts.
  • the promoter would be positioned to transcribe from a 3′ site towards the 5′ end of the CD83 coding region.
  • an RNA that can inhibit the function of an endogenous CD83 RNA is an anti-sense oligonucleotide.
  • the anti-sense oligonucleotide is complementary to at least a portion of the coding sequence of a gene comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10.
  • Such anti-sense oligonucleotides are generally at least six nucleotides in length, but can be about 8, 12, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides long. Longer oligonucleotides can also be used.
  • CD83 anti-sense oligonucleotides can be provided in a DNA construct and introduced into cells whose division is to be decreased, for example, into CD4 + T cells, Th-1 cells, Th-2 cells or lymphocyte precursor cells.
  • Anti-sense oligonucleotides can be composed of deoxyribonucleotides, ribonucleotides, or a combination of both. Oligonucleotides can be synthesized endogenously from transgenic expression cassettes or vectors as described herein.
  • oligonucleotides can be synthesized manually or by an automated synthesizer, by covalently linking the 5′ end of one nucleotide with the 3′ end of another nucleotide with non-phosphodiester internucleotide linkages such alkylphosphonates, phosphorothioates, phosphorodithioates, alkylphosphonothioates, alkylphosphonates, phosphoramidates, phosphate esters, carbamates, acetamidate, carboxymethyl esters, carbonates, and phosphate triesters. See Brown, 1994, Meth. Mol. Biol. 20:1-8; Sonveaux, 1994, Meth. Mol. Biol. 26:1-72; Uhlmann et al., 1990, Chem. Rev. 90:543-583.
  • CD83 anti-sense oligonucleotides can be modified without affecting their ability to hybridize to a CD83 RNA. These modifications can be internal or at one or both ends of the anti-sense molecule.
  • internucleoside phosphate linkages can be modified by adding peptidyl, cholesteryl or diamine moieties with varying numbers of carbon residues between these moieties and the terminal ribose.
  • Modified bases and/or sugars such as arabinose instead of ribose, or a 3′,5′-substituted oligonucleotide in which the 3′ hydroxyl group or the 5′ phosphate group are substituted, can also be employed in a modified anti-sense oligonucleotide.
  • modified oligonucleotides can be prepared by methods available in the art. Agrawal et al., 1992, Trends Biotechnol. 10: 152-158; Uhlmann et al., 1990, Chem. Rev. 90:543-584; Uhlmann et al., 1987, Tetrahedron. Lett. 215:3539-3542.
  • ribozyme is an RNA molecule with catalytic activity. See, e.g., Cech, 1987, Science 236: 1532-1539; Cech, 1990, Ann. Rev. Biochem. 59:543-568; Cech, 1992, Curr. Opin. Struct. Biol. 2: 605-609; Couture and Stinchcomb, 1996, Trends Genet. 12: 510-515. Ribozymes can be used to inhibit gene function by cleaving an RNA sequence, as is known in the art (see, e.g., Haseloff et al., U.S. Pat. No. 5,641,673).
  • CD83 nucleic acids complementary to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10 can be used to generate ribozymes that will specifically bind to mRNA transcribed from a CD83 gene.
  • Methods of designing and constructing ribozymes that can cleave other RNA molecules in trans in a highly sequence specific manner have been developed and described in the art (see Haseloffet al. (1988), Nature 334:585-591).
  • the cleavage activity of ribozymes can be targeted to specific RNAs by engineering a discrete “hybridization” region into the ribozyme.
  • the hybridization region contains a sequence complementary to the target RNA and thus specifically hybridizes with the target (see, for example, Gerlach et al., EP 321,201).
  • the target sequence can be a segment of about 10, 12, 15, 20, or 50 contiguous nucleotides selected from a nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. Longer complementary sequences can be used to increase the affinity of the hybridization sequence for the target.
  • the hybridizing and cleavage regions of the ribozyme can be integrally related; thus, upon hybridizing to the target RNA through the complementary regions, the catalytic region of the ribozyme can cleave the target.
  • compositions and methods for modulating CD83 activity or expression can include these molecules as well as other components. Representative examples that are discussed in more detail below include transcription factors, RNA-binding factors, organic molecules, or peptides.
  • RNA-Binding Factors [0216]
  • RNA binding factors include those described in PCT/EP01/14820 and other sources.
  • the HuR protein (Genbank accession number U38175) has the ability to specifically bind to CD83 RNA at AU-rich elements or sites.
  • Such AU-rich elements comprise sequences such as AUUUA (SEQ ID NO:49), AUUUUA (SEQ ID NO:50) and AUUUUUA (SEQ ID NO:51). Binding by such HuR proteins to CD83 mRNA is thought to increase the stability, transport and translation of CD83 mRNA, and thereby increase the expression of CD83 polypeptides.
  • CD83 expression may be increase by administering HuR proteins or nucleic acids to a mammal.
  • CD83 expression may be decreased by administering factors that block HuR binding to CD83 mRNA.
  • Factors that block HuR binding include proteins or nucleic acids that can bind to the AU-rich elements normally bound by HuR, for example, nucleic acids or anti-sense nucleic acids that are complementary to AU-rich elements.
  • organic molecules may be used to modulate the immune system. These compounds include any compound that can interact with a component of the immune system. Such compounds may interact directly with CD83, indirectly with CD83 or with some other polypeptide, cell or factor that plays a role in the function of the immune system. In some embodiments, the organic molecule can bind to a CD83 polypeptide or a CD83 nucleic acid.
  • Organic molecules can be tested or assayed for their ability to modulate CD83 activity, CD83 function or for their ability to modulate components of the immune system.
  • suitable organic molecules may be selected either from a chemical library, wherein chemicals are assayed individually, or from combinatorial chemical libraries where multiple compounds are assayed at once, then deconvoluted to determine and isolate the most active compounds.
  • combinatorial chemical libraries include those described by Agrafiotis et al., “System and method of automatically generating chemical compounds with desired properties,” U.S. Pat. No. 5,463,564; Armstrong, R. W., “Synthesis of combinatorial arrays of organic compounds through the use of multiple component combinatorial array syntheses,” WO 95/02566; Baldwin, J. J. et al., “Sulfonamide derivatives and their use,” WO 95/24186; Baldwin, J. J.
  • Peptide molecules that modulate the immune system may be obtained through the screening of combinatorial peptide libraries.
  • Such libraries may either be prepared by one of skill in the art (see e.g., U.S. Pat. Nos. 4,528,266 and 4,359,535, and Patent Cooperation Treaty Publication Nos. WO 92/15679, WO 92/15677, WO 90/07862, WO 90/02809, or purchased from commercially available sources (e.g., New England Biolabs Ph.D.TM Phage Display Peptide Library Kit).
  • the invention provides a method for identifying ligands, receptors, therapeutic drugs and other molecules that can modulate the phenotype of the mutant CD83 in vivo.
  • This method involves administering a test compound to the mutant CD83 mouse of the invention and observing whether the compound causes a change in the phenotype of the mutant mouse.
  • Changes in phenotype that are of interest include increases or decreases in T cells (especially CD4+ T cells), increases or decreases in GMCSF, IL-2, IL-4 or IL-10 cytokine production, increases or decreases in inflammation, increases or decreases in dendritic cell function and other T cell responses known to one of skill in the art.
  • Test compounds can be screened in vitro to ascertain whether they interact directly with CD83.
  • In vitro screening can, for example, identify whether a test compound or molecule can bind to the cytoplasmic tail or the membrane-associated portions of CD83.
  • Such information combined with observation of the in vivo phenotype before and after administration of the test compound provides further insight into the function of CD83 and provides targets for manipulation T cell activation and other functions modulated by CD83.
  • the invention is not limited to identification of molecules that directly associate with CD83.
  • the in vivo screening methods provided herein can, also identify test compounds that have an indirect effect on CD83, or that partially or completely replace a function of CD83.
  • T cell numbers can be observed in blood samples or in samples obtained from thymus, spleen or lymph node tissues.
  • dendritic cells can be pulsed with antigens ex vivo and then injected into mice to prime CD4+ T cells in draining lymphoid organs. See Inaba et al., J. Exp. Med. 172: 631-640, 1990; Liu, et al., J. Exp. Med. 177: 1299-1307, 1993; Sornasse et al., J. Exp. Med. 175: 15-21, 1992.
  • Antigens can also be deposited intramuscularly and dendritic cells from the corresponding afferent lymphatics can carry that antigen in a form stimulatory for T cells. Bujdoso et al., J. Exp. Med. 170: 1285-1302, 1989. According to the invention, factors stimulating the interaction of dendritic cells with T cells in vivo can be identified by administering antigens in this manner and then observing how T cell respond, e.g. by observing whether T cell activation occurs.
  • the CD83 nucleic acids, polypeptides and antibodies of the invention, including their salts, are administered so as to achieve a reduction in at least one symptom associated with an infection, indication or disease.
  • the nucleic acid, polypeptide or antibody, a variant thereof or a combination thereof may be administered as single or divided dosages, for example, of at least about 0.01 mg/kg to about 500 to 750 mg/kg, of at least about 0.01 mg/kg to about 300 to 500 mg/kg, at least about 0.1 mg/kg to about 100 to 300 mg/kg or at least about 1 mg/kg to about 50 to 100 mg/kg of body weight, although other dosages may provide beneficial results.
  • the amount administered will vary depending on various factors including, but not limited to, the nucleic acid, polypeptide or antibody chosen, the disease, the weight, the physical condition, the health, the age of the mammal, whether prevention or treatment is to be achieved, and if the nucleic acid, polypeptide or antibody is chemically modified. Such factors can be readily determined by the clinician employing animal models or other test systems that are available in the art.
  • Administration of the therapeutic agents in accordance with the present invention may be in a single dose, in multiple doses, in a continuous or intermittent manner, depending, for example, upon the recipient's physiological condition, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to skilled practitioners.
  • the administration of the CD83 nucleic acids, polypeptides and antibodies of the invention may be essentially continuous over a preselected period of time or may be in a series of spaced doses. Both local and systemic administration is contemplated.
  • CD83 nucleic acids, polypeptides and antibodies are synthesized or otherwise obtained, purified as necessary or desired and then lyophilized and stabilized.
  • the nucleic acid, polypeptide or antibody can then be adjusted to the appropriate concentration, and optionally combined with other agents.
  • the absolute weight of a given nucleic acid, polypeptide or antibody included in a unit dose can vary widely. For example, about 0.01 to about 2 g, or about 0.1 to about 500 mg, of at least one nucleic acid, polypeptide or antibody of the invention, or a plurality of CD83 nucleic acid, polypeptides and antibodies specific for a particular cell type can be administered.
  • the unit dosage can vary from about 0.01 g to about 50 g, from about 0.01 g to about 35 g, from about 0.1 g to about 25 g, from about 0.5 g to about 12 g, from about 0.5 g to about 8 g, from about 0.5 g to about 4 g, or from about 0.5 g to about 2 g.
  • Daily doses of the CD83 nucleic acids, polypeptides or antibodies of the invention can vary as well. Such daily doses can range, for example, from about 0.1 g/day to about 50 g/day, from about 0.1 g/day to about 25 g/day, from about 0.1 g/day to about 12 g/day, from about 0.5 g/day to about 8 g/day, from about 0.5 g/day to about 4 g/day, and from about 0.5 g/day to about 2 g/day.
  • one or more suitable unit dosage forms comprising the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can be administered by a variety of routes including oral, parenteral (including subcutaneous, intravenous, intramuscular and intraperitoneal), rectal, dermal, transdermal, intrathoracic, intrapulmonary and intranasal (respiratory) routes.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies may also be formulated for sustained release (for example, using microencapsulation, see WO 94/07529, and U.S. Pat. No. 4,962,091).
  • the formulations may, where appropriate, be conveniently presented in discrete unit dosage forms and may be prepared by any of the methods well known to the pharmaceutical arts. Such methods may include the step of mixing the therapeutic agent with liquid carriers, solid matrices, semi-solid carriers, finely divided solid carriers or combinations thereof, and then, if necessary, introducing or shaping the product into the desired delivery system.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention are prepared for oral administration, they are generally combined with a pharmaceutically acceptable carrier, diluent or excipient to form a pharmaceutical formulation, or unit dosage form.
  • a pharmaceutically acceptable carrier diluent or excipient
  • the CD83 nucleic acids, polypeptides or antibodies may be present as a powder, a granular formulation, a solution, a suspension, an emulsion or in a natural or synthetic polymer or resin for ingestion of the active ingredients from a chewing gum.
  • the active CD83 nucleic acids, polypeptides or antibodies may also be presented as a bolus, electuary or paste.
  • CD83 nucleic acids, polypeptides or antibodies of the invention can also be formulated for sustained release, e.g., the CD83 nucleic acids, polypeptides or antibodies can be coated, micro-encapsulated, or otherwise placed within a sustained delivery device.
  • the total active ingredients in such formulations comprise from 0.1 to 99.9% by weight of the formulation.
  • pharmaceutically acceptable it is meant a carrier, diluent, excipient, and/or salt that is compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof.
  • nucleic acid, polypeptide or antibodies of the invention can be prepared by procedures known in the art using well-known and readily available ingredients.
  • the nucleic acid, polypeptide or antibody can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, solutions, suspensions, powders, aerosols and the like.
  • excipients, diluents, and carriers that are suitable for such formulations include buffers, as well as fillers and extenders such as starch, cellulose, sugars, mannitol, and silicic derivatives.
  • Binding agents can also be included such as carboxymethyl cellulose, hydroxymethylcellulose, hydroxypropyl methylcellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl-pyrrolidone.
  • Moisturizing agents can be included such as glycerol, disintegrating agents such as calcium carbonate and sodium bicarbonate.
  • Agents for retarding dissolution can also be included such as paraffin.
  • Resorption accelerators such as quaternary ammonium compounds can also be included.
  • Surface active agents such as cetyl alcohol and glycerol monostearate can be included.
  • Adsorptive carriers such as kaolin and bentonite can be added.
  • Lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols can also be included. Preservatives may also be added.
  • the compositions of the invention can also contain thickening agents such as cellulose and/or cellulose derivatives. They may also contain gums such as xanthan, guar or carbo gum or gum arabic, or alternatively polyethylene glycols, bentones and montmorillonites, and the like.
  • tablets or caplets containing the CD83 nucleic acids, polypeptides or antibodies of the invention can include buffering agents such as calcium carbonate, magnesium oxide and magnesium carbonate.
  • Caplets and tablets can also include inactive ingredients such as cellulose, pregelatinized starch, silicon dioxide, hydroxy propyl methyl cellulose, magnesium stearate, microcrystalline cellulose, starch, talc, titanium dioxide, benzoic acid, citric acid, corn starch, mineral oil, polypropylene glycol, sodium phosphate, zinc stearate, and the like.
  • Hard or soft gelatin capsules containing at least one nucleic acid, polypeptide or antibody of the invention can contain inactive ingredients such as gelatin, microcrystalline cellulose, sodium lauryl sulfate, starch, talc, and titanium dioxide, and the like, as well as liquid vehicles such as polyethylene glycols (PEGs) and vegetable oil.
  • enteric-coated caplets or tablets containing one or more CD83 nucleic acids, polypeptides or antibodies of the invention are designed to resist disintegration in the stomach and dissolve in the more neutral to alkaline environment of the duodenum.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can also be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for instance by intramuscular, subcutaneous, intraperitoneal or intravenous routes.
  • the pharmaceutical formulations of the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can also take the form of an aqueous or anhydrous solution or dispersion, or alternatively the form of an emulsion or suspension or salve.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies may be formulated for parenteral administration (e.g., by injection, for example, bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion containers or in multi-dose containers. As noted above, preservatives can be added to help maintain the shelve life of the dosage form.
  • the active CD83 nucleic acids, polypeptides or antibodies and other ingredients may form suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • the active CD83 nucleic acids, polypeptides or antibodies and other ingredients may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.
  • a suitable vehicle e.g., sterile, pyrogen-free water
  • formulations can contain pharmaceutically acceptable carriers, vehicles and adjuvants that are well known in the art. It is possible, for example, to prepare solutions using one or more organic solvent(s) that is/are acceptable from the physiological standpoint, chosen, in addition to water, from solvents such as acetone, ethanol, isopropyl alcohol, glycol ethers such as the products sold under the name “Dowanol,” polyglycols and polyethylene glycols, C 1 -C 4 alkyl esters of short-chain acids, ethyl or isopropyl lactate, fatty acid triglycerides such as the products marketed under the name “Miglyol,” isopropyl myristate, animal, mineral and vegetable oils and polysiloxanes.
  • organic solvent(s) that is/are acceptable from the physiological standpoint, chosen, in addition to water, from solvents such as acetone, ethanol, isopropyl alcohol, glycol ethers such as the products sold under the name “Dowanol,” polyg
  • antioxidants chosen from antioxidants, surfactants, other preservatives, film-forming, keratolytic or comedolytic agents, perfumes, flavorings and colorings.
  • Antioxidants such as t-butylhydroquinone, butylated hydroxyanisole, butylated hydroxytoluene and a-tocopherol and its derivatives can be added.
  • combination products that include one or more CD83 nucleic acids, polypeptides or antibodies of the present invention and one or more other anti-microbial agents.
  • antibiotics can be included in the pharmaceutical compositions of the invention, such as aminoglycosides (e.g., streptomycin, gentamicin, sisomicin, tobramycin and amicacin), ansamycins (e.g. rifamycin), antimycotics (e.g. polyenes and benzofuran derivatives), ⁇ -lactams (e.g.
  • penicillins and cephalosporins include chloramphenical (including thiamphenol and azidamphenicol), linosamides (lincomycin, clindamycin), macrolides (erythromycin, oleandomycin, spiramycin), polymyxins, bacitracins, tyrothycin, capreomycin, vancomycin, tetracyclines (including oxytetracycline, minocycline, doxycycline), phosphomycin and fusidic acid.
  • the CD83 nucleic acids, polypeptides or antibodies are well suited to formulation as sustained release dosage forms and the like.
  • the formulations can be so constituted that they release the active nucleic acids, polypeptide or antibody, for example, in a particular part of the intestinal or respiratory tract, possibly over a period of time.
  • Coatings, envelopes, and protective matrices may be made, for example, from polymeric substances, such as polylactide-glycolates, liposomes, microemulsions, microparticles, nanoparticles, or waxes. These coatings, envelopes, and protective matrices are useful to coat indwelling devices, e.g., stents, catheters, peritoneal dialysis tubing, draining devices and the like.
  • the therapeutic agents may be formulated as is known in the art for direct application to a target area.
  • Forms chiefly conditioned for topical application take the form, for example, of creams, milks, gels, dispersion or microemulsions, lotions thickened to a greater or lesser extent, impregnated pads, ointments or sticks, aerosol formulations (e.g., sprays or foams), soaps, detergents, lotions or cakes of soap.
  • Other conventional forms for this purpose include wound dressings, coated bandages or other polymer coverings, ointments, creams, lotions, pastes, jellies, sprays, and aerosols.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can be delivered via patches or bandages for dermal administration.
  • the nucleic acid, polypeptide or antibody can be formulated to be part of an adhesive polymer, such as polyacrylate or acrylate/vinyl acetate copolymer.
  • an adhesive polymer such as polyacrylate or acrylate/vinyl acetate copolymer.
  • the backing layer can be any appropriate thickness that will provide the desired protective and support functions. A suitable thickness will generally be from about 10 to about 200 microns.
  • Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents.
  • Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents.
  • the active CD83 nucleic acids, polypeptides or antibodies can also be delivered via iontophoresis, e.g., as disclosed in U.S. Pat. Nos. 4,140,122; 4,383,529; or 4,051,842.
  • the percent by weight of a therapeutic agent of the invention present in a topical formulation will depend on various factors, but generally will be from 0.01% to 95% of the total weight of the formulation, and typically 0.1-85% by weight.
  • Drops such as eye drops or nose drops, may be formulated with one or more of the therapeutic CD83 nucleic acids, polypeptides or antibodies in an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilizing agents or suspending agents.
  • Liquid sprays are conveniently delivered from pressurized packs. Drops can be delivered via a simple eye dropper-capped bottle, or via a plastic bottle adapted to deliver liquid contents dropwise, via a specially shaped closure.
  • the therapeutic nucleic acids, polypeptide or antibody may further be formulated for topical administration in the mouth or throat.
  • the active ingredients may be formulated as a lozenge further comprising a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the composition in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the composition of the present invention in a suitable liquid carrier.
  • the pharmaceutical formulations of the present invention may include, as optional ingredients, pharmaceutically acceptable carriers, diluents, solubilizing or emulsifying agents, and salts of the type that are available in the art.
  • pharmaceutically acceptable carriers such as physiologically buffered saline solutions and water.
  • diluents such as phosphate buffered saline solutions pH 7.0-8.0.
  • the CD83 nucleic acids, polypeptides or antibodies of the invention can also be administered to the respiratory tract.
  • the present invention also provides aerosol pharmaceutical formulations and dosage forms for use in the methods of the invention.
  • dosage forms comprise an amount of at least one of the agents of the invention effective to treat or prevent the clinical symptoms of a specific infection, indication or disease. Any statistically significant attenuation of one or more symptoms of an infection, indication or disease that has been treated pursuant to the method of the present invention is considered to be a treatment of such infection, indication or disease within the scope of the invention.
  • the composition may take the form of a dry powder, for example, a powder mix of the therapeutic agent and a suitable powder base such as lactose or starch.
  • the powder composition may be presented in unit dosage form in, for example, capsules or cartridges, or, e.g., gelatin or blister packs from which the powder may be administered with the aid of an inhalator, insufflator, or a metered-dose inhaler (see, for example, the pressurized metered dose inhaler (MDI) and the dry powder inhaler disclosed in Newman, S. P. in Aerosols and the Lung , Clarke, S. W. and Davia, D. eds., pp. 197-224, Butterworths, London, England, 1984).
  • MDI pressurized metered dose inhaler
  • the dry powder inhaler disclosed in Newman, S. P. in Aerosols and the Lung , Clarke, S. W. and Davia, D. eds., pp. 197
  • CD83 nucleic acids, polypeptides or antibodies of the present invention can also be administered in an aqueous solution when administered in an aerosol or inhaled form.
  • other aerosol pharmaceutical formulations may comprise, for example, a physiologically acceptable buffered saline solution containing between about 0.1 mg/ml and about 100 mg/ml of one or more of the CD83 nucleic acids, polypeptides or antibodies of the present invention specific for the indication or disease to be treated.
  • Dry aerosol in the form of finely divided solid nucleic acid, polypeptide or antibody particles that are not dissolved or suspended in a liquid are also useful in the practice of the present invention.
  • CD83 nucleic acids, polypeptides or antibodies of the present invention may be formulated as dusting powders and comprise finely divided particles having an average particle size of between about 1 and 5 ⁇ m, alternatively between 2 and 3 ⁇ m.
  • Finely divided particles may be prepared by pulverization and screen filtration using techniques well known in the art.
  • the particles may be administered by inhaling a predetermined quantity of the finely divided material, which can be in the form of a powder. It will be appreciated that the unit content of active ingredient or ingredients contained in an individual aerosol dose of each dosage form need not in itself constitute an effective amount for treating the particular infection, indication or disease since the necessary effective amount can be reached by administration of a plurality of dosage units. Moreover, the effective amount may be achieved using less than the dose in the dosage form, either individually, or in a series of administrations.
  • the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention are conveniently delivered from a nebulizer or a pressurized pack or other convenient means of delivering an aerosol spray.
  • Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas.
  • the dosage unit may be determined by providing a valve to deliver a metered amount.
  • Nebulizers include, but are not limited to, those described in U.S. Pat. Nos.
  • Aerosol delivery systems of the type disclosed herein are available from numerous commercial sources including Fisons Corporation (Bedford, Mass.), Schering Corp. (Kenilworth, N.J.) and American Pharmoseal Co., (Valencia, Calif.).
  • the therapeutic agent may also be administered via nose drops, a liquid spray, such as via a plastic bottle atomizer or metered-dose inhaler.
  • atomizers are the Mistometer (Wintrop) and the Medihaler (Riker).
  • the active ingredients may also be used in combination with other therapeutic agents, for example, pain relievers, anti-inflammatory agents, antihistamines, bronchodilators and the like, whether for the conditions described or some other condition.
  • other therapeutic agents for example, pain relievers, anti-inflammatory agents, antihistamines, bronchodilators and the like, whether for the conditions described or some other condition.
  • the present invention further pertains to a packaged pharmaceutical composition for controlling microbial infections such as a kit or other container.
  • a packaged pharmaceutical composition for controlling microbial infections such as a kit or other container.
  • the kit or container holds a therapeutically effective amount of a pharmaceutical composition for modulating immune responses and instructions for using the pharmaceutical composition for control of the immune response.
  • the pharmaceutical composition includes at least one nucleic acid, polypeptide or antibody of the present invention, in a therapeutically effective amount such that the selected disease or immunological condition is controlled.
  • mice Male C57BL6 mice received 3 weekly injections of N-ethyl-N-nitrosourea (ENU) at a concentration of 100 mg /kg. N-Ethyl-N-nitrosourea was quantified prior to injection by spectrophotometry. Mice that regained fertility after a minimum period of 12 weeks were then used to generate pedigree founder G1 animals. G1 male mice were crossed to C57BL6J females and their female progeny (G2 animals) crossed back to their fathers to generate G3 animals for screening.
  • ENU N-ethyl-N-nitrosourea
  • G3 mice were weaned at 3 weeks of age. Each animal then underwent a series of screens designed to assess a number of parameters, including immune function, inflammatory response and bone development.
  • 150-200 ⁇ l of whole blood was collected by retro-orbital bleed into heparinized tubes. Cells were pelleted and red blood cells lysed. Samples were then stained with antibodies to cell surface markers expressed on distinct lymphoid and myeloid sub-populations. These samples were analyzed by flow-cytometry.
  • the number of phenodeviants identified (2 from a litter of 9 animals) was suggestive of a trait controlled by a single gene and inherited in a Mendelian fashion.
  • the phenotype was designated LCD4.1 (Low CD4 Mutant # 1) and was used for mapping experiments.
  • DNA samples were prepared from samples of tail tissue collected from these N2 mice and used for a genome scan, using a collection of assembled markers, and performed on the ABI 3100 DNA analyzer.
  • Initial genetic linkage was seen to the tip of chromosome 13, where the closest microsatellite marker was D 13Mit139 with a LOD score of 8.2.
  • the mutant gene was located between 13.4 and 29.6 cM on chromosome 13. Through additional genotyping, this region was reduced to an 11 cM interval on chromosome 13. No significant linkage to other chromosomal regions was seen.
  • CD83 A candidate gene, CD83, was identified for gene-testing based upon its reported position within the interval. CD83 has previously been used as a marker of dendritic cell activation, suggesting that it might play a role in dendritic cell function and hence in regulating T cell development and function.
  • Dendritic cells can be differentiated from bone marrow of wild type mice by culture in GM-CSF. These cells can be characterized by the surface expression of dendritic cell markers, including CD86 and CD 11 c. Both LCD4.1 affected and normal animals were capable of giving rise to CD86+CD11c+cells under these culture conditions. LCD4.1 mutant mice thus were capable of generating dendritic cells under in vitro culture conditions. These data suggest that the phenotype seen in LCD4.1 mice is not due to a failure of dendritic cells to develop but rather may reflect a defect in function.
  • the sensitizing agent FITC was applied to the dorsal surface of the ears of LCD4.1 affected and wild-type mice. FITC was picked up by dendritic cells that then migrated to the draining auricular lymph nodes, where the presence of the FITC label on the dendritic cell surface permitted detection by flow-cytometry. FITC labeled cells expressing CD86 were detected in equal proportions in draining lymph node from normal and affected LCD4.1 mice. These data indicate that LCD4.1 mutant animals are capable of generating dendritic cells in vivo and that these cells are able to pick up antigen in the ear and travel to the draining lymph node.
  • Spleens were removed from wild type and mutant mice and digested with collagenase to liberate dendritic cells. Spleens were stained for surface expression of CD4 (helper T cells) and CD 11c (dendritic cells). Cells expressing these markers were purified by fluorescence activated cell sorting (FACS sorting). CD 11c and CD4+positive cells were also purified from an allogeneic mouse strain, BALBc.
  • CD83 may have a novel requirement for CD83 functionality on T cells during allogeneic activation.
  • CD83 may be influencing the extent of CD4+ T cell activation or altering the duration of the CD4+ T cell proliferative response.
  • the therapeutic manipulation of CD83 may thus represent a mechanism for the specific regulation of T cell function in the treatment of T cell mediated diseases, including autoimmune disorders.
  • Antibodies capable of blocking CD83 function may be used as therapeutics in the treatment of immune diseases whilst the activation of CD83 may-have utility in enhancing immune responses in cancer and other circumstances.
  • CD83 has been described as a marker of dendritic cell activation there has previously been little data describing its function in vivo.
  • the mutation provided by the invention destabilizes or inactivates the protein and leads to impaired surface expression.
  • CD4+ T cell function is impaired.
  • the development of dendritic cells is not inhibited and mutant dendritic cells retain functionality. Nonetheless, the result is impaired development of CD4+ T cells. This impaired ability to activate T cells is also seen in a slight decrease in contact sensitivity responses in LCD4.1 mutant mice.
  • This Example demonstrates that CD4 + T-cells from CD83 mutant animals express higher levels of IL-4 and lower levels of IL-2 compared to CD4 + T-cells from CD83 wild type animals.
  • 20,000 CD4 + T-cells (either wild type or CD83 mutant) were added in a 200 ⁇ L final volume of RPMI containing 10% FBS, 55 ⁇ M ⁇ -mercaptoethanol and antibiotics. The plates were then incubated in a CO 2 incubator at 37° C. for 44 to 72 hours.
  • cytokine levels supernatants were harvested and cytokines were measured using either a Cytometric Bead Array system (Pharmingen) or ELISA (R&D).
  • RNA measurements the cells were harvested and RNA was isolated using Tri reagent (Sigma). IL-10 and IL-4 mRNA levels were measured by reverse transcription and TaqMan (Applied Biosystems) analysis.
  • FIG. 7 shows the IL-2, IL-4, IL-5, TNFa and IFN? levels produced by either wild type or CD83 mutant CD4 + T-cells.
  • Purified cells were incubated as described above in the presence of 1 ⁇ g/mL of anti-CD3 and 0.2 ⁇ g/mL of anti-CD28 antibodies for 72 hours. The supernatants were then simultaneously analyzed for production of IL-2, IL-4, IL-5, TNFa and IFN? using the cytometric bead array system from Pharmingen.
  • FIG. 7 demonstrates that CD4 + T-cells from CD83 mutant animals expressed higher levels of IL-4 and lower levels of IL-2 compared to CD4 + T-cells from CD83 wild type animals.
  • Other cytokines and a new set of stimulation assays were analyzed including the production levels of IL-10 and GMCSF by these cells (FIGS. 8 and 9). In both cases, cells from mutant animals produce larger amounts of IL-10 and GMCSF than did wild type animals.
  • FIG. 10 shows that mRNA levels for both IL-4 and IL-10 were increased in cells from activated mutant CD83, CD4 + T-cells compared with cells from wild type animals.
  • CD4 + T-cells were isolated and activated as described above. Activation was performed in the presence of increasing concentrations of anti-CD83 antibodies.
  • CD4 + T-cells were isolated from an OT2tg mouse. OT2tg mice are transgenic mice with a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide.
  • OVA ovalbumin
  • Dendritic cells were isolated from a C57BL6 mouse by a negative selection using B220 magnetic beads (Miltenyi Biotec) followed by positive selection using CDl 1-c magnetic beads (Milteny Biotec).
  • CD4 + T-cells Five thousand CD4 + T-cells were then mixed with five thousand dendritic cells in a 96 well plate in the presences of 1 ⁇ M OVA peptide using RPMI (55 ⁇ M BME, 10% FBS plus antibiotics) in a final 200uL volume. These cells were then incubated for 48 to 72 hours in a CO 2 incubator at 37° C. and pulsed using [ 3 H] thymidine for 8 hours. Cells were then harvested and [ 3 H] thymidine incorporation was quantified using a top counter.
  • RPMI 55 ⁇ M BME, 10% FBS plus antibiotics
  • anti-CD83 antibodies decreased production of IL-4 by activated CD4 + T-cells in a dose dependent manner. Different antibody preparations did provide somewhat different degrees of inhibition of IL-4 production (FIG. 11). Accordingly, the epitope and/or degree of affinity of the antibodies for the CD83 antigen may influence whether or not IL-4 production is significantly inhibited.
  • CD4 + T-cells derived from these TCR transgenic animals express high levels of a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide and thus have high levels of proliferation when mixed with antigen presenting cells (dendritic cells were used) in the presence of the OVA peptide.
  • OVA ovalbumin
  • anti-CD83 antibodies were able to decrease proliferation of CD4 + T-cells in this system (FIG. 12).
  • different antibody preparations had somewhat different effects on the proliferation of CD4 + T-cells. Accordingly, the CD83 epitope and/or degree of affinity of the antibodies for the CD83 antigen may influence whether or not CD4 + T-cell proliferation is significantly inhibited.
  • a 34.3 kb fragment of normal mouse genomic DNA, including the ⁇ 18 kb coding region of the CD83 gene, as well as ⁇ 10.6 kb of upstream flanking sequences and ⁇ 5.7 kb of downstream sequences was microinjected into normal mouse one-cell embryos.
  • Four individual founder animals were generated.
  • Transgenic mice were then crossed to a male OT2tg mouse.
  • Male offspring carrying both the CD83 and OT2 transgene were used to analyze peptide specific T-cell proliferation.
  • CD4 + T-cells and dendritic cells were isolated from either OT2tg [transgenic mice with a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide] CD83 wild type or from OT2tg CD83 transgenic mice as described above (Example 4).
  • OT2tg CD4 + T-cells from either wild type or CD83 transgenic animals were then mixed with five thousand wild type dendritic cells or five thousand CD83 transgenic dendritic cells in a 96 well plate in the presence of increasing concentrations of OVA peptide using RPMI (55 ⁇ M BME, 110% FBS plus antibiotics) in a final 200uL volume.
  • OT2tg CD4 + T-cells derived from CD83 transgenic mice proliferated at higher rates than the same cell population derived from a CD83 wild type animal (FIG. 13). This increased proliferation was seen at all the concentrations of OVA peptide tested. Whereas OT2tg CD4 + T-cells derived from CD83 transgenic animals exhibited increased proliferation, dendritic cells from CD83 transgenic animals did not exhibit a substantial increase in proliferation. Therefore, it appears that transgenic expression in the CD4 + T-cell, and not in dendritic cells is what led to the increased proliferation of CD4 + T-cells.
  • This Example shows that antibodies raised against the CD83 protein can inhibit proliferation of human peripheral blood mononuclear cells.
  • Rabbit polyclonal sera was raised against mouse CD83 protein by immunizing rabbits using a mouse CD83 external domain protein fused to a rabbit Ig domain (FIG. 14). Pre-immune sera and anti-mouse polyclonal sera were then purified using a protein A column (Pharmacia Biotech) as described by the manufacturer, then dialyzed against PBS and stored at 4° C. To monitor the recognition of mouse CD83 protein by the polyclonal sera, which was obtained at different dates post immunization, a titer was obtained using an antigen specific ELISA (FIG. 15). As illustrated by FIG. 15, a good polyclonal response was obtained against the mouse CD83 protein.
  • PBMCs Human peripheral blood mononuclear cells
  • PBMCs Human peripheral blood mononuclear cells
  • media RPMI, 10% FBS, antibiotics
  • PHA Phaseolus vulgaris leucoagglutinin
  • a Selected Lymphocyte Antibody Method (SLAM) procedure was used to establish monoclonal antibody cell lines from the rabbits used to generate the anti-CD83 antibodies.
  • Antibody forming cells were isolated from the immunized rabbits that produced antibodies capable of binding CD83, the genes encoding antibodies that recognized CD83 and inhibited proliferation of lymphocytes were then cloned by PCR amplification and sequenced. Separate lines of monoclonal antibody producing cells were then established and expanded in culture. Antibodies were purified using Protein A chromatography according to manufacturer's instructions and tested for their ability to recognize CD83 proteins and to inhibit proliferation of PHA stimulated human PBMCs.
  • FIG. 16 illustrates that proliferation of PHA-activated human PBMCs was inhibited by polyclonal antibodies raised against the external region of the mouse CD83 protein. Proliferation of PHA-activated human PBMCs was not affected by addition of increasing concentrations of protein A purified rabbit pre-immune sera. When increasing concentrations of protein A purified rabbit polyclonal sera raised against the mouse CD83 protein was added, a concentration dependent decrease in proliferation was observed.
  • 96G08 antibodies appeared to have reduced affinity for human CD83 protein via the Biacore and ELISA assays, the FACS assay indicated that this antibody preparation could bind to endogenously produced human CD83 (FIGS. 18 and 19). Moreover, the 96G08 antibodies were able to inhibit proliferation of human peripheral blood mononuclear cells (PBMCs), as illustrated in FIG. 20. Hence, some aspect of either the purification or the structure of the isolated recombinant human protein may have influenced the in vitro binding of 96G08 antibodies to the recombinant human CD83.
  • PBMCs peripheral blood mononuclear cells
  • the recombinant human CD83 protein employed for the Biacore and ELISA assays is a chimeric protein that is joined to a portion of an immunoglobulin Fc fragment. Removal of this Fc fragment may improve in vitro binding to the human CD83 protein.
  • FIG. 20 illustrates that the 95F04 and 96G08 antibody preparations can inhibit proliferation of PHA activated human peripheral blood mononuclear cells as detected by incorporation of [ 3 H] thymidine.
  • the 95F04 and 96G08 antibody preparations can inhibit proliferation of PHA activated human peripheral blood mononuclear cells as detected by incorporation of [ 3 H] thymidine.
  • no antibody was present about 10,000 cpm of [ 3 H] thymidine was incorporated into human peripheral blood mononuclear cells.
  • incorporation of [ 3 H] thymidine dropped to about 2000 cpm.
  • the 96G08 antibody preparation had an even greater effect on [ 3 H] thymidine incorporation.
  • This Example shows that antibodies raised against the CD83 protein as described in the previous example are particularly effective at inhibiting proliferation of immune cells after the antibodies are multimerized or multimerized by binding the antibodies to a solid support or by cross-linking in solution.
  • Mouse (C57B 16) spleen cells were isolated and plated in the antibody or control treated wells at 30,000 cells per well. For activation, Concavalin A was added to a final concentration of 1.0 ⁇ g/ml. Cellular proliferation was assessed by measuring the incorporation of tritiated thymidine during the last 6 to 8 hours of a 48 hour incubation. In another experiment, the specificity of the observed antibody-induced inhibition of lymphocyte proliferation was tested by repeating this experiment with addition of mouse CD83 protein before adding the lymphocytes to the antibody coated microtiter wells.
  • the 6G05 antibody preparation was identified as a good inhibitor of lymphocyte proliferation.
  • the 112D08 antibody preparation was identified as having little or no inhibitory activity when bound to microtiter wells.
  • the 112D08 antibody preparation was used as a negative control in some of the subsequent experiments.
  • Plate-bound 6G05 antibodies were prepared as described above. Approximately 30,000 activated lymphocytes were added per well to antibody coated plates or to non-coated plates containing 1.0 or 5.0 ⁇ g/ml soluble 6G05 antibody preparation. A secondary rabbit anti-mouse antibody (10 ⁇ g/ml or 25 ⁇ g/ml) was added to the wells containing the soluble 6G05 antibody preparation to act as a cross-linking reagent for the 6G05 antibodies. Cellular proliferation was assessed by incorporation of tritiated thymidine as described above.
  • FIGS. 25 A-B The results of one screen for anti-CD83 antibody preparations that can inhibit lymphocyte proliferation are shown in FIGS. 25 A-B.
  • many anti-CD83 antibody preparations inhibit proliferation of activated lymphocytes, including the 94c09, 98a02, 94d08, 98d11, 101b08, 6g05, 20d04, 14c12, 11g05, 12g04, 32f12 and 98b11 preparations. Note that some variation in the degree of inhibition obtained is observed. For example, while the 98b 1 preparation is not so effective, the 6g05 antibody preparation is a highly effective inhibitor of lymphocyte proliferation.
  • FIG. 25B further illustrates that some antibody preparations are highly effective inhibitors (e.g. 117G12) but others are not (e.g. 98g08).
  • the 824pb antibody refers to rabbit polyclonal antisera; as shown this polyclonal antisera was not particularly effective at inhibiting lymphocyte proliferation
  • FIG. 26 illustrates that the inhibitory activity of the 6g05 antibody preparation is quenched by soluble mouse CD83 protein.
  • mouse CD83 protein was added to anti-CD83 antibody-coated wells before activated lymphocytes were introduced. Both a highly effective proliferation inhibitor (6g05) and an antibody preparation with little or no inhibitory activity (98g08) were tested. A control having no antibody and no mouse CD83 protein as well as a control with added mouse CD83 and no antibody was included. Cellular proliferation of the activated lymphocytes was assessed by observing the incorporation of tritiated thymidine as described above. As shown in FIG. 26, the 6g05 antibody strongly inhibits lymphocyte proliferation when no mouse CD83 is present.
  • FIGS. 27 and 28 illustrate that anti-CD83 antibodies that are multimerized by use of a rabbit anti-mouse antibody have inhibitory activity that is like that of plate-bound anti-CD83 antibodies.
  • the proliferation of lymphocytes was measured by observing the incorporation of tritiated thymidine with and without anti-CD83 antibodies as described above.
  • plate-bound 6g05 antibodies were used and in another soluble 6g05 antibodies were employed.
  • the soluble 6g05 antibodies were cross-linked by addition of rabbit anti-mouse antibodies that bind to the Fc region of the 6g05 antibodies.
  • a soluble and plate-bound antibody preparation with no inhibitory activity was also tested.
  • a similar series of assays were set up using a panel of soluble anti-CD83 antibodies.
  • both plate-bound and crosslinked 6g05 antibodies were highly effective inhibitors of lymphocyte proliferation.
  • the method of aggregating anti-CD83 antibodies is not particularly important.
  • the multimerization can be achieved by adhering or attaching antibodies to a solid support or by crosslinking the anti-CD83 antibodies through their Fc regions using a rabbit anti-mouse secondary antibody. So long as the anti-CD83 antibodies are in close proximity, they are effective inhibitors of lymphocyte proliferation.
  • FIG. 28 shows that many soluble anti-CD83 antibodies exhibit good inhibition of lymphocyte proliferation when they are cross-linked with the rabbit anti-mouse secondary antibody.
  • the 6g05, 11g04, 12g04, 14c12, 20d04, 32f12, 94c09, 94d08, 98a02, 98d11(3), 101B08(2.7) and 117g12 antibody preparations strongly inhibit lymphocyte multimerization when cross-linked with the rabbit anti-mouse antibodies.
  • This Example shows that multimerized anti-CD83 antibodies inhibit proliferation of lymphocytes in a mixed lymphocyte reaction (MLR) assay.
  • MLR mixed lymphocyte reaction
  • MLR assay employed was a modification of the procedure described in Bradley, pp 162-166 in Mishell et al., eds. Selected Methods in Cellular Immunology (Freeman, San Francisco, 1980); and Battisto, et al., Meth, in Enzymol. 150:83-91 (1987).
  • Spleens were removed from BALBc and C57B 16 mice and digested with collagenase to liberate dendritic and CD4 + cells, respectively. Spleens were stained for surface expression of CD4 (helper T cells) or CD11c (dendritic cells). Cells expressing these markers were purified by using magnetic beads (Miltenyi) according to the manufacturer's instructions.
  • FIG. 29 shows that the conditions employed several monoclonal anti-CD83 antibodies can inhibit lymphocyte proliferation in a mixed lymphocyte reaction assay.
  • the 98a02, 98d11, 20d04, 14c12, 12g04, and 117g12 inhibit lymphocyte proliferation in this assay.
  • FIG. 30 shows that many anti-CD83 antibody preparations can inhibit the recall response of lymphocytes.
  • 94c09, 98a02, 6g05, 20d04, and 117104 antibody preparations inhibited proliferation of activated lymphocytes exposed to an antigen (KLH) to which they had been immunized.
  • KLH antigen
  • anti-CD83 antibodies can quiet the proliferative response of CD4+ T cells after stimulation by allogenic CD11 cells and/or antigen.
  • This Example shows that exposure to anti-CD83 antibodies does not lead to apoptosis of activated lymphocytes.
  • Mouse (C57B 16) spleen cells were isolated and activated by incubation for 24 hours with 1.0 ⁇ g/ml Concavalin A in the presence or absence of anti-CD83 antibodies and rabbit anti-mouse antibodies as a crosslinking reagent as described above. Cells were incubated for 48 hours at 37° C. Proliferative responses were measured by incorporation of tritiated thymidine. Total caspase activity and annexinV expression levels were used as a measure of apoptosis.
  • FIGS. 31 A-B shows that soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations not only inhibit activated lymphocyte cell proliferation (FIG. 31B) but also have very low caspase activity (FIG. 31A).
  • FIG. 32 shows that the percentage of activated lymphocytes that express annexinV is reduced after treatment with soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations.
  • anti-CD83 antibodies inhibit proliferation of ConA activated splenocytes, they do not induce apoptosis of immune cells. Instead, anti-CD83 antibodies actually depress the expression of apoptosis markers. Hence, the reduction in cell proliferation observed when activated lymphocytes are exposed to anti-CD83 antibodies is not due to increased programmed cell death.
  • Mouse (B6) spleen cells were isolated and activated using Concavalin A as described above in the presence or absence of anti-CD83 antibodies and the secondary anti-mouse crosslinking antibodies.
  • FIG. 33 illustrates that splenocytes activated with Concavalin A express the CD69 activation marker even though they were incubated with anti-CD83 antibodies.
  • the star or asterisks in the lower right hand corner of the graph shows the level of CD69 expression observed when splenocytes are not activated with Concavalin A.
  • splenocytes were activated with Concavalin A they expressed high levels of CD69 even after incubation with any of the 6g05, 14c12, 98b11 or 112d08 anti-CD83 antibody preparations.
  • Anti-CD83 Antibodies Arrest the Lymphocyte Cell Cycle in the G0/G1 Stage
  • This Example shows that exposure to anti-CD83 antibodies arrests activated lymphocytes in the G0/G 1 stage of the cell cycle.
  • Mouse (B6) spleen cells were isolated and activated by incubation for 48 hours with 1.0 ⁇ g/ml Concavalin A in the presences of anti-CD83 antibodies with the crosslinking antibodies as described above.
  • cells were fixed and DNA was stained with propidium iodine according to the protocol described for the flowcytometer (Cold Spring Harbor, N.Y.).
  • WinMDI software was used for background subtraction caused by debris in the DNA histogram. Each histogram was further analyzed by cycle red software to obtain the distribution of cells therein.
  • the size and shape of the activated cells was assessed by their forward (FSC) and side (SSC) scatter during this experiment.
  • the anti-CD83 antibody preparations employed were the 6g05 and 14c12 preparations that had been shown to inhibit cellular proliferation and the 112d08 preparation that had little or no effect on cellular proliferation.
  • Cells having 2N complement of DNA were assumed to be in the G 1/G0 phase of the cell cycle; cells having 3N complement of DNA were assumed to be in the G2/M phase of the cell cycle; and cells having 4N complement of DNA were assumed to be in the S phase of the cell cycle.
  • the percentage of cells having G1/G0, G2/M or S phase of the cell cycle was determined and plotted in FIGS. 35 A-C.
  • FIG. 34 shows that a population of activated splenocytes mixed with anti-CD83 antibody preparations have lost the blasting (dividing) cells as detected by FACS sorting. Almost all cells sort as small cells with a 2N content of DNA as illustrated by the high proportion of cells towards the left (smaller) side of the population distribution in FIG. 34.
  • FIGS. 35 A-C show that treatment of Concavalin A activated lymphocytes with either of 6g05 and 14c12 antibody preparations leads to a cellular population that was enriched in cells in the G 1/G0 stage of the cell cycle. Treatment with either the rabbit anti-mouse antibody or the 112d08 antibody preparation that has little or no effect on cell proliferation did not lead to a cellular population that was enriched in cells in the G1/G0 stage of the cell cycle.
  • a host cell includes a plurality (for example, a culture or population) of such host cells, and so forth.
  • a reference to “a host cell” includes a plurality (for example, a culture or population) of such host cells, and so forth.
  • the patent be interpreted to be limited to the specific examples or embodiments or methods specifically disclosed herein.
  • the patent be interpreted to be limited by any statement made by any Examiner or any other official or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or reservation expressly adopted in a responsive writing by Applicants.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention provides methods for modulating the immune system using anti-CD83 antibodies that can influence CD83 function.

Description

  • This application is a continuation under 35 U.S.C. 111(a) of International Application No. PCT/US02/37738 filed Nov. 21, 2002 and published in English as [0001] WO 03/045318 on Jun. 5, 2003, which claimed priority under 35 U.S.C. 119(e) from U.S. Provisional Application Ser. No. 60/331,958 filed Nov. 21, 2001, which applications and publication are incorporated herein by reference.
  • This application also claims priority to U.S. Provisional Application Ser. No. 60/428,130 filed Nov. 21, 2002 and U.S. Provisional Application Ser. No. 60/473,279 filed May 22, 2003 which are incorporate here by reference.[0002]
    • FIELD OF THE INVENTION
  • The invention relates to multimerized antibodies directed against the CD83 gene product, and methods of modulating the immune response of an animal by using such multimerized antibodies. [0003]
    • BACKGROUND OF THE INVENTION
  • CD83 is a 45 kilodalton glycoprotein that is predominantly expressed on the surface of dendritic cells and other cells of the immune system. Structural analysis of the predicted amino acid sequence of CD83 indicates that it is a member of the immunoglobulin superfamily. See, Zhou et al., J. Immunol. 149:735 (1992)). U.S. Pat. No. 5,316,920 and WO 95/29236 disclose further information about CD83. While such information suggests that CD83 plays a role in the immune system, that role is undefined, and the interrelationship of CD83 with cellular factors remains unclear. [0004]
  • Moreover, treatment of many diseases could benefit from more effective methods for increasing or decreasing the immune response. Hence, new reagents and methods are needed for modulating the immune system through the CD83 gene and its gene product. [0005]
    • SUMMARY OF THE INVENTION
  • The invention provides methods for modulating an immune response. In one aspect, the invention relates to the surprising discovery that multimerized antibodies raised against the CD83 gene product can arrest cellular proliferation of immune cells. Hence, the invention provides a method of modulating the immune response by modulating the activity or expression of the CD83 gene products, for example, by using such multimerized antibodies. [0006]
  • Also according to the invention, the production of a cytokine such as interleukin-2, interleukin-4, or interleukin-10 can be modulated by modulating the activity or expression of a CD83 polypeptide. In some embodiments, a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide. For example, the antibody can be administered to the mammal or the immune cell can be contacted with the antibody. In some embodiments, the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4[0007] + T cells.
  • The invention also provides a method of modulating granulocyte macrophage colony stimulating factor production in a mammal or in an immune cell by modulating the activity or expression of CD83 polypeptides. In some embodiments, an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide. For example, the antibody can be administered to the mammal or the immune cell can be contacted with the antibody. In some embodiments, the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4[0008] + T cells.
  • The invention also provides a method of modulating tumor necrosis factor production in a mammal or in a mammalian cell by modulating the activity or expression of CD83 polypeptides. In some embodiments, an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide. For example, the antibody can be administered to the mammal or the mammalian cell can be contacted with the antibody. In some embodiments, the immune cells are T cells or antigen presenting cells. In other embodiments, the immune cells are CD4[0009] + T cells.
  • The invention further provides a method of inhibiting proliferation of a human peripheral blood mononuclear cell by modulating the activity or expression of CD83 polypeptides. In some embodiments, an antibody or a multimerized antibody is used that can modulate the activity or expression of a CD83 polypeptide. For example, the antibody can be administered to the mammal or the human peripheral blood mononuclear cell can be contacted with the antibody. [0010]
  • The invention also provides an antibody that can bind to a CD83 polypeptide comprising SEQ ID NO:4, SEQ ID NO:8 or SEQ ID NO:9, wherein activated CD4[0011] + T-cells produce lower levels of interleukin-4 when the T-cells are contacted with the antibody. The invention further provides an antibody that can bind to a CD83 polypeptide comprising SEQ ID NO:4, SEQ ID NO:8 or SEQ ID NO:9, wherein CD4+ T-cells proliferation is decreased when the T-cells are contacted with the antibody. The antibody can be a multimerized antibody. Such multimerized antibodies can be bound to a solid support, covalently crosslinked or bound together by a second entity such as a secondary antibody. Examples of antibodies of the invention include those that have an amino acid sequence that includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. Nucleic acids encoding such an antibody can have, for example, a sequence that includes SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85 or SEQ ID NO:90.
  • The invention also provides a method for decreasing the activity of a CD83 gene product, comprising contacting the CD83 gene product with an antibody that comprises amino acid sequence includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. The antibody can be a multimerized antibody. The activity of a CD83 gene product can be decreased in a mammal or in a cell that is involved in an immune response, for example, a T cell. [0012]
  • The invention further provides a method for decreasing the translation of a CD83 gene product in a mammalian cell, comprising contacting the mammalian cell with a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. [0013]
  • In another embodiment, the invention provides a method for decreasing the translation of a CD83 gene product in a mammal, comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. [0014]
  • The invention further provides a method for decreasing proliferation of CD4[0015] + T-cells in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9. The antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. The antibody can be a multimerized antibody.
  • The invention also provides a method for decreasing interleukin-2 levels and increasing interleukin-4 levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9. The antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. The antibody can be a multimerized antibody. [0016]
  • The invention further provides a method for decreasing interleukin-2 levels and increasing interleukin-4 levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. In some embodiments the interleukin-2 levels are decreased and the interleukin-4 levels are increased to treat an autoimmune disease. In other embodiments, the interleukin-2 levels are decreased and the interleukin-4 levels are increased to stimulate production of Th2-associated cytokines in transplant recipients, for example, to prolong survival of transplanted tissues. [0017]
  • The invention also provides a method for increasing interleukin-10 levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9. The antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. The antibody can be a multimerized antibody. [0018]
  • The invention further provides a method for increasing interleukin-10 levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. In some embodiments, the interleukin-10 levels are increased to treat neoplastic disease. In other embodiments, the interleukin-10 levels are increased to treat a tumor. [0019]
  • The invention also provides a method for increasing interleukin-2 levels in a mammal comprising administering to the mammal a functional CD83 polypeptide that comprises SEQ ID NO:9. [0020]
  • The invention further provides a method for increasing interleukin-2 levels in a mammal comprising: (a) transforming a T cell from the mammal with a nucleic acid encoding a functional CD83 polypeptide operably linked to a promoter functional in a mammalian cell, to generate a transformed T cell; (b) administering the transformed T cell to the mammal to provide increased levels of interleukin-2. In some embodiments, the CD83 polypeptide has a sequence that comprises SEQ ID NO:9 or the nucleic acid has a sequence that comprises SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. Such methods for increasing interleukin-2 levels can be used to treat an allergy or an infectious disease. [0021]
  • The invention also provides a method for increasing granulocyte macrophage colony stimulating factor levels in a mammal comprising administering to the mammal an antibody that can bind to a CD83 gene product, wherein the CD83 gene product comprises SEQ ID NO:2 or SEQ ID NO:9. [0022]
  • Such an antibody can have a sequence comprising includes SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99. The antibody can be a multimerized antibody. [0023]
  • The invention further provides a method for increasing granulocyte macrophage colony stimulating factor levels in a mammal comprising administering to the mammal a nucleic acid complementary to a CD83 nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:10. [0024]
  • The invention also provides a method for increasing tumor necrosis factor levels at a selected site in a mammal comprising administering to the site a functional CD83 polypeptide. In another embodiment, the invention provides a method for increasing tumor necrosis factor levels in a selected mammalian cell comprising transforming the cell with a nucleic acid encoding a functional CD83 polypeptide. The CD83 polypeptide employed can, for example, have a sequence comprising SEQ ID NO:9. [0025]
  • Animals such as mammals and birds may be treated by the methods and compositions described herein. Such mammals and birds include humans, dogs, cats, and livestock, for example, horses, cattle, sheep, goats, chickens, turkeys and the like. [0026]
  • The invention further provides a mutant mouse that can serve as an animal model of diminished T cell activation or altered cytokine levels. The mutant mouse has an altered CD83 gene that produces a larger gene product, having SEQ ID NO:4 or containing SEQ ID NO:8. Also provided are methods of using the mutant mouse model to study the effects of cytokines on the immune system, inflammation, the function and regulation of CD83, T cell and dendritic cell activity, the immune response and conditions and treatments related thereto. Hence, the invention further provides a mutant mouse whose somatic and germ cells comprise a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8, wherein expression of the mutant CD83 gene reduces CD4+ T cell activation. The mutant CD83 gene can, for example, comprise SEQ ID NO:3. [0027]
  • The invention further provides a method of identifying a compound that can modulate CD4+ T cell activation comprising administering a test compound to a mouse having a mutant or wild type transgenic CD83 gene and observing whether CD4+ T cell activation is decreased or increased. The somatic and/or germ cells of the mutant mouse can comprise a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8. Alternatively, the somatic and/or germ cells of the mouse can contain a wild type CD83 gene, for example, SEQ ID NO:1 or SEQ ID NO:9. [0028]
  • The invention also provides a mutant CD83 gene encoding a polypeptide comprising SEQ ID NO:4 or SEQ ID NO:8. The invention further provides a mutant CD83 gene comprising nucleotide sequence SEQ ID NO:3.[0029]
    • DESCRIPTION OF THE FIGURES
  • FIG. 1 summarizes flow cytometry data for G3 animals. As shown, reduced numbers of CD4+ T cells are seen in two animals from [0030] Pedigree 9, mouse 9.4.1 and mouse 9.4.9. All other animals analyzed on that day exhibit normal numbers of CD4+ T cells.
  • FIG. 2 provides a graph of flow cytometry data for G3 animals that may have a mutant CD83 gene. Each diamond symbol represents an individual animal. As shown, multiple animals from the N2 generation exhibit a reduced percentage of CD4+ T cells. [0031]
  • FIG. 3 provides the nucleotide sequence of wild type mouse CD83 (SEQ ID NO:1). The ATG start codon and the TGA stop codon are underlined. [0032]
  • FIGS. [0033] 4A-B provides the nucleotide sequence of the mutant CD83 gene (SEQ ID NO:3) of the invention derived from the mutant LCD4.1 animal. The ATG start codon, the mutation and the TGA stop codon are underlined.
  • FIG. 5 provides the amino acid sequence for wild type (top, SEQ ID NO:2) and mutant (bottom, SEQ ID NO:4) CD83 coding regions. The additional C-terminal sequences arising because of the CD83 mutation are underlined. [0034]
  • FIG. 6A illustrates that dendritic cells from wild type (?, WT DC) and mutant (¦, mutant DC) mice are capable of the allogeneic activation of CD4+ T cells. CD4+ T cells were stimulated with 10,000, 1000 or 100 dendritic cells for 5 days and proliferation was measured by incorporation of tritiated thymidine. [0035]
  • FIG. 6B illustrates that CD4[0036] + T cells from mutant mice (¦, mutant CD4) fail to respond to allogeneic stimulation with BALBc dendritic cells, although wild type animals (?, WT CD4+) respond normally. CD4+ T cells were stimulated with 10,000, 1000 or 100 dendritic cells for 5 days and proliferation measured by incorporation of tritiated thymidine.
  • FIG. 7 provides a bar graph illustrating IL-2, IL-4, IL-5, TNFa, and IFN? production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with 1 μg/ml of anti-CD3 antibodies and 0.2 μg/ml of anti-CD28 antibodies for 72 hours. As illustrated, IL-2 levels are lower, and IL-4 levels are higher in the CD83 mutant T cells. [0037]
  • FIG. 8 provides a bar graph illustrating IL-10 production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with 0.1 μg/ml of anti-CD28 antibodies and 1 to 10 μg/ml of anti-CD3 antibodies for 72 hours. As illustrated, IL-10 levels are higher in the CD83 mutant T cells. [0038]
  • FIG. 9 provides a bar graph illustrating GM-CSF production from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, GM-CSF production is higher in the CD83 mutant cells than in wild type cells. [0039]
  • FIG. 10A provides a bar graph illustrating IL-4 mRNA levels from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, the IL-4 mRNA levels are higher in the CD83 mutant cells. [0040]
  • FIG. 10B provides a bar graph illustrating IL-10 mRNA levels from wild type CD4+ T cells (white bar) or CD83 mutant CD4+ T cells (dark bar) that had been stimulated with anti-CD3 and anti-CD28 antibodies. As illustrated, the IL-10 mRNA levels are higher in the CD83 mutant cells. [0041]
  • FIG. 11 provides a graph illustrating that various preparations of anti-CD83 antibodies inhibit IL-4 production in anti-CD3 and anti-CD28 antibody stimulated T cells. The amount of IL-4 produced by T cells in pg/ml is plotted versus the concentration of different anti-CD83 antibody preparations, including the 20B08 (?) anti-CD83 preparation, the 20D04 (¦) anti-CD83 preparation, the 14C12 (?) anti-CD83 preparation and the 11 G05 (X) anti-CD83 antibody preparation. [0042]
  • FIG. 12 provides a graph illustrating that various preparations of anti-CD83 antibodies inhibit T cell proliferation. The graph plots the incorporation of radioactive thymidine in cpms, which was used as an indicator of the amount of T cell proliferation, versus the concentration of the different anti-CD83 antibody preparations, including the 20D04 (?) anti-CD83 preparation, the 11G05 (¦) anti-CD83 antibody preparation, the 14C12 (?) anti-CD83 preparation and the 6G05 anti-CD83 preparation (X). [0043]
  • FIG. 13 provides a graph illustrating that transgenic mice that over-express wild type CD83 have increased T cell proliferation. The graph plots the incorporation of radioactive thymidine in cpms, which was used as an indicator of the amount of T cell proliferation, versus the concentration of OVA peptide. The transgenic mice utilized had a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide that can activate T-cells. When mixed with either transgenic or wild type dendritic cells in the presence of OVA peptide, transgenic CD4+ T cells had increased T-cell proliferation. However, transgenic dendritic cells could not substantially increase wild type CD4+ T cell proliferation. Transgenic CD83 CD4+T cells mixed with wild type dendritic cells (?); transgenic CD83 CD4+ T cells mixed with transgenic dendritic cells (¦); wild type CD4+ T cells mixed with transgenic dendritic cells (?); and wild type CD4+ T cells mixed with wild type dendritic cells (X). [0044]
  • FIG. 14 provides a schematic diagram of the structural elements included in the mouse CD83 protein used for generating antibodies. [0045]
  • FIG. 15 provides a graph of ELISA data illustrating the titer obtained for different isolates of polyclonal anti-CD83 anti-sera. The first (?), second (¦) and third (?) isolates had similar titers, though the titer of the second isolate (¦) was somewhat higher. [0046]
  • FIG. 16 illustrates that proliferation of PHA-activated human PBMCs was inhibited by antibodies raised against the external region of the mouse CD83 protein (?). Pre-immune serum (¦) had little effect on the proliferation of human PBMCs. [0047]
  • FIG. 17A provides a sequence alignment of anti-CD83 heavy chain variable regions isolated by the invention. Sequences for isolates 20B08H (SEQ ID NO:52), 6G05H (SEQ ID NO:53), 20D04H (SEQ ID NO:54), 11 G05 (SEQ ID NO:66) and 14C12 (SEQ ID NO:67) are provided. The CDR regions are highlighted in bold. [0048]
  • FIG. 17B provides a sequence alignment of anti-CD83 light chain variable regions isolated by the invention. Sequences for isolates 20B08L (SEQ ID NO:55), 6G05L (SEQ ID NO:56), 20D04L (SEQ ID NO:57), 11G05L (SEQ ID NO:68) and 14C12L (SEQ ID NO:69) are provided. The CDR regions are highlighted in bold. [0049]
  • FIG. 18 graphically illustrates that cells expressing CD83 can be detected and sorted using an anti-CD83 antibody preparation. In this study, a Hodgkin's lymphoma cell line, KMH2, and a commercially available anti-CD83 antibody preparation was used for FACS sorting. [0050]
  • FIGS. [0051] 19A-B shows that two antibody preparations of the invention can bind to endogenously produced human CD83, as detected by FACS sorting of KMH2 cells (see also FIG. 18). Note that “Beer” is another name used for CD83.
  • FIG. 20 illustrates that the 95F04 and 96G08 antibody preparations described herein can inhibit proliferation of human peripheral blood mononuclear cells as detected by [[0052] 3H] thymidine incorporation. As shown, when 30 μg/ml of the 95F04 (?) antibody preparation was present, incorporation of [3H] thymidine dropped to about 2000 cpm. When 30 μg/ml 96G08 antibody preparation (?) was added to human peripheral blood mononuclear cells, [3H] thymidine incorporation was reduced to about 300 cpm. A third antibody preparation (98B 11, ¦) provided slight inhibition of PBMC proliferation. These data indicate that the 95F04 and 96G08 antibody preparations can alter the function of human CD83 in vivo.
  • FIG. 21 provides nucleotide and amino acid sequences for the monoclonal antibody 96G08 light chain. [0053]
  • FIG. 22 provides nucleotide and amino acid sequences for the monoclonal antibody 96G08 heavy chain. [0054]
  • FIG. 23 provides nucleotide and amino acid sequences for the monoclonal antibody 95F04 light chain. [0055]
  • FIG. 24 provides nucleotide and amino acid sequences for the monoclonal antibody 95F04 heavy chain. [0056]
  • FIGS. [0057] 25A-B provides the results of one screen of anti-CD83 antibody preparations that were multimerized by binding them to microtiter plates. The plate-bound antibodies were screened for an ability to inhibit lymphocyte proliferation as measured by tritiated thymidine incorporation. As illustrated in FIG. 25A many plate-bound anti-CD83 antibody preparations inhibit proliferation of activated lymphocytes, including the 94c09, 98a02, 94d08, 98d11, 101b08, 6g05, 20d04, 14c12, 11g05, 12g04, 32f12 and 98b11 preparations. FIG. 25B further illustrates that some antibody preparations are highly effective inhibitors (e.g. 117G12) but others are not (e.g. 824pb and 98g08).
  • FIG. 26 illustrates that the inhibitory activity of the multimerized (plate-bound) 6g05 antibody preparation is quenched by soluble mouse CD83 protein (mCD83rFc). Lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown, the multimerized 6g05 antibody preparation is strongly inhibitory of proliferation when no CD83 protein is added. However, when the mouse CD83 protein is added to assay, little or no inhibition of lymphocyte proliferation is observed. The 98g08 antibody preparation was used as a negative control because it exhibited little or no lymphocyte inhibition in previous tests (see FIG. 25B). [0058]
  • FIG. 27 is a bar graph showing that both plate-bound and cross-linked 6g05 antibodies are highly effective inhibitors of lymphocyte proliferation. Lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown on the left side of the graph above “plate-bound” the presence of plate-bound 6g05 antibodies in the lymphocyte proliferation assay cause little tritiated thymidine incorporation (about 1000 cpm). Similarly, as shown on the right side of the graph above “1[0059] st Ab (1 μg/ml)” soluble cross-linked 6g05 antibodies also cause little tritiated thymidine incorporation (about 1800 cpm).
  • FIG. 28 is a bar graph showing that several preparations of soluble cross-linked anti-CD83 antibodies are highly effective inhibitors of lymphocyte proliferation. Antibody preparations were cross-linked with the rabbit anti-mouse secondary antibody and lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown, soluble cross-linked antibody preparations including the 6g05, 11g04, 12g04, 14c12, 20d04, 32f12, 94c09, 94d08, 98a02, 98d11(3), 101B08(2.7) and 117g12 preparations caused little tritiated thymidine incorporation. [0060]
  • FIG. 29 shows that soluble, multimerized anti-CD83 antibodies exhibit inhibitory activity in mixed lymphocyte reaction assays. A series of anti-CD83 antibody preparations that were cross-linked using a rabbit anti-mouse antibody and then screened for inhibition of CD4[0061] + T cellular proliferation after activation of the CD4+ T cells with CD11 cells in a mixed lymphocyte reaction assay. As shown, the 98a02, 98d11, 20d04, 14c12, 12g04, and 117g12 inhibit lymphocyte proliferation in this assay.
  • FIG. 30 shows that many anti-CD83 antibody preparations can inhibit the recall response of lymphocytes. BALBc mice were first immunized with keyhole limpet hemocyanin (KLH) prior to spleen removal and CD 11 c and CD4+cell isolation. CD11c and CD4+cells were mixed and added to microtiter wells coated with anti-CD83 antibodies. Lymphocyte proliferation was measured by tritiated thymidine incorporation. As shown, the 94c09, 98a02, 6g05, 20d04, and 117104 antibody preparations inhibited proliferation of activated lymphocytes exposed to an antigen (KLH) to which they had been immunized. [0062]
  • FIGS. [0063] 31A-B shows that soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations not only inhibit activated lymphocyte cell proliferation (FIG. 31B) but also have very low caspase activity (FIG. 31A). Caspase activity was determined using a fluorogenic substrate and plotted as relative fluorescent units (RFU) on the y axis.
  • FIG. 32 shows that the percentage of activated lymphocytes that express annexin V is reduced after treatment with soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations. [0064]
  • FIG. 33 shows that the activation marker CD69 is expressed on splenocytes that were activated with Concavalin A and exposed to anti-CD83 antibodies. The anti-CD83 antibodies employed were the 6g05, 14c12, 98b11 and 112d08 anti-CD83 antibody preparations that were shown to inhibit activated splenocyte proliferation. [0065]
  • FIGS. [0066] 34A-E shows that a population of activated splenocytes mixed with anti-CD83 antibody preparations have lost the blasting (dividing) cells as detected by FACS sorting. The antibody preparations employed were the rabbit anti-mouse antibody, called the 2nd Ab (FIG. 34A), the 6g05 antibody preparation (FIG. 34B), the 98b11 antibody preparation (FIG. 34C), the 14c12 antibody preparation (FIG. 34D), and the 112d08 antibody preparation (FIG. 34E). Almost all cells exposed to the 6g05 or 98b11 antibody preparations sort as small cells with a 2N content of DNA as illustrated by the high proportion of cells towards the left (smaller) side of the population distribution compared to cells exposed to the control 2nd Ab, 14c12 and 112d08 preparations in FIGS. 34A, C and E.
  • FIG. 35A shows that the proportion of cells in the G1/G0 phase of the cell cycle is increased when a population of activated splenocytes is treated with anti-CD83 antibody preparations. The antibody preparations employed were the control rabbit anti-mouse antibody, called the 2[0067] nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation. Both of the 6g05 and 14c12 antibody preparations arrest the activated splenocytes in the G1/G0 phase of the cell cycle.
  • FIG. 35B shows the proportion of cells in the G2/M phase of the cell cycle after a population of activated splenocytes is treated with anti-CD83 antibody preparations. The antibody preparations employed were the control rabbit anti-mouse antibody, called the 2[0068] nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation.
  • FIG. 35C shows that the proportion of cells in the S phase of the cell cycle is decreased when a population of activated splenocytes is treated with anti-CD83 antibody preparations. The antibody preparations employed were the control rabbit anti-mouse antibody, called the 2[0069] nd Ab, the 6g05 antibody preparation, the 14c12 antibody preparation, and the negative control 112d08 antibody preparation. Activated splenocytes treated with either of the 6g05 or 14c12 antibody preparations have lesser numbers of cells in the S phase of the cell cycle.
    • DETAILED DESCRIPTION OF THE INVENTION
  • The invention provides methods for modulating the immune system. For example, according to the invention, loss or reduction of CD83 activity in vivo results in decreased numbers of immune cells, for example, decreased numbers of T cells. In some embodiments, binding entities such as monoclonal antibodies that are capable of inhibiting the function of CD83 are provided for use in the invention. In other embodiments the binding entities or antibodies are multimerized. The compositions and methods of the invention can be used for treating conditions involving an inappropriate immune response, for example, autoimmune diseases, inflammation, tissue rejection, arthritis, atherosclerosis and the like. [0070]
  • CD83 [0071]
  • CD83 is a lymphocyte and dendritic cell activation antigen that is expressed by activated lymphocytes and dendritic cells. CD83 is also a single-chain cell-surface glycoprotein with a molecular weight of about 45,000 that is believed to be a member of the Ig superfamily. The structure predicted from the CD83 amino acid sequence indicates that CD83 is a membrane glycoprotein with a single extracellular Ig-like domain, a transmembrane domain and cytoplasmic domain of approximately forty amino acids. The mature CD83 protein has about 186 amino acids and is composed of a single extracellular V type immunoglobulin (Ig)-like domain, a transmembrane domain and a thirty nine amino acid cytoplasmic domain. Northern blot analysis has revealed that CD83 is translated from three mRNA transcripts of about 1.7, 2.0 and 2.5 kb that are expressed by lymphoblastoid cell lines. It is likely that CD83 undergoes extensive post-translational processing because CD83 is expressed as a single chain molecule, but the determined molecular weight is twice the predicted size of the core protein. See U.S. Pat. No. 5,766,570. [0072]
  • An example of a human CD83 gene product that can be used in the invention is provided below (SEQ ID NO:9): [0073]
      1 MSRGLQLLLL SCAYSLAPAT PEVKVACSED VDLPCTAPWD
     41 PQVPYTVSWV KLLEGGEERM ETPQEDHLRG QHYHQKGQNG
     81 SFDAPNERPY SLKIRNTTSC NSGTYRCTLQ DPDGQRNLSG
    121 KVILRVTGCP AQRKEETFKK YRAEIVLLLA LVIFYLTLII
    161 FTCKFARLQS IFPDFSKAGM ERAFLPVTSP NKHLGLVTPH
    201 KTELV
  • Such a CD83 gene product can be encoded by a number of different nucleic acids. One example of a human CD83 nucleic acid is provided below (SEQ ID NO:10). [0074]
       1 CCTGGCGCAG CCGCAGCAGC GACGCGAGCG AACTCGGCCG
      41 GGCCCGGGCG CGCGGGGGCG GGACGCGCAC GCGGCGAGGG
      81 CGGCGGGTGA GCCGGGGGCG GGGACGGGGG CGGGACGGGG
     121 GCGAAGGGGG CGGGGACGGG GGCGCCCGCC GGCCTAACGG
     161 GATTAGGAGG GCGCGCCACC CGCTTCCGCT GCCCGCCGGG
     201 GAATCCCCCG GGTGGCGCCC AGGGAAGTTC CCGAACGGGC
     241 GGGCATAAAA GGGCAGCCGC GCCGGCGCCC CACAGCTCTG
     281 CAGCTCGTGG CAGCGGCGCA GCGCTCCAGC CATGTCGCGC
     321 GGCCTCCAGC TTCTGCTCCT GAGCTGCGCC TACAGCCTGG
     361 CTCCCGCGAC GCCGGAGGTG AAGGTGGCTT GCTCCGAAGA
     401 TGTGGACTTG CCCTGCACCG CCCCCTGGGA TCCGCAGGTT
     441 CCCTACACGG TCTCCTGGGT CAAGTTATTG GAGGGTGGTG
     481 AAGAGAGGAT GGAGACACCC CAGGAAGACC ACCTCAGGGG
     521 ACAGCACTAT CATCAGAAGG GGCAAAATGG TTCTTTCGAC
     561 GCCCCCAATG AAAGGCCCTA TTCCCTGAAG ATCCGAAACA
     601 CTACCAGCTG CAACTCGGGG ACATACAGGT GCACTCTGCA
     641 GGACCCGGAT GGGCAGAGAA ACCTAAGTGG CAAGGTGATC
     681 TTGAGAGTGA CAGGATGCCC TGCACAGCGT AAAGAAGAGA
     721 CTTTTAAGAA ATACAGAGCG GAGATTGTCC TGCTGCTGGC
     761 TCTGGTTATT TTCTACTTAA CACTCATCAT TTTCACTTGT
     801 AAGTTTGCAC GGCTACAGAG TATCTTCCCA GATTTTTCTA
     841 AAGCTGGCAT GGAACGAGCT TTTCTCCCAG TTACCTCCCC
     881 AAATAAGCAT TTAGGGCTAG TGACTCCTCA CAAGACAGAA
     921 CTGGTATGAG CAGGATTTCT GCAGGTTCTT CTTCCTGAAG
     961 CTGAGGCTCA GGGGTGTGCC TGTCTGTTAC ACTGGAGGAG
    1001 AGAAGAATGA GCCTACGCTG AAGATGGCAT CCTGTGAAGT
    1041 CCTTCACCTC ACTGAAAACA TCTGGAAGGG GATCCCACCC
    1081 CATTTTCTGT GGGCAGGCCT CGAAAACCAT CACATGACCA
    1121 CATAGCATGA GGCCACTGCT GCTTCTCCAT GGCCACCTTT
    1161 TCAGCGATGT ATGCAGCTAT CTGGTCAACC TCCTGGACAT
    1201 TTTTTCAGTC ATATAAAAGC TATGGTGAGA TGCAGCTGGA
    1241 AAACGGTCTT GGGAAATATG AATGCCCCCA GCTGGCCCGT
    1281 GACAGACTCC TGAGGACAGC TGTCCTCTTC TGCATCTTGG
    1321 GGACATCTCT TTGAATTTTC TGTGTTTTGC TGTACCAGCC
    1361 CAGATGTTTT ACGTCTGGGA GAAATTGACA GATCAAGCTG
    1401 TGAGACAGTG GGAAATATTT AGCAAATAAT TTCCTGGTGT
    1441 GAAGGTCCTG CTATTACTAA GGAGTAATCT GTGTACAAAG
    1481 AAATAACAAG TCGATGAACT ATTCCCCAGC AGGGTCTTTT
    1521 CATCTGGGAA AGACATCCAT AAAGAAGCAA TAAAGAAGAG
    1561 TGCCACATTT ATTTTTATAT CTATATGTAC TTGTCAAAGA
    1601 AGGTTTGTGT TTTTCTGCTT TTGAAATCTG TATCTGTAGT
    1641 GAGATAGCAT TGTGAACTGA CAGGCAGCCT GGACATAGAG
    1681 AGGGAGAAGA AGTCAGAGAG GGTGACAAGA TAGAGAGCTA
    1721 TTTAATGGCC GGCTGGAAAT GCTGGGCTGA CGGTGCAGTC
    1761 TGGGTGCTCG CCCACTTGTC CCACTATCTG GGTGCATGAT
    1801 CTTGAGCAAG TTCCTTCTGG TGTCTGCTTT CTCCATTGTA
    1841 AACCACAAGG CTGTTGCATG GGCTAATGAA GATCATATAC
    1881 GTGAAAATTA TTTGAAAACA TATAAAGCAC TATACAGATT
    1921 CGAAACTCCA TTGAGTCATT ATCCTTGCTA TGATGATGGT
    1961 GTTTTGGGGA TGAGAGGGTG CTATCCATTT CTCATGTTTT
    2001 CCATTGTTTG AAACAAAGAA GGTTACCAAG AAGCCTTTCC
    2041 TGTAGCCTTC TGTAGGAATT CTTTTGGGGA AGTGAGGAAG
    2081 CCAGGTCCAC GGTCTGTTCT TGAAGCAGTA GCCTAACACA
    2121 CTCCAAGATA TGGACACACG GGAGCCGCTG GCAGAAGGGA
    2161 CTTCACGAAG TGTTGCATGG ATGTTTTAGC CATTGTTGGC
    2201 TTTCCCTTAT CAAACTTGGG CCCTTCCCTT CTTGGTTTCC
    2241 AAAGGCATTT ATTGCTGAGT TATATGTTCA CTGTCCCCCT
    2281 AATATTAGGG AGTAAAACGG ATACCAAGTT GATTTAGTGT
    2321 TTTTACCTCT GTCTTGGCTT TCATGTTATT AAACGTATGC
    2361 ATGTGAAGAA GGGTGTTTTT CTGTTTTATA TTCAACTCAT
    2401 AAGACTTTGG GATAGGAAAA ATGAGTAATG GTTACTAGGC
    2441 TTAATACCTG GGTGATTACA TAATCTGTAC AACGAACCCC
    2481 CATGATGTAA GTTTACCTAT GTAACAAACC TGCACTTATA
    2521 CCCATGAACT TAAAATGAAA GTTAAAAATA AAAAACATAT
    2561 ACAAATAAAA AAAA
  • A sequence of a wild type mouse CD83 gene that can be used in the invention is provided herein as SEQ ID NO:1. SEQ ID NO:1 is provided below with the ATG start codon and the TGA stop codon identified by underlining. [0075]
       1 GCGCTCCAGC CGC ATG TCGC AAGGCCTCCA GCTCCTGTTT
      41 CTAGGCTGCG CCTGCAGCCT GGCACCCGCG ATGGCGATGC
      81 GGGAGGTGAC GGTGGCTTGC TCCGAGACCG CCGACTTGCC
     121 TTGCACAGCG CCCTGGGACC CGCAGCTCTC CTATGCAGTG
     161 TCCTGGGCCA AGGTCTCCGA GAGTGGCACT GAGAGTGTGG
     201 AGCTCCCGGA GAGCAAGCAA AACAGCTCCT TCGAGGCCCC
     241 CAGGAGAAGG GCCTATTCCC TGACGATCCA AAACACTACC
     281 ATCTGCAGCT CGGGCACCTA CAGGTGTGCC CTGCAGGAGC
     321 TCGGAGGGCA GCGCAACTTG AGCGGCACCG TGGTTCTGAA
     361 GGTGACAGGA TGCCCCAAGG AAGCTACAGA GTCAACTTTC
     401 AGGAAGTACA GGGCAGAAGC TGTGTTGCTC TTCTCTCTGG
     441 TTGTTTTCTA CCTGACACTC ATCATTTTCA CCTGCAAATT
     481 TGCACGACTA CAAAGCATTT TCCCAGATAT TTCTAAACCT
     521 GGTACGGAAC AAGCTTTTCT TCCAGTCACC TCCCCAAGCA
     561 AACATTTGGG GCCAGTGACC CTTCCTAAGA CAGAAACGGT
     601 A TGA GTAGGA TCTCCACTGG TTTTTACAAA GCCAAGGGCA
     641 CATCAGATCA GTGTGCCTGA ATGCCACCCG GACAAGAGAA
     681 GAATGAGCTC CATCCTCAGA TGGCAACCTT TCTTTGAAGT
     721 CCTTCACCTG ACAGTGGGCT CCACACTACT CCCTGACACA
     761 GGGTCTTGAG CACCATCATA TGATCACGAA GCATGGAGTA
     801 TCACCGCTTC TCTGTGGCTG TCAGCTTAAT GTTTCATGTG
     841 GCTATCTGGT CAACCTCGTG AGTGCTTTTC AGTCATCTAC
     881 AAGCTATGGT GAGATGCAGG TGAAGCAGGG TCATGGGAAA
     921 TTTGAACACT CTGAGCTGGC CCTGTGACAG ACTCCTGAGG
     961 ACAGCTGTCC TCTCCTACAT CTGGGATACA TCTCTTTGAA
    1001 TTTGTCCTGT TTCGTTGCAC CAGCCCAGAT GTCTCACATC
    1041 TGGCGGAAAT TGACAGGCCA AGCTGTGAGC CAGTGGGAAA
    1081 TATTTAGCAA ATAATTTCCC AGTGCGAAGG TCCTGCTATT
    1121 AGTAAGGAGT ATTATGTGTA CATAGAAATG AGAGGTCAGT
    1161 GAACTATTCC CCAGCAGGGC CTTTTCATCT GGAAAAGACA
    1201 TCCACAAAAG CAGCAATACA GAGGGATGCC ACATTTATTT
    1241 TTTTAATCTT CATGTACTTG TCAAAGAAGA ATTTTTCATG
    1281 TTTTTTCAAA GAAGTGTGTT TCTTTCCTTT TTTAAAATAT
    1321 GAAGGTCTAG TTACATAGCA TTGCTAGCTG ACAAGCAGCC
    1361 TGAGAGAAGA TGGAGAATGT TCCTCAAAAT AGGGACAGCA
    1401 AGCTAGAAGC ACTGTACAGT GCCCTGCTGG GAAGGGCAGA
    1441 CAATGGACTG AGAAACCAGA AGTCTGGCCA CAAGATTGTC
    1481 TGTATGATTC TGGACGAGTC ACTTGTGGTT TTCACTCTCT
    1521 GGTTAGTAAA CCAGATAGTT TAGTCTGGGT TGAATACAAT
    1561 GGATGTGAAG TTGCTTGGGG AAAGCTGAAT GTAGTGAATA
    1601 CATTGGCAAC TCTACTGGGC TGTTACCTTG TTGATATCCT
    1641 AGAGTTCTGG AGCTGAGCGA ATGCCTGTCA TATCTCAGCT
    1681 TGCCCATCAA TCCAAACACA GGAGGCTACA AAAAGGACAT
    1721 GAGCATGGTC TTCTGTGTGA ACTCCTCCTG AGAAACGTGG
    1761 AGACTGGCTC AGCGCTTTGC GCTTGAAGGA CTAATCACAA
    1801 GTTCTTGAAG ATATGGACCT AGGGGAGCTA TTGCGCCACG
    1841 ACAGGAGGAA GTTCTCAGAT GTTGCATTGA TGTAACATTG
    1881 TTGCATTTCT TTAATGAGCT GGGCTCCTTC CTCATTTGCT
    1921 TCCCAAAGAG ATTTTGTCCC ACTAATGGTG TGCCCATCAC
    1961 CCACACTATG AAAGTAAAAG GGATGCTGAG CAGATACAGC
    2001 GTGCTTACCT CTCAGCCATG ACTTTCATGC TATTAAAAGA
    2041 ATGCATGTGA A
  • Nucleic acids having SEQ ID NO:1 encode a mouse polypeptide having SEQ ID NO:2, provided below. [0076]
      1 MSQGLQLLFL GCACSLAPAM AMREVTVACS ETADLPCTAP
     41 WDPQLSYAVS WAKVSESGTE SVELPESKQN SSFEAPRRRA
     81 YSLTIQNTTI CSSGTYRCAL QELGGQRNLS GTVVLKVTGC
    121 PKEATESTFR KYRAEAVLLF SLVVFYLTLI IFTCKFARLQ
    161 SIFPDISKPG TEQAFLPVTS PSKHLGPVTL PKTETV
  • According to the invention, loss or reduction of CD83 activity in vivo results in a decreased immune response, for example, decreased numbers of T cells. The effect of CD83 on the immune response was initially ascertained through use of a mutant mouse that encodes a mutant CD83. Such a mutant mouse has a CD83 gene encoding SEQ ID NO:4, with added C-terminal sequences provided by SEQ ID NO:8. In contrast to these wild type CD83 nucleic acids and polypeptides, the mutant CD83 gene of the invention has SEQ ID NO:3. SEQ ID NO:3 is provided below with the ATG start codon, the mutation, and the TGA stop codon are identified by underlining. [0077]
       1 GCGCTCCAGC CGC ATG TCGC AAGGCCTCCA GCTCCTGTTT
      41 CTAGGCTGCG CCTGCAGCCT GGCACCCGCG ATGGCGATGC
      81 GGGAGGTGAC GGTGGCTTGC TCCGAGACCG CCGACTTGCC
     121 TTGCACAGCG CCCTGGGACC CGCAGCTCTC CTATGCAGTG
     161 TCCTGGGCCA AGGTCTCCGA GAGTGGCACT GAGAGTGTGG
     201 AGCTCCCGGA GAGCAAGCAA AACAGCTCCT TCGAGGCCCC
     241 CAGGAGAAGG GCCTATTCCC TGACGATCCA AAACACTACC
     281 ATCTGCAGCT CGGGCACCTA CAGGTGTGCC CTGCAGGAGC
     321 TCGGAGGGCA GCGCAACTTG AGCGGCACCG TGGTTCTGAA
     361 GGTGACAGGA TGCCCCAAGG AAGCTACAGA GTCAACTTTC
     401 AGGAAGTACA GGGCAGAAGC TGTGTTGCTC TTCTCTCTGG
     441 TTGTTTTCTA CCTGACACTC ATCATTTTCA CCTGCAAATT
     481 TGCACGACTA CAAAGCATTT TCCCAGATAT TTCTAAACCT
     521 GGTACGGAAC AAGCTTTTCT TCCAGTCACC TCCCCAAGCA
     561 AACATTTGGG GCCAGTGACC CTTCCTAAGA CAGAAACGGT
     601 A A GAGTAGGA TCTCCACTGG TTTTTACAAA GCCAAGGGCA
     641 CATCAGATCA GTGTGCCTGA ATGCCACCCG GACAAGAGAA
     681 GAATGAGCTC CATCCTCAGA TGGCAACCTT TCTTTGAAGT
     721 CCTTCACCTG ACAGTGGGCT CCACACTACT CCCTGACACA
     761 GGGTCT TGA G CACCATCATA TGATCACGAA GCATGGAGTA
     801 TCACCGCTTC TCTGTGGCTG TCAGCTTAAT GTTTCATGTG
     841 GCTATCTGGT CAACCTCGTG AGTGCTTTTC AGTCATCTAC
     881 AAGCTATGGT GAGATGCAGG TGAAGCAGGG TCATGGGAAA
     921 TTTGAACACT CTGAGCTGGC CCTGTGACAG ACTCCTGAGG
     961 ACAGCTGTCC TCTCCTACAT CTGGGATACA TCTCTTTGAA
    1001 TTTGTCCTGT TTCGTTGCAC CAGCCCAGAT GTCTCACATC
    1041 TGGCGGAAAT TGACAGGCCA AGCTGTGAGC CAGTGGGAAA
    1081 TATTTAGCAA ATAATTTCCC AGTGCGAAGG TCCTGCTATT
    1121 AGTAAGGAGT ATTATGTGTA CATAGAAATG AGAGGTCAGT
    1161 GAACTATTCC CCAGCAGGGC CTTTTCATCT GGAAAAGACA
    1201 TCCACAAAAG CAGCAATACA GAGGGATGCC ACATTTATTT
    1241 TTTTAATCTT CATGTACTTG TCAAAGAAGA ATTTTTCATG
    1281 TTTTTTCAAA GAAGTGTGTT TCTTTCCTTT TTTAAAATAT
    1321 GAAGGTCTAG TTACATAGCA TTGCTAGCTG ACAAGCAGCC
    1361 TGAGAGAAGA TGGAGAATGT TCCTCAAAAT AGGGACAGCA
    1401 AGCTAGAAGC ACTGTACAGT GCCCTGCTGG GAAGGGCAGA
    1441 CAATGGACTG AGAAACCAGA AGTCTGGCCA CAAGATTGTC
    1481 TGTATGATTC TGGACGAGTC ACTTGTGGTT TTCACTCTCT
    1521 GGTTAGTAAA CCAGATAGTT TAGTCTGGGT TGAATACAAT
    1561 GGATGTGAAG TTGCTTGGGG AAAGCTGAAT GTAGTGAATA
    1601 CATTGGCAAC TCTACTGGGC TGTTACCTTG TTGATATCCT
    1641 AGAGTTCTGG AGCTGAGCGA ATGCCTGTCA TATCTCAGCT
    1681 TGCCCATCAA TCCAAACACA GGAGGCTACA AAAAGGACAT
    1721 GAGCATGGTC TTCTGTGTGA ACTCCTCCTG AGAAACGTGG
    1761 AGACTGGCTC AGCGCTTTGC GCTTGAAGGA CTAATCACAA
    1801 GTTCTTGAAG ATATGGACCT AGGGGAGCTA TTGCGCCACG
    1841 ACAGGAGGAA GTTCTCAGAT GTTGCATTGA TGTAACATTG
    1881 TTGCATTTCT TTAATGAGCT GGGCTCCTTC CTCATTTGCT
    1921 TCCCAAAGAG ATTTTGTCCC ACTAATGGTG TGCCCATCAC
    1961 CCACACTATG AAAGTAAAAG GGATGCTGAG CAGATACAGC
    2001 GTGCTTACCT CTCAGCCATG ACTTTCATGC TATTAAAAGA
    2041 ATGCATGTGA A
  • The change from a thymidine in SEQ ID NO:1 to an adenine in SEQ ID NO:3 at the indicated position (602) leads to read-through translation because the stop codon at positions 602-604 in SEQ ID NO:1 is changed to a codon that encodes an arginine. Accordingly, mutant CD83 nucleic acids having SEQ ID NO:3 encode an elongated polypeptide having SEQ ID NO:4, provided below, where the extra amino acids are underlined. [0078]
      1 MSQGLQLLFL GCACSLAPAM AMREVTVACS ETADLPCTAP
     41 WDPQLSYAVS WAKVSESGTE SVELPESKQN SSFEAPRRRA
     81 YSLTIQNTTI CSSGTYRCAL QELGGQRNLS GTVVLKVTGC
    121 PKEATESTFR KYRAEAVLLF SLVVFYLTLI IFTCKFARLQ
    161 SIFPDISKPG TEQAFLPVTS PSKHLGPVTL PKTETVRVGS
    201 PLVFTKPRAH QISVPECHPD KRRMSSILRW QPFFEVLHLT
    241 VGSTLLPDTG S
  • In another embodiment, the invention provides mutant CD83 nucleic acids that include SEQ ID NO:5. [0079]
      1 ATG TCGCAAG GCCTCCAGCT CCTGTTTCTA GGCTGCGCCT
     41 GCAGCCTGGC ACCCGCGATG GCGATGCGGG AGGTGACGGT
     81 GGCTTGCTCC GAGACCGCCG ACTTGCCTTG CACAGCGCCC
    121 TGGGACCCGC AGCTCTCCTA TGCAGTGTCC TGGGCCAAGG
    161 TCTCCGAGAG TGGCACTGAG AGTGTGGAGC TCCCGGAGAG
    201 CAAGCAAAAC AGCTCCTTCG AGGCCCCCAG GAGAAGGGCC
    241 TATTCCCTGA CGATCCAAAA CACTACCATC TGCAGCTCGG
    281 GCACCTACAG GTGTGCCCTG CAGGAGCTCG GAGGGCAGCG
    321 CAACTTGAGC GGCACCGTGG TTCTGAAGGT GACAGGATGC
    361 CCCAAGGAAG CTACAGAGTC AACTTTCAGG AAGTACAGGG
    401 CAGAAGCTGT GTTGCTCTTC TCTCTGGTTG TTTTCTACCT
    441 GACACTCATC ATTTTCACCT GCAAATTTGC ACGACTACAA
    481 AGCATTTTCC CAGATATTTC TAAACCTGGT ACGGAACAAG
    521 CTTTTCTTCC AGTCACCTCC CCAAGCAAAC ATTTGGGGCC
    561 AGTGACCCTT CCTAAGACAG AAACGGTAAG AGTAGGATCT
    601 CCACTGGTTT TTACAAAGCC AAGGGCACAT CAGATCAGTG
    641 TGCCTGAATG CCACCCGGAC AAGAGAAGAA TGAGCTCCAT
    681 CCTCAGATGG CAACCTTTCT TTGAAGTCCT TCACCTGACA
    721 GTGGGCTCCA CACTACTCCC TGACACAGGG TCT TGA
  • Nucleic acids having SEQ ID NO:5 also encode a polypeptide having SEQ ID NO:4. [0080]
  • In another embodiment, the invention provides mutant CD83 nucleic acids that include SEQ ID NO:7. [0081]
      1 A GAGTAGGAT CTCCACTGGT TTTTACAAAG CCAAGGGCAC
     41 ATCAGATCAG TGTGCCTGAA TGCCACCCGG ACAAGAGAAG
     81 AATGAGCTCC ATCCTCAGAT GGCAACCTTT CTTTGAAGTC
    121 CTTCACCTGA CAGTGGGCTC CACACTACTC CCTGACACAG
    161 GGTCT TGA
  • The invention also provides a mutant CD83 containing SEQ ID NO:8, provided below. [0082]
     1 RVGSPLVFTK PRAHQISVPE CHPDKRRMSS ILRWQPPFEV
    41 LHLTVGSTLL PDTGS
  • SEQ ID NO:8 contains read through sequences that are not present in the wild type CD83 polypeptide but are present in the mutant CD83 gene product provided by the invention. [0083]
  • In some embodiments, the CD83 gene product is used for generating antibodies. While any of the CD83 gene products described herein can be employed for immunization of animal, in some embodiments the extracellular Ig-like domain of the CD83 gene product is used for immunization, or antibodies are screened for reactivity with the extracellular Ig-like domain. The extracellular Ig-like domain of the human CD83 gene product spans amino acids 21-127, and is provided below (SEQ ID NO:97): [0084]
     21                       PEVKVACSED VDLPCTAPWD
     41 PQVPYTVSWV KLLEGGEERM ETPQEDHLRG QHYHQKGQNG
     81 SFDAPNERPY SLKIRNTTSC NSGTYRCTLQ DPDGQRNLSG
    121 KVILRVT
  • CD83 Antibodies [0085]
  • The invention provides antibody preparations directed against the mutant and wild type CD83 polypeptides of the invention, for example, against a polypeptide having SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:8 or SEQ ID NO:9. Other antibodies of interest can bind to the cytoplasmic tail of CD83. [0086]
  • In some embodiments, the anti-CD83 antibodies are multimerized antibodies. According to the invention, multimerized anti-CD83 antibodies are surprisingly effective inhibitors of lymphocyte cell proliferation. As used herein, an “multimerized” anti-CD83 antibody is a collection of anti-CD83 antibodies that are in close proximity. While such multimerized antibodies can be covalently linked, no such covalent linkage is necessary. Instead, multimerization of anti-CD83 antibodies can simply involve bringing the antibodies into close proximity, for example, by attachment to a solid support such as a plate or a bead. Alternatively, the antibodies can be non-covalently linked together through another entity, for example, any convenient non-covalent binding entity or secondary antibody. Hence, any available means for bringing the anti-CD83 antibodies into proximity can be used to generate the multimerized antibodies of the invention. [0087]
  • In some embodiments, the anti-CD83 binding proteins or antibodies can be chemically cross-linked or genetically fused with any available crosslinking reagent. Crosslinking can be achieved using one or a combination of a wide variety of multifunctional reagents. Such crosslinking agents include glutaraldehyde, succinaldehyde, octanedialdehyde and glyoxal. Additional multifunctional crosslinking agents include halo-triazines, e.g., cyanuric chloride; halo-pyrimidines, e.g., 2,4,6-trichloro/bromo-pyrimidine; anhydrides or halides of aliphatic or aromatic mono- or di-carboxylic acids, e.g., maleic anhydride, (meth)acryloyl chloride, chloroacetyl chloride; N-methylol compounds, e.g., N-methylol-chloro acetamide; di-isocyanates or di-isothiocyanates, e.g., phenylene-1,4-di-isocyanate and aziridines. Other crosslinking agents include epoxides, such as, for example, di-epoxides, tri-epoxides and tetra-epoxides. Other crosslinking agents include, for example, [0088] dimethyl 3,3′-dithiobispropionimidate-HCl (DTBP); dithiobis (succinimidylpropionate) (DSP); bismaleimidohexane (BMH); bis[Sulfosuccinimidyl]suberate (BS); 1,5-difluoro-2,4-dinitrobenzene (DFDNB); dimethylsuberimidate-2HCl (DMS); disuccinimidyl glutarate (DSG); disulfosuccinimidyl tartarate (Sulfo-DST); 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC); ethylene glycolbis [sulfo-succinimidylsuccinate] (Sulfo-EGS); N-[?-maleimido-butyryloxy]succinimide ester (GMBS); N-hydroxysulfosuccinimidyl-4-azidobenzoate (Sulfo-HSAB); sulfosuccinimidyl-6-[a-methyl-a-(2-pyridyldithio) toluamido]hexanoate (Sulfo-LC-SMPT); bis-[β-(4-azidosalicylamido) ethyl]disulfide (BASED); and NHS-PEG-Vinylsulfone (NHS-PEG-VS).
  • In some embodiments, crosslinkers useful with various preparations of anti-CD83 antibodies of this invention include (1) those which create covalent links from one cysteine side chain of a protein to another cysteine side chain, (2) those which create covalent links from one lysine side chain of a protein to another, or (3) those which create covalent links from one cysteine side chain of a protein to a lysine side chain. [0089]
  • In other embodiments, the anti-CD83 antibodies are reversibly crosslinked. Such reversibly crosslinked antibodies are useful for short term use, for example, for short term control of the immune response either in vivo or in vitro, or for controlled dissipation of the anti-CD83 antibodies at a localized site after administration for short term therapeutic purposes. Examples of reversible crosslinkers are described in T. W. Green, Protective Groups in Organic Synthesis, John Wiley & Sons (Eds.) (1981). Other types of reversible crosslinkers are disulfide bond-containing crosslinkers. The crosslinks formed by such crosslinkers can be broken by the addition of reducing agent, such as cysteine, to the environment of the crosslinked anti-CD83 antibodies. Disulfide crosslinkers are described in the Pierce Catalog and Handbook (1994-1995). [0090]
  • Examples of crosslinkers that may be used also include: Homobifunctional (Symmetric); DSP—Dithiobis(succinimidylpropionate), also know as Lomant,'s Reagent; DTSSP—3-3′-Dithiobis (sulfosuccinimidyl-propionate), water soluble version of DSP; DTBP—Dimethyl 3,3′-dithiobispropionimidate-HCl; BASED—Bis-(13-[4-azidosalicylamido] ethyl)disulfide; DPDPB—1,4-Di-(3′-[2′-pyridyldithio]-propionamido)butane; Heterobifunctional (Asymmetric); SPDP—N-Succinimidyl-3-(2-pyridyldithio)propionate; LC-SPDP—Succinimidyl-6-(3-[2-pyridyldithio] propionate)hexanoate; Sulfo-LC-SPDP—Sulfosuccinimidyl-6-(3-[2-pyridyldlthio] propionate)hexanoate, water soluble version of LC-SPDP; APDP—N-(4-[p-azidosalicylamido]butyl)-3′-(2′-pyridyldithio) propionamide; SADP—N-Succinimidyl(4-azidophenyl)1,3′-dithiopropionate; Sulfo-SADP—Sulfosuccinimidyl(4-azidophenyl) 1,3′-dithiopropionate, water soluble version of SADP; SAED—Sulfosuccinimidyl-2-(7-azido-4-methycoumarin-3-acetamide)ethyl-1,3′dithiopropionate; SAND—Sulfosuccinimidyl-2-(m-azido-o-nitrobenzamido)ethyl-1,3′-dithiopropionate; SASD—Sulfosuccinimidyl-2-(p-azidosalicylamido)ethyl-1,3′-dithiopropionate; SMPB—Succinimidyl-4-(p-maleimidophenyl)butyrate; Sulfo-SMPB—Sulfosuccinimidyl-4-(p-maleimidophenyl)butyrate; SMPT—4-Succinimidyloxycarbonyl-methyl-a-(2-pyridylthio) toluene; Sulfo-LC-SMPT—Sulfosuccinimidyl-6-(a-methyl-a-(2-pyridylthio)toluamido)hexanoate. [0091]
  • In another embodiment, a fusion protein can be made with a selected anti-CD83 antibody to allow a domain to be attached to one or both of the polypeptides comprising the anti-CD83 antibody to be bound to a solid substrate. For example, glutathione-S-transferase/anti-CD83 fusion proteins can be linked to another anti-CD83 preparation having glutathione attached thereto or the glutathione-S-transferase/anti-CD83 fusion proteins can be adsorbed onto a solid support having glutathione attached thereto, such as glutathione sepharose beads (Sigma Chemical, St. Louis, Mo.) or glutathione derivatized microtiter plate. In another embodiment, DSP-crosslinked antibodies can be linked to protein G agarose beads. Other techniques for immobilizing polypeptides on solid support materials can also be used. For example, an anti-CD83 antibody can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated anti-CD83 polypeptides can be prepared from biotin-NHS(N-hydroxy-succinimide) using techniques known in the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, Ill.), and immobilized with a streptavidin-linked antiCD83 antibody preparation, streptavidin-coated beads or another solid support material. [0092]
  • Therefore, in one embodiment, the invention provides antibodies capable of reducing CD83 activity and decreasing an immune response in a mammal. Such antibodies can be multimerized antibodies. These antibodies may be used as CD83 inhibitory agents in the methods of the invention as described herein. In another embodiment, the antibodies of the invention can activate CD83 activity. Such activating antibodies may be used as CD83 stimulatory agents. [0093]
  • All antibody molecules belong to a family of plasma proteins called immunoglobulins, whose basic building block, the immunoglobulin fold or domain, is used in various forms in many molecules of the immune system and other biological recognition systems. A typical immunoglobulin has four polypeptide chains, containing an antigen binding region known as a variable region and a non-varying region known as the constant region. [0094]
  • Native antibodies and immunoglobulins are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies between the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has at one end a variable domain (VH) followed by a number of constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end. The constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains (Clothia et al., J. Mol. Biol. 186, 651-66, 1985); Novotny and Haber, Proc. Natl. Acad. Sci. USA 82, 4592-4596(1985). [0095]
  • Depending on the amino acid sequences of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are at least five (5) major classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses (isotypes), e.g. IgG-1, IgG-2, IgG-3 and IgG-4; IgA-1 and IgA-2. The heavy chains constant domains that correspond to the different classes of immunoglobulins are called alpha (a), delta (d), epsilon (e), gamma (?) and mu (μ), respectively. The light chains of antibodies can be assigned to one of two clearly distinct types, called kappa (?) and lambda (?), based on the amino sequences of their constant domain. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. [0096]
  • The term “variable” in the context of variable domain of antibodies, refers to the fact that certain portions of the variable domains differ extensively in sequence among antibodies. The variable domains are for binding and determine the specificity of each particular antibody for its particular antigen. However, the variability is not evenly distributed through the variable domains of antibodies. It is concentrated in three segments called complementarity determining regions (CDRs) also known as hypervariable regions both in the light chain and the heavy chain variable domains. [0097]
  • The more highly conserved portions of variable domains are called the framework (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a β-sheet configuration, connected by three CDRs, which form loops connecting, and in some cases forming part of, the β-sheet structure. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site of antibodies. The constant domains are not involved directly in binding an antibody to an antigen, but exhibit various effector function, such as participation of the antibody in antibody-dependent cellular toxicity. [0098]
  • An antibody that is contemplated for use in the present invention thus can be in any of a variety of forms, including a whole immunoglobulin, an antibody fragment such as Fv, Fab, and similar fragments, a single chain antibody that includes the variable domain complementarity determining regions (CDR), and the like forms, all of which fall under the broad term “antibody,” as used herein. Moreover, the multimerized antibodies of the invention can be an aggregation or multimerization of whole immunoglobulins. Alternatively, the multimerized antibodies of the invention can be an aggregation or multimerization of antibody fragments such as Fv, Fab, single chain antibodies that include the variable domain complementarity determining regions (CDR), CDRs and the like. Such intact antibodies or antibody fragments can be multimerized by any convenient means, including covalent linkage or non-covalent association. [0099]
  • The present invention contemplates the use of any specificity of an antibody, polyclonal or monoclonal, and is not limited to antibodies that recognize and immunoreact with a specific epitope. In preferred embodiments, in the context of both the therapeutic and screening methods described below, an antibody or fragment thereof is used that is immunospecific for an extracellular portion of the CD83 protein. [0100]
  • The term “antibody fragment” refers to a portion of a full-length antibody, generally the antigen binding or variable region. Examples of antibody fragments include Fab, Fab′, F(ab′)[0101] 2 and Fv fragments. Papain digestion of antibodies produces two identical antigen binding fragments, called the Fab fragment, each with a single antigen binding site, and a residual “Fc” fragment, so-called for its ability to crystallize readily. Pepsin treatment yields an F(ab′)2 fragment that has two antigen binding fragments, which are capable of cross-linking antigen, and a residual other fragment (which is termed pFc′). Additional fragments can include diabodies, linear antibodies, single-chain antibody molecules, and multispecific antibodies formed from antibody fragments. As used herein, “functional fragment” with respect to antibodies, refers to Fv, F(ab) and F(ab′)2 fragments.
  • Antibody fragments retain some ability to selectively bind with its antigen or receptor and are defined as follows: [0102]
  • (1) Fab is the fragment that contains a monovalent antigen-binding fragment of an antibody molecule. A Fab fragment can be produced by digestion of whole antibody with the enzyme papain to yield an intact light chain and a portion of one heavy chain. [0103]
  • (2) Fab′ is the fragment of an antibody molecule can be obtained by treating whole antibody with pepsin, followed by reduction, to yield an intact light chain and a portion of the heavy chain. Two Fab′ fragments are obtained per antibody molecule. Fab′ fragments differ from Fab fragments by the addition of a few residues at the carboxyl terminus of the [0104] heavy chain CH 1 domain including one or more cysteines from the antibody hinge region.
  • (3) (Fab′)[0105] 2 is the fragment of an antibody that can be obtained by treating whole antibody with the enzyme pepsin without subsequent reduction. F(ab′)2 is a dimer of two Fab′ fragments held together by two disulfide bonds.
  • (4) Fv is the minimum antibody fragment that contains a complete antigen recognition and binding site. This region consists of a dimer of one heavy and one light chain variable domain in a tight, non-covalent association (VH-V L dimer). It is in this configuration that the three CDRs of each variable domain interact to define an antigen binding site on the surface of the VH-V L dimer. Collectively, the six CDRs confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site. [0106]
  • (5) Single chain antibody (“SCA”), defined as a genetically engineered molecule containing the variable region of the light chain, the variable region of the heavy chain, linked by a suitable polypeptide linker as a genetically fused single chain molecule. Such single chain antibodies are also referred to as “single-chain Fv” or “sFv” antibody fragments. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that enables the sFv to form the desired structure for antigen binding. For a review of sFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, N.Y., pp. 269-315 (1994). [0107]
  • The term “diabodies” refers to a small antibody fragments with two antigen-binding sites, which fragments comprise a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites. Diabodies are described more fully in, for example, EP 404,097; WO 93/11161, and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). [0108]
  • The preparation of polyclonal antibodies is well-known to those skilled in the art. See, for example, Green, et al., Production of Polyclonal Antisera, in: [0109] Immunochemical Protocols (Manson, ed.), pages 1-5 (Humana Press); Coligan, et al., Production of Polyclonal Antisera in Rabbits, Rats Mice and Hamsters, in: Current Protocols in Immunology, section 2.4.1 (1992), which are hereby incorporated by reference.
  • The preparation of monoclonal antibodies likewise is conventional. See, for example, Kohler & Milstein, Nature, 256:495 (1975); Coligan, et al., sections 2.5.1-2.6.7; and Harlow, et al., in: [0110] Antibodies: A Laboratory Manual, page 726 (Cold Spring Harbor Pub. (1988)), which are hereby incorporated by reference. Methods of in vitro and in vivo manipulation of monoclonal antibodies are also available to those skilled in the art. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler and Milstein, Nature 256, 495 (1975), or they may be made by recombinant methods, for example, as described in U.S. Pat. No. 4,816,567. The monoclonal antibodies for use with the present invention may also be isolated from antibody libraries using the techniques described in Clackson et al. Nature 352: 624-628 (1991), as well as in Marks et al., J. Mol. Biol. 222: 581-597 (1991).
  • Monoclonal antibodies can be isolated and purified from hybridoma cultures by a variety of well-established techniques. Such isolation techniques include affinity chromatography with Protein-A Sepharose, size-exclusion chromatography, and ion-exchange chromatography. See, e.g., Coligan, et al., sections 2.7.1-2.7.12 and sections 2.9.1-2.9.3; Bames, et al., Purification of Immunoglobulin G (IgG), in: [0111] Methods in Molecular Biology, Vol. 10, pages 79-104 (Humana Press (1992).
  • Another method for generating antibodies involves a Selected Lymphocyte Antibody Method (SLAM). The SLAM technology permits the generation, isolation and manipulation of monoclonal antibodies without the process of hybridoma generation. The methodology principally involves the growth of antibody forming cells, the physical selection of specifically selected antibody forming cells, the isolation of the genes encoding the antibody and the subsequent cloning and expression of those genes. [0112]
  • More specifically, an animal is immunized with a source of specific antigen. The animal can be a rabbit, mouse, rat, or any other convenient animal. This immunization may consist of purified protein, in either native or recombinant form, peptides, DNA encoding the protein of interest or cells expressing the protein of interest. After a suitable period, during which antibodies can be detected in the serum of the animal (usually weeks to months), blood, spleen or other tissues are harvested from the animal. Lymphocytes are isolated from the blood and cultured under specific conditions to generate antibody-forming cells, with antibody being secreted into the culture medium. These cells are detected by any of several means (complement mediated lysis of antigen-bearing cells, fluorescence detection or other) and then isolated using micromanipulation technology. The individual antibody forming cells are then processed for eventual single cell PCR to obtain the expressed Heavy and Light chain genes that encode the specific antibody. Once obtained and sequenced, these genes are cloned into an appropriate expression vector and recombinant, monoclonal antibody produced in a heterologous cell system. These antibodies are then purified via standard methodologies such as the use of protein A affinity columns. These types of methods are further described in Babcook, et al., Proc. Natl. Acad. Sci. (USA) 93: 7843-7848 (1996); U.S. Pat. No. 5,627,052; and PCT WO 92/02551 by Schrader. [0113]
  • Another method involves humanizing a monoclonal antibody by recombinant means to generate antibodies containing human specific and recognizable sequences. See, for review, Holmes, et al., J. Immunol., 158:2192-2201 (1997) and Vaswani, et al., Annals Allergy, Asthma & Immunol., 81:105-115 (1998). The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional polyclonal antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In additional to their specificity, the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other immunoglobulins. The modifier “monoclonal” indicates the antibody is obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. [0114]
  • The monoclonal antibodies herein specifically include “chimeric” antibodies (immunoglobulins) in which a portion of the heavy and/or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Pat. No. 4,816,567); Morrison et al. Proc. Natl. Acad. Sci. 81, 6851-6855 (1984). [0115]
  • Methods of making antibody fragments are also known in the art (see for example, Harlow and Lane, [0116] Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York, (1988), incorporated herein by reference). Antibody fragments of the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. Coli of DNA encoding the fragment. Antibody fragments can be obtained by pepsin or papain digestion of whole antibodies conventional methods. For example, antibody fragments can be produced by enzymatic cleavage of antibodies with pepsin to provide a 5S fragment denoted F(ab′)2. This fragment can be further cleaved using a thiol reducing agent, and optionally a blocking group for the sulfhydryl groups resulting from cleavage of disulfide linkages, to produce 3.5S Fab=monovalent fragments. Alternatively, an enzymatic cleavage using pepsin produces two monovalent Fab′ fragments and an Fc fragment directly. These methods are described, for example, in U.S. Pat. No. 4,036,945 and No. 4,331,647, and references contained therein. These patents are hereby incorporated in their entireties by reference.
  • Other methods of cleaving antibodies, such as separation of heavy chains to form monovalent light-heavy chain fragments, further cleavage of fragments, or other enzymatic, chemical, or genetic techniques may also be used, so long as the fragments bind to the antigen that is recognized by the intact antibody. For example, Fv fragments comprise an association of VH and VL chains. This association may be noncovalent or the variable chains can be linked by an intermolecular disulfide bond or cross-linked by chemicals such as glutaraldehyde. Preferably, the Fv fragments comprise VH and VL chains connected by a peptide linker. These single-chain antigen binding proteins (sFv) are prepared by constructing a structural gene comprising DNA sequences encoding the VH and VL domains connected by an oligonucleotide. The structural gene is inserted into an expression vector, which is subsequently introduced into a host cell such as [0117] E. coli. The recombinant host cells synthesize a single polypeptide chain with a linker peptide bridging the two V domains. Methods for producing sFvs are described, for example, by Whitlow, et al., Methods: a Companion to Methods in Enzmmology, Vol. 2, page 97 (1991); Bird, et al., Science 242:423-426 (1988); Ladner, et al, U.S. Pat. No. 4,946,778; and Pack, et al., Bio/Technology 11:1271-77 (1993).
  • Another form of an antibody fragment is a peptide coding for a single complementarity-determining region (CDR). CDR peptides (“minimal recognition units”) can be obtained by constructing genes encoding the CDR of an antibody of interest. Such genes are prepared, for example, by using the polymerase chain reaction to synthesize the variable region from RNA of antibody-producing cells. See, for example, Larrick, et al., [0118] Methods: a Companion to Methods in Enzymology, Vol. 2, page 106 (1991).
  • The invention further contemplates human and humanized forms of non-human (e.g. murine) antibodies. Such humanized antibodies can be chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab′, F(ab′)[0119] 2 or other antigen-binding subsequences of antibodies) that contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a nonhuman species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity.
  • In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications are made to further refine and optimize antibody performance. In general, humanized antibodies can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the Fv regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see: Jones et al., Nature 321, 522-525 (1986); Reichmann et al., Nature 332, 323-329 (1988); Presta, Curr. Op. Struct. Biol. 2, 593-596 (1992); Holmes, et al., J. Immunol., 158:2192-2201 (1997) and Vaswani, et al., Annals Allergy, Asthma & Immunol., 81:105-115 (1998); U.S. Pat. Nos. 4,816,567 and 6,331,415; PCT/GB84/00094; PCT/US86/02269; PCT/US89/00077; PCT/US88/02514; and WO91/09967, each of which is incorporated herein by reference in its entirety. [0120]
  • The invention also provides methods of mutating antibodies to optimize their affinity, selectivity, binding strength or other desirable property. A mutant antibody refers to an amino acid sequence variant of an antibody. In general, one or more of the amino acid residues in the mutant antibody is different from what is present in the reference antibody. Such mutant antibodies necessarily have less than 100% sequence identity or similarity with the reference amino acid sequence. In general, mutant antibodies have at least 75% amino acid sequence identity or similarity with the amino acid sequence of either the heavy or light chain variable domain of the reference antibody. Preferably, mutant antibodies have at least 80%, more preferably at least 85%, even more preferably at least 90%, and most preferably at least 95% amino acid sequence identity or similarity with the amino acid sequence of either the heavy or light chain variable domain of the reference antibody. [0121]
  • The antibodies of the invention are isolated antibodies. An isolated antibody is one that has been identified and separated and/or recovered from a component of the environment in which it was produced. Contaminant components of its production environment are materials that would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes. The term “isolated antibody” also includes antibodies within recombinant cells because at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step. [0122]
  • If desired, the antibodies of the invention can be purified by any available procedure. For example, the antibodies can be affinity purified by binding an antibody preparation to a solid support to which the antigen used to raise the antibodies is bound. After washing off contaminants, the antibody can be eluted by known procedures. Those of skill in the art will know of various techniques common in the immunology arts for purification and/or concentration of polyclonal antibodies, as well as monoclonal antibodies (see for example, Coligan, et al., [0123] Unit 9, Current Protocols in Immunology, Wiley Interscience, 1991, incorporated by reference).
  • In preferred embodiments, the antibody will be purified as measurable by at least three different methods: 1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight; 2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator; or 3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomasie blue or, preferably, silver stain. [0124]
  • The invention also provides antibodies that can bind to CD83 polypeptides. Sequences of complementarity determining regions (CDRs) or hypervariable regions from light and heavy chains of these anti-CD83 antibodies are provided. For example, a heavy chain variable region having a CDR1 sequence of SYDMT (SEQ ID NO:23), SYDMS (SEQ ID NO:24), DYDLS (SEQ ID NO:25) or SYDMS (SEQ ID NO:26) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products and/or modulate the immune response. In other embodiments, a heavy chain variable region having a CDR2 sequence of YASGSTYY (SEQ ID NO:27), SSSGTTYY (SEQ ID NO:28), YASGSTYY (SEQ ID NO:29), AIDGNPYY (SEQ ID NO:30) or STAYNSHY (SEQ ID NO:31) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system. In further embodiments of the invention, a heavy chain variable region having a CDR3 sequence of EHAGYSGDTGH (SEQ ID NO:32), EGAGVSMT (SEQ ID NO:33), EDAGFSNA (SEQ ID NO:34), GAGD (SEQ ID NO:35) or GGSWLD (SEQ ID NO:36) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system. [0125]
  • Moreover, a light chain variable region having a CDR1 sequence of RCAYD (SEQ ID NO:37), RCADVV (SEQ ID NO:38), or RCALV (SEQ ID NO:39) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system. In other embodiments, a light chain variable region having a CDR2 sequence of QSISTY (SEQ ID NO:40), QSVSSY (SEQ ID NO:41), ESISNY (SEQ ID NO:42), KNVYNNNW (SEQ ID NO:43), or QSVYDNDE (SEQ ID NO:98) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products or modulate the immune system. In further embodiments, a light chain variable region having a CDR3 sequence of QQGYTHSNVDNV (SEQ ID NO:44), QQGYSISDIDNA (SEQ ID NO:45), QCTSGGKFISDGAA (SEQ ID NO:46), AGDYSSSSDNG (SEQ ID NO:47), or QATHYSSDWLTY (SEQ ID NO:48) can be used in an antibody, multimerized antibody or other single- or multi-valent binding moiety to bind to CD83 gene products. [0126]
  • Light and heavy chains that can bind CD83 polypeptides are also provided by the invention. For example, in one embodiment, the invention provides a 20D04 light chain that can bind to CD83 polypeptides. The amino acid sequence for this 20D04 light chain is provided below (SEQ ID NO:11). [0127]
      1 MDMRAPTQLL GLLLLWLPGA RCADVVMTQT PASVSAAVGG
     41 TVTINCQASE SISNYLSWYQ QKPGQPPKLL IYRTSTLASG
     81 VSSRFKGSGS GTEYTLTISG VQCDDVATYY CQCTSGGKFI
    121 SDGAAFGGGT EVVVKGDPVA PTVLLFPPSS DEVATGTVTI
    161 VCVANKYFPD VTVTWEVDGT TQTTGIENSK TPQNSADCTY
    201 NLSSTLTLTS TQYNSHKEYT CKVTQGTTSV VQSFSRKNC
  • A nucleic acid sequence for this 20D04 anti-CD83 light chain is provided below (SEQ ID NO:12). [0128]
      1 ATGGACATGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCG ATGTCGTGAT
     81 GACCCAGACT CCAGCCTCCG TGTCTGCAGC TGTGGGAGGC
    121 ACAGTCACCA TCAATTGCCA GGCCAGTGAA AGCATTAGCA
    161 ACTACTTATC CTGGTATCAG CAGAAACCAG GGCAGCCTCC
    201 CAAGCTCCTG ATCTACAGGA CATCCACTCT GGCATCTGGG
    241 GTCTCATCGC GGTTCAAAGG CAGTGGATCT GGGACAGAGT
    281 ACACTCTCAC CATCAGCGGC GTGCAGTGTG ACGATGTTGC
    321 CACTTACTAC TGTCAATGCA CTTCTGGTGG GAAGTTCATT
    361 AGTGATGGTG CTGCTTTCGG CGGAGGGACC GAGGTGGTGG
    401 TCAAAGGTGA TCCAGTTGCA CCTACTGTCC TCCTCTTCCC
    441 ACCATCTAGC GATGAGGTGG CAACTGGAAC AGTCACCATC
    481 GTGTGTGTGG CGAATAAATA CTTTCCCGAT GTCACCGTCA
    521 CCTGGGAGGT GGATGGCACC ACCCAAACAA CTGGCATCGA
    561 GAACAGTAAA ACACCGCAGA ATTCTGCAGA TTGTACCTAC
    601 AACCTCAGCA GCACTCTGAC ACTGACCAGC ACACAGTACA
    641 ACAGCCACAA AGAGTACACC TGCAAGGTGA CCCAGGGCAC
    681 GACCTCAGTC GTCCAGAGCT TCAGTAGGAA GAACTGTTAA
  • In another embodiment, the invention provides a 20D04 heavy chain that can bind to CD83 polypeptides. The amino acid sequence for this 20D04 heavy chain is provided below (SEQ ID NO:13). [0129]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGFSLSNN AINWVRQAPG KGLEWIGYIW SGGLTYYANW
     81 AEGRFTISKT STTVDLKMTS PTIEDTATYF CARGINNSAL
    121 WGPGTLVTVS SGQPKAPSVF PLAPCCGDTP SSTVTLGCLV
    161 KGYLPEPVTV TWNSGTLTNG VRTFPSVRQS SGLYSLSSVV
    201 SVTSSSQPVT CNVAHPATNT KVDKTVAPST CSKPTCPPPE
    241 LLGGPSVFIF PPKPKDTLMI SRTPEVTCVV VDVSQDDPEV
    281 QFTWYINNEQ VRTARPPLRE QQFNSTIRVV STLPIAHQDW
    321 LRGKEFKCKV HNKALPAPIE KTISKARGQP LEPKVYTMGP
    361 PREELSSRSV SLTCMINGFY PSDISVEWEK NGKAEDNYKT
    401 TPAVLDSDGS YFLYNKLSVP TSEWQRGDVF TCSVMHEALH
    441 NHYTQKSISR SPGK
  • A nucleic acid sequence for this 20D04 anti-CD83 heavy chain is provided below (SEQ ID NO:14). [0130]
       1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACCGTCTCTG GATTCTCCCT CAGTAACAAT GCAATAAACT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTAG AGTGGATCGG
     201 ATACATTTGG AGTGGTGGGC TTACATACTA CGCGAACTGG
     241 GCGGAAGGCC GATTCACCAT CTCCAAAACC TCGACTACGG
     281 TGGATCTGAA GATGACCAGT CCGACAATCG AGGACACGGC
     321 CACCTATTTC TGTGCCAGAG GGATTAATAA CTCCGCTTTG
     361 TGGGGCCCAG GCACCCTGGT CACCGTCTCC TCAGGGCAAC
     401 CTAAGGCTCC ATCAGTCTTC CCACTGGCCC CCTGCTGCGG
     441 GGACACACCC TCTAGCACGG TGACCTTGGG CTGCCTGGTC
     481 AAAGGCTACC TCCCGGAGCC AGTGACCGTG ACCTGGAACT
     521 CGGGCACCCT CACCAATGGG GTACGCACCT TCCCGTCCGT
     561 CCGGCAGTCC TCAGGCCTCT ACTCGCTGAG CAGCGTGGTG
     601 AGCGTGACCT CAAGCAGCCA GCCCGTCACC TGCAACGTGG
     641 CCCACCCAGC CACCAACACC AAAGTGGACA AGACCGTTGC
     681 GCCCTCGACA TGCAGCAAGC CCACGTGCCC ACCCCCTGAA
     721 CTCCTGGGGG GACCGTCTGT CTTCATCTTC CCCCCAAAAC
     761 CCAAGGACAC CCTCATGATC TCACGCACCC CCGAGGTCAC
     801 ATGCGTGGTG GTGGACGTGA GCCAGGATGA CCCCGAGGTG
     841 CAGTTCACAT GGTACATAAA CAACGAGCAG GTGCGCACCG
     881 CCCGGCCGCC GCTACGGGAG CAGCAGTTCA ACAGCACGAT
     921 CCGCGTGGTC AGCACCCTCC CCATCGCGCA CCAGGACTGG
     961 CTGAGGGGCA AGGAGTTCAA GTGCAAAGTC CACAACAAGG
    1001 CACTCCCGGC CCCCATCGAG AAAACCATCT CCAAAGCCAG
    1041 AGGGCAGCCC CTGGAGCCGA AGGTCTACAC CATGGGCCCT
    1081 CCCCGGGAGG AGCTGAGCAG CAGGTCGGTC AGCCTGACCT
    1121 GCATGATCAA CGGCTTCTAC CCTTCCGACA TCTCGGTGGA
    1161 GTGGGAGAAG AACGGGAAGG CAGAGGACAA CTACAAGACC
    1201 ACGCCGGCCG TGCTGGACAG CGACGGCTCC TACTTCCTCT
    1241 ACAACAAGCT CTCAGTGCCC ACGAGTGAGT GGCAGCGGGG
    1281 CGACGTCTTC ACCTGCTCCG TGATGCACGA GGCCTTGCAC
    1321 AACCACTACA CGCAGAAGTC CATCTCCCGC TCTCCGGGTA
    1361 AA
  • In another embodiment, the invention provides a 111 G05 light chain that can bind to CD83 polypeptides. The amino acid sequence for this 11 G05 light chain is provided below (SEQ ID NO:15). [0131]
      1 MDTRAPTQLL GLLLLWLPGA RCADVVMTQT PASVSAAVGG
     41 TVTINCQSSK NVYNNNWLSW FQQKPGQPPK LLIYYASTLA
     81 SGVPSRFRGS GSGTQFTLTI SDVQCDDAAT YYCAGDYSSS
    121 SDNGFGGGTE VVVKGDPVAP TVLLFPPSSD EVATGTVTIV
    161 CVANKYFPDV TVTWEVDGTT QTTGIENSKT PQNSADCTYN
    201 LSSTLTLTST QYNSHKEYTC KVTQGTTSVV QSFSRKNC
  • A nucleic acid sequence for this 11G05 anti-CD83 light chain is provided below (SEQ ID NO:16). [0132]
      1 ATGGACACCA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCG ACGTCGTGAT
     81 GACCCAGACT CCAGCCTCCG TGTCTGCAGC TGTGGGAGGC
    121 ACAGTCACCA TCAATTGCCA GTCCAGTAAG AATGTTTATA
    161 ATAACAACTG GTTATCCTGG TTTCAGCAGA AACCAGGGCA
    201 GCCTCCCAAG CTCCTGATCT ATTATGCATC CACTCTGGCA
    241 TCTGGGGTCC CATCGCGGTT CAGAGGCAGT GGATCTGGGA
    281 CACAGTTCAC TCTCACCATT AGCGACGTGC AGTGTGACGA
    321 TGCTGCCACT TACTACTGTG CAGGCGATTA TAGTAGTAGT
    361 AGTGATAATG GTTTCGGCGG AGGGACCGAG GTGGTGGTCA
    401 AAGGTGATCC AGTTGCACCT ACTGTCCTCC TCTTCCCACC
    441 ATCTAGCGAT GAGGTGGCAA CTGGAACAGT CACCATCGTG
    481 TGTGTGGCGA ATAAATACTT TCCCGATGTC ACCGTCACCT
    521 GGGAGGTGGA TGGCACCACC CAAACAACTG GCATCGAGAA
    561 CAGTAAAACA CCGCAGAATT CTGCAGATTG TACCTACAAC
    601 CTCAGCAGCA CTCTGACACT GACCAGCACA CAGTACAACA
    641 GCCACAAAGA GTACACCTGC AAGGTGACCC AGGGCACGAC
    681 CTCAGTCGTC CAGAGCTTCA GTAGGAAGAA CTGTTAA
  • In another embodiment, the invention provides a 111 G05 heavy chain that can bind to CD83 polypeptides. The amino acid sequence for this 11G05 heavy chain is provided below (SEQ ID NO:17). [0133]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGFTISDY DLSWVRQAPG EGLKYIGFIA IDGNPYYATW
     81 AKGRFTISKT STTVDLKITA PTTEDTATYF CARGAGDLWG
    121 PGTLVTVSSG QPKAPSVFPL APCCGDTPSS TVTLGCLVKG
    161 YLPEPVTVTW NSGTLTNGVR TFPSVRQSSG LYSLSSVVSV
    201 TSSSQPVTCN VAHPATNTKV DKTVAPSTCS KPTCPPPELL
    241 GGPSVFIFPP KPKDTLMISR TPEVTCVVVD VSQDDPEVQF
    281 TWYINNEQVR TARPPLREQQ FNSTIRVVST LPIAHQDWLR
    321 GKEFKCKVHN KALPAPIEKT ISKARGQPLE PKVYTMGPPR
    361 EELSSRSVSL TCMINGFYPS DISVEWEKNG KAEDNYKTTP
    401 AVLDSDGSYF LYNKLSVPTS EWQRGDVFTC SVMHEALHNH
    441 YTQKSISRSP GK
  • A nucleic acid sequence for this 11 G05 anti-CD83 heavy chain is provided below (SEQ ID NO:18). [0134]
       1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTCTCTG GATTCACCAT CAGTGACTAC GACTTGAGCT
     161 GGGTCCGCCA GGCTCCAGGG GAGGGGCTGA AATACATCGG
     201 ATTCATTGCT ATTGATGGTA ACCCATACTA CGCGACCTGG
     241 GCAAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG
     281 TGGATCTGAA AATCACCGCT CCGACAACCG AAGACACGGC
     321 CACGTATTTC TGTGCCAGAG GGGCAGGGGA CCTCTGGGGC
     361 CCAGGGACCC TCGTCACCGT CTCTTCAGGG CAACCTAAGG
     401 CTCCATCAGT CTTCCCACTG GCCCCCTGCT GCGGGGACAC
     441 ACCCTCTAGC ACGGTGACCT TGGGCTGCCT GGTCAAAGGC
     481 TACCTCCCGG AGCCAGTGAC CGTGACCTGG AACTCGGGCA
     521 CCCTCACCAA TGGGGTACGC ACCTTCCCGT CCGTCCGGCA
     561 GTCCTCAGGC CTCTACTCGC TGAGCAGCGT GGTGAGCGTG
     601 ACCTCAAGCA GCCAGCCCGT CACCTGCAAC GTGGCCCACC
     641 CAGCCACCAA CACCAAAGTG GACAAGACCG TTGCGCCCTC
     681 GACATGCAGC AAGCCCACGT GCCCACCCCC TGAACTCCTG
     721 GGGGGACCGT CTGTCTTCAT CTTCCCCCCA AAACCCAAGG
     761 ACACCCTCAT GATCTCACGC ACCCCCGAGG TCACATGCGT
     801 GGTGGTGGAC GTGAGCCAGG ATGACCCCGA GGTGCAGTTC
     841 ACATGGTACA TAAACAACGA GCAGGTGCGC ACCGCCCGGC
     881 CGCCGCTACG GGAGCAGCAG TTCAACAGCA CGATCCGCGT
     921 GGTCAGCACC CTCCCCATCG CGCACCAGGA CTGGCTGAGG
     961 GGCAAGGAGT TCAAGTGCAA AGTCCACAAC AAGGCACTCC
    1001 CGGCCCCCAT CGAGAAAACC ATCTCCAAAG CCAGAGGGCA
    1041 GCCCCTGGAG CCGAAGGTCT ACACCATGGG CCCTCCCCGG
    1081 GAGGAGCTGA GCAGCAGGTC GGTCAGCCTG ACCTGCATGA
    1120 TCAACGGCTT CTACCCTTCC GACATCTCGG TGGAGTGGGA
    1161 GAAGAACGGG AAGGCAGAGG ACAACTACAA GACCACGCCG
    1201 GCCGTGCTGG ACAGCGACGG CTCCTACTTC CTCTACAACA
    1241 AGCTCTCAGT GCCCACGAGT GAGTGGCAGC GGGGCGACGT
    1281 CTTCACCTGC TCCGTGATGC ACGAGGCCTT GCACAACCAC
    1321 TACACGCAGA AGTCCATCTC CCGCTCTCCG GGTAAA
  • In another embodiment, the invention provides a [0135] 14C 12 light chain that can bind to CD83 polypeptides. The amino acid sequence for this 14C12 light chain is provided below (SEQ ID NO:19).
      1 MDXRAPTQLL GLLLLWLPGA RCALVMTQTP ASVSAAVGGT
     41 VTINCQSSQS VYDNDELSWY QQKPGQPPKL LIYLASKLAS
     81 GVPSRFKGSG SGTQFALTIS GVQCDDAATY YCQATHYSSD
    121 WYLTFGGGTE VVVKGDPVAP TVLLFPPSSD EVATGTVTIV
    161 CVANKYFPDV TVTWEVDGTT QTTGIENSKT PQNSADCTYN
    201 LSSTLTLTST QYNSHKEYTC KVTQGTTSVV QSFSRKNC
  • A nucleic acid sequence for this 14C12 anti-CD83 light chain is provided below (SEQ ID NO:20). [0136]
      1 ATGGACATRA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCC TTGTGATGAC
     81 CCAGACTCCA GCCTCCGTGT CTGCAGCTGT GGGAGGCACA
    121 GTCACCATCA ATTGCCAGTC CAGTCAGAGT GTTTATGATA
    161 ACGACGAATT ATCCTGGTAT CAGCAGAAAC CAGGGCAGCC
    201 TCCCAAGCTC CTGATCTATC TGGCATCCAA GTTGGCATCT
    241 GGGGTCCCAT CCCGATTCAA AGGCAGTGGA TCTGGGACAC
    281 AGTTCGCTCT CACCATCAGC GGCGTGCAGT GTGACGATGC
    321 TGCCACTTAC TACTGTCAAG CCACTCATTA TAGTAGTGAT
    361 TGGTATCTTA CTTTCGGCGG AGGGACCGAG GTGGTGGTCA
    401 AAGGTGATCC AGTTGCACCT ACTGTCCTCC TCTTCCCACC
    441 ATCTAGCGAT GAGGTGGCAA CTGGAACAGT CACCATCGTG
    481 TGTGTGGCGA ATAAATACTT TCCCGATGTC ACCGTCACCT
    521 GGGAGGTGGA TGGCACCACC CAAACAACTG GCATCGAGAA
    561 CAGTAAAACA CCGCAGAATT CTGCAGATTG TACCTACAAC
    601 CTCAGCAGCA CTCTGACACT GACCAGCACA CAGTACAACA
    641 GCCACAAAGA GTACACCTGC AAGGTGACCC AGGGCACGAC
    681 CTCAGTCGTC CAGAGCTTCA GTAGGAAGAA CTGTTAA
  • In another embodiment, the invention provides a [0137] 14C 12 heavy chain that can bind to CD83 polypeptides. The amino acid sequence for this 14C 12 heavy chain is provided below (SEQ ID NO:21).
      1 METGLRWLLL VAVLKGVHCQ SVEESGGRLV TPGTPLTLTC
     41 TASGFSRSSY DMSWVRQAPG KGLEWVGVIS TAYNSHYASW
     81 AKGRFTISRT STTVDLKMTS LTTEDTATYF CARGGSWLDL
    121 WGQGTLVTVS SGQPKAPSVF PLAPCCGDTP SSTVTLGCLV
    161 KGYLPEPVTV TWNSGTLTNG VRTFPSVRQS SGLYSLSSVV
    201 SVTSSSQPVT CNVAHPATNT KVDKTVAPST CSKPTCPPPE
    241 LLGGPSVFIF PPKPKDTLMI SRTPEVTCVV VDVSQDDPEV
    281 QFTWYINNEQ VRTARPPLRE QQFNSTIRVV STLPIAHQDW
    321 LRGKEFKCKV HNKALPAPIE KTISKARGQP LEPKVYTMGP
    361 PREELSSRSV SLTCMINGFY PSDISVEWEK NGKAEDNYKT
    401 TPAVLDSDGS YFLYNKLSVP TSEWQRGDVF TCSVMHEALH
    441 NHYTQKSISR SPGK
  • A nucleic acid sequence for this 14C12 anti-CD83 heavy chain is provided below (SEQ ID NO:22). [0138]
       1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCACTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGCCTCTG GATTCTCCCG CAGCAGCTAC GACATGAGCT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGGTCGG
     201 AGTCATTAGT ACTGCTTATA ACTCACACTA CGCGAGCTGG
     241 GCAAAAGGCC GATTCACCAT CTCCAGAACC TCGACCACGG
     281 TGGATCTGAA AATGACCAGT CTGACAACCG AAGACACGGC
     321 CACCTATTTC TGTGCCAGAG GGGGTAGTTG GTTGGATCTC
     361 TGGGGCCAGG GCACCCTGGT CACCGTCTCC TCAGGGCAAC
     401 CTAAGGCTCC ATCAGTCTTC CCACTGGCCC CCTGCTGCGG
     441 GGACACACCC TCTAGCACGG TGACCTTGGG CTGCCTGGTC
     481 AAAGGCTACC TCCCGGAGCC AGTGACCGTG ACCTGGAACT
     521 CGGGCACCCT CACCAATGGG GTACGCACCT TCCCGTCCGT
     561 CCGGCAGTCC TCAGGCCTCT ACTCGCTGAG CAGCGTGGTG
     601 AGCGTGACCT CAAGCAGCCA GCCCGTCACC TGCAACGTGG
     641 CCCACCCAGC CACCAACACC AAAGTGGACA AGACCGTTGC
     681 GCCCTCGACA TGCAGCAAGC CCACGTGCCC ACCCCCTGAA
     721 CTCCTGGGGG GACCGTCTGT CTTCATCTTC CCCCCAAAAC
     761 CCAAGGACAC CCTCATGATC TCACGCACCC CCGAGGTCAC
     801 ATGCGTGGTG GTGGACGTGA GCCAGGATGA CCCCGAGGTG
     841 CAGTTCACAT GGTACATAAA CAACGAGCAG GTGCGCACCG
     881 CCCGGCCGCC GCTACGGGAG CAGCAGTTCA ACAGCACGAT
     921 CCGCGTGGTC AGCACCCTCC CCATCGCGCA CCAGGACTGG
     961 CTGAGGGGCA AGGAGTTCAA GTGCAAAGTC CACAACAAGG
    1001 CACTCCCGGC CCCCATCGAG AAAACCATCT CCAAAGCCAG
    1041 AGGGCAGCCC CTGGAGCCGA AGGTCTACAC CATGGGCCCT
    1081 CCCCGGGAGG AGCTGAGCAG CAGGTCGGTC AGCCTGACCT
    1121 GCATGATCAA CGGCTTCTAC CCTTCCGACA TCTCGGTGGA
    1161 GTGGGAGAAG AACGGGAAGG CAGAGGACAA CTACAAGACC
    1200 ACGCCGGCCG TGCTGGACAG CGACGGCTCC TACTTCCTCT
    1241 ACAACAAGCT CTCAGTGCCC ACGAGTGAGT GGCAGCGGGG
    1281 CGACGTCTTC ACCTGCTCCG TGATGCACGA GGCCTTGCAC
    1321 AACCACTACA CGCAGAAGTC CATCTCCCGC TCTCCGGGTA
    1361 AA
  • In another embodiment, the invention provides a M83 020B08L light chain that can bind to CD83 polypeptides. The amino acid sequence for this M83 020B08L light chain is provided below (SEQ ID NO:58). [0139]
      1 MDMRAPTQLL GLLLLWLPGA RCAYDMTQTP ASVEVAVGGT
     41 VTIKCQASQS ISTYLDWYQQ KPGQPPKLLI YDASDLASGV
     81 PSRFKGSGSG TQFTLTTSDL ECADAATYYC QQGYTHSNVD
    121 NVFGGGTEVV VKGDPVAPTV LLFPPSSDEV ATGTVTIVCV
    161 ANKYFPDVTV TWEVDGTTQT TGIENSKTPQ NSADCTYNLS
    201 STLTLTSTQY NSHKEYTCKV TQGTTSVVQS FSRKNC
  • A nucleic acid sequence for this M83 020B08L anti-CD83 light chain is provided below (SEQ ID NO:59). [0140]
      1 ATGGACATGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCT ATGATATGAC
     81 CCAGACTCCA GCCTCTGTGG AGGTAGCTGT GGGAGGCACA
    121 GTCACCATCA AGTGCCAGGC CAGTCAGAGC ATTAGTACCT
    161 ACTTAGACTG GTATCAGCAG AAACCAGGGC AGCCTCCCAA
    201 GCTCCTGATC TATGATGCAT CCGATCTGGC ATCTGGGGTC
    241 CCATCGCGGT TCAAAGGCAG TGGATCTGGG ACACAGTTCA
    281 CTCTCACCAT CAGCGACCTG GAGTGTGCCG ATGCTGCCAC
    321 TTACTACTGT CAACAGGGTT ATACACATAG TAATGTTGAT
    361 AATGTTTTCG GCGGAGGGAC CGAGGTGGTG GTCAAAGGTG
    401 ATCCAGTTGC ACCTACTGTC CTCCTCTTCC CACCATCTAG
    441 CGATGAGGTG GCAACTGGAA CAGTCACCAT CGTGTGTGTG
    481 GCGAATAAAT ACTTTCCCGA TGTCACCGTC ACCTGGGAGG
    521 TGGATGGCAC CACCCAAACA ACTGGCATCG AGAACAGTAA
    561 AACACCGCAG AATTCTGCAG ATTGTACCTA CAACCTCAGC
    601 AGCACTCTGA CACTGACCAG CACACAGTAC AACAGCCACA
    641 AAGAGTACAC CTGCAAGGTG ACCCAGGGCA CGACCTCAGT
    681 CGTCCAGAGC TTCAGTAGGA AGAACTGTTA A
  • In another embodiment, the invention provides a M83 020B08H heavy chain that can bind to CD83 polypeptides. The amino acid sequence for this M83 020B08H heavy chain is provided below (SEQ ID NO:60). [0141]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGFSLSSY DMTWVRQAPG KGLEWIGIIY ASGTTYYANW
     81 AKGRFTISKT STTVDLKVTS PTIGDTATYF CAREGAGVSM
    121 TLWGPGTLVT VSSGQPKAPS VFPLAPCCGD TPSSTVTLGC
    161 LVKGYLPEPV TVTWNSGTLT NGVRTFPSVR QSSGLYSLSS
    201 VVSVTSSSQP VTCNVAHPAT NTKVDKTVAP STCSKPTCPP
    241 PELLGGPSVF IFPPKPKDTL MISRTPEVTC VVVDVSQDDP
    281 EVQFTWYINN EQVRTARPPL REQQFNSTIR VVSTLPIAHQ
    321 DWLRGKEFKC KVHNKALPAP IEKTISKARG QPLEPKVYTM
    361 GPPREELSSR SVSLTCMING FYPSDISVEW EKNGKAEDNY
    401 KTTPAVLDSD GSYFLYNKLS VPTSEWQRGD VFTCSVMHEA
    441 LHNHYTQKSI SRSPGK
  • A nucleic acid sequence for this M83 020B08H anti-CD83 heavy chain is provided below (SEQ ID NO:61). [0142]
       1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTCTCTG GATTCTCCCT CAGCAGCTAC GACATGACCT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGATCGG
     201 AATCATTTAT GCTAGTGGTA CCACATACTA CGCGAACTGG
     241 GCGAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG
     281 TGGATCTGAA AGTCACCAGT CCGACAATCG GGGACACGGC
     321 CACCTATTTC TGTGCCAGAG AGGGGGCTGG TGTTAGTATG
     361 ACCTTGTGGG GCCCAGGCAC CCTGGTCACC GTCTCCTCAG
     401 GGCAACCTAA GGCTCCATCA GTCTTCCCAC TGGCCCCCTG
     441 CTGCGGGGAC ACACCCTCTA GCACGGTGAC CTTGGGCTGC
     481 CTGGTCAAAG GCTACCTCCC GGAGCCAGTG ACCGTGACCT
     521 GGAACTCGGG CACCCTCACC AATGGGGTAC GCACCTTCCC
     561 GTCCGTCCGG CAGTCCTCAG GCCTCTACTC GCTGAGCAGC
     601 GTGGTGAGCG TGACCTCAAG CAGCCAGCCC GTCACCTGCA
     641 ACGTGGCCCA CCCAGCCACC AACACCAAAG TGGACAAGAC
     681 CGTTGCGCCC TCGACATGCA GCAAGCCCAC GTGCCCACCC
     721 CCTGAACTCC TGGGGGGACC GTCTGTCTTC ATCTTCCCCC
     761 CAAAACCCAA GGACACCCTC ATGATCTCAC GCACCCCCGA
     801 GGTCACATGC GTGGTGGTGG ACGTGAGCCA GGATGACCCC
     841 GAGGTGCAGT TCACATGGTA CATAAACAAC GAGCAGGTGC
     881 GCACCGCCCG GCCGCCGCTA CGGGAGCAGC AGTTCAACAG
     921 CACGATCCGC GTGGTCAGCA CCCTCCCCAT CGCGCACCAG
     961 GACTGGCTGA GGGGCAAGGA GTTCAAGTGC AAAGTCCACA
    1001 ACAAGGCACT CCCGGCCCCC ATCGAGAAAA CCATCTCCAA
    1041 AGCCAGAGGG CAGCCCCTGG AGCCGAAGGT CTACACCATG
    1081 GGCCCTCCCC GGGAGGAGCT GAGCAGCAGG TCGGTCAGCC
    1121 TGACCTGCAT GATCAACGGC TTCTACCCTT CCGACATCTC
    1161 GGTGGAGTGG GAGAAGAACG GGAAGGCAGA GGACAACTAC
    1201 AAGACCACGC CGGCCGTGCT GGACAGCGAC GGCTCCTACT
    1241 TCCTCTACAA CAAGCTCTCA GTGCCCACGA GTGAGTGGCA
    1281 GCGGGGCGAC GTCTTCACCT GCTCCGTGAT GCACGAGGCC
    1321 TTGCACAACC ACTACACGCA GAAGTCCATC TCCCGCTCTC
    1361 CGGGTAAA
  • In another embodiment, the invention provides a M83 006G05L light chain that can bind to CD83 polypeptides. The amino acid sequence for this M83 006G05L light chain is provided below (SEQ ID NO:62). [0143]
      1 MDMRAPTQLL GLLLLWLPGA RCAYDMTQTP ASVEVAVGGT
     41 VAIKCQASQS VSSYLAWYQQ KPGQPPKPLI YEASMLAAGV
     81 SSRFKGSGSG TDFTLTISDL ECDDAATYYC QQGYSISDID
    121 NAFGGGTEVV VKGDPVAPTV LLFPPSSDEV ATGTVTIVCV
    161 ANKYFPDVTV TWEVDGTTQT TGIENSKTPQ NSADCTYNLS
    201 STLTLTSTQY NSHKEYTCKV TQGTTSVVQS FSRKNC
  • A nucleic acid sequence for M83 006G05L anti-CD83 light chain is provided below (SEQ ID NO:63). [0144]
      1 ATGGACATGA GGGCCCCCAC TCAACTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC AGATGTGCCT ATGATATGAC
     81 CCAGACTCCA GCCTCTGTGG AGGTAGCTGT GGGAGGCACA
    121 GTCGCCATCA AGTGCCAGGC CAGTCAGAGC GTTAGTAGTT
    161 ACTTAGCCTG GTATCAGCAG AAACCAGGGC AGCCTCCCAA
    201 GCCCCTGATC TACGAAGCAT CCATGCTGGC GGCTGGGGTC
    241 TCATCGCGGT TCAAAGGCAG TGGATCTGGG ACAGACTTCA
    281 CTCTCACCAT CAGCGACCTG GAGTGTGACG ATGCTGCCAC
    321 TTACTATTGT CAACAGGGTT ATTCTATCAG TGATATTGAT
    361 AATGCTTTCG GCGGAGGGAC CGAGGTGGTG GTCAAAGGTG
    401 ATCCAGTTGC ACCTACTGTC CTCCTCTTCC CACCATCTAG
    441 CGATGAGGTG GCAACTGGAA CAGTCACCAT CGTGTGTGTG
    481 GCGAATAAAT ACTTTCCCGA TGTCACCGTC ACCTGGGAGG
    521 TGGATGGCAC CACCCAAACA ACTGGCATCG AGAACAGTAA
    561 AACACCGCAG AATTCTGCAG ATTGTACCTA CAACCTCAGC
    601 AGCACTCTGA CACTGACCAG CACACAGTAC AACAGCCACA
    641 AAGAGTACAC CTGCAAGGTG ACCCAGGGCA CGACCTCAGT
    681 CGTCCAGAGC TTCAGTAGGA AGAACTGTTA A
  • In another embodiment, the invention provides a M83 006G05L heavy chain that can bind to CD83 polypeptides. The amino acid sequence for this M83 006G05L heavy chain is provided below (SEQ ID NO:64). [0145]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV SPGTPLTLTC
     41 TASGFSLSSY DMSWVRQAPG KGLEYIGIIS SSGSTYYASW
     81 AKGRFTISKT STTVDLEVTS LTTEDTATYF CSREHAGYSG
    121 DTGHLWGPGT LVTVSSGQPK APSVFPLAPC CGDTPSSTVT
    161 LGCLVKGYLP EPVTVTWNSG TLTNGVRTFP SVRQSSGLYS
    201 LSSVVSVTSS SQPVTCNVAH PATNTKVDKT VAPSTCSKPT
    241 CPPPELLGGP SVFIFPPKPK DTLMISRTPE VTCVVVDVSQ
    281 DDPEVQFTWY INNEQVRTAR PPLREQQFNS TIRVVSTLPI
    321 AHQDWLRGKE FKCKVHNKAL PAPIEKTISK ARGQPLEPKV
    361 YTMGPPREEL SSRSVSLTCM INGFYPSDIS VEWEKNGKAE
    401 DNYKTTPAVL DSDGSYFLYN KLSVPTSEWQ RGDVFTCSVM
    441 HEALHNHYTQ KSISRSPGK
  • A nucleic acid sequence for this M83 006G05L anti-CD83 heavy chain is provided below (SEQ ID NO:65). [0146]
       1 ATGGAGACAG GCCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC TCGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGCCTCTG GATTCTCCCT CAGTAGCTAC GACATGAGCT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATACATCGG
     201 AATCATTAGT AGTAGTGGTA GCACATACTA CGCGAGCTGG
     241 GCGAAAGGCC GATTCACCAT CTCCAAAACC TCGACCACGG
     281 TGGATCTGGA AGTGACCAGT CTGACAACCG AGGACACGGC
     321 CACCTATTTC TGTAGTAGAG AACATGCTGG TTATAGTGGT
     361 GATACGGGTC ACTTGTGGGG CCCAGGCACC CTGGTCACCG
     401 TCTCCTCGGG GCAACCTAAG GCTCCATCAG TCTTCCCACT
     441 GGCCCCCTGC TGCGGGGACA CACCCTCTAG CACGGTGACC
     481 TTGGGCTGCC TGGTCAAAGG CTACCTCCCG GAGCCAGTGA
     521 CCGTGACCTG GAACTCGGGC ACCCTCACCA ATGGGGTACG
     561 CACCTTCCCG TCCGTCCGGC AGTCCTCAGG CCTCTACTCG
     601 CTGAGCAGCG TGGTGAGCGT GACCTCAAGC AGCCAGCCCG
     641 TCACCTGCAA CGTGGCCCAC CCAGCCACCA ACACCAAAGT
     681 GGACAAGACC GTTGCGCCCT CGACATGCAG CAAGCCCACG
     721 TGCCCACCCC CTGAACTCCT GGGGGGACCG TCTGTCTTCA
     761 TCTTCCCCCC AAAACCCAAG GACACCCTCA TGATCTCACG
     801 CACCCCCGAG GTCACATGCG TGGTGGTGGA CGTGAGCCAG
     841 GATGACCCCG AGGTGCAGTT CACATGGTAC ATAAACAACG
     881 AGCAGGTGCG CACCGCCCGG CCGCCGCTAC GGGAGCAGCA
     921 GTTCAACAGC ACGATCCGCG TGGTCAGCAC CCTCCCCATC
     961 GCGCACCAGG ACTGGCTGAG GGGCAAGGAG TTCAAGTGCA
    1001 AAGTCCACAA CAAGGCACTC CCGGCCCCCA TCGAGAAAAC
    1041 CATCTCCAAA GCCAGAGGGC AGCCCCTGGA GCCGAAGGTC
    1081 TACACCATGG GCCCTCCCCG GGAGGAGCTG AGCAGCAGGT
    1121 CGGTCAGCCT GACCTGCATG ATCAACGGCT TCTACCCTTC
    1162 CGACATCTCG GTGGAGTGGG AGAAGAACGG GAAGGCAGAG
    1201 GACAACTACA AGACCACGCC GGCCGTGCTG GACAGCGACG
    1241 GCTCCTACTT CCTCTACAAC AAGCTCTCAG TGCCCACGAG
    1281 TGAGTGGCAG CGGGGCGACG TCTTCACCTG CTCCGTGATG
    1321 CACGAGGCCT TGCACAACCA CTACACGCAG AAGTCCATCT
    1361 CCCGCTCTCC GGGTAAA
  • In another embodiment, the invention provides a 96G08 light chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs). The amino acid sequence for this 96G08 light chain is provided below (SEQ ID NO:70). [0147]
      1 MDTRAPTQLL GLLLLWLPGA TFAQVLTQTA SPVSAPVGGT
     41 VTINCQSSQS VYNNDFLSWY QQKPGQPPKL LIYYASTLAS
     81 GVPSRFKGSG SGTQFTLTIS DLECDDAATY YCTGTYGNSA
    121 WYEDAFGGGT EVVVKRTPVA PTVLLFPPSS AELATGTATI
    161 VCVANKYFPD GTVTWKVDGI TQSSGINNSR TPQNSADCTY
    201 NLSSTLTLSS DEYNSHDEYT CQVAQDSGSP VVQSFSRKSC
  • The amino acid sequence for this 96G08 light chain with the CDR regions identified by underlining is provided below (SEQ ID NO:70). [0148]
      1 MDTRAPTQLL GLLLLWLPGA TFAQVLTQTA SPVSAPVGGT
     41 VTINCQSSQS VYNNDFLSWY QQKPGQPPKL LIYYASTLAS
     81 GVPSRFKGSG SGTQFTLTIS DLECDDAATY YCTGTYGNSA
    121 WYEDAFGGGT EVVVKRTPVA PTVLLFPPSS AELATGTATI
    161 VCVANKYFPD GTVTWKVDGI TQSSGINNSR TPQNSADCTY
    201 NLSSTLTLSS DEYNSHDEYT CQVAQDSGSP VVQSFSRKSC
  • Hence, the CDR regions in the 96G08 light chain include amino acid sequences QSSQSVYNNDFLS (SEQ ID NO:71), YASTLAS (SEQ ID NO:72), and TGTYGNSAWYEDA (SEQ ID NO:73). [0149]
  • A nucleic acid sequence for this 96G08 anti-CD83 light chain is provided below (SEQ ID NO:74). [0150]
      1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC ACATTTGCGC AAGTGCTGAC
     81 CCAGACTGCA TCGCCCGTGT CTGCACCTGT GGGAGGCACA
    121 GTCACCATCA ATTGCCAGTC CAGTCAGAGT GTTTATAATA
    161 ACGACTTCTT ATCCTGGTAT CAGCAGAAAC CAGGGCAGCC
    201 TCCCAAACTC CTGATCTATT ATGCATCCAC TCTGGCATCT
    241 GGGGTCCCAT CCCGGTTCAA AGGCAGTGGA TCTGGGACAC
    281 AGTTCACTCT CACCATCAGC GACCTGGAGT GTGACGATGC
    321 TGCCACTTAC TACTGTACAG GCACTTATGG TAATAGTGCT
    361 TGGTACGAGG ATGCTTTCGG CGGAGGGACC GAGGTGGTGG
    401 TCAAACGTAC GCCAGTTGCA CCTACTGTCC TCCTCTTCCC
    441 ACCATCTAGC GCTGAGCTGG CAACTGGAAC AGCCACCATC
    481 GTGTGCGTGG CGAATAAATA CTTTCCCGAT GGCACCGTCA
    521 CCTGGAAGGT GGATGGCATC ACCCAAAGCA GCGGCATCAA
    561 TAACAGTAGA ACACCGCAGA ATTCTGCAGA TTGTACCTAC
    601 AACCTCAGCA GTACTCTGAC ACTGAGCAGC GACGAGTACA
    641 ACAGCCACGA CGAGTACACC TGCCAGGTGG CCCAGGACTC
    681 AGGCTCACCG GTCGTCCAGA GCTTCAGTAG GAAGAGCTGT
    721 TAG
  • This nucleic acid sequence for the 96G08 anti-CD83 light chain with CDR regions identified by underlining is provided below (SEQ ID NO:99). [0151]
      1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC ACATTTGCGC AAGTGCTGAC
     81 CCAGACTGCA TCGCCCGTGT CTGCACCTGT GGGAGGCACA
    121 GTCACCATCA ATTGCCAGTC CAGTCACAGT GTTTATAATA
    161 ACGACTTCTT ATCC TGGTAT CAGCAGAAAC CAGGGCAGCC
    201 TCCCAAACTC CTGATCTATT ATGCATCCAC TCTGGCATCT
    241 GGGGTCCCAT CCCGGTTCAA AGGCAGTGGA TCTGGGACAC
    281 AGTTCACTCT CACCATCAGC GACCTGGAGT GTGACGATGC
    321 GCCACTTACT ACTGTACAGG CACTTATGGT AATAGTGCTT
    361 GGTACGAGGA TGCT TTCGGC GGAGGGACCG AGGTGGTGGT
    401 CAAACGTACG CCAGTTGCAC CTACTGTCCT CCTCTTCCCA
    441 CCATCTAGCG CTGAGCTGGC AACTGGAACA GCCACCATCG
    481 TGTGCGTGGC GAATAAATAC TTTCCCGATG GCACCGTCAC
    521 CTGGAAGGTG GATGGCATCA CCCAAAGCAG CGGCATCAAT
    561 AACAGTAGAA CACCGCAGAA TTCTGCAGAT TGTACCTACA
    601 ACCTCAGCAG TACTCTGACA CTGAGCAGCG ACGAGTACAA
    641 CAGCCACGAC GAGTACACCT GCCAGGTGGC CCAGGACTCA
    681 GGCTCACCGG TCGTCCAGAG CTTCAGTAGG AAGAGCTGTT
  • Hence, the CDR regions in the 96G08 light chain include nucleic acid sequences CAGTCCAGTCAGAGTGTTTATAATA (SEQ ID NO:75), ATGCATCCACTCTGGCATCT (SEQ ID NO:76), and ACAGGCACTTATGGT AATAGTGCTT (SEQ ID NO:77). [0152]
  • In another embodiment, the invention provides a 96G08 heavy chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs). The amino acid sequence for this 96G08 heavy chain is provided below (SEQ ID NO:78). [0153]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGIDLSSD GISWVRQAPG KGLEWIGIIS SGGNTYYASW
     81 AKGRFTISRT STTVDLKMTS LTTEDTATYF CARVVGGTYS
    121 IWGQGTLVTV SSASTKGPSV YPLAPGSAAQ TNSMVTLGCL
    161 VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ SDLYTLSSSV
    201 TVPSSTWPSE TVTCNVAHPA SSTKVDKKIV PRDCGCKPCI
    241 CTVPEVSSVF IFPPKPKDVL TITLTPKVTC VVVDISKDDP
    281 EVQFSWFVDD VEVHTAQTQP REEQFNSTFR SVSELPIMHQ
    321 DWLNGKEFKC RVNSAAFPAP IEKTISKTKG RPKAPQVYTI
    361 PPPKEQMAKD KVSLTCMITD FFPEDITVEW QWNGQPAENY
    401 KNTQPIMDTD GSYFVYSKLN VQKSNWEAGN TFTCSVLHEG
    441 LHNHHTEKSL SHSPGK
  • The amino acid sequence for the 96G08 heavy chain with the CDR regions identified by underlining is provided below (SEQ ID NO:78). [0154]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGIDLSSD GISWVRQAPG KGLEWIGIIS SGGNTYYASW
     81 AKGRFTISRT STTVDLKMTS LTTEDTATYF CARVVGGTYS
    121 IWGQGTLVTV SSASTKGPSV YPLAPGSAAQ TNSMVTLGCL
    161 VKGYFPEPVT VTWNSGSLSS GVHTFPAVLQ SDLYTLSSSV
    201 TVPSSTWPSE TVTCNVAHPA SSTKVDKKIV PRDCGCKPCI
    241 CTVPEVSSVF IFPPKPKDVL TITLTPKVTC VVVDISKDDP
    281 EVQFSWFVDD VEVHTAQTQP REEQFNSTFR SVSELPIMHQ
    321 DWLNGKEFKC RVNSAAFPAP IEKTISKTKG RPKAPQVYTI
    361 PPPKEQMAKD KVSLTCMITD FFPEDITVEW QWNGQPAENY
    401 KNTQPIMDTD GSYFVYSKLN VQKSNWEAGN TFTCSVLHEG
    441 LHNHHTEKSL SHSPGK
  • Hence, the CDR regions in the 96G08 heavy chain include amino acid sequences SDGIS (SEQ ID NO:79), IISSGGNTYYASWAKG (SEQ ID NO:80) and VVGGTYSI (SEQ ID NO:81). [0155]
  • A nucleic acid sequence for the 96G08 anti-CD83 heavy chain is provided below (SEQ ID NO:82). [0156]
       1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACACCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTGTCTG GAATCGACCT CAGTAGCGAT GGAATAAGCT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGATCGG
     201 AATCATTAGT AGTGGTGGTA ACACATACTA CGCGAGCTGG
     241 GCAAAAGGCC GATTCACCAT CTCCAGAACC TCGACCACGG
     281 TGGATCTGAA GATGACCAGT CTGACAACCG AGGACACGGC
     321 CACCTATTTC TGTGCCAGAG TTGTTGGTGG TACTTATAGC
     361 ATCTGGGGCC AGGGCACCCT CGTCACCGTC TCGAGCGCTT
     401 CTACAAAGGG CCCATCTGTC TATCCACTGG CCCCTGGATC
     441 TGCTGCCCAA ACTAACTCCA TGGTGACCCT GGGATGCCTG
     481 GTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA
     521 ACTCTGGATC CCTGTCCAGC GGTGTGCACA CCTTCCCAGC
     561 TGTCCTGCAG TCTGACCTCT ACACTCTGAG CAGCTCAGTG
     601 ACTGTCCCCT CCAGCACCTG GCCCAGCGAG ACCGTCACCT
     641 GCAACGTTGC CCACCCGGCC AGCAGCACCA AGGTGGACAA
     681 GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA
     721 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC
     761 CAAAGCCCAA GGATGTGCTC ACCATTACTC TGACTCCTAA
     801 GGTCACGTGT GTTGTGGTAG ACATCAGCAA GGATGATCCC
     841 GAGGTCCAGT TCAGCTGGTT TGTAGATGAT GTGGAGGTGC
     881 ACACAGCTCA GACGCAACCC CGGGAGGAGC AGTTCAACAG
     921 CACTTTCCGC TCAGTCAGTG AACTTCCCAT CATGCACCAG
     961 GACTGGCTCA ATGGCAAGGA GTTCAAATGC AGGGTCAACA
    1001 GTGCAGCTTT CCCTGCCCCC ATCGAGAAAA CCATCTCCAA
    1041 AACCAAAGGC AGACCGAAGG CTCCACAGGT GTACACCATT
    1081 CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC
    1121 TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC
    1161 TGTGGAGTGG CAGTGGAATG GGCAGCCAGC GGAGAACTAC
    1201 AAGAACACTC AGCCCATCAT GGACACAGAT GGCTCTTACT
    1241 TCGTCTACAG CAAGCTCAAT GTGCAGAAGA GCAACTGGGA
    1281 GGCAGGAAAT ACTTTCACCT GCTCTGTGTT ACATGAGGGC
    1321 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTC
    1361 CTGGTAAATG A
  • The nucleic acid sequence for the 96G08 anti-CD83 heavy chain with CDR regions identified by underlining is provided below is provided below (SEQ ID NO:82). [0157]
       1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACACCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTGTCTG GAATCGACCT CAGTAGCGAT GGAATAAGCT
     161 GGGTCCGCCA GGCTCCAGGG AAGGGGCTGG AATGGATCGG
     201 AATCATTAGT AGTGGTGGTA ACACATACTA CGCGAGCTGG
     241 GCAAAAGGCC GATTCACCAT CTCCAGAACC TCGACCACGG
     281 TGGATCTGAA GATGACCAGT CTGACAACCG AGGACACGGC
     321 CACCTATTTC TGTGCCAGAG TTGTTGGTGG TACTTATAGC
     361 ATCTGGGGCC AGGGCACCCT CGTCACCGTC TCGAGCGCTT
     401 CTACAAAGGG CCCATCTGTC TATCCACTGG CCCCTGGATC
     441 TGCTCCCCAA ACTAACTCCA TGGTGACCCT GGGATGCCTG
     481 GTCAAGGGCT ATTTCCCTGA GCCAGTGACA GTGACCTGGA
     521 ACTCTGGATC CCTGTCCAGC GGTGTGCACA CCTTCCCAGC
     561 TGTCCTGCAG TCTGACCTCT ACACTCTGAG CAGCTCAGTG
     601 ACTGTCCCCT CCAGCACCTG GCCCAGCGAG ACCGTCACCT
     641 GCAACGTTGC CCACCCGGCC AGCAGCACCA AGGTGGACAA
     681 GAAAATTGTG CCCAGGGATT GTGGTTGTAA GCCTTGCATA
     721 TGTACAGTCC CAGAAGTATC ATCTGTCTTC ATCTTCCCCC
     761 CAAAGCCCAA GGATGTGCTC ACCATTACTC TGACTCCTAA
     801 GGTCACGTGT GTTGTGGTAG ACATCAGCAA GGATGATCCC
     841 GAGGTCCAGT TCAGCTGGTT TGTAGATGAT GTGGAGGTGC
     881 ACACAGCTCA GACGCAACCC CGGGAGGAGC AGTTCAACAG
     921 CACTTTCCGC TCAGTCAGTG AACTTCCCAT CATGCACCAG
     961 GACTGGCTCA ATGGCAAGGA GTTCAAATGC AGGGTCAACA
    1001 GTGCAGCTTT CCCTGCCCCC ATCGAGAAAA CCATCTCCAA
    1041 AACCAAAGGC AGACCGAAGG CTCCACAGGT GTACACCATT
    1081 CCACCTCCCA AGGAGCAGAT GGCCAAGGAT AAAGTCAGTC
    1121 TGACCTGCAT GATAACAGAC TTCTTCCCTG AAGACATTAC
    1161 TGTGGAGTGG CAGTGGAATG GGCAGCCAGC GGAGAACTAC
    1201 AAGAACACTC AGCCCATCAT GGACACAGAT GGCTCTTACT
    1241 TCGTCTACAG CAAGCTCAAT GTGCAGAAGA GCAACTGGGA
    1281 GGCAGGAAAT ACTTTCACCT GCTCTGTGTT ACATGAGGGC
    1321 CTGCACAACC ACCATACTGA GAAGAGCCTC TCCCACTCTC
    1361 CTGGTAAATG A
  • Hence, the CDR regions in the 96G08 anti-CD83 heavy chain include AGCGATGGAATAAGC (SEQ ID NO:83), ATCATTAGTAGTGGTGGTA ACACATACTACGCGAGCTGGGCAAAAGGC (SEQ ID NO:84), and G TTGTTGGTGG TACTTATAGC ATC (SEQ ID NO:85). [0158]
  • In another embodiment, the invention provides a 95F04 light chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs). The amino acid sequence for this 95F04 light chain is provided below (SEQ ID NO:86). [0159]
      1 MDTRAPTQLL GLLLLWLPGA TFAQAVVTQT TSPVSAPVGG
     41 TVTINCQSSQ SVYGNNELSW YQQKPGQPPK LLIYQASSLA
     81 SGVPSRFKGS GSGTQFTLTI SDLECDDAAT YYCLGEYSIS
    121 ADNHFGGGTE VVVKRTPVAP TVLLFPPSSA ELATGTATIV
    161 CVANKYFPDG TVTWKVDGIT QSSGINNSRT PQNSADCTYN
    201 LSSTLTLSSD EYNSHDEYTC QVAQDSGSPV VQSFSRKSC
  • The amino acid sequence for the 95F04 anti-CD83 light chain with the CDR regions identified by underlining is provided below (SEQ ID NO:86). [0160]
      1 MDTRAPTQLL GLLLLWLPGA TFAQAVVTQT TSPVSAPVGG
     41 TVTINCQSSQ SVYGNNELSW YQQKPGQPPK LLIYQASSLA
     81 SGVPSRFKGS GSGTQFTLTI SDLECDDAAT YYCLGEYSIS
    121 ADNHFGGGTE VVVKRTPVAP TVLLFPPSSA ELATGTATIV
    161 CVANKYFPDG TVTWKVDGIT QSSGINNSRT PQNSADCTYN
    201 LSSTLTLSSD EYNSHDEYTC QVAQDSGSPV VQSFSRKSC
  • Hence, the CDR regions in the 95F04 anti-CD83 light chain include amino acid sequences QSSQSVYGNNELS (SEQ ID NO:87), QASSLAS (SEQ ID NO:88) and LGEYSISADNH (SEQ ID NO:89). [0161]
  • A nucleic acid sequence for this 95F04 anti-CD83 light chain is provided below (SEQ ID NO:90). [0162]
      1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC ACATTTGCCC AAGCCGTGGT
     81 GACCCAGACT ACATCGCCCG TGTCTGCACC TGTGGGAGGC
    121 ACAGTCACCA TCAATTGCCA GTCCAGTCAG AGTGTTTATG
    161 GTAACAACGA ATTATCCTGG TATCAGCAGA AACCAGGGCA
    201 GCCTCCCAAG CTCCTGATCT ACCAGGCATC CAGCCTGGCA
    241 TCTGGGGTCC CATCGCGGTT CAAAGGCAGT GGATCTGGGA
    281 CACAGTTCAC TCTCACCATC AGCGACCTGG AGTGTGACGA
    321 TGCTGCCACT TACTACTGTC TAGGCGAATA TAGCATTAGT
    361 GCTGATAATC ATTTCGGCGG AGGGACCGAG GTGGTGGTCA
    401 AACGTACGCC AGTTGCACCT ACTGTCCTCC TCTTCCCACC
    441 ATCTAGCGCT GAGCTGGCAA CTGGAACAGC CACCATCGTG
    481 TGCGTGGCGA ATAAATACTT TCCCGATGGC ACCGTCACCT
    521 GGAAGGTGGA TGGCATCACC CAAAGCAGCG GCATCAATAA
    561 CAGTAGAACA CCGCAGAATT CTGCAGATTG TACCTACAAC
    601 CTCAGCAGTA CTCTGACACT GAGCAGCGAC GAGTACAACA
    641 GCCACGACGA GTACACCTGC CAGGTGGCCC AGGACTCAGG
    681 CTCACCGGTC GTCCAGAGCT TCAGTAGGAA GAGCTGTTAG
  • The nucleic acid sequence for the 95F04 anti-CD83 light chain with CDR regions identified by underlining is provided below (SEQ ID NO:90). [0163]
      1 ATGGACACGA GGGCCCCCAC TCAGCTGCTG GGGCTCCTGC
     41 TGCTCTGGCT CCCAGGTGCC ACATTTGCCC AAGCCGTGGT
     81 GACCCAGACT ACATCGCCCG TGTCTGCACC TGTGGGAGGC
    121 ACAGTCACCA TCAATTGCCA GTCCAGTCAG AGTGTTTATG
    161 GTAACAACGA ATTATCC TGG TATCAGCAGA AACCAGGGCA
    201 GCCTCCCAAG CTCCTGATCT ACCAGGCATC CAGCCTGGCA
    241 TCTGGGGTCC CATCGCGGTT CAAAGGCAGT GGATCTGGGA
    281 CACAGTTCAC TCTCACCATC AGCGACCTGG AGTGTGACGA
    321 TGCTGCCACT TACTACTGTC TAGGCGAATA TAGCATTAGT
    361 GCTGATAATC ATTTCGGCGG AGGGACCGAG GTGGTGGTCA
    401 AACGTACGCC AGTTGCACCT ACTGTCCTCC TCTTCCCACC
    441 ATCTAGCGCT GAGCTGGCAA CTGGAACAGC CACCATCGTG
    481 TGCGTGGCGA ATAAATACTT TCCCGATGGC ACCGTCACCT
    521 GGAAGGTGGA TGGCATCACC CAAAGCAGCG GCATCAATAA
    561 CAGTAGAACA CCGCAGAATT CTGCAGATTG TACCTACAAC
    601 CTCAGCAGTA CTCTGACACT GAGCAGCGAC GAGTACAACA
    641 GCCACGACGA GTACACCTGC CAGGTGGCCC AGGACTCAGG
    681 CTCACCGGTC GTCCAGAGCT TCAGTAGGAA GAGCTGTTAG
  • In another embodiment, the invention provides a 95F04 heavy chain that can bind to CD83 polypeptides and can inhibit proliferation of human peripheral blood mononuclear cells (PBMCs). The amino acid sequence for this 95F04 heavy chain is provided below (SEQ ID NO:91). [0164]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGIDLSSN AMIWVRQAPR EGLEWIGAND SNSRTYYATW
     81 AKGRFTISRT SSITVDLKIT SPTTEDTATY FCARGDGGSS
    121 DYTEMWGPGT LVTVSSASTK GPSVYPLAPG SAAQTNSMVT
    161 LGCLVKGYFP EPVTVTWNSG SLSSGVHTFP AVLQSDLYTL
    201 SSSVTVPSST WPSETVTCNV AHPASSTKVD KKIVPRDCGC
    241 KPCICTVPEV SSVFIFPPKP KDVLTITLTP KVTCVVVDIS
    281 KDDPEVQFSW FVDDVEVHTA QTQPREEQFN STFRSVSELP
    321 IMHQDWLNGK EFKCRVNSAA FPAPIEKTIS KTKGRPKAPQ
    361 VYTIPPPKEQ MAKDKVSLTC MITDFFPEDI TVEWQWNGQP
    401 AENYKNTQPI MDTDGSYFVY SKLNVQKSNW EAGNTFTCSV
    441 LHEGLHNHHT EKSLSHSPGK
  • The amino acid sequence for the 95F04 anti-CD83 heavy chain with the CDR regions identified by underlining is provided below (SEQ ID NO:91). [0165]
      1 METGLRWLLL VAVLKGVQCQ SVEESGGRLV TPGTPLTLTC
     41 TVSGIDLSSN AMIWVRQAPR EGLEWIGAMD SNSRTYYATW
     81 AKGRFTISRT SSITVDLKIT SPTTEDTATY FCARGDGGSS
    121 DYTEMWGPGT LVTVSSASTK GPSVYPLAPG SAAQTNSMVT
    161 LGCLVKGYFP EPVTVTWNSG SLSSGVHTFP AVLQSDLYTL
    201 SSSVTVPSST WPSETVTCNV AHPASSTKVD KKIVPRDCGC
    241 KPCICTVPEV SSVFIFPPKP KDVLTITLTP KVTCVVVDIS
    281 KDDPEVQFSW FVDDVEVHTA QTQPREEQFN STFRSVSELP
    321 IMHQDWLNGK EFKCRVNSAA FPAPIEKTIS KTKGRPKAPQ
    361 VYTIPPPKEQ MAKDKVSLTC MITDFFPEDI TVEWQWNGQP
    401 AENYKNTQPI MDTDGSYFVY SKLNVQKSNW EAGNTFTCSV
    441 LHEGLHNHHT EKSLSHSPGK
  • Hence, the CDR regions in the 95F04 anti-CD83 heavy chain include amino acid sequences SNAMI (SEQ ID NO:92), AMDSNSRTYYATWAKG (SEQ ID NO:93), and GDGGSSDYTEM (SEQ ID NO:94). [0166]
  • A nucleic acid sequence for this 95F04 anti-CD83 heavy chain is provided below (SEQ ID NO:95). [0167]
       1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTCTCTG GAATCGACCT CAGTAGCAAT GCAATGATCT
     161 GGGTCCGCCA GGCTCCAAGG GAGGGGCTGG AATGGATCGG
     201 AGCCATGGAT AGTAATAGTA GGACGTACTA CGCGACCTGG
     241 GCGAAAGGCC GATTCACCAT CTCCAGAACC TCGTCGATTA
     281 CGGTGGATCT GAAAATCACC AGTCCGACAA CCGAGGACAC
     321 GGCCACCTAT TTCTGTGCCA GAGGGGATGG TGGCAGTAGT
     361 GATTATACAG AGATGTGGGG CCCAGGGACC CTCGTCACCG
     401 TCTCGAGCGC TTCTACAAAG GGCCCATCTG TCTATCCACT
     441 GGCCCCTGGA TCTGCTGCCC AAACTAACTC CATGGTGACC
     481 CTGGGATGCC TGGTCAAGGG CTATTTCCCT GAGCCAGTGA
     521 CAGTGACCTG GAACTCTGGA TCCCTGTCCA GCGGTGTGCA
     561 CACCTTCCCA GCTGTCCTGC AGTCTGACCT CTACACTCTG
     601 AGCAGCTCAG TGACTGTCCC CTCCAGCACC TGGCCCAGCG
     641 AGACCGTCAC CTGCAACGTT GCCCACCCGG CCAGCAGCAC
     681 CAAGGTGGAC AAGAAAATTG TGCCCAGGGA TTGTGGTTGT
     721 AAGCCTTGCA TATGTACAGT CCCAGAAGTA TCATCTGTCT
     761 TCATCTTCCC CCCAAAGCCC AAGGATGTGC TCACCATTAC
     801 TCTGACTCCT AAGGTCACGT GTGTTGTGGT AGACATCAGC
     841 AAGGATGATC CCGAGGTCCA GTTCAGCTGG TTTGTAGATG
     881 ATGTGGAGGT GCACACAGCT CAGACGCAAC CCCGGGAGGA
     921 GCAGTTCAAC AGCACTTTCC GCTCAGTCAG TGAACTTCCC
     961 ATCATGCACC AGGACTGGCT CAATGGCAAG GAGTTCAAAT
    1001 GCAGGGTCAA CAGTGCAGCT TTCCCTGCCC CCATCGAGAA
    1041 AACCATCTCC AAAACCAAAG GCAGACCGAA GGCTCCACAG
    1081 GTGTACACCA TTCCACCTCC CAAGGAGCAG ATGGCCAAGG
    1141 ATAAAGTCAG TCTGACCTGC ATGATAACAG ACTTCTTCCC
    1161 TGAAGACATT ACTGTGGAGT GGCAGTGGAA TGGGCAGCCA
    1201 GCGGAGAACT ACAAGAACAC TCAGCCCATC ATGGACACAG
    1241 ATGGCTCTTA CTTCGTCTAC AGCAAGCTCA ATGTGCAGAA
    1281 GAGCAACTGG GAGGCAGGAA ATACTTTCAC CTGCTCTGTG
    1321 TTACATGAGG GCCTGCACAA CCACCATACT GAGAAGAGCC
    1361 TCTCCCACTC TCCTGGTAAA TGA
  • A related nucleic acid sequence for the 95F04 anti-CD83 light chain is provided below (SEQ ID NO:96). [0168]
       1 ATGGAGACTG GGCTGCGCTG GCTTCTCCTG GTCGCTGTGC
      41 TCAAAGGTGT CCAGTGTCAG TCGGTGGAGG AGTCCGGGGG
      81 TCGCCTGGTC ACGCCTGGGA CACCCCTGAC ACTCACCTGC
     121 ACAGTCTCTG GAATCGACCT CAGTAGCAAT GCAATGATCT
     161 GGGTCCGCCA GGCTCCAAGG GAGGGGCTGG AATGGATCGG
     201 AGCCATGGAT AGTAATAGTA GGACGTACTA CGCGACCTGG
     241 GCGAAAGGCC GATTCACCAT CTCCAGAACC TCGTCGATTA
     281 CGGTGGATCT GAAAATCACC AGTCCGACAA CCGAGGACAC
     321 GGCCACCTAT TTCTGTGCCA GAGGGGATGG TGGCAGTAGT
     361 GATTATACAG AGATGTGGGG CCCAGGGACC CTCGTCACCG
     401 TCTCGAGCGC TTCTACAAAG GGCCCATCTG TCTATCCACT
     441 GGCCCCTGGA TCTGCTGCCC AAACTAACTC CATGGTGACC
     481 CTGGGATGCC TGGTCAAGGG CTATTTCCCT GAGCCAGTGA
     521 CAGTGACCTG GAACTCTGGA TCCCTGTCCA GCGGTGTGCA
     561 CACCTTCCCA GCTGTCCTGC AGTCTGACCT CTACACTCTG
     601 AGCAGCTCAG TGACTGTCCC CTCCAGCACC TGGCCCAGCG
     641 AGACCGTCAC CTGCAACGTT GCCCACCCGG CCAGCAGCAC
     681 CAAGGTGGAC AAGAAAATTG TGCCCAGGGA TTGTGGTTGT
     721 AAGCCTTGCA TATGTACAGT CCCAGAAGTA TCATCTGTCT
     761 TCATCTTCCC CCCAAAGCCC AAGGATGTGC TCACCATTAC
     801 TCTGACTCCT AAGGTCACGT GTGTTGTGGT AGACATCAGC
     841 AAGGATGATC CCGAGGTCCA GTTCAGCTGG TTTGTAGATG
     881 ATGTGGAGGT GCACACAGCT CAGACGCAAC CCCGGGAGGA
     921 GCAGTTCAAC AGCACTTTCC GCTCAGTCAG TGAACTTCCC
     961 ATCATGCACC AGGACTGGCT CAATGGCAAG GAGTTCAAAT
    1001 GCAGGGTCAA CAGTGCAGCT TTCCCTGCCC CCATCGAGAA
    1041 AACCATCTCC AAAACCAAAG GCAGACCGAA GGCTCCACAG
    1081 GTGTACACCA TTCCACCTCC CAAGGAGCAG ATGGCCAAGG
    1121 ATAAAGTCAG TCTGACCTGC ATGATAACAG ACTTCTTCCC
    1161 TGAAGACATT ACTGTGGAGT GGCAGTGGAA TGGGCAGCCA
    1201 GCGGAGAACT ACAAGAACAC TCAGCCCATC ATGGACACAG
    1241 ATGGCTCTTA CTTCGTCTAC AGCAAGCTCA ATGTGCAGAA
    1281 GAGCAACTGG GAGGCAGGAA ATACTTTCAC CTGCTCTGTG
    1321 TTACATGAGG GCCTGCACAA CCACCATACT GAGAAGAGCC
    1361 TCTCCCACTC TCCTGGTAAA TGA
  • CD83 Modulation of the Immune System [0169]
  • The invention also provides compositions and methods for decreasing inappropriate immune responses in animals, including humans. According to the invention, the CD83 gene has a profound effect upon T cell activity. In particular, CD4+ T cells require CD83-related functions. Without CD83, CD4+ T cell activation and/or proliferation is impaired. The therapeutic manipulation of CD83 may thus represent a mechanism for the specific regulation of T cell function in the treatment of T cell mediated diseases, including autoimmune disorders. For example, antibodies capable of blocking CD83 function can be used as therapeutics in the treatment of immune diseases. [0170]
  • In some embodiments, the CD83-related compositions and methods of the invention can be used in the treatment of autoimmune diseases. Many autoimmune disorders are the result of inappropriate activation of T cells that are reactive against “self tissues” and that promote the production of cytokines and auto-antibodies involved in the pathology of the diseases. Modulation of T cell activity by modulating CD83 can have an effect on the course of the autoimmune disease. [0171]
  • Non-limiting examples of autoimmune diseases and disorders having an autoimmune component that may be treated according to the invention include diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiple sclerosis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjogren's Syndrome, including keratoconjunctivitis sicca secondary to Sjogren's Syndrome, alopecia areata, allergic responses due to arthropod bite reactions, Crohn's disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Crohn's disease, Graves ophthalmopathy, sarcoidosis, primary biliary cirrhosis, uveitis posterior, and interstitial lung fibrosis. [0172]
  • As illustrated and provided herein, anti-CD83 antibodies can inhibit T cell proliferation. The efficacy of anti-CD83-related compositions for treating autoimmune diseases can be tested in the animal models provided herein or other models of human diseases (e.g., EAE as a model of multiple sclerosis and the NOD mice as a model for diabetes). Such animal models include the mrl/lpr/lpr mouse as a model for lupus erythematosus, murine collagen-induced arthritis as a model for rheumatoid arthritis, and murine experimental myasthenia gravis (see Paul ed., Fundamental Immunology, Raven Press, New York, 1989, pp. 840-856). A CD83-modulatory (e.g., inhibitory) agent of the invention is administered to test animals and the course of the disease in the test animals is then monitored by the standard methods for the particular model being used. Effectiveness of the modulatory agent is evidenced by amelioration of the disease condition in animals treated with the agent as compared to untreated animals (or animals treated with a control agent). [0173]
  • Similarly, the compositions and methods of the invention that involve decreasing CD83 function can be used to decrease transplant rejection and prolong survival of the tissue graft. These methods can be used both in solid organ transplantation and in bone marrow transplantation (e.g., to inhibit graft-versus-host disease). These methods can involve either direct administration of a CD83 inhibitory agent to the transplant recipient or ex vivo treatment of cells obtained from the subject (e.g., T cells, Th1 cells, B cells, non-lymphoid cells) with an inhibitory agent followed by re-administration of the cells to the subject. [0174]
  • According to the invention, any agent that can modulate CD83 or to further decrease T cell levels can also be used in the compositions and methods of the invention. In some embodiments, anti-CD83 antibodies of the invention are used to either activate or inhibit CD83 activity. [0175]
  • Stimulating or Inhibiting CD83 [0176]
  • According to the invention, any agent that can inhibit CD83 from performing its natural functions can be used in the compositions and methods of the invention as a CD83 inhibitory agent. Indicators that CD83 activity is inhibited include decreased T cell counts, increased IL-4 cytokine levels, increased IL-10 levels, decreased IL-2 production, and decreased TNF levels relative to uninhibited levels in wild type CD83 cells. [0177]
  • Examples of CD83 inhibitors include anti-CD83 antibodies, CD83 anti-sense nucleic acids (e.g. nucleic acids that can hybridize to CD83 nucleic acids), organic compounds, peptides and agents that can mutate an endogenous CD83 gene. [0178]
  • In some embodiments, the CD83 stimulatory or inhibitory agents are proteins, for example, CD83 gene products, anti-CD83 antibody preparations, CD83 inhibitors, peptides and protein factors that can promote CD83 transcription or translation. In other embodiments, the CD83 stimulatory or inhibitory agents are peptides or organic molecules. Such proteins, organic molecules and organic molecules can be prepared and/or purified as described herein or by methods available in the art, and administered as provided herein. [0179]
  • In other embodiments, the CD83 inhibitory agents can be nucleic acids including recombinant expression vectors or expression cassettes encoding CD83 anti-sense nucleic acid, intracellular antibodies capable of binding to CD83 or dominant negative CD83 inhibitors. Such nucleic acids can be operably linked to a promoter that is functional in a mammalian cell, and then introduced into cells of the subject mammal using methods known in the art for introducing nucleic acid (e.g., DNA) into cells. [0180]
  • The “promoter functional in a mammalian cell” or “mammalian promoter” is capable of directing transcription of a polypeptide coding sequence operably linked to the promoter. The promoter should generally be active in T cells and antigen presenting cells and may be obtained from a gene that is expressed in T cells or antigen presenting cells. However, it need not be a T cell-specific or an antigen presenting cell specific-promoter. Instead, the promoter may be selected from any mammalian or viral promoter that can function in a T cell. Hence the promoter may be an actin promoter, an immunoglobulin promoter, a heat-shock promoter, or a viral promoter obtained from the genome of viruses such as adenoviruses, retroviruses, lentiviruses, herpes viruses, including but not limited to, polyoma virus, fowlpox virus, [0181] adenovirus 2, bovine papilloma virus, avian sarcoma virus, cytomegalovirus (CMV), hepatitis-B virus, Simian Virus 40 (SV40), Epstein Barr virus (EBV), feline immunodeficiency virus (FIV), and Sra, or are respiratory synsitial viral promoters (RSV) or long terminal repeats (LTRs) of a retrovirus, i.e., a Moloney Murine Leukemia Virus (MoMuLv) (Cepko et al. (1984) Cell 37:1053-1062). The promoter functional in a mammalian cell can be inducible or constitutive.
  • Any cloning procedure used by one of skill in the art can be employed to make the expression vectors or expression that comprise a promoter operably linked to a CD83 nucleic acid, CD83 transcription factor or a nucleic acid encoding an anti-CD83 antibody. See, e.g., Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y., 1989; Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, N.Y., 2001. [0182]
  • After constructing an expression vector or an expression cassette encoding CD83 transcription factors, CD83 anti-sense nucleic acid, intracellular antibodies capable of binding to CD83 or dominant negative CD83 inhibitors, mammalian cells can be transformed with the vector or cassette. Examples of such methods include: [0183]
  • Direct Injection: Naked DNA can be introduced into cells in vivo by directly injecting the DNA into the cells (see e.g., Acsadi et al. (1991) Nature 332:815-818; Wolff et al. (1990) Science 247:1465-1468). For example, a delivery apparatus (e.g., a “gene gun”) for injecting DNA into cells in vivo can be used. Such an apparatus is commercially available (e.g., from BioRad). [0184]
  • Receptor-Mediated DNA Uptake: Naked DNA can also be introduced into cells in vivo by complexing the DNA to a cation, such as polylysine, which is coupled to a ligand for a cell-surface receptor (see for example Wu, G. and Wu, C. H. (1988) J. Biol. Chem. 263:14621; Wilson et al. (1992) J. Biol. Chem. 267:963-967; and U.S. Pat. No. 5,166,320). Binding of the DNA-ligand complex to the receptor facilitates uptake of the DNA by receptor-mediated endocytosis. A DNA-ligand complex linked to adenovirus capsids that naturally disrupt endosomes, thereby releasing material into the cytoplasm can be used to avoid degradation of the complex by intracellular lysosomes (see for example Curiel et al. (1991) Proc. Natl. Acad. Sci. USA 88:8850; Cristiano et al. (1993) Proc. Natl. Acad. Sci. USA 90:2122-2126). [0185]
  • Retroviruses: Defective retroviruses are well characterized for use in gene transfer for gene therapy purposes (for a review see Miller, A. D. (1990) Blood 76:271). A recombinant retrovirus can be constructed having nucleotide sequences of interest incorporated into the retroviral genome. Additionally, portions of the retroviral genome can be removed to render the retrovirus replication defective. The replication defective retrovirus is then packaged into virions that can be used to infect a target cell through the use of a helper virus by standard techniques. Protocols for producing recombinant retroviruses and for infecting cells in vitro or in vivo with such viruses can be found in Current Protocols in Molecular Biology, Ausubel, F. M. et al. (eds.) Greene Publishing Associates, (1989), Sections 9.10-9.14 and other standard laboratory manuals. Examples of suitable retroviruses include pLJ, pZIP, pWE and pEM which are available to those skilled in the art. Examples of suitable packaging virus lines include ? Crip, ? Cre, ? 2 and ? Am. Retroviruses have been used to introduce a variety of genes into many different cell types, including epithelial cells, endothelial cells, lymphocytes, myoblasts, hepatocytes, bone marrow cells, in vitro and/or in vivo (see for example Eglitis, et al. (1985) Science 230:1395-1398; Danos and Mulligan (1988) Proc. Natl. Acad. Sci. USA 85:6460-6464; Wilson et al. (1988) Proc. Natl. Acad. Sci. USA 85:3014-3018; Armentano et al. (1990) Proc. Natl. Acad. Sci. USA 87:6141-6145; Huber et al. (1991) Proc. Natl. Acad. Sci. USA 88:8039-8043; Ferry et al. (1991) Proc. Natl. Acad. Sci. USA 88:8377-8381; Chowdhury et al. (1991) Science 254:1802-1805; van Beusechem et al. (1992) Proc. Natl. Acad. Sci. USA 89:7640-7644; Kay et al. (1992) Human Gene Therapy 3:641-647; Dai et al. (1992) Proc. Natl. Acad. Sci USA 89:10892-10895; Hwu et al. (1993) J. Immunol. 150:4104-4115; U.S. Pat. Nos. 4,868,116; 4,980,286; PCT Application WO 89/07136; PCT Application WO 89/02468; PCT Application WO 89/05345; and PCT Application WO 92/07573). Retroviral vectors require target cell division in order for the retroviral genome (and foreign nucleic acid inserted into it) to be integrated into the host genome to stably introduce nucleic acid into the cell. Thus, it may be necessary to stimulate replication of the target cell. [0186]
  • Adenoviruses: The genome of an adenovirus can be manipulated such that it encodes and expresses a gene product of interest but is inactivated in terms of its ability to replicate in a normal lytic viral life cycle. See, for example, Berkner et al. (1988) BioTechniques 6:616; Rosenfeld et al. (1991) Science 252:431-434; and Rosenfeld et al. (1992) Cell 68:143-155. Suitable adenoviral vectors derived from the adenovirus [0187] strain Ad type 5 dl 324 or other strains of adenovirus (e.g., Ad2, Ad3, Ad7 etc.) are available to those skilled in the art. Recombinant adenoviruses are advantageous in that they do not require dividing cells to be effective gene delivery vehicles and can be used to infect a wide variety of cell types, including airway epithelium (Rosenfeld et al. (1992) cited supra), endothelial cells (Lemarchand et al. (1992) Proc. Natl. Acad. Sci. USA 89:6482-6486), hepatocytes (Herz and Gerard (1993) Proc. Natl. Acad. Sci. USA 90:2812-2816) and muscle cells (Quantin et al. (1992) Proc. Natl. Acad. Sci. USA 89:2581-2584). Additionally, introduced adenoviral DNA (and foreign DNA contained therein) is not integrated into the genome of a host cell but remains episomal, thereby avoiding potential problems that can occur as a result of insertional mutagenesis in situations where introduced DNA becomes integrated into the host genome (e.g., retroviral DNA). Moreover, the carrying capacity of the adenoviral genome for foreign DNA is large (up to 8 kilobases) relative to other gene delivery vectors (Berkner et al. cited supra; Haj-Ahmand and Graham (1986) J. Virol. 57:267). Most replication-defective adenoviral vectors currently in use are deleted for all or parts of the viral E1 and E3 genes but retain as much as 80% of the adenoviral genetic material.
  • Adeno-Associated Viruses: Adeno-associated virus (AAV) is a naturally occurring defective virus that requires another virus, such as an adenovirus or a herpes virus, as a helper virus for efficient replication and a productive life cycle. (For a review see Muzyczka et al. Curr. Topics in Micro. and Immunol. (1992) 158:97-129). It is also one of the few viruses that may integrate its DNA into non-dividing cells, and exhibits a high frequency of stable integration (see for example Flotte et al. (1992) Am. J. Respir. Cell. Mol. Biol. 7:349-356; Samulski et al. (1989) J. Virol. 63:3822-3828; and McLaughlin et al. (1989) J. Virol. 62:1963-1973). Vectors containing as little as 300 base pairs of AAV can be packaged and can integrate. Space for exogenous DNA is limited to about 4.5 kb. An AAV vector such as that described in Tratschin et al. (1985) Mol. Cell. Biol. 5:3251-3260 can be used to introduce DNA into cells. A variety of nucleic acids have been introduced into different cell types using AAV vectors (see for example Hermonat et al. (1984) Proc. Natl. Acad. Sci. USA 81:6466-6470; Tratschin et al. (1985) Mol. Cell. Biol. 4:2072-2081; Wondisford et al. (1988) Mol. Endocrinol. 2:32-39; Tratschin et al. (1984) J. Virol. 51:611-619; and Flotte et al. (1993) J. Biol. Chem. 268:3781-3790). [0188]
  • Transformed mammalian cells can then be identified and administered to the mammal from whence they came to permit expression of a CD83 transcription factor, CD83 anti-sense nucleic acid, intracellular antibody capable of binding to CD83 proteins, or dominant negative CD83 inhibitors. The efficacy of a particular expression vector system and method of introducing nucleic acid into a cell can be assessed by standard approaches routinely used in the art. For example, DNA introduced into a cell can be detected by a filter hybridization technique (e.g., Southern blotting). RNA produced by transcription of an introduced DNA can be detected, for example, by Northern blotting, RNase protection or reverse transcriptase-polymerase chain reaction (RT-PCR). The CD83 gene product can be detected by an appropriate assay, for example, by immunological detection of a produced CD83 protein, such as with a CD83-specific antibody. [0189]
  • Anti-sense Nucleic Acids [0190]
  • Anti-sense nucleic acids can be used to inhibit the function of CD83. In general, the function of CD83 RNA is inhibited, for example, by administering to a mammal a nucleic acid that can inhibit the functioning of CD83 RNA. Nucleic acids that can inhibit the function of a CD83 RNA can be generated from coding and non-coding regions of the CD83 gene. However, nucleic acids that can inhibit the function of a CD83 RNA are often selected to be complementary to CD83 nucleic acids that are naturally expressed in the mammalian cell to be treated with the methods of the invention. In some embodiments, the nucleic acids that can inhibit CD83 RNA functions are complementary to CD83 sequences found near the 5′ end of the CD83 coding region. For example, nucleic acids that can inhibit the function of a CD83 RNA can be complementary to the 5′ region of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. [0191]
  • A nucleic acid that can inhibit the functioning of a CD83 RNA need not be 100% complementary to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. Instead, some variability the sequence of the nucleic acid that can inhibit the functioning of a CD83 RNA is permitted. For example, a nucleic acid that can inhibit the functioning of a CD83 RNA from a human can be complementary to a nucleic acid encoding either a human or a mouse CD83 gene product. [0192]
  • Moreover, nucleic acids that can hybridize under moderately or highly stringent hybridization conditions to a nucleic acid comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10 are sufficiently complementary to inhibit the functioning of a CD83 RNA and can be utilized in the methods of the invention. [0193]
  • “Stringent hybridization conditions” and “stringent hybridization wash conditions” in the context of nucleic acid hybridization are somewhat sequence dependent, and may differ depending upon the environmental conditions of the nucleic acid. For example, longer sequences tend to hybridize specifically at higher temperatures. An extensive guide to the hybridization of nucleic acids is found in Tijssen, Laboratory Techniques in Biochemistry and Molecular biology-Hybridization with Nucleic Acid Probes, [0194] page 1, chapter 2 “Overview of principles of hybridization and the strategy of nucleic acid probe assays” Elsevier, New York (1993). See also, J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, N.Y., pp 9.31-9.58 (1989); J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, N.Y. (3rd ed. 2001).
  • Generally, highly stringent hybridization and wash conditions are selected to be about 5° C. lower than the thermal melting point (T[0195] m) for the specific double-stranded sequence at a defined ionic strength and pH. For example, under “highly stringent conditions” or “highly stringent hybridization conditions” a nucleic acid will hybridize to its complement to a detectably greater degree than to other sequences (e.g., at least 2- fold over background). By controlling the stringency of the hybridization and/or washing conditions nucleic acids that are 100% complementary can be hybridized.
  • For DNA-DNA Hybrids, the T[0196] m can be Approximated from the Equation of Meinkoth and Wahl Anal. Biochem. 138:267-284 (1984):
  • T m81.5°C.+16.6 (log M)+0.41 (% GC)−0.61 (% form)−500/L
  • where M is the molarity of monovalent cations, % GC is the percentage of guanosine and cytosine nucleotides in the DNA, % form is the percentage of formamide in the hybridization solution, and L is the length of the hybrid in base pairs. The T[0197] m is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe.
  • Very stringent conditions are selected to be equal to the T[0198] m for a particular probe.
  • Alternatively, stringency conditions can be adjusted to allow some mismatching in sequences so that lower degrees of similarity can hybridize. Typically, stringent conditions will be those in which the salt concentration is less than about 1.5 M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30° C. for short probes (e.g., 10 to 50 nucleotides) and at least about 60° C. for long probes (e.g., greater than 50 nucleotides). Stringent conditions may also be achieved with the addition of destabilizing agents such as formamide. [0199]
  • Exemplary low stringency conditions include hybridization with a buffer solution of 30 to 35% formamide, 1 M NaCl, 1% SDS (sodium dodecyl sulphate) at 37° C., and a wash in 1X to 2X SSC (20X SSC=3.0 M NaCl and 0.3 M trisodium citrate) at 50 to 55° C. Exemplary moderate stringency conditions include hybridization in 40 to 45% formamide, 1.0 M NaCl, 1% SDS at 37° C., and a wash in 0.5X to 1X SSC at 55 to 60° C. Exemplary high stringency conditions include hybridization in 50% formamide, 1 M NaCl, 1% SDS at 37° C., and a wash in 0.1X SSC at 60 to 65° C. [0200]
  • The degree of complementarity or sequence identity of hybrids obtained during hybridization is typically a function of post-hybridization washes, the critical factors being the ionic strength and temperature of the final wash solution. The type and length of hybridizing nucleic acids also affects whether hybridization will occur and whether any hybrids formed will be stable under a given set of hybridization and wash conditions. [0201]
  • An example of stringent hybridization conditions for hybridization of complementary nucleic acids that have more than 100 complementary residues on a filter in a Southern or Northern blot is 50% formamide with 1 mg of heparin at 42° C., with the hybridization being carried out overnight. An example of highly stringent conditions is 0.1 5 M NaCl at 72° C. for about 15 minutes. An example of stringent wash conditions is a 0.2x SSC wash at 65° C. for 15 minutes (see also, Sambrook, infra). Often, a high stringency wash is preceded by a low stringency wash to remove background probe signal. An example of medium stringency for a duplex of, e.g., more than 100 nucleotides, is 1x SSC at 45° C. for 15 minutes. An example low stringency wash for a duplex of, e.g., more than 100 nucleotides, is 4-6x SSC at 40° C. for 15 minutes. For short probes (e.g., about 10 to 50 nucleotides), stringent conditions typically involve salt concentrations of less than about 1.0M Na ion, typically about 0.01 to 1.0 M Na ion concentration (or other salts) at pH 7.0 to 8.3, and the temperature is typically at least about 30° C. [0202]
  • Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide. In general, a signal to noise ratio of 2x (or higher) than that observed for an unrelated probe in the particular hybridization assay indicates detection of a specific hybridization. Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the proteins that they encode are substantially identical. This occurs, e.g., when a copy of a nucleic acid is created using the maximum codon degeneracy permitted by the genetic code. [0203]
  • The following are examples of sets of hybridization/wash conditions that may be used to detect and isolate homologous nucleic acids that are substantially identical to reference nucleic acids of the present invention: a reference nucleotide sequence preferably hybridizes to the reference nucleotide sequence in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO[0204] 4, 1 mM EDTA at 50° C. with washing in 2X SSC, 0.1% SDS at 50° C., more desirably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 1X SSC, 0.1% SDS at 50° C., more desirably still in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.5X SSC, 0.1% SDS at 50° C., preferably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.1X SSC, 0.1% SDS at 50° C., more preferably in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 0.1X SSC, 0.1% SDS at 65° C.
  • In general, T[0205] m is reduced by about 1° C. for each 1% of mismatching. Thus, Tm, hybridization, and/or wash conditions can be adjusted to hybridize to sequences of the desired sequence identity. For example, if sequences with >90% identity are sought, the Tm can be decreased 10° C. Generally, stringent conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence and its complement at a defined ionic strength and pH. However, severely stringent conditions can utilize a hybridization and/or wash at 1, 2, 3, or 4° C. lower than the thermal melting point (Tm); moderately stringent conditions can utilize a hybridization and/or wash at 6, 7, 8, 9, or 10° C. lower than the thermal melting point (Tm); low stringency conditions can utilize a hybridization and/or wash at 11, 12, 13, 14, 15, or 20° C. lower than the thermal melting point (Tm).
  • If the desired degree of mismatching results in a T[0206] m of less than 45° C. (aqueous solution) or 32° C. (formamide solution), it is preferred to increase the SSC concentration so that a higher temperature can be used. An extensive guide to the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in Biochemistry and Molecular Biology-Hybridization with Nucleic Acid Probes, Part 1, Chapter 2 (Elsevier, N.Y.); and Ausubel et al., eds. (1995) Current Protocols in Molecular Biology, Chapter 2 (Greene Publishing and Wiley-Interscience, New York). See Sambrook et al. (1989) Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Plainview, N.Y.). Using these references and the teachings herein on the relationship between Tm, mismatch, and hybridization and wash conditions, those of ordinary skill can generate variants of the present homocysteine S-methyltransferase nucleic acids.
  • Precise complementarity is therefore not required for successful duplex formation between a nucleic acid that can inhibit a CD83 RNA and the complementary coding sequence of a CD83 RNA. Inhibitory nucleic acid molecules that comprise, for example, 2, 3, 4, or 5 or more stretches of contiguous nucleotides that are precisely complementary to a CD83 coding sequence, each separated by a stretch of contiguous nucleotides that are not complementary to adjacent CD83 coding sequences, can inhibit the function of CD83 RNA. In general, each stretch of contiguous nucleotides is at least 4, 5, 6, 7, or 8 or more nucleotides in length. Non-complementary intervening sequences are preferably 1, 2, 3, or 4 nucleotides in length. One skilled in the art can easily use the calculated melting point of an anti-sense nucleic acid hybridized to a sense nucleic acid to determine the degree of mismatching that will be tolerated between a particular anti-sense nucleic acid and a particular CD83 RNA. [0207]
  • Nucleic acids that complementary a CD83 RNA can be administered to a mammal or to directly to the site of the inappropriate immune system activity. Alternatively, nucleic acids that are complementary to a CD83 RNA can be generated by transcription from an expression cassette that has been administered to a mammal. For example, a complementary RNA can be transcribed from a CD83 nucleic acid that has been inserted into an expression cassette in the 3′ to 5′ orientation, that is, opposite to the usual orientation employed to generate sense RNA transcripts. Hence, to generate a complementary RNA that can inhibit the function of an endogenous CD83 RNA, the promoter would be positioned to transcribe from a 3′ site towards the 5′ end of the CD83 coding region. [0208]
  • In some embodiments an RNA that can inhibit the function of an endogenous CD83 RNA is an anti-sense oligonucleotide. The anti-sense oligonucleotide is complementary to at least a portion of the coding sequence of a gene comprising SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. Such anti-sense oligonucleotides are generally at least six nucleotides in length, but can be about 8, 12, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides long. Longer oligonucleotides can also be used. CD83 anti-sense oligonucleotides can be provided in a DNA construct and introduced into cells whose division is to be decreased, for example, into CD4[0209] + T cells, Th-1 cells, Th-2 cells or lymphocyte precursor cells.
  • Anti-sense oligonucleotides can be composed of deoxyribonucleotides, ribonucleotides, or a combination of both. Oligonucleotides can be synthesized endogenously from transgenic expression cassettes or vectors as described herein. Alternatively, such oligonucleotides can be synthesized manually or by an automated synthesizer, by covalently linking the 5′ end of one nucleotide with the 3′ end of another nucleotide with non-phosphodiester internucleotide linkages such alkylphosphonates, phosphorothioates, phosphorodithioates, alkylphosphonothioates, alkylphosphonates, phosphoramidates, phosphate esters, carbamates, acetamidate, carboxymethyl esters, carbonates, and phosphate triesters. See Brown, 1994, Meth. Mol. Biol. 20:1-8; Sonveaux, 1994, Meth. Mol. Biol. 26:1-72; Uhlmann et al., 1990, Chem. Rev. 90:543-583. [0210]
  • CD83 anti-sense oligonucleotides can be modified without affecting their ability to hybridize to a CD83 RNA. These modifications can be internal or at one or both ends of the anti-sense molecule. For example, internucleoside phosphate linkages can be modified by adding peptidyl, cholesteryl or diamine moieties with varying numbers of carbon residues between these moieties and the terminal ribose. Modified bases and/or sugars, such as arabinose instead of ribose, or a 3′,5′-substituted oligonucleotide in which the 3′ hydroxyl group or the 5′ phosphate group are substituted, can also be employed in a modified anti-sense oligonucleotide. These modified oligonucleotides can be prepared by methods available in the art. Agrawal et al., 1992, Trends Biotechnol. 10: 152-158; Uhlmann et al., 1990, Chem. Rev. 90:543-584; Uhlmann et al., 1987, Tetrahedron. Lett. 215:3539-3542. [0211]
  • In one embodiment of the invention, expression of a CD83 gene is decreased using a ribozyme. A ribozyme is an RNA molecule with catalytic activity. See, e.g., Cech, 1987, Science 236: 1532-1539; Cech, 1990, Ann. Rev. Biochem. 59:543-568; Cech, 1992, Curr. Opin. Struct. Biol. 2: 605-609; Couture and Stinchcomb, 1996, Trends Genet. 12: 510-515. Ribozymes can be used to inhibit gene function by cleaving an RNA sequence, as is known in the art (see, e.g., Haseloff et al., U.S. Pat. No. 5,641,673). [0212]
  • CD83 nucleic acids complementary to SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10 can be used to generate ribozymes that will specifically bind to mRNA transcribed from a CD83 gene. Methods of designing and constructing ribozymes that can cleave other RNA molecules in trans in a highly sequence specific manner have been developed and described in the art (see Haseloffet al. (1988), Nature 334:585-591). For example, the cleavage activity of ribozymes can be targeted to specific RNAs by engineering a discrete “hybridization” region into the ribozyme. The hybridization region contains a sequence complementary to the target RNA and thus specifically hybridizes with the target (see, for example, Gerlach et al., EP 321,201). The target sequence can be a segment of about 10, 12, 15, 20, or 50 contiguous nucleotides selected from a nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:10. Longer complementary sequences can be used to increase the affinity of the hybridization sequence for the target. The hybridizing and cleavage regions of the ribozyme can be integrally related; thus, upon hybridizing to the target RNA through the complementary regions, the catalytic region of the ribozyme can cleave the target. [0213]
  • Other CD83 Modulating Molecules [0214]
  • A wide variety of molecules may be used to modulate CD83 activity or function. Such molecules can also be used to modulate the immune system independent of CD83. Compositions and methods for modulating CD83 activity or expression can include these molecules as well as other components. Representative examples that are discussed in more detail below include transcription factors, RNA-binding factors, organic molecules, or peptides. [0215]
  • RNA-Binding Factors: [0216]
  • One class of molecules that can be used to modulate the CD83 gene is the RNA binding factors. Such factors include those described in PCT/EP01/14820 and other sources. [0217]
  • For example, the HuR protein (Genbank accession number U38175) has the ability to specifically bind to CD83 RNA at AU-rich elements or sites. Such AU-rich elements comprise sequences such as AUUUA (SEQ ID NO:49), AUUUUA (SEQ ID NO:50) and AUUUUUA (SEQ ID NO:51). Binding by such HuR proteins to CD83 mRNA is thought to increase the stability, transport and translation of CD83 mRNA, and thereby increase the expression of CD83 polypeptides. Hence, CD83 expression may be increase by administering HuR proteins or nucleic acids to a mammal. [0218]
  • Conversely, CD83 expression may be decreased by administering factors that block HuR binding to CD83 mRNA. Factors that block HuR binding include proteins or nucleic acids that can bind to the AU-rich elements normally bound by HuR, for example, nucleic acids or anti-sense nucleic acids that are complementary to AU-rich elements. [0219]
  • Organic Molecules: [0220]
  • Numerous organic molecules may be used to modulate the immune system. These compounds include any compound that can interact with a component of the immune system. Such compounds may interact directly with CD83, indirectly with CD83 or with some other polypeptide, cell or factor that plays a role in the function of the immune system. In some embodiments, the organic molecule can bind to a CD83 polypeptide or a CD83 nucleic acid. [0221]
  • Organic molecules can be tested or assayed for their ability to modulate CD83 activity, CD83 function or for their ability to modulate components of the immune system. For example, within one embodiment of the invention suitable organic molecules may be selected either from a chemical library, wherein chemicals are assayed individually, or from combinatorial chemical libraries where multiple compounds are assayed at once, then deconvoluted to determine and isolate the most active compounds. [0222]
  • Representative examples of such combinatorial chemical libraries include those described by Agrafiotis et al., “System and method of automatically generating chemical compounds with desired properties,” U.S. Pat. No. 5,463,564; Armstrong, R. W., “Synthesis of combinatorial arrays of organic compounds through the use of multiple component combinatorial array syntheses,” WO 95/02566; Baldwin, J. J. et al., “Sulfonamide derivatives and their use,” WO 95/24186; Baldwin, J. J. et al., “Combinatorial dihydrobenzopyran library,” WO 95/30642; Brenner, S., “New kit for preparing combinatorial libraries,” WO 95/16918; Chenera, B. et al., “Preparation of library of resin-bound aromatic carbocyclic compounds,” WO 95/16712; Ellman, J. A., “Solid phase and combinatorial synthesis of benzodiazepine compounds on a solid support,” U.S. Pat. No. 5,288,514; Felder, E. et al., “Novel combinatorial compound libraries,” WO 95/16209; Lemer, R. et al., “Encoded combinatorial chemical libraries,” WO 93/20242; Pavia, M. R. et al., “A method for preparing and selecting pharmaceutically useful non-peptide compounds from a structurally diverse universal library,” WO 95/04277; Summerton, J. E. and D. D. Weller, “Morpholino-subunit combinatorial library and method,” U.S. Pat. No. 5,506,337; Holmes, C., “Methods for the Solid Phase Synthesis of Thiazolidinones, Metathiazanones, and Derivatives thereof,” WO 96/00148; Phillips, G. B. and G. P. Wei, “Solid-phase Synthesis of Benzimidazoles,” [0223] Tet. Letters 37:4887-90, 1996; Ruhland, B. et al., “Solid-supported Combinatorial Synthesis of Structurally Diverse-Lactams,” J. Amer. Chem. Soc. 111:253-4, 1996; Look, G. C. et al., “The Indentification of Cyclooxygenase-1 Inhibitors from 4-Thiazolidinone Combinatorial Libraries,” Bioorg and Med. Chem. Letters 6:707-12, 1996.
  • Peptides: [0224]
  • Peptide molecules that modulate the immune system may be obtained through the screening of combinatorial peptide libraries. Such libraries may either be prepared by one of skill in the art (see e.g., U.S. Pat. Nos. 4,528,266 and 4,359,535, and Patent Cooperation Treaty Publication Nos. WO 92/15679, WO 92/15677, WO 90/07862, WO 90/02809, or purchased from commercially available sources (e.g., New England Biolabs Ph.D.™ Phage Display Peptide Library Kit). [0225]
  • Methods of Using the CD83 Mutant Mouse [0226]
  • In one embodiment, the invention provides a method for identifying ligands, receptors, therapeutic drugs and other molecules that can modulate the phenotype of the mutant CD83 in vivo. This method involves administering a test compound to the mutant CD83 mouse of the invention and observing whether the compound causes a change in the phenotype of the mutant mouse. Changes in phenotype that are of interest include increases or decreases in T cells (especially CD4+ T cells), increases or decreases in GMCSF, IL-2, IL-4 or IL-10 cytokine production, increases or decreases in inflammation, increases or decreases in dendritic cell function and other T cell responses known to one of skill in the art. [0227]
  • Test compounds can be screened in vitro to ascertain whether they interact directly with CD83. In vitro screening can, for example, identify whether a test compound or molecule can bind to the cytoplasmic tail or the membrane-associated portions of CD83. Such information, combined with observation of the in vivo phenotype before and after administration of the test compound provides further insight into the function of CD83 and provides targets for manipulation T cell activation and other functions modulated by CD83. [0228]
  • The invention is not limited to identification of molecules that directly associate with CD83. The in vivo screening methods provided herein can, also identify test compounds that have an indirect effect on CD83, or that partially or completely replace a function of CD83. [0229]
  • Increases or decreases in T cell numbers can be observed in blood samples or in samples obtained from thymus, spleen or lymph node tissues. In order to observe the activation of T cells and/or the interaction of T cells and dendritic cells, dendritic cells can be pulsed with antigens ex vivo and then injected into mice to prime CD4+ T cells in draining lymphoid organs. See Inaba et al., J. Exp. Med. 172: 631-640, 1990; Liu, et al., J. Exp. Med. 177: 1299-1307, 1993; Sornasse et al., J. Exp. Med. 175: 15-21, 1992. Antigens can also be deposited intramuscularly and dendritic cells from the corresponding afferent lymphatics can carry that antigen in a form stimulatory for T cells. Bujdoso et al., J. Exp. Med. 170: 1285-1302, 1989. According to the invention, factors stimulating the interaction of dendritic cells with T cells in vivo can be identified by administering antigens in this manner and then observing how T cell respond, e.g. by observing whether T cell activation occurs. [0230]
  • Increases or decreases in cytokine levels can be observed by methods provided herein or by other methods available in the art. [0231]
  • Compositions [0232]
  • The CD83 nucleic acids, polypeptides and antibodies of the invention, including their salts, are administered so as to achieve a reduction in at least one symptom associated with an infection, indication or disease. [0233]
  • To achieve the desired effect(s), the nucleic acid, polypeptide or antibody, a variant thereof or a combination thereof, may be administered as single or divided dosages, for example, of at least about 0.01 mg/kg to about 500 to 750 mg/kg, of at least about 0.01 mg/kg to about 300 to 500 mg/kg, at least about 0.1 mg/kg to about 100 to 300 mg/kg or at least about 1 mg/kg to about 50 to 100 mg/kg of body weight, although other dosages may provide beneficial results. The amount administered will vary depending on various factors including, but not limited to, the nucleic acid, polypeptide or antibody chosen, the disease, the weight, the physical condition, the health, the age of the mammal, whether prevention or treatment is to be achieved, and if the nucleic acid, polypeptide or antibody is chemically modified. Such factors can be readily determined by the clinician employing animal models or other test systems that are available in the art. [0234]
  • Administration of the therapeutic agents in accordance with the present invention may be in a single dose, in multiple doses, in a continuous or intermittent manner, depending, for example, upon the recipient's physiological condition, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to skilled practitioners. The administration of the CD83 nucleic acids, polypeptides and antibodies of the invention may be essentially continuous over a preselected period of time or may be in a series of spaced doses. Both local and systemic administration is contemplated. [0235]
  • To prepare the composition, CD83 nucleic acids, polypeptides and antibodies are synthesized or otherwise obtained, purified as necessary or desired and then lyophilized and stabilized. The nucleic acid, polypeptide or antibody can then be adjusted to the appropriate concentration, and optionally combined with other agents. The absolute weight of a given nucleic acid, polypeptide or antibody included in a unit dose can vary widely. For example, about 0.01 to about 2 g, or about 0.1 to about 500 mg, of at least one nucleic acid, polypeptide or antibody of the invention, or a plurality of CD83 nucleic acid, polypeptides and antibodies specific for a particular cell type can be administered. Alternatively, the unit dosage can vary from about 0.01 g to about 50 g, from about 0.01 g to about 35 g, from about 0.1 g to about 25 g, from about 0.5 g to about 12 g, from about 0.5 g to about 8 g, from about 0.5 g to about 4 g, or from about 0.5 g to about 2 g. [0236]
  • Daily doses of the CD83 nucleic acids, polypeptides or antibodies of the invention can vary as well. Such daily doses can range, for example, from about 0.1 g/day to about 50 g/day, from about 0.1 g/day to about 25 g/day, from about 0.1 g/day to about 12 g/day, from about 0.5 g/day to about 8 g/day, from about 0.5 g/day to about 4 g/day, and from about 0.5 g/day to about 2 g/day. [0237]
  • Thus, one or more suitable unit dosage forms comprising the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can be administered by a variety of routes including oral, parenteral (including subcutaneous, intravenous, intramuscular and intraperitoneal), rectal, dermal, transdermal, intrathoracic, intrapulmonary and intranasal (respiratory) routes. The therapeutic CD83 nucleic acids, polypeptides or antibodies may also be formulated for sustained release (for example, using microencapsulation, see WO 94/07529, and U.S. Pat. No. 4,962,091). The formulations may, where appropriate, be conveniently presented in discrete unit dosage forms and may be prepared by any of the methods well known to the pharmaceutical arts. Such methods may include the step of mixing the therapeutic agent with liquid carriers, solid matrices, semi-solid carriers, finely divided solid carriers or combinations thereof, and then, if necessary, introducing or shaping the product into the desired delivery system. [0238]
  • When the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention are prepared for oral administration, they are generally combined with a pharmaceutically acceptable carrier, diluent or excipient to form a pharmaceutical formulation, or unit dosage form. For oral administration, the CD83 nucleic acids, polypeptides or antibodies may be present as a powder, a granular formulation, a solution, a suspension, an emulsion or in a natural or synthetic polymer or resin for ingestion of the active ingredients from a chewing gum. The active CD83 nucleic acids, polypeptides or antibodies may also be presented as a bolus, electuary or paste. Orally administered therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can also be formulated for sustained release, e.g., the CD83 nucleic acids, polypeptides or antibodies can be coated, micro-encapsulated, or otherwise placed within a sustained delivery device. The total active ingredients in such formulations comprise from 0.1 to 99.9% by weight of the formulation. [0239]
  • By “pharmaceutically acceptable” it is meant a carrier, diluent, excipient, and/or salt that is compatible with the other ingredients of the formulation, and not deleterious to the recipient thereof. [0240]
  • Pharmaceutical formulations containing the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can be prepared by procedures known in the art using well-known and readily available ingredients. For example, the nucleic acid, polypeptide or antibody can be formulated with common excipients, diluents, or carriers, and formed into tablets, capsules, solutions, suspensions, powders, aerosols and the like. Examples of excipients, diluents, and carriers that are suitable for such formulations include buffers, as well as fillers and extenders such as starch, cellulose, sugars, mannitol, and silicic derivatives. Binding agents can also be included such as carboxymethyl cellulose, hydroxymethylcellulose, hydroxypropyl methylcellulose and other cellulose derivatives, alginates, gelatin, and polyvinyl-pyrrolidone. Moisturizing agents can be included such as glycerol, disintegrating agents such as calcium carbonate and sodium bicarbonate. Agents for retarding dissolution can also be included such as paraffin. Resorption accelerators such as quaternary ammonium compounds can also be included. Surface active agents such as cetyl alcohol and glycerol monostearate can be included. Adsorptive carriers such as kaolin and bentonite can be added. Lubricants such as talc, calcium and magnesium stearate, and solid polyethyl glycols can also be included. Preservatives may also be added. The compositions of the invention can also contain thickening agents such as cellulose and/or cellulose derivatives. They may also contain gums such as xanthan, guar or carbo gum or gum arabic, or alternatively polyethylene glycols, bentones and montmorillonites, and the like. [0241]
  • For example, tablets or caplets containing the CD83 nucleic acids, polypeptides or antibodies of the invention can include buffering agents such as calcium carbonate, magnesium oxide and magnesium carbonate. Caplets and tablets can also include inactive ingredients such as cellulose, pregelatinized starch, silicon dioxide, hydroxy propyl methyl cellulose, magnesium stearate, microcrystalline cellulose, starch, talc, titanium dioxide, benzoic acid, citric acid, corn starch, mineral oil, polypropylene glycol, sodium phosphate, zinc stearate, and the like. Hard or soft gelatin capsules containing at least one nucleic acid, polypeptide or antibody of the invention can contain inactive ingredients such as gelatin, microcrystalline cellulose, sodium lauryl sulfate, starch, talc, and titanium dioxide, and the like, as well as liquid vehicles such as polyethylene glycols (PEGs) and vegetable oil. Moreover, enteric-coated caplets or tablets containing one or more CD83 nucleic acids, polypeptides or antibodies of the invention are designed to resist disintegration in the stomach and dissolve in the more neutral to alkaline environment of the duodenum. [0242]
  • The therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can also be formulated as elixirs or solutions for convenient oral administration or as solutions appropriate for parenteral administration, for instance by intramuscular, subcutaneous, intraperitoneal or intravenous routes. The pharmaceutical formulations of the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can also take the form of an aqueous or anhydrous solution or dispersion, or alternatively the form of an emulsion or suspension or salve. [0243]
  • Thus, the therapeutic CD83 nucleic acids, polypeptides or antibodies may be formulated for parenteral administration (e.g., by injection, for example, bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion containers or in multi-dose containers. As noted above, preservatives can be added to help maintain the shelve life of the dosage form. The active CD83 nucleic acids, polypeptides or antibodies and other ingredients may form suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active CD83 nucleic acids, polypeptides or antibodies and other ingredients may be in powder form, obtained by aseptic isolation of sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use. [0244]
  • These formulations can contain pharmaceutically acceptable carriers, vehicles and adjuvants that are well known in the art. It is possible, for example, to prepare solutions using one or more organic solvent(s) that is/are acceptable from the physiological standpoint, chosen, in addition to water, from solvents such as acetone, ethanol, isopropyl alcohol, glycol ethers such as the products sold under the name “Dowanol,” polyglycols and polyethylene glycols, C[0245] 1-C4 alkyl esters of short-chain acids, ethyl or isopropyl lactate, fatty acid triglycerides such as the products marketed under the name “Miglyol,” isopropyl myristate, animal, mineral and vegetable oils and polysiloxanes.
  • It is possible to add, if necessary, an adjuvant chosen from antioxidants, surfactants, other preservatives, film-forming, keratolytic or comedolytic agents, perfumes, flavorings and colorings. Antioxidants such as t-butylhydroquinone, butylated hydroxyanisole, butylated hydroxytoluene and a-tocopherol and its derivatives can be added. [0246]
  • Also contemplated are combination products that include one or more CD83 nucleic acids, polypeptides or antibodies of the present invention and one or more other anti-microbial agents. For example, a variety of antibiotics can be included in the pharmaceutical compositions of the invention, such as aminoglycosides (e.g., streptomycin, gentamicin, sisomicin, tobramycin and amicacin), ansamycins (e.g. rifamycin), antimycotics (e.g. polyenes and benzofuran derivatives), β-lactams (e.g. penicillins and cephalosporins), chloramphenical (including thiamphenol and azidamphenicol), linosamides (lincomycin, clindamycin), macrolides (erythromycin, oleandomycin, spiramycin), polymyxins, bacitracins, tyrothycin, capreomycin, vancomycin, tetracyclines (including oxytetracycline, minocycline, doxycycline), phosphomycin and fusidic acid. [0247]
  • Additionally, the CD83 nucleic acids, polypeptides or antibodies are well suited to formulation as sustained release dosage forms and the like. The formulations can be so constituted that they release the active nucleic acids, polypeptide or antibody, for example, in a particular part of the intestinal or respiratory tract, possibly over a period of time. Coatings, envelopes, and protective matrices may be made, for example, from polymeric substances, such as polylactide-glycolates, liposomes, microemulsions, microparticles, nanoparticles, or waxes. These coatings, envelopes, and protective matrices are useful to coat indwelling devices, e.g., stents, catheters, peritoneal dialysis tubing, draining devices and the like. [0248]
  • For topical administration, the therapeutic agents may be formulated as is known in the art for direct application to a target area. Forms chiefly conditioned for topical application take the form, for example, of creams, milks, gels, dispersion or microemulsions, lotions thickened to a greater or lesser extent, impregnated pads, ointments or sticks, aerosol formulations (e.g., sprays or foams), soaps, detergents, lotions or cakes of soap. Other conventional forms for this purpose include wound dressings, coated bandages or other polymer coverings, ointments, creams, lotions, pastes, jellies, sprays, and aerosols. Thus, the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention can be delivered via patches or bandages for dermal administration. Alternatively, the nucleic acid, polypeptide or antibody can be formulated to be part of an adhesive polymer, such as polyacrylate or acrylate/vinyl acetate copolymer. For long-term applications it might be desirable to use microporous and/or breathable backing laminates, so hydration or maceration of the skin can be minimized. The backing layer can be any appropriate thickness that will provide the desired protective and support functions. A suitable thickness will generally be from about 10 to about 200 microns. [0249]
  • Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents. The active CD83 nucleic acids, polypeptides or antibodies can also be delivered via iontophoresis, e.g., as disclosed in U.S. Pat. Nos. 4,140,122; 4,383,529; or 4,051,842. The percent by weight of a therapeutic agent of the invention present in a topical formulation will depend on various factors, but generally will be from 0.01% to 95% of the total weight of the formulation, and typically 0.1-85% by weight. [0250]
  • Drops, such as eye drops or nose drops, may be formulated with one or more of the therapeutic CD83 nucleic acids, polypeptides or antibodies in an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilizing agents or suspending agents. Liquid sprays are conveniently delivered from pressurized packs. Drops can be delivered via a simple eye dropper-capped bottle, or via a plastic bottle adapted to deliver liquid contents dropwise, via a specially shaped closure. [0251]
  • The therapeutic nucleic acids, polypeptide or antibody may further be formulated for topical administration in the mouth or throat. For example, the active ingredients may be formulated as a lozenge further comprising a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the composition in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the composition of the present invention in a suitable liquid carrier. [0252]
  • The pharmaceutical formulations of the present invention may include, as optional ingredients, pharmaceutically acceptable carriers, diluents, solubilizing or emulsifying agents, and salts of the type that are available in the art. Examples of such substances include normal saline solutions such as physiologically buffered saline solutions and water. Specific non-limiting examples of the carriers and/or diluents that are useful in the pharmaceutical formulations of the present invention include water and physiologically acceptable buffered saline solutions such as phosphate buffered saline solutions pH 7.0-8.0. [0253]
  • The CD83 nucleic acids, polypeptides or antibodies of the invention can also be administered to the respiratory tract. Thus, the present invention also provides aerosol pharmaceutical formulations and dosage forms for use in the methods of the invention. In general, such dosage forms comprise an amount of at least one of the agents of the invention effective to treat or prevent the clinical symptoms of a specific infection, indication or disease. Any statistically significant attenuation of one or more symptoms of an infection, indication or disease that has been treated pursuant to the method of the present invention is considered to be a treatment of such infection, indication or disease within the scope of the invention. [0254]
  • Alternatively, for administration by inhalation or insufflation, the composition may take the form of a dry powder, for example, a powder mix of the therapeutic agent and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form in, for example, capsules or cartridges, or, e.g., gelatin or blister packs from which the powder may be administered with the aid of an inhalator, insufflator, or a metered-dose inhaler (see, for example, the pressurized metered dose inhaler (MDI) and the dry powder inhaler disclosed in Newman, S. P. in [0255] Aerosols and the Lung, Clarke, S. W. and Davia, D. eds., pp. 197-224, Butterworths, London, England, 1984).
  • Therapeutic CD83 nucleic acids, polypeptides or antibodies of the present invention can also be administered in an aqueous solution when administered in an aerosol or inhaled form. Thus, other aerosol pharmaceutical formulations may comprise, for example, a physiologically acceptable buffered saline solution containing between about 0.1 mg/ml and about 100 mg/ml of one or more of the CD83 nucleic acids, polypeptides or antibodies of the present invention specific for the indication or disease to be treated. Dry aerosol in the form of finely divided solid nucleic acid, polypeptide or antibody particles that are not dissolved or suspended in a liquid are also useful in the practice of the present invention. CD83 nucleic acids, polypeptides or antibodies of the present invention may be formulated as dusting powders and comprise finely divided particles having an average particle size of between about 1 and 5 μm, alternatively between 2 and 3 μm. Finely divided particles may be prepared by pulverization and screen filtration using techniques well known in the art. The particles may be administered by inhaling a predetermined quantity of the finely divided material, which can be in the form of a powder. It will be appreciated that the unit content of active ingredient or ingredients contained in an individual aerosol dose of each dosage form need not in itself constitute an effective amount for treating the particular infection, indication or disease since the necessary effective amount can be reached by administration of a plurality of dosage units. Moreover, the effective amount may be achieved using less than the dose in the dosage form, either individually, or in a series of administrations. [0256]
  • For administration to the upper (nasal) or lower respiratory tract by inhalation, the therapeutic CD83 nucleic acids, polypeptides or antibodies of the invention are conveniently delivered from a nebulizer or a pressurized pack or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Nebulizers include, but are not limited to, those described in U.S. Pat. Nos. 4,624,251; 3,703,173; 3,561,444; and 4,635,627. Aerosol delivery systems of the type disclosed herein are available from numerous commercial sources including Fisons Corporation (Bedford, Mass.), Schering Corp. (Kenilworth, N.J.) and American Pharmoseal Co., (Valencia, Calif.). For intra-nasal administration, the therapeutic agent may also be administered via nose drops, a liquid spray, such as via a plastic bottle atomizer or metered-dose inhaler. Typical of atomizers are the Mistometer (Wintrop) and the Medihaler (Riker). [0257]
  • Furthermore, the active ingredients may also be used in combination with other therapeutic agents, for example, pain relievers, anti-inflammatory agents, antihistamines, bronchodilators and the like, whether for the conditions described or some other condition. [0258]
  • The present invention further pertains to a packaged pharmaceutical composition for controlling microbial infections such as a kit or other container. The kit or container holds a therapeutically effective amount of a pharmaceutical composition for modulating immune responses and instructions for using the pharmaceutical composition for control of the immune response. The pharmaceutical composition includes at least one nucleic acid, polypeptide or antibody of the present invention, in a therapeutically effective amount such that the selected disease or immunological condition is controlled. [0259]
  • The invention will be further described by reference to the following detailed examples, which are given for illustration of the invention, and are not intended to be limiting thereof. [0260]
    • EXAMPLE 1 Mouse Mutation and Characterization Mutant Generation
  • Male C57BL6 mice received 3 weekly injections of N-ethyl-N-nitrosourea (ENU) at a concentration of [0261] 100 mg/kg. N-Ethyl-N-nitrosourea was quantified prior to injection by spectrophotometry. Mice that regained fertility after a minimum period of 12 weeks were then used to generate pedigree founder G1 animals. G1 male mice were crossed to C57BL6J females and their female progeny (G2 animals) crossed back to their fathers to generate G3 animals for screening.
  • G3 mice were weaned at 3 weeks of age. Each animal then underwent a series of screens designed to assess a number of parameters, including immune function, inflammatory response and bone development. In the initial screen, conducted at 6 weeks of age, 150-200 μl of whole blood was collected by retro-orbital bleed into heparinized tubes. Cells were pelleted and red blood cells lysed. Samples were then stained with antibodies to cell surface markers expressed on distinct lymphoid and myeloid sub-populations. These samples were analyzed by flow-cytometry. [0262]
  • Mutant Identification [0263]
  • A group of 27 G3 mice from 2 different pedigrees, [0264] pedigree 9 and pedigree 57 (i.e. derived from 2 distinct G1 fathers) were analyzed in this screen. Two animals from pedigree 9 were identified as having a reduced (>2 standard deviation from normal) percentage of CD4+ T cells in peripheral blood (FIG. 1). Both animals were descended from the same G1 and shared the same mother. All other animals screened on that day had a normal percentage of CD4+ T cells. The number of phenodeviants identified (2 from a litter of 9 animals) was suggestive of a trait controlled by a single gene and inherited in a Mendelian fashion.
  • A second litter generated from [0265] Pedigree 9 bred to G2 daughter #4 exhibited an identical phenotype with reduced numbers of CD4+ T cells, further suggesting that the trait had a genetic basis. The phenotype was designated LCD4.1 (Low CD4 Mutant # 1) and was used for mapping experiments.
  • Mutation Mapping [0266]
  • In order to map the LCD4.1 mutant phenotype, affected G3 male mice (presumptive homozygous for the mutation) were bred to female animals from the C3HeB/FeJ strain to generate [0267] F 1 progeny. These F 1 females (presumptively heterozygous for the mutation) were then mated back to their affected father to generate N2 progeny.
  • Blood was collected from N2 animals and flow cytometric analysis was performed to identify CD4+ T cells. For a phenotype controlled by a single gene, breeding homozygous fathers to heterozygous daughters should yield 50% normal N2 animals and 50% affected N2 animals. This ratio of normal to affected animals was observed in the N2 generation: Multiple N2 animals exhibited a reduced percentage of CD4+ T cells, indicating that the phenotype was heritable (FIG. 2). [0268]
  • DNA samples were prepared from samples of tail tissue collected from these N2 mice and used for a genome scan, using a collection of assembled markers, and performed on the ABI 3100 DNA analyzer. Initial genetic linkage was seen to the tip of chromosome 13, where the closest microsatellite marker was D 13Mit139 with a LOD score of 8.2. By calculating upper and lower confidence limits, the mutant gene was located between 13.4 and 29.6 cM on chromosome 13. Through additional genotyping, this region was reduced to an 11 cM interval on chromosome 13. No significant linkage to other chromosomal regions was seen. [0269]
  • Mutation Identification [0270]
  • A candidate gene, CD83, was identified for gene-testing based upon its reported position within the interval. CD83 has previously been used as a marker of dendritic cell activation, suggesting that it might play a role in dendritic cell function and hence in regulating T cell development and function. [0271]
  • Sequence analysis of the mutant DNA revealed a mutation in the stop codon of CD83. All affected animals were homozygous for this mutation while non-affected animals carried one wild-type allele and one mutant allele (FIG. 3 and FIG. 4). The mutation destroyed the stop codon and resulted in the addition of a unique 55 amino acid tail to the C-terminus of CD83 (FIG. 5). [0272]
  • Additional Functional Data [0273]
  • A reduction in CD4+ T cells was seen in peripheral blood, spleen tissues and lymph nodes from homozygous LCD4.1 mice. Although there were a reduced number of CD4+ T cells in the thymus there is no overt block in the developmental process and there was substantially no alteration in B cell development in the bone marrow. Histological evaluation of thymus, spleen and lymph nodes from affected mice revealed no gross alteration in tissue architecture. [0274]
  • Dendritic cells can be differentiated from bone marrow of wild type mice by culture in GM-CSF. These cells can be characterized by the surface expression of dendritic cell markers, including CD86 and CD 11 c. Both LCD4.1 affected and normal animals were capable of giving rise to CD86+CD11c+cells under these culture conditions. LCD4.1 mutant mice thus were capable of generating dendritic cells under in vitro culture conditions. These data suggest that the phenotype seen in LCD4.1 mice is not due to a failure of dendritic cells to develop but rather may reflect a defect in function. [0275]
  • To track dendritic cells, the sensitizing agent FITC was applied to the dorsal surface of the ears of LCD4.1 affected and wild-type mice. FITC was picked up by dendritic cells that then migrated to the draining auricular lymph nodes, where the presence of the FITC label on the dendritic cell surface permitted detection by flow-cytometry. FITC labeled cells expressing CD86 were detected in equal proportions in draining lymph node from normal and affected LCD4.1 mice. These data indicate that LCD4.1 mutant animals are capable of generating dendritic cells in vivo and that these cells are able to pick up antigen in the ear and travel to the draining lymph node. [0276]
    • EXAMPLE 2 CD83 and CD4+ T Cell Function Materials and Methods
  • Spleens were removed from wild type and mutant mice and digested with collagenase to liberate dendritic cells. Spleens were stained for surface expression of CD4 (helper T cells) and CD 11c (dendritic cells). Cells expressing these markers were purified by fluorescence activated cell sorting (FACS sorting). CD 11c and CD4+positive cells were also purified from an allogeneic mouse strain, BALBc. [0277]
  • Mixed lymphocyte cultures were set up using purified cell populations. Dendritic cells from BALBc animals were used to stimulate CD4+ T cells from wild type and mutant mice. In a reciprocal experiment dendritic cells prepared from wild type and mutant mice were used to stimulate BALBc CD4+ T cells. After 5 days in culture proliferative responses were measured by incorporation of tritiated thymidine. [0278]
  • Dendritic cells from wild type and mutant mice were both capable of activating allogeneic T cells, suggesting that dendritic cell function was unimpaired in the mutant animal (FIG. 6[0279] a). In contrast CD4+ T cells from mutant animals exhibited a diminished response after 5 days of stimulation (FIG. 6b).
  • These data suggest that the mutation in the CD83 gene has minimal effect on dendritic cells intrinsic function but rather has a profound effect upon T cell activity. The CD4+ T cell therefore may have a novel requirement for CD83 functionality on T cells during allogeneic activation. CD83 may be influencing the extent of CD4+ T cell activation or altering the duration of the CD4+ T cell proliferative response. The therapeutic manipulation of CD83 may thus represent a mechanism for the specific regulation of T cell function in the treatment of T cell mediated diseases, including autoimmune disorders. Antibodies capable of blocking CD83 function may be used as therapeutics in the treatment of immune diseases whilst the activation of CD83 may-have utility in enhancing immune responses in cancer and other circumstances. [0280]
    • CONCLUSION
  • Although CD83 has been described as a marker of dendritic cell activation there has previously been little data describing its function in vivo. However, the mutation provided by the invention destabilizes or inactivates the protein and leads to impaired surface expression. As a consequence, CD4+ T cell function is impaired. However, the development of dendritic cells is not inhibited and mutant dendritic cells retain functionality. Nonetheless, the result is impaired development of CD4+ T cells. This impaired ability to activate T cells is also seen in a slight decrease in contact sensitivity responses in LCD4.1 mutant mice. [0281]
    • EXAMPLE 3 Mutant CD83 Have Different Cytokine Levels than Wild Type Mice
  • This Example demonstrates that CD4[0282] + T-cells from CD83 mutant animals express higher levels of IL-4 and lower levels of IL-2 compared to CD4+ T-cells from CD83 wild type animals.
  • Methods for Cell Activation and Cytokine Measurements [0283]
  • Spleens cells from 6-8-week-old homozygous CD83 wild type or CD83 mutant (LCD4.1) mice were used to isolate CD4[0284] + T-cells by positive selection using magnetic beads (Miltenyi Biotec). A 96 round bottom plate was coated with 50 μL per well of a solution containing either 1 or 10 μg/mL of anti-CD3 and 0.1 or 0.2 μg/mL of anti-CD28 antibodies (both from Pharmingen) in PBS overnight. This plate was then washed using 150 μL of PBS three times. To this pre-coated plate, 20,000 CD4+ T-cells (either wild type or CD83 mutant) were added in a 200 μL final volume of RPMI containing 10% FBS, 55 μM β-mercaptoethanol and antibiotics. The plates were then incubated in a CO2 incubator at 37° C. for 44 to 72 hours. For determination of cytokine levels, supernatants were harvested and cytokines were measured using either a Cytometric Bead Array system (Pharmingen) or ELISA (R&D). For RNA measurements, the cells were harvested and RNA was isolated using Tri reagent (Sigma). IL-10 and IL-4 mRNA levels were measured by reverse transcription and TaqMan (Applied Biosystems) analysis.
  • Results: [0285]
  • FIG. 7 shows the IL-2, IL-4, IL-5, TNFa and IFN? levels produced by either wild type or CD83 mutant CD4[0286] + T-cells. Purified cells were incubated as described above in the presence of 1 μg/mL of anti-CD3 and 0.2 μg/mL of anti-CD28 antibodies for 72 hours. The supernatants were then simultaneously analyzed for production of IL-2, IL-4, IL-5, TNFa and IFN? using the cytometric bead array system from Pharmingen.
  • FIG. 7 demonstrates that CD4[0287] + T-cells from CD83 mutant animals expressed higher levels of IL-4 and lower levels of IL-2 compared to CD4+ T-cells from CD83 wild type animals. Other cytokines and a new set of stimulation assays were analyzed including the production levels of IL-10 and GMCSF by these cells (FIGS. 8 and 9). In both cases, cells from mutant animals produce larger amounts of IL-10 and GMCSF than did wild type animals. FIG. 10 shows that mRNA levels for both IL-4 and IL-10 were increased in cells from activated mutant CD83, CD4+ T-cells compared with cells from wild type animals.
    • EXAMPLE 4 Anti-CD83 Antibodies Mimic the Effects of the CD83 Mutation
  • Methods for antibody testing: [0288]
  • For modulation of cytokine production by anti-CD83 antibodies, CD4[0289] + T-cells were isolated and activated as described above. Activation was performed in the presence of increasing concentrations of anti-CD83 antibodies. For proliferation assays, CD4+ T-cells were isolated from an OT2tg mouse. OT2tg mice are transgenic mice with a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide. Dendritic cells were isolated from a C57BL6 mouse by a negative selection using B220 magnetic beads (Miltenyi Biotec) followed by positive selection using CDl 1-c magnetic beads (Milteny Biotec). Five thousand CD4+ T-cells were then mixed with five thousand dendritic cells in a 96 well plate in the presences of 1 μM OVA peptide using RPMI (55 μM BME, 10% FBS plus antibiotics) in a final 200uL volume. These cells were then incubated for 48 to 72 hours in a CO2 incubator at 37° C. and pulsed using [3H] thymidine for 8 hours. Cells were then harvested and [3H] thymidine incorporation was quantified using a top counter.
  • Results: [0290]
  • In some assays, anti-CD83 antibodies decreased production of IL-4 by activated CD4[0291] + T-cells in a dose dependent manner. Different antibody preparations did provide somewhat different degrees of inhibition of IL-4 production (FIG. 11). Accordingly, the epitope and/or degree of affinity of the antibodies for the CD83 antigen may influence whether or not IL-4 production is significantly inhibited.
  • The effects of anti CD83 antibodies on proliferation of a peptide specific T-cell proliferation assay using the OT2 T-cell receptor (TCR) transgenic system were also observed. CD4[0292] + T-cells derived from these TCR transgenic animals express high levels of a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide and thus have high levels of proliferation when mixed with antigen presenting cells (dendritic cells were used) in the presence of the OVA peptide. In such assays, anti-CD83 antibodies were able to decrease proliferation of CD4+ T-cells in this system (FIG. 12). However, different antibody preparations had somewhat different effects on the proliferation of CD4+ T-cells. Accordingly, the CD83 epitope and/or degree of affinity of the antibodies for the CD83 antigen may influence whether or not CD4+ T-cell proliferation is significantly inhibited.
    • EXAMPLE 5 Increased T-Cell Proliferation by Transgenic Expression of CD83
  • This Example illustrates that over expression of CD83 in transgenic mice leads to increased T-cell proliferation. [0293]
  • Materials and Methods [0294]
  • A 34.3 kb fragment of normal mouse genomic DNA, including the ˜18 kb coding region of the CD83 gene, as well as ˜10.6 kb of upstream flanking sequences and ˜5.7 kb of downstream sequences was microinjected into normal mouse one-cell embryos. Four individual founder animals were generated. Transgenic mice were then crossed to a male OT2tg mouse. Male offspring carrying both the CD83 and OT2 transgene were used to analyze peptide specific T-cell proliferation. [0295]
  • For proliferation assays, CD4[0296] + T-cells and dendritic cells were isolated from either OT2tg [transgenic mice with a T-cell receptor specific for chicken ovalbumin (OVA) 323-339 peptide] CD83 wild type or from OT2tg CD83 transgenic mice as described above (Example 4). Five thousand OT2tg CD4+ T-cells from either wild type or CD83 transgenic animals were then mixed with five thousand wild type dendritic cells or five thousand CD83 transgenic dendritic cells in a 96 well plate in the presence of increasing concentrations of OVA peptide using RPMI (55 μM BME, 110% FBS plus antibiotics) in a final 200uL volume. These cells were then incubated for 48 to 72 hours in a CO2 incubator at 37C and pulsed using [3H] thymidine for 8 hours. Cells were then harvested and [3H] thymidine incorporation was quantified using a top counter.
  • Results: [0297]
  • OT2tg CD4[0298] + T-cells derived from CD83 transgenic mice proliferated at higher rates than the same cell population derived from a CD83 wild type animal (FIG. 13). This increased proliferation was seen at all the concentrations of OVA peptide tested. Whereas OT2tg CD4+ T-cells derived from CD83 transgenic animals exhibited increased proliferation, dendritic cells from CD83 transgenic animals did not exhibit a substantial increase in proliferation. Therefore, it appears that transgenic expression in the CD4+ T-cell, and not in dendritic cells is what led to the increased proliferation of CD4+ T-cells.
    • EXAMPLE 6 Inhibition of Proliferation of PHA Activated Human PBMCs by Protein A Purified Rabbit Anti-Mouse CD83 Antibodies
  • This Example shows that antibodies raised against the CD83 protein can inhibit proliferation of human peripheral blood mononuclear cells. [0299]
  • Materials and Methods [0300]
  • Rabbit polyclonal sera was raised against mouse CD83 protein by immunizing rabbits using a mouse CD83 external domain protein fused to a rabbit Ig domain (FIG. 14). Pre-immune sera and anti-mouse polyclonal sera were then purified using a protein A column (Pharmacia Biotech) as described by the manufacturer, then dialyzed against PBS and stored at 4° C. To monitor the recognition of mouse CD83 protein by the polyclonal sera, which was obtained at different dates post immunization, a titer was obtained using an antigen specific ELISA (FIG. 15). As illustrated by FIG. 15, a good polyclonal response was obtained against the mouse CD83 protein. [0301]
  • Human peripheral blood mononuclear cells (PBMCs) were isolated using a Ficoll gradient (Ficoll Paque Plus, Pharmacia) and washed with PBS buffer. For activation and proliferation studies, five thousand cells were incubated in 200 μL of media (RPMI, 10% FBS, antibiotics) and 5 μg/mL of [0302] Phaseolus vulgaris leucoagglutinin (PHA) in the presence or absence of increasing concentrations of Protein A purified pre-immune sera or with similarly purified anti-CD83 polyclonal antibodies. After 48 hours at 37° C. in a CO2 incubator the cells were pulsed with [3H] thymidine for 8 hours and harvested. Thymidine incorporation into the PBMCs was measured using a top counter for analysis.
  • A Selected Lymphocyte Antibody Method (SLAM) procedure was used to establish monoclonal antibody cell lines from the rabbits used to generate the anti-CD83 antibodies. Antibody forming cells were isolated from the immunized rabbits that produced antibodies capable of binding CD83, the genes encoding antibodies that recognized CD83 and inhibited proliferation of lymphocytes were then cloned by PCR amplification and sequenced. Separate lines of monoclonal antibody producing cells were then established and expanded in culture. Antibodies were purified using Protein A chromatography according to manufacturer's instructions and tested for their ability to recognize CD83 proteins and to inhibit proliferation of PHA stimulated human PBMCs. [0303]
  • Results [0304]
  • FIG. 16 illustrates that proliferation of PHA-activated human PBMCs was inhibited by polyclonal antibodies raised against the external region of the mouse CD83 protein. Proliferation of PHA-activated human PBMCs was not affected by addition of increasing concentrations of protein A purified rabbit pre-immune sera. When increasing concentrations of protein A purified rabbit polyclonal sera raised against the mouse CD83 protein was added, a concentration dependent decrease in proliferation was observed. [0305]
  • These data indicate that antibodies raised against the mouse protein are able to cross-react with the human protein. Moreover, antibodies raised against the mouse protein are able to inhibit proliferation of PHA-activated human PBMCs. [0306]
  • A summary of the characteristics of two monoclonal antibody preparations having functional activity is shown in Table 1. Isolated recombinant mouse and human CD83 protein preparations were used for the BIACORE and ELISA assays. Endogenous human CD83 protein expressed in a human KMH2 cell line was used for FACS assays. [0307]
    TABLE 1
    Monoclonal Antibody Functionality and Reactivity
    with Mouse and Human CD83
    Assay 95F04 Antibodies 96G08 Antibodies
    Inhibition of human PBMC ++ +++
    proliferation
    Biacore - mouse CD83 +++ +++
    Biacore - human CD83 ++ −
    ELISA - mouse CD83 +++ +++
    ELISA - human CD83 ++ −
    FACS - human CD83 ND ++
  • While the 96G08 antibodies appeared to have reduced affinity for human CD83 protein via the Biacore and ELISA assays, the FACS assay indicated that this antibody preparation could bind to endogenously produced human CD83 (FIGS. 18 and 19). Moreover, the 96G08 antibodies were able to inhibit proliferation of human peripheral blood mononuclear cells (PBMCs), as illustrated in FIG. 20. Hence, some aspect of either the purification or the structure of the isolated recombinant human protein may have influenced the in vitro binding of 96G08 antibodies to the recombinant human CD83. For example, the recombinant human CD83 protein employed for the Biacore and ELISA assays is a chimeric protein that is joined to a portion of an immunoglobulin Fc fragment. Removal of this Fc fragment may improve in vitro binding to the human CD83 protein. [0308]
  • FIG. 20 illustrates that the 95F04 and 96G08 antibody preparations can inhibit proliferation of PHA activated human peripheral blood mononuclear cells as detected by incorporation of [[0309] 3H] thymidine. As shown, when no antibody was present about 10,000 cpm of [3H] thymidine was incorporated into human peripheral blood mononuclear cells. However, when 30 μg/ml of the 95F04 antibody preparation was present, incorporation of [3H] thymidine dropped to about 2000 cpm. The 96G08 antibody preparation had an even greater effect on [3H] thymidine incorporation. When 30 μg/ml 96G08 antibody preparation was added to human peripheral blood mononuclear cells, [3H] thymidine incorporation was reduced to about 300 cpm. These data indicate that the 95F04 and 96G08 antibody preparations can alter the function of human CD83 in vitro.
    • EXAMPLE 7 Multimerized Anti-CD83 Antibodies Inhibit Proliferation of Immune Cells
  • This Example shows that antibodies raised against the CD83 protein as described in the previous example are particularly effective at inhibiting proliferation of immune cells after the antibodies are multimerized or multimerized by binding the antibodies to a solid support or by cross-linking in solution. [0310]
  • Materials and Methods [0311]
  • Round bottom microtiter plates were coated with different preparations of anti-CD83 antibody preparations by incubating the plates with 50 μl of 50 μg/ml antibody preparation per well either for 2 hours at 37° C. or overnight at 4° C. As a positive control, some wells were coated with anti-LFA antibodies that are known to inhibit proliferation of lymphocytes. After coating, the wells were then washed thoroughly with PBS. [0312]
  • Mouse (C57B 16) spleen cells were isolated and plated in the antibody or control treated wells at 30,000 cells per well. For activation, Concavalin A was added to a final concentration of 1.0 μg/ml. Cellular proliferation was assessed by measuring the incorporation of tritiated thymidine during the last 6 to 8 hours of a 48 hour incubation. In another experiment, the specificity of the observed antibody-induced inhibition of lymphocyte proliferation was tested by repeating this experiment with addition of mouse CD83 protein before adding the lymphocytes to the antibody coated microtiter wells. [0313]
  • As described in more detail below, the 6G05 antibody preparation was identified as a good inhibitor of lymphocyte proliferation. In contrast, the 112D08 antibody preparation was identified as having little or no inhibitory activity when bound to microtiter wells. The 112D08 antibody preparation was used as a negative control in some of the subsequent experiments. [0314]
  • The inhibitory activities of plate-bound versus soluble, cross-linked 6G05 antibodies were compared in another experiment. Plate-bound 6G05 antibodies were prepared as described above. Approximately 30,000 activated lymphocytes were added per well to antibody coated plates or to non-coated plates containing 1.0 or 5.0 μg/ml soluble 6G05 antibody preparation. A secondary rabbit anti-mouse antibody (10 μg/ml or 25 μg/ml) was added to the wells containing the soluble 6G05 antibody preparation to act as a cross-linking reagent for the 6G05 antibodies. Cellular proliferation was assessed by incorporation of tritiated thymidine as described above. [0315]
  • Results [0316]
  • The results of one screen for anti-CD83 antibody preparations that can inhibit lymphocyte proliferation are shown in FIGS. [0317] 25A-B. As illustrated in FIG. 25A many anti-CD83 antibody preparations inhibit proliferation of activated lymphocytes, including the 94c09, 98a02, 94d08, 98d11, 101b08, 6g05, 20d04, 14c12, 11g05, 12g04, 32f12 and 98b11 preparations. Note that some variation in the degree of inhibition obtained is observed. For example, while the 98b 1 preparation is not so effective, the 6g05 antibody preparation is a highly effective inhibitor of lymphocyte proliferation.
  • FIG. 25B further illustrates that some antibody preparations are highly effective inhibitors (e.g. 117G12) but others are not (e.g. 98g08). The 824pb antibody refers to rabbit polyclonal antisera; as shown this polyclonal antisera was not particularly effective at inhibiting lymphocyte proliferation [0318]
  • FIG. 26 illustrates that the inhibitory activity of the 6g05 antibody preparation is quenched by soluble mouse CD83 protein. In this assay, mouse CD83 protein was added to anti-CD83 antibody-coated wells before activated lymphocytes were introduced. Both a highly effective proliferation inhibitor (6g05) and an antibody preparation with little or no inhibitory activity (98g08) were tested. A control having no antibody and no mouse CD83 protein as well as a control with added mouse CD83 and no antibody was included. Cellular proliferation of the activated lymphocytes was assessed by observing the incorporation of tritiated thymidine as described above. As shown in FIG. 26, the 6g05 antibody strongly inhibits lymphocyte proliferation when no mouse CD83 is present. However, when mouse CD83 is added before the lymphocytes, the 6g05 antibody exhibits little or no inhibition of lymphocyte proliferation. These data indicate that the inhibitory activity of the 6g05 antibody preparation operates through the CD83 gene product, rather than through some non-specific interaction with lymphocytes. [0319]
  • FIGS. 27 and 28 illustrate that anti-CD83 antibodies that are multimerized by use of a rabbit anti-mouse antibody have inhibitory activity that is like that of plate-bound anti-CD83 antibodies. The proliferation of lymphocytes was measured by observing the incorporation of tritiated thymidine with and without anti-CD83 antibodies as described above. In one set of assays plate-bound 6g05 antibodies were used and in another soluble 6g05 antibodies were employed. The soluble 6g05 antibodies were cross-linked by addition of rabbit anti-mouse antibodies that bind to the Fc region of the 6g05 antibodies. For comparison, a soluble and plate-bound antibody preparation with no inhibitory activity (the 112D08 antibody preparation was also tested. A similar series of assays were set up using a panel of soluble anti-CD83 antibodies. [0320]
  • As shown in FIG. 27, both plate-bound and crosslinked 6g05 antibodies were highly effective inhibitors of lymphocyte proliferation. These data indicate that the method of aggregating anti-CD83 antibodies is not particularly important. In other words the multimerization can be achieved by adhering or attaching antibodies to a solid support or by crosslinking the anti-CD83 antibodies through their Fc regions using a rabbit anti-mouse secondary antibody. So long as the anti-CD83 antibodies are in close proximity, they are effective inhibitors of lymphocyte proliferation. [0321]
  • FIG. 28 shows that many soluble anti-CD83 antibodies exhibit good inhibition of lymphocyte proliferation when they are cross-linked with the rabbit anti-mouse secondary antibody. For example, the 6g05, 11g04, 12g04, 14c12, 20d04, 32f12, 94c09, 94d08, 98a02, 98d11(3), 101B08(2.7) and 117g12 antibody preparations strongly inhibit lymphocyte multimerization when cross-linked with the rabbit anti-mouse antibodies. [0322]
    • EXAMPLE 8 Multimerized Anti-CD83 Antibodies Inhibit Proliferation of Immune Cells in a Mixed Lymphocyte Reaction
  • This Example shows that multimerized anti-CD83 antibodies inhibit proliferation of lymphocytes in a mixed lymphocyte reaction (MLR) assay. [0323]
  • Materials and Methods [0324]
  • The MLR assay employed was a modification of the procedure described in Bradley, pp 162-166 in Mishell et al., eds. Selected Methods in Cellular Immunology (Freeman, San Francisco, 1980); and Battisto, et al., Meth, in Enzymol. 150:83-91 (1987). [0325]
  • Spleens were removed from BALBc and [0326] C57B 16 mice and digested with collagenase to liberate dendritic and CD4+ cells, respectively. Spleens were stained for surface expression of CD4 (helper T cells) or CD11c (dendritic cells). Cells expressing these markers were purified by using magnetic beads (Miltenyi) according to the manufacturer's instructions.
  • Mixed lymphocyte cultures were set up using purified cell populations. Plates with different anti-CD83 antibody preparations bound thereto were prepared as described in the previous examples. Approximately 1250 CD11c dendritic cells were used to stimulate approximately 20,000 CD4+ T cells. After 4 days in culture, proliferative responses were measured by incorporation of tritiated thymidine. A positive control antibody, the anti-LFA antibody, was also used for comparison purposes in this assay because it is known to inhibit lymphocyte proliferation in MLR assays. [0327]
  • A similar experiment was performed to assess the recall response of lymphocytes exposed to 100 μg/ml anti-CD83 antibodies. Prior to spleen removal and CD 11 c and CD4+ cell isolation, BALBc mice were first immunized with keyhole limpet hemocyanin (KLH) in a 1:1 ratio with complete Freund's adjuvant close to the lymph node area. Lymph nodes were harvested and challenged in vitro with KLH at a final concentration of 2.5 μg/ml and the proliferative response of the cells was assayed as described above by observing incorporation of tritiated thymidine. [0328]
  • Results [0329]
  • FIG. 29 shows that the conditions employed several monoclonal anti-CD83 antibodies can inhibit lymphocyte proliferation in a mixed lymphocyte reaction assay. For example, the 98a02, 98d11, 20d04, 14c12, 12g04, and 117g12 inhibit lymphocyte proliferation in this assay. [0330]
  • FIG. 30 shows that many anti-CD83 antibody preparations can inhibit the recall response of lymphocytes. For example, 94c09, 98a02, 6g05, 20d04, and 117104 antibody preparations inhibited proliferation of activated lymphocytes exposed to an antigen (KLH) to which they had been immunized. [0331]
  • These data suggest that anti-CD83 antibodies can quiet the proliferative response of CD4+ T cells after stimulation by allogenic CD11 cells and/or antigen. [0332]
    • EXAMPLE 9
  • Exposure to Anti-CD83 Antibodies Does Not Cause Apoptosis of Activated Lymphocytes [0333]
  • This Example shows that exposure to anti-CD83 antibodies does not lead to apoptosis of activated lymphocytes. [0334]
  • Materials and Methods [0335]
  • Mouse (C57B 16) spleen cells were isolated and activated by incubation for 24 hours with 1.0 μg/ml Concavalin A in the presence or absence of anti-CD83 antibodies and rabbit anti-mouse antibodies as a crosslinking reagent as described above. Cells were incubated for 48 hours at 37° C. Proliferative responses were measured by incorporation of tritiated thymidine. Total caspase activity and annexinV expression levels were used as a measure of apoptosis. [0336]
  • Homogeneous total caspase activity was measured using a kit (Roche( following the manufacturer's instructions. [0337]
  • To test for apoptosis using annexinV expression, cells were incubated with annexin-FITC and propidium iodide (AnnexinV-FITC kit, Calbiochem) and the percentage of positive Annexin V-FITC labeled cells was determined by Fluorescence Activated Cell sorting (FACs). [0338]
  • Results [0339]
  • FIGS. [0340] 31A-B shows that soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations not only inhibit activated lymphocyte cell proliferation (FIG. 31B) but also have very low caspase activity (FIG. 31A). Similarly, FIG. 32 shows that the percentage of activated lymphocytes that express annexinV is reduced after treatment with soluble but cross-linked 6g05 and 14c12 anti-CD83 antibody preparations.
  • These data indicate that while anti-CD83 antibodies inhibit proliferation of ConA activated splenocytes, they do not induce apoptosis of immune cells. Instead, anti-CD83 antibodies actually depress the expression of apoptosis markers. Hence, the reduction in cell proliferation observed when activated lymphocytes are exposed to anti-CD83 antibodies is not due to increased programmed cell death. [0341]
    • EXAMPLE 10
  • : Exposure to Anti-CD83 Antibodies Does Not Inhibit Activation of Lymphocytes [0342]
  • This Example shows that exposure to anti-CD83 antibodies does not inhibit activation of lymphocytes. [0343]
  • Materials and Methods [0344]
  • Mouse (B6) spleen cells were isolated and activated using Concavalin A as described above in the presence or absence of anti-CD83 antibodies and the secondary anti-mouse crosslinking antibodies. The anti-CD83 antibody preparations employed included the 6g05, 14c12, 98b11 and 112d08 preparations. Activation of the cells was assessed using CD69 expression as a marker of cell activation. [0345]
  • Results [0346]
  • FIG. 33 illustrates that splenocytes activated with Concavalin A express the CD69 activation marker even though they were incubated with anti-CD83 antibodies. In particular, the star or asterisks in the lower right hand corner of the graph shows the level of CD69 expression observed when splenocytes are not activated with Concavalin A. However, when splenocytes were activated with Concavalin A they expressed high levels of CD69 even after incubation with any of the 6g05, 14c12, 98b11 or 112d08 anti-CD83 antibody preparations. [0347]
  • These results indicate that while cellular proliferation of lymphocytes exposed to anti-CD83 antibodies is arrested, the lymphocytes still undergo activation. [0348]
    • EXAMPLE 11 Anti-CD83 Antibodies Arrest the Lymphocyte Cell Cycle in the G0/G1 Stage
  • This Example shows that exposure to anti-CD83 antibodies arrests activated lymphocytes in the G0/[0349] G 1 stage of the cell cycle.
  • Materials and Methods [0350]
  • Mouse (B6) spleen cells were isolated and activated by incubation for 48 hours with 1.0 μg/ml Concavalin A in the presences of anti-CD83 antibodies with the crosslinking antibodies as described above. To analyze cell cycle distribution, cells were fixed and DNA was stained with propidium iodine according to the protocol described for the flowcytometer (Cold Spring Harbor, N.Y.). WinMDI software was used for background subtraction caused by debris in the DNA histogram. Each histogram was further analyzed by cycle red software to obtain the distribution of cells therein. In addition, the size and shape of the activated cells was assessed by their forward (FSC) and side (SSC) scatter during this experiment. [0351]
  • The anti-CD83 antibody preparations employed were the 6g05 and 14c12 preparations that had been shown to inhibit cellular proliferation and the 112d08 preparation that had little or no effect on cellular proliferation. Cells having 2N complement of DNA were assumed to be in the [0352] G 1/G0 phase of the cell cycle; cells having 3N complement of DNA were assumed to be in the G2/M phase of the cell cycle; and cells having 4N complement of DNA were assumed to be in the S phase of the cell cycle. The percentage of cells having G1/G0, G2/M or S phase of the cell cycle was determined and plotted in FIGS. 35A-C.
  • Results [0353]
  • FIG. 34 shows that a population of activated splenocytes mixed with anti-CD83 antibody preparations have lost the blasting (dividing) cells as detected by FACS sorting. Almost all cells sort as small cells with a 2N content of DNA as illustrated by the high proportion of cells towards the left (smaller) side of the population distribution in FIG. 34. [0354]
  • FIGS. [0355] 35A-C show that treatment of Concavalin A activated lymphocytes with either of 6g05 and 14c12 antibody preparations leads to a cellular population that was enriched in cells in the G 1/G0 stage of the cell cycle. Treatment with either the rabbit anti-mouse antibody or the 112d08 antibody preparation that has little or no effect on cell proliferation did not lead to a cellular population that was enriched in cells in the G1/G0 stage of the cell cycle.
  • These data indicate that exposure to anti-CD83 antibodies arrests lymphocytes in the G1/G0 stage. Taken together with the data in preceding Examples, these data indicate that anti-CD83 antibodies can cause lymphocytes to enter a state of antigen specific unresponsiveness or anergy. [0356]
  • All patents and publications referenced or mentioned herein are indicative of the levels of skill of those skilled in the art to which the invention pertains, and each such referenced patent or publication is hereby incorporated by reference to the same extent as if it had been incorporated by reference in its entirety individually or set forth herein in its entirety. Applicants reserve the right to physically incorporate into this specification any and all materials and information from any such cited patents or publications. [0357]
  • The specific methods and compositions described herein are representative of preferred embodiments and are exemplary and not intended as limitations on the scope of the invention. Other objects, aspects, and embodiments will occur to those skilled in the art upon consideration of this specification, and are encompassed within the spirit of the invention as defined by the scope of the claims. It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The invention illustratively described herein suitably may be practiced in the absence of any element or elements, or limitation or limitations, which is not specifically disclosed herein as essential. The methods and processes illustratively described herein suitably may be practiced in differing orders of steps, and that they are not necessarily restricted to the orders of steps indicated herein or in the claims. As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to “a host cell” includes a plurality (for example, a culture or population) of such host cells, and so forth. Under no circumstances may the patent be interpreted to be limited to the specific examples or embodiments or methods specifically disclosed herein. Under no circumstances may the patent be interpreted to be limited by any statement made by any Examiner or any other official or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or reservation expressly adopted in a responsive writing by Applicants. [0358]
  • The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intent in the use of such terms and expressions to exclude any equivalent of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention as claimed. Thus, it will be understood that although the present invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims. [0359]
  • The invention has been described broadly and generically herein. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the invention. This includes the generic description of the invention with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein. [0360]
  • Other embodiments are within the following claims. In addition, where features or aspects of the invention are described in terms of Markush groups, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group. [0361]
  • 0
    SEQUENCE LISTING
    <160> NUMBER OF SEQ ID NOS: 99
    <210> SEQ ID NO 1
    <211> LENGTH: 2051
    <212> TYPE: DNA
    <213> ORGANISM: Mus musculus
    <400> SEQUENCE: 1
    gcgctccagc cgcatgtcgc aaggcctcca gctcctgttt ctaggctgcg cctgcagcct 60
    ggcacccgcg atggcgatgc gggaggtgac ggtggcttgc tccgagaccg ccgacttgcc 120
    ttgcacagcg ccctgggacc cgcagctctc ctatgcagtg tcctgggcca aggtctccga 180
    gagtggcact gagagtgtgg agctcccgga gagcaagcaa aacagctcct tcgaggcccc 240
    caggagaagg gcctattccc tgacgatcca aaacactacc atctgcagct cgggcaccta 300
    caggtgtgcc ctgcaggagc tcggagggca gcgcaacttg agcggcaccg tggttctgaa 360
    ggtgacagga tgccccaagg aagctacaga gtcaactttc aggaagtaca gggcagaagc 420
    tgtgttgctc ttctctctgg ttgttttcta cctgacactc atcattttca cctgcaaatt 480
    tgcacgacta caaagcattt tcccagatat ttctaaacct ggtacggaac aagcttttct 540
    tccagtcacc tccccaagca aacatttggg gccagtgacc cttcctaaga cagaaacggt 600
    atgagtagga tctccactgg tttttacaaa gccaagggca catcagatca gtgtgcctga 660
    atgccacccg gacaagagaa gaatgagctc catcctcaga tggcaacctt tctttgaagt 720
    ccttcacctg acagtgggct ccacactact ccctgacaca gggtcttgag caccatcata 780
    tgatcacgaa gcatggagta tcaccgcttc tctgtggctg tcagcttaat gtttcatgtg 840
    gctatctggt caacctcgtg agtgcttttc agtcatctac aagctatggt gagatgcagg 900
    tgaagcaggg tcatgggaaa tttgaacact ctgagctggc cctgtgacag actcctgagg 960
    acagctgtcc tctcctacat ctgggataca tctctttgaa tttgtcctgt ttcgttgcac 1020
    cagcccagat gtctcacatc tggcggaaat tgacaggcca agctgtgagc cagtgggaaa 1080
    tatttagcaa ataatttccc agtgcgaagg tcctgctatt agtaaggagt attatgtgta 1140
    catagaaatg agaggtcagt gaactattcc ccagcagggc cttttcatct ggaaaagaca 1200
    tccacaaaag cagcaataca gagggatgcc acatttattt ttttaatctt catgtacttg 1260
    tcaaagaaga atttttcatg ttttttcaaa gaagtgtgtt tctttccttt tttaaaatat 1320
    gaaggtctag ttacatagca ttgctagctg acaagcagcc tgagagaaga tggagaatgt 1380
    tcctcaaaat agggacagca agctagaagc actgtacagt gccctgctgg gaagggcaga 1440
    caatggactg agaaaccaga agtctggcca caagattgtc tgtatgattc tggacgagtc 1500
    acttgtggtt ttcactctct ggttagtaaa ccagatagtt tagtctgggt tgaatacaat 1560
    ggatgtgaag ttgcttgggg aaagctgaat gtagtgaata cattggcaac tctactgggc 1620
    tgttaccttg ttgatatcct agagttctgg agctgagcga atgcctgtca tatctcagct 1680
    tgcccatcaa tccaaacaca ggaggctaca aaaaggacat gagcatggtc ttctgtgtga 1740
    actcctcctg agaaacgtgg agactggctc agcgctttgc gcttgaagga ctaatcacaa 1800
    gttcttgaag atatggacct aggggagcta ttgcgccacg acaggaggaa gttctcagat 1860
    gttgcattga tgtaacattg ttgcatttct ttaatgagct gggctccttc ctcatttgct 1920
    tcccaaagag attttgtccc actaatggtg tgcccatcac ccacactatg aaagtaaaag 1980
    ggatgctgag cagatacagc gtgcttacct ctcagccatg actttcatgc tattaaaaga 2040
    atgcatgtga a 2051
    <210> SEQ ID NO 2
    <211> LENGTH: 196
    <212> TYPE: PRT
    <213> ORGANISM: Mus musculus
    <400> SEQUENCE: 2
    Met Ser Gln Gly Leu Gln Leu Leu Phe Leu Gly Cys Ala Cys Ser Leu
    1 5 10 15
    Ala Pro Ala Met Ala Met Arg Glu Val Thr Val Ala Cys Ser Glu Thr
    20 25 30
    Ala Asp Leu Pro Cys Thr Ala Pro Trp Asp Pro Gln Leu Ser Tyr Ala
    35 40 45
    Val Ser Trp Ala Lys Val Ser Glu Ser Gly Thr Glu Ser Val Glu Leu
    50 55 60
    Pro Glu Ser Lys Gln Asn Ser Ser Phe Glu Ala Pro Arg Arg Arg Ala
    65 70 75 80
    Tyr Ser Leu Thr Ile Gln Asn Thr Thr Ile Cys Ser Ser Gly Thr Tyr
    85 90 95
    Arg Cys Ala Leu Gln Glu Leu Gly Gly Gln Arg Asn Leu Ser Gly Thr
    100 105 110
    Val Val Leu Lys Val Thr Gly Cys Pro Lys Glu Ala Thr Glu Ser Thr
    115 120 125
    Phe Arg Lys Tyr Arg Ala Glu Ala Val Leu Leu Phe Ser Leu Val Val
    130 135 140
    Phe Tyr Leu Thr Leu Ile Ile Phe Thr Cys Lys Phe Ala Arg Leu Gln
    145 150 155 160
    Ser Ile Phe Pro Asp Ile Ser Lys Pro Gly Thr Glu Gln Ala Phe Leu
    165 170 175
    Pro Val Thr Ser Pro Ser Lys His Leu Gly Pro Val Thr Leu Pro Lys
    180 185 190
    Thr Glu Thr Val
    195
    <210> SEQ ID NO 3
    <211> LENGTH: 2051
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic mutant CD83 sequence
    <400> SEQUENCE: 3
    gcgctccagc cgcatgtcgc aaggcctcca gctcctgttt ctaggctgcg cctgcagcct 60
    ggcacccgcg atggcgatgc gggaggtgac ggtggcttgc tccgagaccg ccgacttgcc 120
    ttgcacagcg ccctgggacc cgcagctctc ctatgcagtg tcctgggcca aggtctccga 180
    gagtggcact gagagtgtgg agctcccgga gagcaagcaa aacagctcct tcgaggcccc 240
    caggagaagg gcctattccc tgacgatcca aaacactacc atctgcagct cgggcaccta 300
    caggtgtgcc ctgcaggagc tcggagggca gcgcaacttg agcggcaccg tggttctgaa 360
    ggtgacagga tgccccaagg aagctacaga gtcaactttc aggaagtaca gggcagaagc 420
    tgtgttgctc ttctctctgg ttgttttcta cctgacactc atcattttca cctgcaaatt 480
    tgcacgacta caaagcattt tcccagatat ttctaaacct ggtacggaac aagcttttct 540
    tccagtcacc tccccaagca aacatttggg gccagtgacc cttcctaaga cagaaacggt 600
    aagagtagga tctccactgg tttttacaaa gccaagggca catcagatca gtgtgcctga 660
    atgccacccg gacaagagaa gaatgagctc catcctcaga tggcaacctt tctttgaagt 720
    ccttcacctg acagtgggct ccacactact ccctgacaca gggtcttgag caccatcata 780
    tgatcacgaa gcatggagta tcaccgcttc tctgtggctg tcagcttaat gtttcatgtg 840
    gctatctggt caacctcgtg agtgcttttc agtcatctac aagctatggt gagatgcagg 900
    tgaagcaggg tcatgggaaa tttgaacact ctgagctggc cctgtgacag actcctgagg 960
    acagctgtcc tctcctacat ctgggataca tctctttgaa tttgtcctgt ttcgttgcac 1020
    cagcccagat gtctcacatc tggcggaaat tgacaggcca agctgtgagc cagtgggaaa 1080
    tatttagcaa ataatttccc agtgcgaagg tcctgctatt agtaaggagt attatgtgta 1140
    catagaaatg agaggtcagt gaactattcc ccagcagggc cttttcatct ggaaaagaca 1200
    tccacaaaag cagcaataca gagggatgcc acatttattt ttttaatctt catgtacttg 1260
    tcaaagaaga atttttcatg ttttttcaaa gaagtgtgtt tctttccttt tttaaaatat 1320
    gaaggtctag ttacatagca ttgctagctg acaagcagcc tgagagaaga tggagaatgt 1380
    tcctcaaaat agggacagca agctagaagc actgtacagt gccctgctgg gaagggcaga 1440
    caatggactg agaaaccaga agtctggcca caagattgtc tgtatgattc tggacgagtc 1500
    acttgtggtt ttcactctct ggttagtaaa ccagatagtt tagtctgggt tgaatacaat 1560
    ggatgtgaag ttgcttgggg aaagctgaat gtagtgaata cattggcaac tctactgggc 1620
    tgttaccttg ttgatatcct agagttctgg agctgagcga atgcctgtca tatctcagct 1680
    tgcccatcaa tccaaacaca ggaggctaca aaaaggacat gagcatggtc ttctgtgtga 1740
    actcctcctg agaaacgtgg agactggctc agcgctttgc gcttgaagga ctaatcacaa 1800
    gttcttgaag atatggacct aggggagcta ttgcgccacg acaggaggaa gttctcagat 1860
    gttgcattga tgtaacattg ttgcatttct ttaatgagct gggctccttc ctcatttgct 1920
    tcccaaagag attttgtccc actaatggtg tgcccatcac ccacactatg aaagtaaaag 1980
    ggatgctgag cagatacagc gtgcttacct ctcagccatg actttcatgc tattaaaaga 2040
    atgcatgtga a 2051
    <210> SEQ ID NO 4
    <211> LENGTH: 251
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic mutant CD83 sequence
    <400> SEQUENCE: 4
    Met Ser Gln Gly Leu Gln Leu Leu Phe Leu Gly Cys Ala Cys Ser Leu
    1 5 10 15
    Ala Pro Ala Met Ala Met Arg Glu Val Thr Val Ala Cys Ser Glu Thr
    20 25 30
    Ala Asp Leu Pro Cys Thr Ala Pro Trp Asp Pro Gln Leu Ser Tyr Ala
    35 40 45
    Val Ser Trp Ala Lys Val Ser Glu Ser Gly Thr Glu Ser Val Glu Leu
    50 55 60
    Pro Glu Ser Lys Gln Asn Ser Ser Phe Glu Ala Pro Arg Arg Arg Ala
    65 70 75 80
    Tyr Ser Leu Thr Ile Gln Asn Thr Thr Ile Cys Ser Ser Gly Thr Tyr
    85 90 95
    Arg Cys Ala Leu Gln Glu Leu Gly Gly Gln Arg Asn Leu Ser Gly Thr
    100 105 110
    Val Val Leu Lys Val Thr Gly Cys Pro Lys Glu Ala Thr Glu Ser Thr
    115 120 125
    Phe Arg Lys Tyr Arg Ala Glu Ala Val Leu Leu Phe Ser Leu Val Val
    130 135 140
    Phe Tyr Leu Thr Leu Ile Ile Phe Thr Cys Lys Phe Ala Arg Leu Gln
    145 150 155 160
    Ser Ile Phe Pro Asp Ile Ser Lys Pro Gly Thr Glu Gln Ala Phe Leu
    165 170 175
    Pro Val Thr Ser Pro Ser Lys His Leu Gly Pro Val Thr Leu Pro Lys
    180 185 190
    Thr Glu Thr Val Arg Val Gly Ser Pro Leu Val Phe Thr Lys Pro Arg
    195 200 205
    Ala His Gln Ile Ser Val Pro Glu Cys His Pro Asp Lys Arg Arg Met
    210 215 220
    Ser Ser Ile Leu Arg Trp Gln Pro Phe Phe Glu Val Leu His Leu Thr
    225 230 235 240
    Val Gly Ser Thr Leu Leu Pro Asp Thr Gly Ser
    245 250
    <210> SEQ ID NO 5
    <211> LENGTH: 756
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic mutant CD83 sequence
    <400> SEQUENCE: 5
    atgtcgcaag gcctccagct cctgtttcta ggctgcgcct gcagcctggc acccgcgatg 60
    gcgatgcggg aggtgacggt ggcttgctcc gagaccgccg acttgccttg cacagcgccc 120
    tgggacccgc agctctccta tgcagtgtcc tgggccaagg tctccgagag tggcactgag 180
    agtgtggagc tcccggagag caagcaaaac agctccttcg aggcccccag gagaagggcc 240
    tattccctga cgatccaaaa cactaccatc tgcagctcgg gcacctacag gtgtgccctg 300
    caggagctcg gagggcagcg caacttgagc ggcaccgtgg ttctgaaggt gacaggatgc 360
    cccaaggaag ctacagagtc aactttcagg aagtacaggg cagaagctgt gttgctcttc 420
    tctctggttg ttttctacct gacactcatc attttcacct gcaaatttgc acgactacaa 480
    agcattttcc cagatatttc taaacctggt acggaacaag cttttcttcc agtcacctcc 540
    ccaagcaaac atttggggcc agtgaccctt cctaagacag aaacggtaag agtaggatct 600
    ccactggttt ttacaaagcc aagggcacat cagatcagtg tgcctgaatg ccacccggac 660
    aagagaagaa tgagctccat cctcagatgg caacctttct ttgaagtcct tcacctgaca 720
    gtgggctcca cactactccc tgacacaggg tcttga 756
    <210> SEQ ID NO 6
    <400> SEQUENCE: 6
    000
    <210> SEQ ID NO 7
    <211> LENGTH: 168
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic mutant CD83 sequence
    <400> SEQUENCE: 7
    agagtaggat ctccactggt ttttacaaag ccaagggcac atcagatcag tgtgcctgaa 60
    tgccacccgg acaagagaag aatgagctcc atcctcagat ggcaaccttt ctttgaagtc 120
    cttcacctga cagtgggctc cacactactc cctgacacag ggtcttga 168
    <210> SEQ ID NO 8
    <211> LENGTH: 55
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic mutant CD83 sequence
    <400> SEQUENCE: 8
    Arg Val Gly Ser Pro Leu Val Phe Thr Lys Pro Arg Ala His Gln Ile
    1 5 10 15
    Ser Val Pro Glu Cys His Pro Asp Lys Arg Arg Met Ser Ser Ile Leu
    20 25 30
    Arg Trp Gln Pro Phe Phe Glu Val Leu His Leu Thr Val Gly Ser Thr
    35 40 45
    Leu Leu Pro Asp Thr Gly Ser
    50 55
    <210> SEQ ID NO 9
    <211> LENGTH: 205
    <212> TYPE: PRT
    <213> ORGANISM: Homo sapiens
    <400> SEQUENCE: 9
    Met Ser Arg Gly Leu Gln Leu Leu Leu Leu Ser Cys Ala Tyr Ser Leu
    1 5 10 15
    Ala Pro Ala Thr Pro Glu Val Lys Val Ala Cys Ser Glu Asp Val Asp
    20 25 30
    Leu Pro Cys Thr Ala Pro Trp Asp Pro Gln Val Pro Tyr Thr Val Ser
    35 40 45
    Trp Val Lys Leu Leu Glu Gly Gly Glu Glu Arg Met Glu Thr Pro Gln
    50 55 60
    Glu Asp His Leu Arg Gly Gln His Tyr His Gln Lys Gly Gln Asn Gly
    65 70 75 80
    Ser Phe Asp Ala Pro Asn Glu Arg Pro Tyr Ser Leu Lys Ile Arg Asn
    85 90 95
    Thr Thr Ser Cys Asn Ser Gly Thr Tyr Arg Cys Thr Leu Gln Asp Pro
    100 105 110
    Asp Gly Gln Arg Asn Leu Ser Gly Lys Val Ile Leu Arg Val Thr Gly
    115 120 125
    Cys Pro Ala Gln Arg Lys Glu Glu Thr Phe Lys Lys Tyr Arg Ala Glu
    130 135 140
    Ile Val Leu Leu Leu Ala Leu Val Ile Phe Tyr Leu Thr Leu Ile Ile
    145 150 155 160
    Phe Thr Cys Lys Phe Ala Arg Leu Gln Ser Ile Phe Pro Asp Phe Ser
    165 170 175
    Lys Ala Gly Met Glu Arg Ala Phe Leu Pro Val Thr Ser Pro Asn Lys
    180 185 190
    His Leu Gly Leu Val Thr Pro His Lys Thr Glu Leu Val
    195 200 205
    <210> SEQ ID NO 10
    <211> LENGTH: 2574
    <212> TYPE: DNA
    <213> ORGANISM: Homo sapiens
    <400> SEQUENCE: 10
    cctggcgcag ccgcagcagc gacgcgagcg aactcggccg ggcccgggcg cgcgggggcg 60
    ggacgcgcac gcggcgaggg cggcgggtga gccgggggcg gggacggggg cgggacgggg 120
    gcgaaggggg cggggacggg ggcgcccgcc ggcctaacgg gattaggagg gcgcgccacc 180
    cgcttccgct gcccgccggg gaatcccccg ggtggcgccc agggaagttc ccgaacgggc 240
    gggcataaaa gggcagccgc gccggcgccc cacagctctg cagctcgtgg cagcggcgca 300
    gcgctccagc catgtcgcgc ggcctccagc ttctgctcct gagctgcgcc tacagcctgg 360
    ctcccgcgac gccggaggtg aaggtggctt gctccgaaga tgtggacttg ccctgcaccg 420
    ccccctggga tccgcaggtt ccctacacgg tctcctgggt caagttattg gagggtggtg 480
    aagagaggat ggagacaccc caggaagacc acctcagggg acagcactat catcagaagg 540
    ggcaaaatgg ttctttcgac gcccccaatg aaaggcccta ttccctgaag atccgaaaca 600
    ctaccagctg caactcgggg acatacaggt gcactctgca ggacccggat gggcagagaa 660
    acctaagtgg caaggtgatc ttgagagtga caggatgccc tgcacagcgt aaagaagaga 720
    cttttaagaa atacagagcg gagattgtcc tgctgctggc tctggttatt ttctacttaa 780
    cactcatcat tttcacttgt aagtttgcac ggctacagag tatcttccca gatttttcta 840
    aagctggcat ggaacgagct tttctcccag ttacctcccc aaataagcat ttagggctag 900
    tgactcctca caagacagaa ctggtatgag caggatttct gcaggttctt cttcctgaag 960
    ctgaggctca ggggtgtgcc tgtctgttac actggaggag agaagaatga gcctacgctg 1020
    aagatggcat cctgtgaagt ccttcacctc actgaaaaca tctggaaggg gatcccaccc 1080
    cattttctgt gggcaggcct cgaaaaccat cacatgacca catagcatga ggccactgct 1140
    gcttctccat ggccaccttt tcagcgatgt atgcagctat ctggtcaacc tcctggacat 1200
    tttttcagtc atataaaagc tatggtgaga tgcagctgga aaagggtctt gggaaatatg 1260
    aatgccccca gctggcccgt gacagactcc tgaggacagc tgtcctcttc tgcatcttgg 1320
    ggacatctct ttgaattttc tgtgttttgc tgtaccagcc cagatgtttt acgtctggga 1380
    gaaattgaca gatcaagctg tgagacagtg ggaaatattt agcaaataat ttcctggtgt 1440
    gaaggtcctg ctattactaa ggagtaatct gtgtacaaag aaataacaag tcgatgaact 1500
    attccccagc agggtctttt catctgggaa agacatccat aaagaagcaa taaagaagag 1560
    tgccacattt atttttatat ctatatgtac ttgtcaaaga aggtttgtgt ttttctgctt 1620
    ttgaaatctg tatctgtagt gagatagcat tgtgaactga caggcagcct ggacatagag 1680
    agggagaaga agtcagagag ggtgacaaga tagagagcta tttaatggcc ggctggaaat 1740
    gctgggctga cggtgcagtc tgggtgctcg cccacttgtc ccactatctg ggtgcatgat 1800
    cttgagcaag ttccttctgg tgtctgcttt ctccattgta aaccacaagg ctgttgcatg 1860
    ggctaatgaa gatcatatac gtgaaaatta tttgaaaaca tataaagcac tatacagatt 1920
    cgaaactcca ttgagtcatt atccttgcta tgatgatggt gttttgggga tgagagggtg 1980
    ctatccattt ctcatgtttt ccattgtttg aaacaaagaa ggttaccaag aagcctttcc 2040
    tgtagccttc tgtaggaatt cttttgggga agtgaggaag ccaggtccac ggtctgttct 2100
    tgaagcagta gcctaacaca ctccaagata tggacacacg ggagccgctg gcagaaggga 2160
    cttcacgaag tgttgcatgg atgttttagc cattgttggc tttcccttat caaacttggg 2220
    cccttccctt cttggtttcc aaaggcattt attgctgagt tatatgttca ctgtccccct 2280
    aatattaggg agtaaaacgg ataccaagtt gatttagtgt ttttacctct gtcttggctt 2340
    tcatgttatt aaacgtatgc atgtgaagaa gggtgttttt ctgttttata ttcaactcat 2400
    aagactttgg gataggaaaa atgagtaatg gttactaggc ttaatacctg ggtgattaca 2460
    taatctgtac aacgaacccc catgatgtaa gtttacctat gtaacaaacc tgcacttata 2520
    cccatgaact taaaatgaaa gttaaaaata aaaaacatat acaaataaaa aaaa 2574
    <210> SEQ ID NO 11
    <211> LENGTH: 239
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 20D04 light chain sequence
    <400> SEQUENCE: 11
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Asp Val Val Met Thr Gln Thr Pro Ala
    20 25 30
    Ser Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ala
    35 40 45
    Ser Glu Ser Ile Ser Asn Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly
    50 55 60
    Gln Pro Pro Lys Leu Leu Ile Tyr Arg Thr Ser Thr Leu Ala Ser Gly
    65 70 75 80
    Val Ser Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu
    85 90 95
    Thr Ile Ser Gly Val Gln Cys Asp Asp Val Ala Thr Tyr Tyr Cys Gln
    100 105 110
    Cys Thr Ser Gly Gly Lys Phe Ile Ser Asp Gly Ala Ala Phe Gly Gly
    115 120 125
    Gly Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu
    130 135 140
    Leu Phe Pro Pro Ser Ser Asp Glu Val Ala Thr Gly Thr Val Thr Ile
    145 150 155 160
    Val Cys Val Ala Asn Lys Tyr Phe Pro Asp Val Thr Val Thr Trp Glu
    165 170 175
    Val Asp Gly Thr Thr Gln Thr Thr Gly Ile Glu Asn Ser Lys Thr Pro
    180 185 190
    Gln Asn Ser Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu
    195 200 205
    Thr Ser Thr Gln Tyr Asn Ser His Lys Glu Tyr Thr Cys Lys Val Thr
    210 215 220
    Gln Gly Thr Thr Ser Val Val Gln Ser Phe Ser Arg Lys Asn Cys
    225 230 235
    <210> SEQ ID NO 12
    <211> LENGTH: 720
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 20D04 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 12
    atggacatga gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    agatgtgccg atgtcgtgat gacccagact ccagcctccg tgtctgcagc tgtgggaggc 120
    acagtcacca tcaattgcca ggccagtgaa agcattagca actacttatc ctggtatcag 180
    cagaaaccag ggcagcctcc caagctcctg atctacagga catccactct ggcatctggg 240
    gtctcatcgc ggttcaaagg cagtggatct gggacagagt acactctcac catcagcggc 300
    gtgcagtgtg acgatgttgc cacttactac tgtcaatgca cttctggtgg gaagttcatt 360
    agtgatggtg ctgctttcgg cggagggacc gaggtggtgg tcaaaggtga tccagttgca 420
    cctactgtcc tcctcttccc accatctagc gatgaggtgg caactggaac agtcaccatc 480
    gtgtgtgtgg cgaataaata ctttcccgat gtcaccgtca cctgggaggt ggatggcacc 540
    acccaaacaa ctggcatcga gaacagtaaa acaccgcaga attctgcaga ttgtacctac 600
    aacctcagca gcactctgac actgaccagc acacagtaca acagccacaa agagtacacc 660
    tgcaaggtga cccagggcac gacctcagtc gtccagagct tcagtaggaa gaactgttaa 720
    <210> SEQ ID NO 13
    <211> LENGTH: 454
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 20D04 heavy chain sequence
    <400> SEQUENCE: 13
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser
    35 40 45
    Asn Asn Ala Ile Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Ile Gly Tyr Ile Trp Ser Gly Gly Leu Thr Tyr Tyr Ala Asn Trp
    65 70 75 80
    Ala Glu Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Met Thr Ser Pro Thr Ile Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Gly Ile Asn Asn Ser Ala Leu Trp Gly Pro Gly Thr Leu Val Thr
    115 120 125
    Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu Ala Pro
    130 135 140
    Cys Cys Gly Asp Thr Pro Ser Ser Thr Val Thr Leu Gly Cys Leu Val
    145 150 155 160
    Lys Gly Tyr Leu Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Thr
    165 170 175
    Leu Thr Asn Gly Val Arg Thr Phe Pro Ser Val Arg Gln Ser Ser Gly
    180 185 190
    Leu Tyr Ser Leu Ser Ser Val Val Ser Val Thr Ser Ser Ser Gln Pro
    195 200 205
    Val Thr Cys Asn Val Ala His Pro Ala Thr Asn Thr Lys Val Asp Lys
    210 215 220
    Thr Val Ala Pro Ser Thr Cys Ser Lys Pro Thr Cys Pro Pro Pro Glu
    225 230 235 240
    Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
    245 250 255
    Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
    260 265 270
    Val Ser Gln Asp Asp Pro Glu Val Gln Phe Thr Trp Tyr Ile Asn Asn
    275 280 285
    Glu Gln Val Arg Thr Ala Arg Pro Pro Leu Arg Glu Gln Gln Phe Asn
    290 295 300
    Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile Ala His Gln Asp Trp
    305 310 315 320
    Leu Arg Gly Lys Glu Phe Lys Cys Lys Val His Asn Lys Ala Leu Pro
    325 330 335
    Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Arg Gly Gln Pro Leu Glu
    340 345 350
    Pro Lys Val Tyr Thr Met Gly Pro Pro Arg Glu Glu Leu Ser Ser Arg
    355 360 365
    Ser Val Ser Leu Thr Cys Met Ile Asn Gly Phe Tyr Pro Ser Asp Ile
    370 375 380
    Ser Val Glu Trp Glu Lys Asn Gly Lys Ala Glu Asp Asn Tyr Lys Thr
    385 390 395 400
    Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Asn Lys
    405 410 415
    Leu Ser Val Pro Thr Ser Glu Trp Gln Arg Gly Asp Val Phe Thr Cys
    420 425 430
    Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Ile
    435 440 445
    Ser Arg Ser Pro Gly Lys
    450
    <210> SEQ ID NO 14
    <211> LENGTH: 1362
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 20D04 anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 14
    atggagacag gcctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acgcctggga cacccctgac actcacctgc 120
    accgtctctg gattctccct cagtaacaat gcaataaact gggtccgcca ggctccaggg 180
    aaggggctag agtggatcgg atacatttgg agtggtgggc ttacatacta cgcgaactgg 240
    gcggaaggcc gattcaccat ctccaaaacc tcgactacgg tggatctgaa gatgaccagt 300
    ccgacaatcg aggacacggc cacctatttc tgtgccagag ggattaataa ctccgctttg 360
    tggggcccag gcaccctggt caccgtctcc tcagggcaac ctaaggctcc atcagtcttc 420
    ccactggccc cctgctgcgg ggacacaccc tctagcacgg tgaccttggg ctgcctggtc 480
    aaaggctacc tcccggagcc agtgaccgtg acctggaact cgggcaccct caccaatggg 540
    gtacgcacct tcccgtccgt ccggcagtcc tcaggcctct actcgctgag cagcgtggtg 600
    agcgtgacct caagcagcca gcccgtcacc tgcaacgtgg cccacccagc caccaacacc 660
    aaagtggaca agaccgttgc gccctcgaca tgcagcaagc ccacgtgccc accccctgaa 720
    ctcctggggg gaccgtctgt cttcatcttc cccccaaaac ccaaggacac cctcatgatc 780
    tcacgcaccc ccgaggtcac atgcgtggtg gtggacgtga gccaggatga ccccgaggtg 840
    cagttcacat ggtacataaa caacgagcag gtgcgcaccg cccggccgcc gctacgggag 900
    cagcagttca acagcacgat ccgcgtggtc agcaccctcc ccatcgcgca ccaggactgg 960
    ctgaggggca aggagttcaa gtgcaaagtc cacaacaagg cactcccggc ccccatcgag 1020
    aaaaccatct ccaaagccag agggcagccc ctggagccga aggtctacac catgggccct 1080
    ccccgggagg agctgagcag caggtcggtc agcctgacct gcatgatcaa cggcttctac 1140
    ccttccgaca tctcggtgga gtgggagaag aacgggaagg cagaggacaa ctacaagacc 1200
    acgccggccg tgctggacag cgacggctcc tacttcctct acaacaagct ctcagtgccc 1260
    acgagtgagt ggcagcgggg cgacgtcttc acctgctccg tgatgcacga ggccttgcac 1320
    aaccactaca cgcagaagtc catctcccgc tctccgggta aa 1362
    <210> SEQ ID NO 15
    <211> LENGTH: 238
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 11G05 light chain sequence
    <400> SEQUENCE: 15
    Met Asp Thr Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Asp Val Val Met Thr Gln Thr Pro Ala
    20 25 30
    Ser Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser
    35 40 45
    Ser Lys Asn Val Tyr Asn Asn Asn Trp Leu Ser Trp Phe Gln Gln Lys
    50 55 60
    Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Tyr Ala Ser Thr Leu Ala
    65 70 75 80
    Ser Gly Val Pro Ser Arg Phe Arg Gly Ser Gly Ser Gly Thr Gln Phe
    85 90 95
    Thr Leu Thr Ile Ser Asp Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr
    100 105 110
    Cys Ala Gly Asp Tyr Ser Ser Ser Ser Asp Asn Gly Phe Gly Gly Gly
    115 120 125
    Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu
    130 135 140
    Phe Pro Pro Ser Ser Asp Glu Val Ala Thr Gly Thr Val Thr Ile Val
    145 150 155 160
    Cys Val Ala Asn Lys Tyr Phe Pro Asp Val Thr Val Thr Trp Glu Val
    165 170 175
    Asp Gly Thr Thr Gln Thr Thr Gly Ile Glu Asn Ser Lys Thr Pro Gln
    180 185 190
    Asn Ser Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu Thr
    195 200 205
    Ser Thr Gln Tyr Asn Ser His Lys Glu Tyr Thr Cys Lys Val Thr Gln
    210 215 220
    Gly Thr Thr Ser Val Val Gln Ser Phe Ser Arg Lys Asn Cys
    225 230 235
    <210> SEQ ID NO 16
    <211> LENGTH: 717
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 11G05 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 16
    atggacacca gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    agatgtgccg acgtcgtgat gacccagact ccagcctccg tgtctgcagc tgtgggaggc 120
    acagtcacca tcaattgcca gtccagtaag aatgtttata ataacaactg gttatcctgg 180
    tttcagcaga aaccagggca gcctcccaag ctcctgatct attatgcatc cactctggca 240
    tctggggtcc catcgcggtt cagaggcagt ggatctggga cacagttcac tctcaccatt 300
    agcgacgtgc agtgtgacga tgctgccact tactactgtg caggcgatta tagtagtagt 360
    agtgataatg gtttcggcgg agggaccgag gtggtggtca aaggtgatcc agttgcacct 420
    actgtcctcc tcttcccacc atctagcgat gaggtggcaa ctggaacagt caccatcgtg 480
    tgtgtggcga ataaatactt tcccgatgtc accgtcacct gggaggtgga tggcaccacc 540
    caaacaactg gcatcgagaa cagtaaaaca ccgcagaatt ctgcagattg tacctacaac 600
    ctcagcagca ctctgacact gaccagcaca cagtacaaca gccacaaaga gtacacctgc 660
    aaggtgaccc agggcacgac ctcagtcgtc cagagcttca gtaggaagaa ctgttaa 717
    <210> SEQ ID NO 17
    <211> LENGTH: 452
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 11G05 heavy chain sequence
    <400> SEQUENCE: 17
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Thr Ile Ser
    35 40 45
    Asp Tyr Asp Leu Ser Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Lys
    50 55 60
    Tyr Ile Gly Phe Ile Ala Ile Asp Gly Asn Pro Tyr Tyr Ala Thr Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Ile Thr Ala Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Gly Ala Gly Asp Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser
    115 120 125
    Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu Ala Pro Cys Cys
    130 135 140
    Gly Asp Thr Pro Ser Ser Thr Val Thr Leu Gly Cys Leu Val Lys Gly
    145 150 155 160
    Tyr Leu Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Thr Leu Thr
    165 170 175
    Asn Gly Val Arg Thr Phe Pro Ser Val Arg Gln Ser Ser Gly Leu Tyr
    180 185 190
    Ser Leu Ser Ser Val Val Ser Val Thr Ser Ser Ser Gln Pro Val Thr
    195 200 205
    Cys Asn Val Ala His Pro Ala Thr Asn Thr Lys Val Asp Lys Thr Val
    210 215 220
    Ala Pro Ser Thr Cys Ser Lys Pro Thr Cys Pro Pro Pro Glu Leu Leu
    225 230 235 240
    Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Thr Leu
    245 250 255
    Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
    260 265 270
    Gln Asp Asp Pro Glu Val Gln Phe Thr Trp Tyr Ile Asn Asn Glu Gln
    275 280 285
    Val Arg Thr Ala Arg Pro Pro Leu Arg Glu Gln Gln Phe Asn Ser Thr
    290 295 300
    Ile Arg Val Val Ser Thr Leu Pro Ile Ala His Gln Asp Trp Leu Arg
    305 310 315 320
    Gly Lys Glu Phe Lys Cys Lys Val His Asn Lys Ala Leu Pro Ala Pro
    325 330 335
    Ile Glu Lys Thr Ile Ser Lys Ala Arg Gly Gln Pro Leu Glu Pro Lys
    340 345 350
    Val Tyr Thr Met Gly Pro Pro Arg Glu Glu Leu Ser Ser Arg Ser Val
    355 360 365
    Ser Leu Thr Cys Met Ile Asn Gly Phe Tyr Pro Ser Asp Ile Ser Val
    370 375 380
    Glu Trp Glu Lys Asn Gly Lys Ala Glu Asp Asn Tyr Lys Thr Thr Pro
    385 390 395 400
    Ala Val Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Asn Lys Leu Ser
    405 410 415
    Val Pro Thr Ser Glu Trp Gln Arg Gly Asp Val Phe Thr Cys Ser Val
    420 425 430
    Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Ile Ser Arg
    435 440 445
    Ser Pro Gly Lys
    450
    <210> SEQ ID NO 18
    <211> LENGTH: 1356
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 11G05 anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 18
    atggagacag gcctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acgcctggga cacccctgac actcacctgc 120
    acagtctctg gattcaccat cagtgactac gacttgagct gggtccgcca ggctccaggg 180
    gaggggctga aatacatcgg attcattgct attgatggta acccatacta cgcgacctgg 240
    gcaaaaggcc gattcaccat ctccaaaacc tcgaccacgg tggatctgaa aatcaccgct 300
    ccgacaaccg aagacacggc cacgtatttc tgtgccagag gggcagggga cctctggggc 360
    ccagggaccc tcgtcaccgt ctcttcaggg caacctaagg ctccatcagt cttcccactg 420
    gccccctgct gcggggacac accctctagc acggtgacct tgggctgcct ggtcaaaggc 480
    tacctcccgg agccagtgac cgtgacctgg aactcgggca ccctcaccaa tggggtacgc 540
    accttcccgt ccgtccggca gtcctcaggc ctctactcgc tgagcagcgt ggtgagcgtg 600
    acctcaagca gccagcccgt cacctgcaac gtggcccacc cagccaccaa caccaaagtg 660
    gacaagaccg ttgcgccctc gacatgcagc aagcccacgt gcccaccccc tgaactcctg 720
    gggggaccgt ctgtcttcat cttcccccca aaacccaagg acaccctcat gatctcacgc 780
    acccccgagg tcacatgcgt ggtggtggac gtgagccagg atgaccccga ggtgcagttc 840
    acatggtaca taaacaacga gcaggtgcgc accgcccggc cgccgctacg ggagcagcag 900
    ttcaacagca cgatccgcgt ggtcagcacc ctccccatcg cgcaccagga ctggctgagg 960
    ggcaaggagt tcaagtgcaa agtccacaac aaggcactcc cggcccccat cgagaaaacc 1020
    atctccaaag ccagagggca gcccctggag ccgaaggtct acaccatggg ccctccccgg 1080
    gaggagctga gcagcaggtc ggtcagcctg acctgcatga tcaacggctt ctacccttcc 1140
    gacatctcgg tggagtggga gaagaacggg aaggcagagg acaactacaa gaccacgccg 1200
    gccgtgctgg acagcgacgg ctcctacttc ctctacaaca agctctcagt gcccacgagt 1260
    gagtggcagc ggggcgacgt cttcacctgc tccgtgatgc acgaggcctt gcacaaccac 1320
    tacacgcaga agtccatctc ccgctctccg ggtaaa 1356
    <210> SEQ ID NO 19
    <211> LENGTH: 238
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <221> NAME/KEY: SITE
    <222> LOCATION: (1)...(238)
    <223> OTHER INFORMATION: Xaa = any amino acid
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 14C12 light chain sequence
    <400> SEQUENCE: 19
    Met Asp Xaa Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Leu Val Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser
    35 40 45
    Gln Ser Val Tyr Asp Asn Asp Glu Leu Ser Trp Tyr Gln Gln Lys Pro
    50 55 60
    Gly Gln Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser Lys Leu Ala Ser
    65 70 75 80
    Gly Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Ala
    85 90 95
    Leu Thr Ile Ser Gly Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys
    100 105 110
    Gln Ala Thr His Tyr Ser Ser Asp Trp Tyr Leu Thr Phe Gly Gly Gly
    115 120 125
    Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu
    130 135 140
    Phe Pro Pro Ser Ser Asp Glu Val Ala Thr Gly Thr Val Thr Ile Val
    145 150 155 160
    Cys Val Ala Asn Lys Tyr Phe Pro Asp Val Thr Val Thr Trp Glu Val
    165 170 175
    Asp Gly Thr Thr Gln Thr Thr Gly Ile Glu Asn Ser Lys Thr Pro Gln
    180 185 190
    Asn Ser Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu Thr
    195 200 205
    Ser Thr Gln Tyr Asn Ser His Lys Glu Tyr Thr Cys Lys Val Thr Gln
    210 215 220
    Gly Thr Thr Ser Val Val Gln Ser Phe Ser Arg Lys Asn Cys
    225 230 235
    <210> SEQ ID NO 20
    <211> LENGTH: 717
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 14C12 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 20
    atggacatra gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    agatgtgccc ttgtgatgac ccagactcca gcctccgtgt ctgcagctgt gggaggcaca 120
    gtcaccatca attgccagtc cagtcagagt gtttatgata acgacgaatt atcctggtat 180
    cagcagaaac cagggcagcc tcccaagctc ctgatctatc tggcatccaa gttggcatct 240
    ggggtcccat cccgattcaa aggcagtgga tctgggacac agttcgctct caccatcagc 300
    ggcgtgcagt gtgacgatgc tgccacttac tactgtcaag ccactcatta tagtagtgat 360
    tggtatctta ctttcggcgg agggaccgag gtggtggtca aaggtgatcc agttgcacct 420
    actgtcctcc tcttcccacc atctagcgat gaggtggcaa ctggaacagt caccatcgtg 480
    tgtgtggcga ataaatactt tcccgatgtc accgtcacct gggaggtgga tggcaccacc 540
    caaacaactg gcatcgagaa cagtaaaaca ccgcagaatt ctgcagattg tacctacaac 600
    ctcagcagca ctctgacact gaccagcaca cagtacaaca gccacaaaga gtacacctgc 660
    aaggtgaccc agggcacgac ctcagtcgtc cagagcttca gtaggaagaa ctgttaa 717
    <210> SEQ ID NO 21
    <211> LENGTH: 454
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 14C12 heavy chain sequence
    <400> SEQUENCE: 21
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val His Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Arg Ser
    35 40 45
    Ser Tyr Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Val Gly Val Ile Ser Thr Ala Tyr Asn Ser His Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Met Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Gly Gly Ser Trp Leu Asp Leu Trp Gly Gln Gly Thr Leu Val Thr
    115 120 125
    Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu Ala Pro
    130 135 140
    Cys Cys Gly Asp Thr Pro Ser Ser Thr Val Thr Leu Gly Cys Leu Val
    145 150 155 160
    Lys Gly Tyr Leu Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly Thr
    165 170 175
    Leu Thr Asn Gly Val Arg Thr Phe Pro Ser Val Arg Gln Ser Ser Gly
    180 185 190
    Leu Tyr Ser Leu Ser Ser Val Val Ser Val Thr Ser Ser Ser Gln Pro
    195 200 205
    Val Thr Cys Asn Val Ala His Pro Ala Thr Asn Thr Lys Val Asp Lys
    210 215 220
    Thr Val Ala Pro Ser Thr Cys Ser Lys Pro Thr Cys Pro Pro Pro Glu
    225 230 235 240
    Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp
    245 250 255
    Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
    260 265 270
    Val Ser Gln Asp Asp Pro Glu Val Gln Phe Thr Trp Tyr Ile Asn Asn
    275 280 285
    Glu Gln Val Arg Thr Ala Arg Pro Pro Leu Arg Glu Gln Gln Phe Asn
    290 295 300
    Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile Ala His Gln Asp Trp
    305 310 315 320
    Leu Arg Gly Lys Glu Phe Lys Cys Lys Val His Asn Lys Ala Leu Pro
    325 330 335
    Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Arg Gly Gln Pro Leu Glu
    340 345 350
    Pro Lys Val Tyr Thr Met Gly Pro Pro Arg Glu Glu Leu Ser Ser Arg
    355 360 365
    Ser Val Ser Leu Thr Cys Met Ile Asn Gly Phe Tyr Pro Ser Asp Ile
    370 375 380
    Ser Val Glu Trp Glu Lys Asn Gly Lys Ala Glu Asp Asn Tyr Lys Thr
    385 390 395 400
    Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr Asn Lys
    405 410 415
    Leu Ser Val Pro Thr Ser Glu Trp Gln Arg Gly Asp Val Phe Thr Cys
    420 425 430
    Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Ile
    435 440 445
    Ser Arg Ser Pro Gly Lys
    450
    <210> SEQ ID NO 22
    <211> LENGTH: 1362
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 14C12 anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 22
    Ala Thr Gly Gly Ala Gly Ala Cys Ala Gly Gly Cys Cys Thr Gly Cys
    1 5 10 15
    Gly Cys Thr Gly Gly Cys Thr Thr Cys Thr Cys Cys Thr Gly Gly Thr
    20 25 30
    Cys Gly Cys Thr Gly Thr Gly Cys Thr Cys Ala Ala Ala Gly Gly Thr
    35 40 45
    Gly Thr Cys Cys Ala Cys Thr Gly Thr Cys Ala Gly Thr Cys Gly Gly
    50 55 60
    Thr Gly Gly Ala Gly Gly Ala Gly Thr Cys Cys Gly Gly Gly Gly Gly
    65 70 75 80
    Thr Cys Gly Cys Cys Thr Gly Gly Thr Cys Ala Cys Gly Cys Cys Thr
    85 90 95
    Gly Gly Gly Ala Cys Ala Cys Cys Cys Cys Thr Gly Ala Cys Ala Cys
    100 105 110
    Thr Cys Ala Cys Cys Thr Gly Cys Ala Cys Ala Gly Cys Cys Thr Cys
    115 120 125
    Thr Gly Gly Ala Thr Thr Cys Thr Cys Cys Cys Gly Cys Ala Gly Cys
    130 135 140
    Ala Gly Cys Thr Ala Cys Gly Ala Cys Ala Thr Gly Ala Gly Cys Thr
    145 150 155 160
    Gly Gly Gly Thr Cys Cys Gly Cys Cys Ala Gly Gly Cys Thr Cys Cys
    165 170 175
    Ala Gly Gly Gly Ala Ala Gly Gly Gly Gly Cys Thr Gly Gly Ala Ala
    180 185 190
    Thr Gly Gly Gly Thr Cys Gly Gly Ala Gly Thr Cys Ala Thr Thr Ala
    195 200 205
    Gly Thr Ala Cys Thr Gly Cys Thr Thr Ala Thr Ala Ala Cys Thr Cys
    210 215 220
    Ala Cys Ala Cys Thr Ala Cys Gly Cys Gly Ala Gly Cys Thr Gly Gly
    225 230 235 240
    Gly Cys Ala Ala Ala Ala Gly Gly Cys Cys Gly Ala Thr Thr Cys Ala
    245 250 255
    Cys Cys Ala Thr Cys Thr Cys Cys Ala Gly Ala Ala Cys Cys Thr Cys
    260 265 270
    Gly Ala Cys Cys Ala Cys Gly Gly Thr Gly Gly Ala Thr Cys Thr Gly
    275 280 285
    Ala Ala Ala Ala Thr Gly Ala Cys Cys Ala Gly Thr Cys Thr Gly Ala
    290 295 300
    Cys Ala Ala Cys Cys Gly Ala Ala Gly Ala Cys Ala Cys Gly Gly Cys
    305 310 315 320
    Cys Ala Cys Cys Thr Ala Thr Thr Thr Cys Thr Gly Thr Gly Cys Cys
    325 330 335
    Ala Gly Ala Gly Gly Gly Gly Gly Thr Ala Gly Thr Thr Gly Gly Thr
    340 345 350
    Thr Gly Gly Ala Thr Cys Thr Cys Thr Gly Gly Gly Gly Cys Cys Ala
    355 360 365
    Gly Gly Gly Cys Ala Cys Cys Cys Thr Gly Gly Thr Cys Ala Cys Cys
    370 375 380
    Gly Thr Cys Thr Cys Cys Thr Cys Ala Gly Gly Gly Cys Ala Ala Cys
    385 390 395 400
    Cys Thr Ala Ala Gly Gly Cys Thr Cys Cys Ala Thr Cys Ala Gly Thr
    405 410 415
    Cys Thr Thr Cys Cys Cys Ala Cys Thr Gly Gly Cys Cys Cys Cys Cys
    420 425 430
    Thr Gly Cys Thr Gly Cys Gly Gly Gly Gly Ala Cys Ala Cys Ala Cys
    435 440 445
    Cys Cys Thr Cys Thr Ala Gly Cys Ala Cys Gly Gly Thr Gly Ala Cys
    450 455 460
    Cys Thr Thr Gly Gly Gly Cys Thr Gly Cys Cys Thr Gly Gly Thr Cys
    465 470 475 480
    Ala Ala Ala Gly Gly Cys Thr Ala Cys Cys Thr Cys Cys Cys Gly Gly
    485 490 495
    Ala Gly Cys Cys Ala Gly Thr Gly Ala Cys Cys Gly Thr Gly Ala Cys
    500 505 510
    Cys Thr Gly Gly Ala Ala Cys Thr Cys Gly Gly Gly Cys Ala Cys Cys
    515 520 525
    Cys Thr Cys Ala Cys Cys Ala Ala Thr Gly Gly Gly Gly Thr Ala Cys
    530 535 540
    Gly Cys Ala Cys Cys Thr Thr Cys Cys Cys Gly Thr Cys Cys Gly Thr
    545 550 555 560
    Cys Cys Gly Gly Cys Ala Gly Thr Cys Cys Thr Cys Ala Gly Gly Cys
    565 570 575
    Cys Thr Cys Thr Ala Cys Thr Cys Gly Cys Thr Gly Ala Gly Cys Ala
    580 585 590
    Gly Cys Gly Thr Gly Gly Thr Gly Ala Gly Cys Gly Thr Gly Ala Cys
    595 600 605
    Cys Thr Cys Ala Ala Gly Cys Ala Gly Cys Cys Ala Gly Cys Cys Cys
    610 615 620
    Gly Thr Cys Ala Cys Cys Thr Gly Cys Ala Ala Cys Gly Thr Gly Gly
    625 630 635 640
    Cys Cys Cys Ala Cys Cys Cys Ala Gly Cys Cys Ala Cys Cys Ala Ala
    645 650 655
    Cys Ala Cys Cys Ala Ala Ala Gly Thr Gly Gly Ala Cys Ala Ala Gly
    660 665 670
    Ala Cys Cys Gly Thr Thr Gly Cys Gly Cys Cys Cys Thr Cys Gly Ala
    675 680 685
    Cys Ala Thr Gly Cys Ala Gly Cys Ala Ala Gly Cys Cys Cys Ala Cys
    690 695 700
    Gly Thr Gly Cys Cys Cys Ala Cys Cys Cys Cys Cys Thr Gly Ala Ala
    705 710 715 720
    Cys Thr Cys Cys Thr Gly Gly Gly Gly Gly Gly Ala Cys Cys Gly Thr
    725 730 735
    Cys Thr Gly Thr Cys Thr Thr Cys Ala Thr Cys Thr Thr Cys Cys Cys
    740 745 750
    Cys Cys Cys Ala Ala Ala Ala Cys Cys Cys Ala Ala Gly Gly Ala Cys
    755 760 765
    Ala Cys Cys Cys Thr Cys Ala Thr Gly Ala Thr Cys Thr Cys Ala Cys
    770 775 780
    Gly Cys Ala Cys Cys Cys Cys Cys Gly Ala Gly Gly Thr Cys Ala Cys
    785 790 795 800
    Ala Thr Gly Cys Gly Thr Gly Gly Thr Gly Gly Thr Gly Gly Ala Cys
    805 810 815
    Gly Thr Gly Ala Gly Cys Cys Ala Gly Gly Ala Thr Gly Ala Cys Cys
    820 825 830
    Cys Cys Gly Ala Gly Gly Thr Gly Cys Ala Gly Thr Thr Cys Ala Cys
    835 840 845
    Ala Thr Gly Gly Thr Ala Cys Ala Thr Ala Ala Ala Cys Ala Ala Cys
    850 855 860
    Gly Ala Gly Cys Ala Gly Gly Thr Gly Cys Gly Cys Ala Cys Cys Gly
    865 870 875 880
    Cys Cys Cys Gly Gly Cys Cys Gly Cys Cys Gly Cys Thr Ala Cys Gly
    885 890 895
    Gly Gly Ala Gly Cys Ala Gly Cys Ala Gly Thr Thr Cys Ala Ala Cys
    900 905 910
    Ala Gly Cys Ala Cys Gly Ala Thr Cys Cys Gly Cys Gly Thr Gly Gly
    915 920 925
    Thr Cys Ala Gly Cys Ala Cys Cys Cys Thr Cys Cys Cys Cys Ala Thr
    930 935 940
    Cys Gly Cys Gly Cys Ala Cys Cys Ala Gly Gly Ala Cys Thr Gly Gly
    945 950 955 960
    Cys Thr Gly Ala Gly Gly Gly Gly Cys Ala Ala Gly Gly Ala Gly Thr
    965 970 975
    Thr Cys Ala Ala Gly Thr Gly Cys Ala Ala Ala Gly Thr Cys Cys Ala
    980 985 990
    Cys Ala Ala Cys Ala Ala Gly Gly Cys Ala Cys Thr Cys Cys Cys Gly
    995 1000 1005
    Gly Cys Cys Cys Cys Cys Ala Thr Cys Gly Ala Gly Ala Ala Ala Ala
    1010 1015 1020
    Cys Cys Ala Thr Cys Thr Cys Cys Ala Ala Ala Gly Cys Cys Ala Gly
    1025 1030 1035 1040
    Ala Gly Gly Gly Cys Ala Gly Cys Cys Cys Cys Thr Gly Gly Ala Gly
    1045 1050 1055
    Cys Cys Gly Ala Ala Gly Gly Thr Cys Thr Ala Cys Ala Cys Cys Ala
    1060 1065 1070
    Thr Gly Gly Gly Cys Cys Cys Thr Cys Cys Cys Cys Gly Gly Gly Ala
    1075 1080 1085
    Gly Gly Ala Gly Cys Thr Gly Ala Gly Cys Ala Gly Cys Ala Gly Gly
    1090 1095 1100
    Thr Cys Gly Gly Thr Cys Ala Gly Cys Cys Thr Gly Ala Cys Cys Thr
    1105 1110 1115 1120
    Gly Cys Ala Thr Gly Ala Thr Cys Ala Ala Cys Gly Gly Cys Thr Thr
    1125 1130 1135
    Cys Thr Ala Cys Cys Cys Thr Thr Cys Cys Gly Ala Cys Ala Thr Cys
    1140 1145 1150
    Thr Cys Gly Gly Thr Gly Gly Ala Gly Thr Gly Gly Gly Ala Gly Ala
    1155 1160 1165
    Ala Gly Ala Ala Cys Gly Gly Gly Ala Ala Gly Gly Cys Ala Gly Ala
    1170 1175 1180
    Gly Gly Ala Cys Ala Ala Cys Thr Ala Cys Ala Ala Gly Ala Cys Cys
    1185 1190 1195 1200
    Ala Cys Gly Cys Cys Gly Gly Cys Cys Gly Thr Gly Cys Thr Gly Gly
    1205 1210 1215
    Ala Cys Ala Gly Cys Gly Ala Cys Gly Gly Cys Thr Cys Cys Thr Ala
    1220 1225 1230
    Cys Thr Thr Cys Cys Thr Cys Thr Ala Cys Ala Ala Cys Ala Ala Gly
    1235 1240 1245
    Cys Thr Cys Thr Cys Ala Gly Thr Gly Cys Cys Cys Ala Cys Gly Ala
    1250 1255 1260
    Gly Thr Gly Ala Gly Thr Gly Gly Cys Ala Gly Cys Gly Gly Gly Gly
    1265 1270 1275 1280
    Cys Gly Ala Cys Gly Thr Cys Thr Thr Cys Ala Cys Cys Thr Gly Cys
    1285 1290 1295
    Thr Cys Cys Gly Thr Gly Ala Thr Gly Cys Ala Cys Gly Ala Gly Gly
    1300 1305 1310
    Cys Cys Thr Thr Gly Cys Ala Cys Ala Ala Cys Cys Ala Cys Thr Ala
    1315 1320 1325
    Cys Ala Cys Gly Cys Ala Gly Ala Ala Gly Thr Cys Cys Ala Thr Cys
    1330 1335 1340
    Thr Cys Cys Cys Gly Cys Thr Cys Thr Cys Cys Gly Gly Gly Thr Ala
    1345 1350 1355 1360
    Ala Ala
    <210> SEQ ID NO 23
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 23
    Ser Tyr Asp Met Thr
    1 5
    <210> SEQ ID NO 24
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 24
    Ser Tyr Asp Met Ser
    1 5
    <210> SEQ ID NO 25
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 25
    Asp Tyr Asp Leu Ser
    1 5
    <210> SEQ ID NO 26
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 26
    Ser Tyr Asp Met Ser
    1 5
    <210> SEQ ID NO 27
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 27
    Tyr Ala Ser Gly Ser Thr Tyr Tyr
    1 5
    <210> SEQ ID NO 28
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 28
    Ser Ser Ser Gly Thr Thr Tyr Tyr
    1 5
    <210> SEQ ID NO 29
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 29
    Tyr Ala Ser Gly Ser Thr Tyr Tyr
    1 5
    <210> SEQ ID NO 30
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 30
    Ala Ile Asp Gly Asn Pro Tyr Tyr
    1 5
    <210> SEQ ID NO 31
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 31
    Ser Thr Ala Tyr Asn Ser His Tyr
    1 5
    <210> SEQ ID NO 32
    <211> LENGTH: 11
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 32
    Glu His Ala Gly Tyr Ser Gly Asp Thr Gly His
    1 5 10
    <210> SEQ ID NO 33
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 33
    Glu Gly Ala Gly Val Ser Met Thr
    1 5
    <210> SEQ ID NO 34
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 34
    Glu Asp Ala Gly Phe Ser Asn Ala
    1 5
    <210> SEQ ID NO 35
    <211> LENGTH: 4
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 35
    Gly Ala Gly Asp
    1
    <210> SEQ ID NO 36
    <211> LENGTH: 6
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 36
    Gly Gly Ser Trp Leu Asp
    1 5
    <210> SEQ ID NO 37
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 37
    Arg Cys Ala Tyr Asp
    1 5
    <210> SEQ ID NO 38
    <211> LENGTH: 6
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 38
    Arg Cys Ala Asp Val Val
    1 5
    <210> SEQ ID NO 39
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 39
    Arg Cys Ala Leu Val
    1 5
    <210> SEQ ID NO 40
    <211> LENGTH: 6
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 40
    Gln Ser Ile Ser Thr Tyr
    1 5
    <210> SEQ ID NO 41
    <211> LENGTH: 6
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 41
    Gln Ser Val Ser Ser Tyr
    1 5
    <210> SEQ ID NO 42
    <211> LENGTH: 6
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 42
    Glu Ser Ile Ser Asn Tyr
    1 5
    <210> SEQ ID NO 43
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 43
    Lys Asn Val Tyr Asn Asn Asn Trp
    1 5
    <210> SEQ ID NO 44
    <211> LENGTH: 12
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 44
    Gln Gln Gly Tyr Thr His Ser Asn Val Asp Asn Val
    1 5 10
    <210> SEQ ID NO 45
    <211> LENGTH: 12
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 45
    Gln Gln Gly Tyr Ser Ile Ser Asp Ile Asp Asn Ala
    1 5 10
    <210> SEQ ID NO 46
    <211> LENGTH: 14
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 46
    Gln Cys Thr Ser Gly Gly Lys Phe Ile Ser Asp Gly Ala Ala
    1 5 10
    <210> SEQ ID NO 47
    <211> LENGTH: 11
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 47
    Ala Gly Asp Tyr Ser Ser Ser Ser Asp Asn Gly
    1 5 10
    <210> SEQ ID NO 48
    <211> LENGTH: 12
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 48
    Gln Ala Thr His Tyr Ser Ser Asp Trp Leu Thr Tyr
    1 5 10
    <210> SEQ ID NO 49
    <211> LENGTH: 5
    <212> TYPE: RNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 49
    auuua 5
    <210> SEQ ID NO 50
    <211> LENGTH: 6
    <212> TYPE: RNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 50
    auuuua 6
    <210> SEQ ID NO 51
    <211> LENGTH: 7
    <212> TYPE: RNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 51
    auuuuua 7
    <210> SEQ ID NO 52
    <211> LENGTH: 157
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 heavy chain variable
    region sequence
    <400> SEQUENCE: 52
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser
    35 40 45
    Ser Tyr Asp Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Ile Gly Ile Ile Tyr Ala Ser Gly Ser Thr Tyr Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Glu Val Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ser
    100 105 110
    Arg Glu His Ala Gly Tyr Ser Gly Asp Thr Gly His Leu Trp Gly Pro
    115 120 125
    Gly Thr Leu Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val
    130 135 140
    Phe Pro Leu Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser
    145 150 155
    <210> SEQ ID NO 53
    <211> LENGTH: 154
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 heavy chain variable
    region sequence
    <400> SEQUENCE: 53
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Ser Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser
    35 40 45
    Ser Tyr Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Tyr Ile Gly Ile Ile Ser Ser Ser Gly Thr Thr Tyr Tyr Ala Asn Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Val Thr Ser Pro Thr Ile Gly Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Glu Gly Ala Gly Val Ser Met Thr Leu Trp Gly Pro Gly Thr Leu
    115 120 125
    Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu
    130 135 140
    Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser
    145 150
    <210> SEQ ID NO 54
    <211> LENGTH: 154
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 heavy chain variable
    region sequence
    <400> SEQUENCE: 54
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser
    35 40 45
    Ser Tyr Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Ile Gly Ile Ile Tyr Ala Ser Gly Ser Thr Tyr Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Val Ala Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Ile Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Glu Asp Ala Gly Phe Ser Asn Ala Leu Trp Gly Pro Gly Thr Leu
    115 120 125
    Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu
    130 135 140
    Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser
    145 150
    <210> SEQ ID NO 55
    <211> LENGTH: 147
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 light chain variable
    region sequence
    <400> SEQUENCE: 55
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Tyr Asp Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Glu Val Ala Val Gly Gly Thr Val Thr Ile Lys Cys Gln Ala Ser
    35 40 45
    Gln Ser Ile Ser Thr Tyr Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln
    50 55 60
    Pro Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asp Leu Ala Ser Gly Val
    65 70 75 80
    Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr
    85 90 95
    Ile Ser Asp Leu Glu Cys Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
    100 105 110
    Gly Tyr Thr His Ser Asn Val Asp Asn Val Phe Gly Gly Gly Thr Glu
    115 120 125
    Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu Phe Pro
    130 135 140
    Pro Ser Ser
    145
    <210> SEQ ID NO 56
    <211> LENGTH: 147
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 light chain variable
    region sequence
    <400> SEQUENCE: 56
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Tyr Asp Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Glu Val Ala Val Gly Gly Thr Val Ala Ile Lys Cys Gln Ala Ser
    35 40 45
    Gln Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
    50 55 60
    Pro Pro Lys Pro Leu Ile Tyr Glu Ala Ser Met Leu Ala Ala Gly Val
    65 70 75 80
    Ser Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
    85 90 95
    Ile Ser Asp Leu Glu Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
    100 105 110
    Gly Tyr Ser Ile Ser Asp Ile Asp Asn Ala Phe Gly Gly Gly Thr Glu
    115 120 125
    Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu Phe Pro
    130 135 140
    Pro Ser Ser
    145
    <210> SEQ ID NO 57
    <211> LENGTH: 150
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 light chain variable
    region sequence
    <400> SEQUENCE: 57
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Asp Val Val Met Thr Gln Thr Pro Ala
    20 25 30
    Ser Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ala
    35 40 45
    Ser Glu Ser Ile Ser Asn Tyr Leu Ser Trp Tyr Gln Gln Lys Pro Gly
    50 55 60
    Gln Pro Pro Lys Leu Leu Ile Tyr Arg Thr Ser Thr Leu Ala Ser Gly
    65 70 75 80
    Val Ser Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Glu Tyr Thr Leu
    85 90 95
    Thr Ile Ser Gly Val Gln Cys Asp Asp Val Ala Thr Tyr Tyr Cys Gln
    100 105 110
    Cys Thr Ser Gly Gly Lys Phe Ile Ser Asp Gly Ala Ala Phe Gly Gly
    115 120 125
    Gly Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu
    130 135 140
    Leu Phe Pro Pro Ser Ser
    145 150
    <210> SEQ ID NO 58
    <211> LENGTH: 236
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 020B08L light chain sequence
    <400> SEQUENCE: 58
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Tyr Asp Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Glu Val Ala Val Gly Gly Thr Val Thr Ile Lys Cys Gln Ala Ser
    35 40 45
    Gln Ser Ile Ser Thr Tyr Leu Asp Trp Tyr Gln Gln Lys Pro Gly Gln
    50 55 60
    Pro Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asp Leu Ala Ser Gly Val
    65 70 75 80
    Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr Leu Thr
    85 90 95
    Ile Ser Asp Leu Glu Cys Ala Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
    100 105 110
    Gly Tyr Thr His Ser Asn Val Asp Asn Val Phe Gly Gly Gly Thr Glu
    115 120 125
    Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu Phe Pro
    130 135 140
    Pro Ser Ser Asp Glu Val Ala Thr Gly Thr Val Thr Ile Val Cys Val
    145 150 155 160
    Ala Asn Lys Tyr Phe Pro Asp Val Thr Val Thr Trp Glu Val Asp Gly
    165 170 175
    Thr Thr Gln Thr Thr Gly Ile Glu Asn Ser Lys Thr Pro Gln Asn Ser
    180 185 190
    Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu Thr Ser Thr
    195 200 205
    Gln Tyr Asn Ser His Lys Glu Tyr Thr Cys Lys Val Thr Gln Gly Thr
    210 215 220
    Thr Ser Val Val Gln Ser Phe Ser Arg Lys Asn Cys
    225 230 235
    <210> SEQ ID NO 59
    <211> LENGTH: 711
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 020B08L anti-CD83 light chain
    sequence
    <400> SEQUENCE: 59
    atggacatga gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    agatgtgcct atgatatgac ccagactcca gcctctgtgg aggtagctgt gggaggcaca 120
    gtcaccatca agtgccaggc cagtcagagc attagtacct acttagactg gtatcagcag 180
    aaaccagggc agcctcccaa gctcctgatc tatgatgcat ccgatctggc atctggggtc 240
    ccatcgcggt tcaaaggcag tggatctggg acacagttca ctctcaccat cagcgacctg 300
    gagtgtgccg atgctgccac ttactactgt caacagggtt atacacatag taatgttgat 360
    aatgttttcg gcggagggac cgaggtggtg gtcaaaggtg atccagttgc acctactgtc 420
    ctcctcttcc caccatctag cgatgaggtg gcaactggaa cagtcaccat cgtgtgtgtg 480
    gcgaataaat actttcccga tgtcaccgtc acctgggagg tggatggcac cacccaaaca 540
    actggcatcg agaacagtaa aacaccgcag aattctgcag attgtaccta caacctcagc 600
    agcactctga cactgaccag cacacagtac aacagccaca aagagtacac ctgcaaggtg 660
    acccagggca cgacctcagt cgtccagagc ttcagtagga agaactgtta a 711
    <210> SEQ ID NO 60
    <211> LENGTH: 456
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 020B08H heavy chain sequence
    <400> SEQUENCE: 60
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Ser Leu Ser
    35 40 45
    Ser Tyr Asp Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Ile Gly Ile Ile Tyr Ala Ser Gly Thr Thr Tyr Tyr Ala Asn Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Val Thr Ser Pro Thr Ile Gly Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Glu Gly Ala Gly Val Ser Met Thr Leu Trp Gly Pro Gly Thr Leu
    115 120 125
    Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu
    130 135 140
    Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser Thr Val Thr Leu Gly Cys
    145 150 155 160
    Leu Val Lys Gly Tyr Leu Pro Glu Pro Val Thr Val Thr Trp Asn Ser
    165 170 175
    Gly Thr Leu Thr Asn Gly Val Arg Thr Phe Pro Ser Val Arg Gln Ser
    180 185 190
    Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Ser Val Thr Ser Ser Ser
    195 200 205
    Gln Pro Val Thr Cys Asn Val Ala His Pro Ala Thr Asn Thr Lys Val
    210 215 220
    Asp Lys Thr Val Ala Pro Ser Thr Cys Ser Lys Pro Thr Cys Pro Pro
    225 230 235 240
    Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Pro
    245 250 255
    Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
    260 265 270
    Val Asp Val Ser Gln Asp Asp Pro Glu Val Gln Phe Thr Trp Tyr Ile
    275 280 285
    Asn Asn Glu Gln Val Arg Thr Ala Arg Pro Pro Leu Arg Glu Gln Gln
    290 295 300
    Phe Asn Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile Ala His Gln
    305 310 315 320
    Asp Trp Leu Arg Gly Lys Glu Phe Lys Cys Lys Val His Asn Lys Ala
    325 330 335
    Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Arg Gly Gln Pro
    340 345 350
    Leu Glu Pro Lys Val Tyr Thr Met Gly Pro Pro Arg Glu Glu Leu Ser
    355 360 365
    Ser Arg Ser Val Ser Leu Thr Cys Met Ile Asn Gly Phe Tyr Pro Ser
    370 375 380
    Asp Ile Ser Val Glu Trp Glu Lys Asn Gly Lys Ala Glu Asp Asn Tyr
    385 390 395 400
    Lys Thr Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Tyr Phe Leu Tyr
    405 410 415
    Asn Lys Leu Ser Val Pro Thr Ser Glu Trp Gln Arg Gly Asp Val Phe
    420 425 430
    Thr Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys
    435 440 445
    Ser Ile Ser Arg Ser Pro Gly Lys
    450 455
    <210> SEQ ID NO 61
    <211> LENGTH: 1368
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 020B08H anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 61
    atggagacag gcctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acgcctggga cacccctgac actcacctgc 120
    acagtctctg gattctccct cagcagctac gacatgacct gggtccgcca ggctccaggg 180
    aaggggctgg aatggatcgg aatcatttat gctagtggta ccacatacta cgcgaactgg 240
    gcgaaaggcc gattcaccat ctccaaaacc tcgaccacgg tggatctgaa agtcaccagt 300
    ccgacaatcg gggacacggc cacctatttc tgtgccagag agggggctgg tgttagtatg 360
    accttgtggg gcccaggcac cctggtcacc gtctcctcag ggcaacctaa ggctccatca 420
    gtcttcccac tggccccctg ctgcggggac acaccctcta gcacggtgac cttgggctgc 480
    ctggtcaaag gctacctccc ggagccagtg accgtgacct ggaactcggg caccctcacc 540
    aatggggtac gcaccttccc gtccgtccgg cagtcctcag gcctctactc gctgagcagc 600
    gtggtgagcg tgacctcaag cagccagccc gtcacctgca acgtggccca cccagccacc 660
    aacaccaaag tggacaagac cgttgcgccc tcgacatgca gcaagcccac gtgcccaccc 720
    cctgaactcc tggggggacc gtctgtcttc atcttccccc caaaacccaa ggacaccctc 780
    atgatctcac gcacccccga ggtcacatgc gtggtggtgg acgtgagcca ggatgacccc 840
    gaggtgcagt tcacatggta cataaacaac gagcaggtgc gcaccgcccg gccgccgcta 900
    cgggagcagc agttcaacag cacgatccgc gtggtcagca ccctccccat cgcgcaccag 960
    gactggctga ggggcaagga gttcaagtgc aaagtccaca acaaggcact cccggccccc 1020
    atcgagaaaa ccatctccaa agccagaggg cagcccctgg agccgaaggt ctacaccatg 1080
    ggccctcccc gggaggagct gagcagcagg tcggtcagcc tgacctgcat gatcaacggc 1140
    ttctaccctt ccgacatctc ggtggagtgg gagaagaacg ggaaggcaga ggacaactac 1200
    aagaccacgc cggccgtgct ggacagcgac ggctcctact tcctctacaa caagctctca 1260
    gtgcccacga gtgagtggca gcggggcgac gtcttcacct gctccgtgat gcacgaggcc 1320
    ttgcacaacc actacacgca gaagtccatc tcccgctctc cgggtaaa 1368
    <210> SEQ ID NO 62
    <211> LENGTH: 236
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 006G05L light chain sequence
    <400> SEQUENCE: 62
    Met Asp Met Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Tyr Asp Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Glu Val Ala Val Gly Gly Thr Val Ala Ile Lys Cys Gln Ala Ser
    35 40 45
    Gln Ser Val Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
    50 55 60
    Pro Pro Lys Pro Leu Ile Tyr Glu Ala Ser Met Leu Ala Ala Gly Val
    65 70 75 80
    Ser Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
    85 90 95
    Ile Ser Asp Leu Glu Cys Asp Asp Ala Ala Thr Tyr Tyr Cys Gln Gln
    100 105 110
    Gly Tyr Ser Ile Ser Asp Ile Asp Asn Ala Phe Gly Gly Gly Thr Glu
    115 120 125
    Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu Phe Pro
    130 135 140
    Pro Ser Ser Asp Glu Val Ala Thr Gly Thr Val Thr Ile Val Cys Val
    145 150 155 160
    Ala Asn Lys Tyr Phe Pro Asp Val Thr Val Thr Trp Glu Val Asp Gly
    165 170 175
    Thr Thr Gln Thr Thr Gly Ile Glu Asn Ser Lys Thr Pro Gln Asn Ser
    180 185 190
    Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu Thr Ser Thr
    195 200 205
    Gln Tyr Asn Ser His Lys Glu Tyr Thr Cys Lys Val Thr Gln Gly Thr
    210 215 220
    Thr Ser Val Val Gln Ser Phe Ser Arg Lys Asn Cys
    225 230 235
    <210> SEQ ID NO 63
    <211> LENGTH: 711
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 006G05L anti-CD83 light chain
    sequence
    <400> SEQUENCE: 63
    atggacatga gggcccccac tcaactgctg gggctcctgc tgctctggct cccaggtgcc 60
    agatgtgcct atgatatgac ccagactcca gcctctgtgg aggtagctgt gggaggcaca 120
    gtcgccatca agtgccaggc cagtcagagc gttagtagtt acttagcctg gtatcagcag 180
    aaaccagggc agcctcccaa gcccctgatc tacgaagcat ccatgctggc ggctggggtc 240
    tcatcgcggt tcaaaggcag tggatctggg acagacttca ctctcaccat cagcgacctg 300
    gagtgtgacg atgctgccac ttactattgt caacagggtt attctatcag tgatattgat 360
    aatgctttcg gcggagggac cgaggtggtg gtcaaaggtg atccagttgc acctactgtc 420
    ctcctcttcc caccatctag cgatgaggtg gcaactggaa cagtcaccat cgtgtgtgtg 480
    gcgaataaat actttcccga tgtcaccgtc acctgggagg tggatggcac cacccaaaca 540
    actggcatcg agaacagtaa aacaccgcag aattctgcag attgtaccta caacctcagc 600
    agcactctga cactgaccag cacacagtac aacagccaca aagagtacac ctgcaaggtg 660
    acccagggca cgacctcagt cgtccagagc ttcagtagga agaactgtta a 711
    <210> SEQ ID NO 64
    <211> LENGTH: 459
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 006G05L heavy chain sequence
    <400> SEQUENCE: 64
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Ser Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Leu Ser
    35 40 45
    Ser Tyr Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Tyr Ile Gly Ile Ile Ser Ser Ser Gly Ser Thr Tyr Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Glu Val Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ser
    100 105 110
    Arg Glu His Ala Gly Tyr Ser Gly Asp Thr Gly His Leu Trp Gly Pro
    115 120 125
    Gly Thr Leu Val Thr Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val
    130 135 140
    Phe Pro Leu Ala Pro Cys Cys Gly Asp Thr Pro Ser Ser Thr Val Thr
    145 150 155 160
    Leu Gly Cys Leu Val Lys Gly Tyr Leu Pro Glu Pro Val Thr Val Thr
    165 170 175
    Trp Asn Ser Gly Thr Leu Thr Asn Gly Val Arg Thr Phe Pro Ser Val
    180 185 190
    Arg Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Ser Val Thr
    195 200 205
    Ser Ser Ser Gln Pro Val Thr Cys Asn Val Ala His Pro Ala Thr Asn
    210 215 220
    Thr Lys Val Asp Lys Thr Val Ala Pro Ser Thr Cys Ser Lys Pro Thr
    225 230 235 240
    Cys Pro Pro Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro
    245 250 255
    Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
    260 265 270
    Cys Val Val Val Asp Val Ser Gln Asp Asp Pro Glu Val Gln Phe Thr
    275 280 285
    Trp Tyr Ile Asn Asn Glu Gln Val Arg Thr Ala Arg Pro Pro Leu Arg
    290 295 300
    Glu Gln Gln Phe Asn Ser Thr Ile Arg Val Val Ser Thr Leu Pro Ile
    305 310 315 320
    Ala His Gln Asp Trp Leu Arg Gly Lys Glu Phe Lys Cys Lys Val His
    325 330 335
    Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Arg
    340 345 350
    Gly Gln Pro Leu Glu Pro Lys Val Tyr Thr Met Gly Pro Pro Arg Glu
    355 360 365
    Glu Leu Ser Ser Arg Ser Val Ser Leu Thr Cys Met Ile Asn Gly Phe
    370 375 380
    Tyr Pro Ser Asp Ile Ser Val Glu Trp Glu Lys Asn Gly Lys Ala Glu
    385 390 395 400
    Asp Asn Tyr Lys Thr Thr Pro Ala Val Leu Asp Ser Asp Gly Ser Tyr
    405 410 415
    Phe Leu Tyr Asn Lys Leu Ser Val Pro Thr Ser Glu Trp Gln Arg Gly
    420 425 430
    Asp Val Phe Thr Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
    435 440 445
    Thr Gln Lys Ser Ile Ser Arg Ser Pro Gly Lys
    450 455
    <210> SEQ ID NO 65
    <211> LENGTH: 1377
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic M83 006G05L anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 65
    atggagacag gcctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc tcgcctggga cacccctgac actcacctgc 120
    acagcctctg gattctccct cagtagctac gacatgagct gggtccgcca ggctccaggg 180
    aaggggctgg aatacatcgg aatcattagt agtagtggta gcacatacta cgcgagctgg 240
    gcgaaaggcc gattcaccat ctccaaaacc tcgaccacgg tggatctgga agtgaccagt 300
    ctgacaaccg aggacacggc cacctatttc tgtagtagag aacatgctgg ttatagtggt 360
    gatacgggtc acttgtgggg cccaggcacc ctggtcaccg tctcctcggg gcaacctaag 420
    gctccatcag tcttcccact ggccccctgc tgcggggaca caccctctag cacggtgacc 480
    ttgggctgcc tggtcaaagg ctacctcccg gagccagtga ccgtgacctg gaactcgggc 540
    accctcacca atggggtacg caccttcccg tccgtccggc agtcctcagg cctctactcg 600
    ctgagcagcg tggtgagcgt gacctcaagc agccagcccg tcacctgcaa cgtggcccac 660
    ccagccacca acaccaaagt ggacaagacc gttgcgccct cgacatgcag caagcccacg 720
    tgcccacccc ctgaactcct ggggggaccg tctgtcttca tcttcccccc aaaacccaag 780
    gacaccctca tgatctcacg cacccccgag gtcacatgcg tggtggtgga cgtgagccag 840
    gatgaccccg aggtgcagtt cacatggtac ataaacaacg agcaggtgcg caccgcccgg 900
    ccgccgctac gggagcagca gttcaacagc acgatccgcg tggtcagcac cctccccatc 960
    gcgcaccagg actggctgag gggcaaggag ttcaagtgca aagtccacaa caaggcactc 1020
    ccggccccca tcgagaaaac catctccaaa gccagagggc agcccctgga gccgaaggtc 1080
    tacaccatgg gccctccccg ggaggagctg agcagcaggt cggtcagcct gacctgcatg 1140
    atcaacggct tctacccttc cgacatctcg gtggagtggg agaagaacgg gaaggcagag 1200
    gacaactaca agaccacgcc ggccgtgctg gacagcgacg gctcctactt cctctacaac 1260
    aagctctcag tgcccacgag tgagtggcag cggggcgacg tcttcacctg ctccgtgatg 1320
    cacgaggcct tgcacaacca ctacacgcag aagtccatct cccgctctcc gggtaaa 1377
    <210> SEQ ID NO 66
    <211> LENGTH: 150
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 heavy chain variable
    region sequence
    <400> SEQUENCE: 66
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Phe Thr Ile Ser
    35 40 45
    Asp Tyr Asp Leu Ser Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Lys
    50 55 60
    Tyr Ile Gly Phe Ile Ala Ile Asp Gly Asn Pro Tyr Tyr Ala Thr Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Lys Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Ile Thr Ala Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Gly Ala Gly Asp Leu Trp Gly Pro Gly Thr Leu Val Thr Val Ser
    115 120 125
    Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu Ala Pro Cys Cys
    130 135 140
    Gly Asp Thr Pro Ser Ser
    145 150
    <210> SEQ ID NO 67
    <211> LENGTH: 152
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 heavy chain variable
    region sequence
    <400> SEQUENCE: 67
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val His Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Ala Ser Gly Phe Ser Arg Ser
    35 40 45
    Ser Tyr Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Val Gly Val Ile Ser Thr Ala Tyr Asn Ser His Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Met Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Gly Gly Ser Trp Leu Asp Leu Trp Gly Gln Gly Thr Leu Val Thr
    115 120 125
    Val Ser Ser Gly Gln Pro Lys Ala Pro Ser Val Phe Pro Leu Ala Pro
    130 135 140
    Cys Cys Gly Asp Thr Pro Ser Ser
    145 150
    <210> SEQ ID NO 68
    <211> LENGTH: 149
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 light chain variable
    region sequence
    <400> SEQUENCE: 68
    Met Asp Thr Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Asp Val Val Met Thr Gln Thr Pro Ala
    20 25 30
    Ser Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser
    35 40 45
    Ser Lys Asn Val Tyr Asn Asn Asn Trp Leu Ser Trp Phe Gln Gln Lys
    50 55 60
    Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Tyr Ala Ser Thr Leu Ala
    65 70 75 80
    Ser Gly Val Pro Ser Arg Phe Arg Gly Ser Gly Ser Gly Thr Gln Phe
    85 90 95
    Thr Leu Thr Ile Ser Asp Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr
    100 105 110
    Cys Ala Gly Asp Tyr Ser Ser Ser Ser Asp Asn Gly Phe Gly Gly Gly
    115 120 125
    Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu
    130 135 140
    Phe Pro Pro Ser Ser
    145
    <210> SEQ ID NO 69
    <211> LENGTH: 149
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <221> NAME/KEY: SITE
    <222> LOCATION: (1)...(149)
    <223> OTHER INFORMATION: Xaa = any amino acid
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic anti-CD83 light chain variable
    region sequence
    <400> SEQUENCE: 69
    Met Asp Xaa Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Arg Cys Ala Leu Val Met Thr Gln Thr Pro Ala Ser
    20 25 30
    Val Ser Ala Ala Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser
    35 40 45
    Gln Ser Val Tyr Asp Asn Asp Glu Leu Ser Trp Tyr Gln Gln Lys Pro
    50 55 60
    Gly Gln Pro Pro Lys Leu Leu Ile Tyr Leu Ala Ser Lys Leu Ala Ser
    65 70 75 80
    Gly Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Ala
    85 90 95
    Leu Thr Ile Ser Gly Val Gln Cys Asp Asp Ala Ala Thr Tyr Tyr Cys
    100 105 110
    Gln Ala Thr His Tyr Ser Ser Asp Trp Tyr Leu Thr Phe Gly Gly Gly
    115 120 125
    Thr Glu Val Val Val Lys Gly Asp Pro Val Ala Pro Thr Val Leu Leu
    130 135 140
    Phe Pro Pro Ser Ser
    145
    <210> SEQ ID NO 70
    <211> LENGTH: 240
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 96G08 light chain sequence
    <400> SEQUENCE: 70
    Met Asp Thr Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Thr Phe Ala Gln Val Leu Thr Gln Thr Ala Ser Pro
    20 25 30
    Val Ser Ala Pro Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser Ser
    35 40 45
    Gln Ser Val Tyr Asn Asn Asp Phe Leu Ser Trp Tyr Gln Gln Lys Pro
    50 55 60
    Gly Gln Pro Pro Lys Leu Leu Ile Tyr Tyr Ala Ser Thr Leu Ala Ser
    65 70 75 80
    Gly Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe Thr
    85 90 95
    Leu Thr Ile Ser Asp Leu Glu Cys Asp Asp Ala Ala Thr Tyr Tyr Cys
    100 105 110
    Thr Gly Thr Tyr Gly Asn Ser Ala Trp Tyr Glu Asp Ala Phe Gly Gly
    115 120 125
    Gly Thr Glu Val Val Val Lys Arg Thr Pro Val Ala Pro Thr Val Leu
    130 135 140
    Leu Phe Pro Pro Ser Ser Ala Glu Leu Ala Thr Gly Thr Ala Thr Ile
    145 150 155 160
    Val Cys Val Ala Asn Lys Tyr Phe Pro Asp Gly Thr Val Thr Trp Lys
    165 170 175
    Val Asp Gly Ile Thr Gln Ser Ser Gly Ile Asn Asn Ser Arg Thr Pro
    180 185 190
    Gln Asn Ser Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu
    195 200 205
    Ser Ser Asp Glu Tyr Asn Ser His Asp Glu Tyr Thr Cys Gln Val Ala
    210 215 220
    Gln Asp Ser Gly Ser Pro Val Val Gln Ser Phe Ser Arg Lys Ser Cys
    225 230 235 240
    <210> SEQ ID NO 71
    <211> LENGTH: 13
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 71
    Gln Ser Ser Gln Ser Val Tyr Asn Asn Asp Phe Leu Ser
    1 5 10
    <210> SEQ ID NO 72
    <211> LENGTH: 7
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 72
    Tyr Ala Ser Thr Leu Ala Ser
    1 5
    <210> SEQ ID NO 73
    <211> LENGTH: 13
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 73
    Thr Gly Thr Tyr Gly Asn Ser Ala Trp Tyr Glu Asp Ala
    1 5 10
    <210> SEQ ID NO 74
    <211> LENGTH: 723
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 96G08 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 74
    atggacacga gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    acatttgcgc aagtgctgac ccagactgca tcgcccgtgt ctgcacctgt gggaggcaca 120
    gtcaccatca attgccagtc cagtcagagt gtttataata acgacttctt atcctggtat 180
    cagcagaaac cagggcagcc tcccaaactc ctgatctatt atgcatccac tctggcatct 240
    ggggtcccat cccggttcaa aggcagtgga tctgggacac agttcactct caccatcagc 300
    gacctggagt gtgacgatgc tgccacttac tactgtacag gcacttatgg taatagtgct 360
    tggtacgagg atgctttcgg cggagggacc gaggtggtgg tcaaacgtac gccagttgca 420
    cctactgtcc tcctcttccc accatctagc gctgagctgg caactggaac agccaccatc 480
    gtgtgcgtgg cgaataaata ctttcccgat ggcaccgtca cctggaaggt ggatggcatc 540
    acccaaagca gcggcatcaa taacagtaga acaccgcaga attctgcaga ttgtacctac 600
    aacctcagca gtactctgac actgagcagc gacgagtaca acagccacga cgagtacacc 660
    tgccaggtgg cccaggactc aggctcaccg gtcgtccaga gcttcagtag gaagagctgt 720
    tag 723
    <210> SEQ ID NO 75
    <211> LENGTH: 25
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 75
    cagtccagtc agagtgttta taata 25
    <210> SEQ ID NO 76
    <211> LENGTH: 20
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 76
    atgcatccac tctggcatct 20
    <210> SEQ ID NO 77
    <211> LENGTH: 25
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 77
    acaggcactt atggtaatag tgctt 25
    <210> SEQ ID NO 78
    <211> LENGTH: 456
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 96G08 heavy chain sequence
    <400> SEQUENCE: 78
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser
    35 40 45
    Ser Asp Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
    50 55 60
    Trp Ile Gly Ile Ile Ser Ser Gly Gly Asn Thr Tyr Tyr Ala Ser Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Arg Thr Ser Thr Thr Val Asp Leu
    85 90 95
    Lys Met Thr Ser Leu Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys Ala
    100 105 110
    Arg Val Val Gly Gly Thr Tyr Ser Ile Trp Gly Gln Gly Thr Leu Val
    115 120 125
    Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Tyr Pro Leu Ala
    130 135 140
    Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr Leu Gly Cys Leu
    145 150 155 160
    Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr Trp Asn Ser Gly
    165 170 175
    Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Asp
    180 185 190
    Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser Ser Thr Trp Pro
    195 200 205
    Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala Ser Ser Thr Lys
    210 215 220
    Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys Lys Pro Cys Ile
    225 230 235 240
    Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe Pro Pro Lys Pro
    245 250 255
    Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val Thr Cys Val Val
    260 265 270
    Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe Ser Trp Phe Val
    275 280 285
    Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro Arg Glu Glu Gln
    290 295 300
    Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro Ile Met His Gln
    305 310 315 320
    Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val Asn Ser Ala Ala
    325 330 335
    Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Arg Pro
    340 345 350
    Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys Glu Gln Met Ala
    355 360 365
    Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp Phe Phe Pro Glu
    370 375 380
    Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro Ala Glu Asn Tyr
    385 390 395 400
    Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser Tyr Phe Val Tyr
    405 410 415
    Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala Gly Asn Thr Phe
    420 425 430
    Thr Cys Ser Val Leu His Glu Gly Leu His Asn His His Thr Glu Lys
    435 440 445
    Ser Leu Ser His Ser Pro Gly Lys
    450 455
    <210> SEQ ID NO 79
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 79
    Ser Asp Gly Ile Ser
    1 5
    <210> SEQ ID NO 80
    <211> LENGTH: 16
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 80
    Ile Ile Ser Ser Gly Gly Asn Thr Tyr Tyr Ala Ser Trp Ala Lys Gly
    1 5 10 15
    <210> SEQ ID NO 81
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 81
    Val Val Gly Gly Thr Tyr Ser Ile
    1 5
    <210> SEQ ID NO 82
    <211> LENGTH: 1371
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 96G08 anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 82
    atggagactg ggctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acacctggga cacccctgac actcacctgc 120
    acagtgtctg gaatcgacct cagtagcgat ggaataagct gggtccgcca ggctccaggg 180
    aaggggctgg aatggatcgg aatcattagt agtggtggta acacatacta cgcgagctgg 240
    gcaaaaggcc gattcaccat ctccagaacc tcgaccacgg tggatctgaa gatgaccagt 300
    ctgacaaccg aggacacggc cacctatttc tgtgccagag ttgttggtgg tacttatagc 360
    atctggggcc agggcaccct cgtcaccgtc tcgagcgctt ctacaaaggg cccatctgtc 420
    tatccactgg cccctggatc tgctgcccaa actaactcca tggtgaccct gggatgcctg 480
    gtcaagggct atttccctga gccagtgaca gtgacctgga actctggatc cctgtccagc 540
    ggtgtgcaca ccttcccagc tgtcctgcag tctgacctct acactctgag cagctcagtg 600
    actgtcccct ccagcacctg gcccagcgag accgtcacct gcaacgttgc ccacccggcc 660
    agcagcacca aggtggacaa gaaaattgtg cccagggatt gtggttgtaa gccttgcata 720
    tgtacagtcc cagaagtatc atctgtcttc atcttccccc caaagcccaa ggatgtgctc 780
    accattactc tgactcctaa ggtcacgtgt gttgtggtag acatcagcaa ggatgatccc 840
    gaggtccagt tcagctggtt tgtagatgat gtggaggtgc acacagctca gacgcaaccc 900
    cgggaggagc agttcaacag cactttccgc tcagtcagtg aacttcccat catgcaccag 960
    gactggctca atggcaagga gttcaaatgc agggtcaaca gtgcagcttt ccctgccccc 1020
    atcgagaaaa ccatctccaa aaccaaaggc agaccgaagg ctccacaggt gtacaccatt 1080
    ccacctccca aggagcagat ggccaaggat aaagtcagtc tgacctgcat gataacagac 1140
    ttcttccctg aagacattac tgtggagtgg cagtggaatg ggcagccagc ggagaactac 1200
    aagaacactc agcccatcat ggacacagat ggctcttact tcgtctacag caagctcaat 1260
    gtgcagaaga gcaactggga ggcaggaaat actttcacct gctctgtgtt acatgagggc 1320
    ctgcacaacc accatactga gaagagcctc tcccactctc ctggtaaatg a 1371
    <210> SEQ ID NO 83
    <211> LENGTH: 15
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 83
    agcgatggaa taagc 15
    <210> SEQ ID NO 84
    <211> LENGTH: 48
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 84
    atcattagta gtggtggtaa cacatactac gcgagctggg caaaaggc 48
    <210> SEQ ID NO 85
    <211> LENGTH: 24
    <212> TYPE: DNA
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 85
    gttgttggtg gtacttatag catc 24
    <210> SEQ ID NO 86
    <211> LENGTH: 239
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 95F04 light chain sequence
    <400> SEQUENCE: 86
    Met Asp Thr Arg Ala Pro Thr Gln Leu Leu Gly Leu Leu Leu Leu Trp
    1 5 10 15
    Leu Pro Gly Ala Thr Phe Ala Gln Ala Val Val Thr Gln Thr Thr Ser
    20 25 30
    Pro Val Ser Ala Pro Val Gly Gly Thr Val Thr Ile Asn Cys Gln Ser
    35 40 45
    Ser Gln Ser Val Tyr Gly Asn Asn Glu Leu Ser Trp Tyr Gln Gln Lys
    50 55 60
    Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Gln Ala Ser Ser Leu Ala
    65 70 75 80
    Ser Gly Val Pro Ser Arg Phe Lys Gly Ser Gly Ser Gly Thr Gln Phe
    85 90 95
    Thr Leu Thr Ile Ser Asp Leu Glu Cys Asp Asp Ala Ala Thr Tyr Tyr
    100 105 110
    Cys Leu Gly Glu Tyr Ser Ile Ser Ala Asp Asn His Phe Gly Gly Gly
    115 120 125
    Thr Glu Val Val Val Lys Arg Thr Pro Val Ala Pro Thr Val Leu Leu
    130 135 140
    Phe Pro Pro Ser Ser Ala Glu Leu Ala Thr Gly Thr Ala Thr Ile Val
    145 150 155 160
    Cys Val Ala Asn Lys Tyr Phe Pro Asp Gly Thr Val Thr Trp Lys Val
    165 170 175
    Asp Gly Ile Thr Gln Ser Ser Gly Ile Asn Asn Ser Arg Thr Pro Gln
    180 185 190
    Asn Ser Ala Asp Cys Thr Tyr Asn Leu Ser Ser Thr Leu Thr Leu Ser
    195 200 205
    Ser Asp Glu Tyr Asn Ser His Asp Glu Tyr Thr Cys Gln Val Ala Gln
    210 215 220
    Asp Ser Gly Ser Pro Val Val Gln Ser Phe Ser Arg Lys Ser Cys
    225 230 235
    <210> SEQ ID NO 87
    <211> LENGTH: 13
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 87
    Gln Ser Ser Gln Ser Val Tyr Gly Asn Asn Glu Leu Ser
    1 5 10
    <210> SEQ ID NO 88
    <211> LENGTH: 7
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 88
    Gln Ala Ser Ser Leu Ala Ser
    1 5
    <210> SEQ ID NO 89
    <211> LENGTH: 11
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 89
    Leu Gly Glu Tyr Ser Ile Ser Ala Asp Asn His
    1 5 10
    <210> SEQ ID NO 90
    <211> LENGTH: 720
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 95F04 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 90
    atggacacga gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    acatttgccc aagccgtggt gacccagact acatcgcccg tgtctgcacc tgtgggaggc 120
    acagtcacca tcaattgcca gtccagtcag agtgtttatg gtaacaacga attatcctgg 180
    tatcagcaga aaccagggca gcctcccaag ctcctgatct accaggcatc cagcctggca 240
    tctggggtcc catcgcggtt caaaggcagt ggatctggga cacagttcac tctcaccatc 300
    agcgacctgg agtgtgacga tgctgccact tactactgtc taggcgaata tagcattagt 360
    gctgataatc atttcggcgg agggaccgag gtggtggtca aacgtacgcc agttgcacct 420
    actgtcctcc tcttcccacc atctagcgct gagctggcaa ctggaacagc caccatcgtg 480
    tgcgtggcga ataaatactt tcccgatggc accgtcacct ggaaggtgga tggcatcacc 540
    caaagcagcg gcatcaataa cagtagaaca ccgcagaatt ctgcagattg tacctacaac 600
    ctcagcagta ctctgacact gagcagcgac gagtacaaca gccacgacga gtacacctgc 660
    caggtggccc aggactcagg ctcaccggtc gtccagagct tcagtaggaa gagctgttag 720
    <210> SEQ ID NO 91
    <211> LENGTH: 460
    <212> TYPE: PRT
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 95F04 heavy chain sequence
    <400> SEQUENCE: 91
    Met Glu Thr Gly Leu Arg Trp Leu Leu Leu Val Ala Val Leu Lys Gly
    1 5 10 15
    Val Gln Cys Gln Ser Val Glu Glu Ser Gly Gly Arg Leu Val Thr Pro
    20 25 30
    Gly Thr Pro Leu Thr Leu Thr Cys Thr Val Ser Gly Ile Asp Leu Ser
    35 40 45
    Ser Asn Ala Met Ile Trp Val Arg Gln Ala Pro Arg Glu Gly Leu Glu
    50 55 60
    Trp Ile Gly Ala Met Asp Ser Asn Ser Arg Thr Tyr Tyr Ala Thr Trp
    65 70 75 80
    Ala Lys Gly Arg Phe Thr Ile Ser Arg Thr Ser Ser Ile Thr Val Asp
    85 90 95
    Leu Lys Ile Thr Ser Pro Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys
    100 105 110
    Ala Arg Gly Asp Gly Gly Ser Ser Asp Tyr Thr Glu Met Trp Gly Pro
    115 120 125
    Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
    130 135 140
    Tyr Pro Leu Ala Pro Gly Ser Ala Ala Gln Thr Asn Ser Met Val Thr
    145 150 155 160
    Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Val Thr
    165 170 175
    Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala Val
    180 185 190
    Leu Gln Ser Asp Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Pro Ser
    195 200 205
    Ser Thr Trp Pro Ser Glu Thr Val Thr Cys Asn Val Ala His Pro Ala
    210 215 220
    Ser Ser Thr Lys Val Asp Lys Lys Ile Val Pro Arg Asp Cys Gly Cys
    225 230 235 240
    Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser Ser Val Phe Ile Phe
    245 250 255
    Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr Leu Thr Pro Lys Val
    260 265 270
    Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp Pro Glu Val Gln Phe
    275 280 285
    Ser Trp Phe Val Asp Asp Val Glu Val His Thr Ala Gln Thr Gln Pro
    290 295 300
    Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser Val Ser Glu Leu Pro
    305 310 315 320
    Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu Phe Lys Cys Arg Val
    325 330 335
    Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr
    340 345 350
    Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr Ile Pro Pro Pro Lys
    355 360 365
    Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr Cys Met Ile Thr Asp
    370 375 380
    Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln Trp Asn Gly Gln Pro
    385 390 395 400
    Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met Asp Thr Asp Gly Ser
    405 410 415
    Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys Ser Asn Trp Glu Ala
    420 425 430
    Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu Gly Leu His Asn His
    435 440 445
    His Thr Glu Lys Ser Leu Ser His Ser Pro Gly Lys
    450 455 460
    <210> SEQ ID NO 92
    <211> LENGTH: 5
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 92
    Ser Asn Ala Met Ile
    1 5
    <210> SEQ ID NO 93
    <211> LENGTH: 16
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 93
    Ala Met Asp Ser Asn Ser Arg Thr Tyr Tyr Ala Thr Trp Ala Lys Gly
    1 5 10 15
    <210> SEQ ID NO 94
    <211> LENGTH: 11
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 94
    Gly Asp Gly Gly Ser Ser Asp Tyr Thr Glu Met
    1 5 10
    <210> SEQ ID NO 95
    <211> LENGTH: 1383
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 95F04 anti-CD83 heavy chain
    sequence
    <400> SEQUENCE: 95
    atggagactg ggctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acgcctggga cacccctgac actcacctgc 120
    acagtctctg gaatcgacct cagtagcaat gcaatgatct gggtccgcca ggctccaagg 180
    gaggggctgg aatggatcgg agccatggat agtaatagta ggacgtacta cgcgacctgg 240
    gcgaaaggcc gattcaccat ctccagaacc tcgtcgatta cggtggatct gaaaatcacc 300
    agtccgacaa ccgaggacac ggccacctat ttctgtgcca gaggggatgg tggcagtagt 360
    gattatacag agatgtgggg cccagggacc ctcgtcaccg tctcgagcgc ttctacaaag 420
    ggcccatctg tctatccact ggcccctgga tctgctgccc aaactaactc catggtgacc 480
    ctgggatgcc tggtcaaggg ctatttccct gagccagtga cagtgacctg gaactctgga 540
    tccctgtcca gcggtgtgca caccttccca gctgtcctgc agtctgacct ctacactctg 600
    agcagctcag tgactgtccc ctccagcacc tggcccagcg agaccgtcac ctgcaacgtt 660
    gcccacccgg ccagcagcac caaggtggac aagaaaattg tgcccaggga ttgtggttgt 720
    aagccttgca tatgtacagt cccagaagta tcatctgtct tcatcttccc cccaaagccc 780
    aaggatgtgc tcaccattac tctgactcct aaggtcacgt gtgttgtggt agacatcagc 840
    aaggatgatc ccgaggtcca gttcagctgg tttgtagatg atgtggaggt gcacacagct 900
    cagacgcaac cccgggagga gcagttcaac agcactttcc gctcagtcag tgaacttccc 960
    atcatgcacc aggactggct caatggcaag gagttcaaat gcagggtcaa cagtgcagct 1020
    ttccctgccc ccatcgagaa aaccatctcc aaaaccaaag gcagaccgaa ggctccacag 1080
    gtgtacacca ttccacctcc caaggagcag atggccaagg ataaagtcag tctgacctgc 1140
    atgataacag acttcttccc tgaagacatt actgtggagt ggcagtggaa tgggcagcca 1200
    gcggagaact acaagaacac tcagcccatc atggacacag atggctctta cttcgtctac 1260
    agcaagctca atgtgcagaa gagcaactgg gaggcaggaa atactttcac ctgctctgtg 1320
    ttacatgagg gcctgcacaa ccaccatact gagaagagcc tctcccactc tcctggtaaa 1380
    tga 1383
    <210> SEQ ID NO 96
    <211> LENGTH: 1383
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 95F04 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 96
    atggagactg ggctgcgctg gcttctcctg gtcgctgtgc tcaaaggtgt ccagtgtcag 60
    tcggtggagg agtccggggg tcgcctggtc acgcctggga cacccctgac actcacctgc 120
    acagtctctg gaatcgacct cagtagcaat gcaatgatct gggtccgcca ggctccaagg 180
    gaggggctgg aatggatcgg agccatggat agtaatagta ggacgtacta cgcgacctgg 240
    gcgaaaggcc gattcaccat ctccagaacc tcgtcgatta cggtggatct gaaaatcacc 300
    agtccgacaa ccgaggacac ggccacctat ttctgtgcca gaggggatgg tggcagtagt 360
    gattatacag agatgtgggg cccagggacc ctcgtcaccg tctcgagcgc ttctacaaag 420
    ggcccatctg tctatccact ggcccctgga tctgctgccc aaactaactc catggtgacc 480
    ctgggatgcc tggtcaaggg ctatttccct gagccagtga cagtgacctg gaactctgga 540
    tccctgtcca gcggtgtgca caccttccca gctgtcctgc agtctgacct ctacactctg 600
    agcagctcag tgactgtccc ctccagcacc tggcccagcg agaccgtcac ctgcaacgtt 660
    gcccacccgg ccagcagcac caaggtggac aagaaaattg tgcccaggga ttgtggttgt 720
    aagccttgca tatgtacagt cccagaagta tcatctgtct tcatcttccc cccaaagccc 780
    aaggatgtgc tcaccattac tctgactcct aaggtcacgt gtgttgtggt agacatcagc 840
    aaggatgatc ccgaggtcca gttcagctgg tttgtagatg atgtggaggt gcacacagct 900
    cagacgcaac cccgggagga gcagttcaac agcactttcc gctcagtcag tgaacttccc 960
    atcatgcacc aggactggct caatggcaag gagttcaaat gcagggtcaa cagtgcagct 1020
    ttccctgccc ccatcgagaa aaccatctcc aaaaccaaag gcagaccgaa ggctccacag 1080
    gtgtacacca ttccacctcc caaggagcag atggccaagg ataaagtcag tctgacctgc 1140
    atgataacag acttcttccc tgaagacatt actgtggagt ggcagtggaa tgggcagcca 1200
    gcggagaact acaagaacac tcagcccatc atggacacag atggctctta cttcgtctac 1260
    agcaagctca atgtgcagaa gagcaactgg gaggcaggaa atactttcac ctgctctgtg 1320
    ttacatgagg gcctgcacaa ccaccatact gagaagagcc tctcccactc tcctggtaaa 1380
    tga 1383
    <210> SEQ ID NO 97
    <211> LENGTH: 107
    <212> TYPE: PRT
    <213> ORGANISM: Homo sapiens
    <400> SEQUENCE: 97
    Pro Glu Val Lys Val Ala Cys Ser Glu Asp Val Asp Leu Pro Cys Thr
    1 5 10 15
    Ala Pro Trp Asp Pro Gln Val Pro Tyr Thr Val Ser Trp Val Lys Leu
    20 25 30
    Leu Glu Gly Gly Glu Glu Arg Met Glu Thr Pro Gln Glu Asp His Leu
    35 40 45
    Arg Gly Gln His Tyr His Gln Lys Gly Gln Asn Gly Ser Phe Asp Ala
    50 55 60
    Pro Asn Glu Arg Pro Tyr Ser Leu Lys Ile Arg Asn Thr Thr Ser Cys
    65 70 75 80
    Asn Ser Gly Thr Tyr Arg Cys Thr Leu Gln Asp Pro Asp Gly Gln Arg
    85 90 95
    Asn Leu Ser Gly Lys Val Ile Leu Arg Val Thr
    100 105
    <210> SEQ ID NO 98
    <211> LENGTH: 8
    <212> TYPE: PRT
    <213> ORGANISM: Oryctolagus cuniculus
    <400> SEQUENCE: 98
    Gln Ser Val Tyr Asp Asn Asp Glu
    1 5
    <210> SEQ ID NO 99
    <211> LENGTH: 720
    <212> TYPE: DNA
    <213> ORGANISM: Artificial Sequence
    <220> FEATURE:
    <223> OTHER INFORMATION: A synthetic 96G08 anti-CD83 light chain
    sequence
    <400> SEQUENCE: 99
    atggacacga gggcccccac tcagctgctg gggctcctgc tgctctggct cccaggtgcc 60
    acatttgcgc aagtgctgac ccagactgca tcgcccgtgt ctgcacctgt gggaggcaca 120
    gtcaccatca attgccagtc cagtcagagt gtttataata acgacttctt atcctggtat 180
    cagcagaaac cagggcagcc tcccaaactc ctgatctatt atgcatccac tctggcatct 240
    ggggtcccat cccggttcaa aggcagtgga tctgggacac agttcactct caccatcagc 300
    gacctggagt gtgacgatgc gccacttact actgtacagg cacttatggt aatagtgctt 360
    ggtacgagga tgctttcggc ggagggaccg aggtggtggt caaacgtacg ccagttgcac 420
    ctactgtcct cctcttccca ccatctagcg ctgagctggc aactggaaca gccaccatcg 480
    tgtgcgtggc gaataaatac tttcccgatg gcaccgtcac ctggaaggtg gatggcatca 540
    cccaaagcag cggcatcaat aacagtagaa caccgcagaa ttctgcagat tgtacctaca 600
    acctcagcag tactctgaca ctgagcagcg acgagtacaa cagccacgac gagtacacct 660
    gccaggtggc ccaggactca ggctcaccgg tcgtccagag cttcagtagg aagagctgtt 720

Claims (36)

What is claimed:
1. An isolated multimerized antibody that can bind to a CD83 polypeptide comprising amino acid sequence SEQ ID NO:97.
2. The isolated antibody of claim 1, wherein proliferation of a lymphocyte is decreased when the lymphocyte is contacted with the multimerized antibody.
3. The isolated antibody claim 1, wherein the multimerized antibody comprises amino acid sequence SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99.
4. An isolated nucleic acid encoding an antibody that can be multimerized and that can bind to a CD83 polypeptide, wherein the antibody comprises any one of amino acid sequences SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99.
5. A nucleic acid encoding an anti-cd83 antibody wherein the nucleic acid comprises any one of amino acid sequences SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85 or SEQ ID NO:90.
6. A method of modulating lymphocyte proliferation in a mammal comprising administering to the mammal a multimerized antibody that is directed against an extracellular domain of CD83 polypeptide, wherein the multimerized antibody can modulate lymphocyte proliferation.
7. The method of claim 6, wherein the multimerized antibody can bind to an extracellular domain of CD83 polypeptide that comprises amino acid sequence SEQ ID NO:97.
8. The method of claims 6, wherein the multimerized antibody comprises amino acid sequence SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99.
9. The method of claim 6, wherein the multimerized antibody is non-covalently multimerized.
10. The method claim 6, wherein the multimerized antibody is covalently multimerized.
11. The method of claim 6, wherein lymphocyte proliferation is modulated at a localized site in the mammal.
12. The method of claim 11, wherein the localized site in the mammal is a joint, a site in a lung, a site in a muscle, a site in a stomach, a site in an intestine, a site in a thyroid, a site on the skin, a site in a bladder, a site in a vagina, a site in the brain, or a site in the prostate.
13. A method for decreasing proliferation of CD4+ T-cells in a mammal comprising administering to the mammal a multimerized antibody that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
14. A method of modulating cytokine production by a lymphocyte by contacting the lymphocyte with a multimerized antibody that can modulate cytokine production and wherein the multimerized antibody can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
15. A method of modulating granulocyte macrophage colony stimulating factor production in a mammal by administering to the mammal a multimerized antibody that can modulate the activity or expression of CD83 polypeptides, wherein the multimerized antibody can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
16. A method of modulating granulocyte macrophage colony stimulating factor production by a lymphocyte by contacting the lymphocyte with a multimerized antibody that can modulate the activity or expression of a CD83 polypeptide, wherein the multimerized antibody can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
17. A method of modulating tumor necrosis factor production in a mammal by administering to the mammal a multimerized antibody that can modulate the activity or expression of CD83 polypeptides, and wherein the multimerized antibody can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
18. A method of inhibiting proliferation of a human peripheral blood mononuclear cell in a mammal by administering to the mammal a multimerized antibody that can modulate the activity or expression of CD83 polypeptides, and wherein the multimerized antibody can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
19. A method for placing an immune cell into anergy, comprising contacting the immune cell that expresses CD83 gene product with a multimerized antibody that can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
20. A method for decreasing the activity of a CD83 gene product in a mammal, comprising administering to the mammal a multimerized antibody that can bind to a polypeptide that comprises amino acid sequence SEQ ID NO:97.
21. A method for modulating cytokine levels in a mammal comprising administering to the mammal a multimerized that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
22. A method for increasing interleukin-10 levels in a mammal comprising administering to the mammal a multimerized antibody that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
23. A method for increasing interleukin-4 levels in a mammal comprising administering to the mammal a multimerized antibody that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
24. A method for increasing granulocyte macrophage colony stimulating factor levels in a mammal comprising administering to the mammal a multimerized antibody that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
25. A method for treating an inappropriate immune response in a mammal comprising administering to the mammal a multimerized antibody that can bind to an extracellular domain of a CD83 gene product that comprises amino acid sequence SEQ ID NO:97.
26. The method of claim 25, wherein the inappropriate immune response is diabetes mellitus, arthritis, rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis, atopic dermatitis, eczematous dermatitis, psoriasis, Sjogren's Syndrome, keratoconjunctivitis sicca secondary to Sjogren's Syndrome, alopecia areata, allergic responses due to arthropod bite reactions, Crohn's disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Crohn's disease, Graves ophthalmopathy, sarcoidosis, primary biliary cirrhosis, uveitis posterior, or interstitial lung fibrosis.
27. The method of claim 25, wherein the inappropriate immune response is tissue rejection of a transplanted tissue.
28. The method of claim 25, wherein the transplanted tissue is skin, cardiac or bone marrow.
29. The method of claim 13, wherein the multimerized antibody comprises amino acid sequence SEQ ID NO:1, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:67, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71 SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90; SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:98 or SEQ ID NO:99.
30. The method of claim 13, wherein the multimerized antibody is non-covalently multimerized.
31. The method of claim 13, wherein the multimerized antibody is covalently multimerized.
32. The method of claim 13, wherein lymphocyte proliferation is modulated at a localized site in the mammal.
33. The method of claim 32, wherein the localized site in the mammal is a joint, a site in a lung, a site in a muscle, a site in a stomach, a site in an intestine, a site in a thyroid, a site on the skin, a site in a bladder, a site in a vagina, brain or prostate.
34. The method of claim 22, wherein the interleukin-10 levels are modulated to treat neoplastic disease.
35. The method of claim 22, wherein the interleukin-10 levels are modulated to treat a tumor.
36. The method of claim 13, 15, 17, 20, 21, 22, 23, 24 or 25 wherein the mammal is a human.
US10/719,642 2001-11-21 2003-11-21 Modulating immune responses Abandoned US20040185040A1 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
US10/719,642 US20040185040A1 (en) 2001-11-21 2003-11-21 Modulating immune responses
US11/805,841 US7700740B2 (en) 2001-11-21 2007-05-23 Antibodies to CD83
US12/699,636 US7872103B2 (en) 2001-11-21 2010-02-03 Modulating immune responses

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US33195801P 2001-11-21 2001-11-21
US42813002P 2002-11-21 2002-11-21
PCT/US2002/037738 WO2003045318A2 (en) 2001-11-21 2002-11-21 Manipulation of cytokine levels using cd83 gene products
US47327903P 2003-05-22 2003-05-22
US10/719,642 US20040185040A1 (en) 2001-11-21 2003-11-21 Modulating immune responses

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/037738 Continuation WO2003045318A2 (en) 2001-11-21 2002-11-21 Manipulation of cytokine levels using cd83 gene products

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US11/805,841 Continuation US7700740B2 (en) 2001-11-21 2007-05-23 Antibodies to CD83

Publications (1)

Publication Number Publication Date
US20040185040A1 true US20040185040A1 (en) 2004-09-23

Family

ID=32996330

Family Applications (3)

Application Number Title Priority Date Filing Date
US10/719,642 Abandoned US20040185040A1 (en) 2001-11-21 2003-11-21 Modulating immune responses
US11/805,841 Expired - Fee Related US7700740B2 (en) 2001-11-21 2007-05-23 Antibodies to CD83
US12/699,636 Expired - Fee Related US7872103B2 (en) 2001-11-21 2010-02-03 Modulating immune responses

Family Applications After (2)

Application Number Title Priority Date Filing Date
US11/805,841 Expired - Fee Related US7700740B2 (en) 2001-11-21 2007-05-23 Antibodies to CD83
US12/699,636 Expired - Fee Related US7872103B2 (en) 2001-11-21 2010-02-03 Modulating immune responses

Country Status (1)

Country Link
US (3) US20040185040A1 (en)

Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040110673A1 (en) * 2002-12-04 2004-06-10 Alexander Steinkasserer Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US20040116338A1 (en) * 2002-12-04 2004-06-17 Alexander Steinkasserer Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US20070167607A1 (en) * 2002-11-19 2007-07-19 Alexander Steinkasserer Use of soluble forms of cd83 and nucleic acids encoding them for the treatment or prevention of diseases
US20070269868A1 (en) * 2006-05-19 2007-11-22 Carvalho Jensen Anne E Culture method for obtaining a clonal population of antigen-specific B cells
WO2008144753A2 (en) * 2007-05-21 2008-11-27 Alder Biopharmaceuticals, Inc. Antibodies to tnf alpha and use thereof
US20090028784A1 (en) * 2007-05-21 2009-01-29 Alder Biopharmaceuticals, Inc. Antibodies to IL-6 and use thereof
US20090104187A1 (en) * 2007-05-21 2009-04-23 Alder Biopharmaceuticals, Inc. Novel Rabbit Antibody Humanization Methods and Humanized Rabbit Antibodies
WO2009092383A2 (en) * 2008-01-22 2009-07-30 Multimerics Aps Products and methods to prevent infection
WO2009120178A1 (en) * 2008-03-26 2009-10-01 Epitomics, Inc. Anti-vegf antibody
US20090291089A1 (en) * 2007-05-21 2009-11-26 Smith Jeffrey T L Antagonists of IL-6 to prevent or treat Thrombosis
US20100150829A1 (en) * 2008-11-25 2010-06-17 Leon Garcia-Martinez Antibodies to IL-6 and use thereof
US20110182903A1 (en) * 2008-05-23 2011-07-28 Stephen Brand Novel soluble cd83 polypeptides, formulations and methods of use
US20110217303A1 (en) * 2007-05-21 2011-09-08 Smith Jeffrey T L Antagonists of il-6 to prevent or treat cachexia, weakness, fatigue, and/or fever
EP2726509A1 (en) * 2011-07-01 2014-05-07 Genentech, Inc. Use of anti-cd83 agonist antibodies for treating autoimmune diseases
US8992920B2 (en) 2008-11-25 2015-03-31 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of arthritis
US8992908B2 (en) 2010-11-23 2015-03-31 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of oral mucositis
WO2015061209A1 (en) * 2013-10-21 2015-04-30 Genentech, Inc. ANTI-Ly6E ANTIBODIES AND METHODS OF USE
US9085615B2 (en) 2008-11-25 2015-07-21 Alderbio Holdings Llc Antibodies to IL-6 to inhibit or treat inflammation
US9187560B2 (en) 2008-11-25 2015-11-17 Alderbio Holdings Llc Antagonists of IL-6 to treat cachexia, weakness, fatigue, and/or fever
US9212223B2 (en) 2008-11-25 2015-12-15 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US9241990B2 (en) 2007-05-21 2016-01-26 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRIP
WO2016018562A1 (en) * 2014-07-31 2016-02-04 Salk Institute For Biological Studies Generation and use of polyclonal and monoclonal antibodies specific for 3-phosphohistidine
US9265825B2 (en) 2008-11-25 2016-02-23 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRP
WO2016090163A3 (en) * 2014-12-03 2016-08-25 Board Of Trustees Of The University Of Arkansas Anti-acetaminophen antibodies and acetaminophen protein adducts
US9468676B2 (en) 2009-11-24 2016-10-18 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
WO2016191643A3 (en) * 2015-05-28 2017-01-12 Oncomed Pharmaceuticals, Inc. Tigit-binding agents and uses thereof
US9701747B2 (en) 2007-05-21 2017-07-11 Alderbio Holdings Llc Method of improving patient survivability and quality of life by anti-IL-6 antibody administration
US9725509B2 (en) 2007-05-21 2017-08-08 Alderbio Holdings Llc Expression vectors containing isolated nucleic acids encoding anti-human IL-6 antibody
US9775921B2 (en) 2009-11-24 2017-10-03 Alderbio Holdings Llc Subcutaneously administrable composition containing anti-IL-6 antibody
US9834603B2 (en) 2007-05-21 2017-12-05 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US9840559B2 (en) 2013-02-01 2017-12-12 The Regents Of The University Of California Anti-CD83 antibodies and use thereof
US10351897B2 (en) 2014-02-14 2019-07-16 Bioventures, Llc Acetaminophen adducts and methods of use thereof
US10736976B2 (en) 2016-12-01 2020-08-11 Regeneron Pharmaceuticals, Inc. Radiolabeled anti-PD-L1 antibodies for immuno-PET imaging
US10870704B2 (en) 2014-10-23 2020-12-22 Kira Biotech Pty Limited CD83 binding proteins and uses thereof
US11136384B2 (en) 2016-11-30 2021-10-05 Mereo Biopharma 5, Inc. Methods for treatment of cancer comprising TIGIT-binding agents
EA039540B1 (en) * 2016-03-25 2022-02-08 Онкомед Фармасьютикалс, Инк. Tigit-binding agents and uses thereof

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2002950779A0 (en) * 2002-08-15 2002-09-12 The Corporation Of The Trustees Of The Order Of The Sisters Of Mercy In Queensland A method of immunomodulation
WO2008061537A2 (en) * 2006-11-23 2008-05-29 Querdenker Aps Oligonucleotides for modulating target rna activity
WO2013059885A2 (en) 2011-10-28 2013-05-02 Cephalon Australia Pty Ltd Polypeptide constructs and uses thereof
US10280215B2 (en) * 2013-01-31 2019-05-07 Seoul National University R&Db Foundation Anti-C5 antibodies and methods of treating complement-related diseases
WO2015200790A2 (en) * 2014-06-26 2015-12-30 Yale University Compositions and methods to regulate renalase in the treatment of diseases and disorders
WO2019126466A1 (en) * 2017-12-22 2019-06-27 Rootpath Genomics, Inc. Compositions and methods for barcoding
MA51917A (en) * 2018-02-23 2021-06-02 H Lee Moffitt Cancer Center & Res Institute Inc CHEMERICAL ANTIGENIC RECEPTORS BINDING TO CD83
CN114615992A (en) * 2019-08-16 2022-06-10 H.李.莫菲特癌症中心和研究所股份有限公司 anti-CD 83 chimeric antigen receptor expressing T regulatory cells

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5316920A (en) * 1992-04-17 1994-05-31 Dana-Faber Cancer Institute, Inc. Lymphocyte activation antigen HB15, a member of the immunoglobulin superfamily
US5766570A (en) * 1992-04-17 1998-06-16 Dana-Farber Cancer Institute, Inc. Lymphocyte activation antigens and antibodies thereto
US6238664B1 (en) * 1997-11-22 2001-05-29 Boehringer Mannheim Gmbh Process for stabilizing proteins
US20060083740A1 (en) * 2001-11-21 2006-04-20 Celltech R&D, Inc. Manipulation of cytokine levels using cd83 gene products

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5316320A (en) * 1992-03-18 1994-05-31 Corrosion Control Corp. Isolation gasket for critical service flow line applications
AU2273397A (en) 1996-02-15 1997-09-02 Immunex Corporation Methods and compositions for modulating an immune response
US6329798B1 (en) * 1998-12-11 2001-12-11 Honeywell International Inc Voltage controller for minimizing the utility power provided to a local load site having a local power source
US6900016B1 (en) * 2000-09-08 2005-05-31 Applera Corporation Polymorphisms in known genes associated with inflammatory autoimmune disease, methods of detection and uses thereof
AU2002257064A1 (en) 2001-03-19 2002-10-03 Cellular Genomics Inc. Methods for isolating proteins expressed by dendritic cells
WO2003038072A1 (en) 2001-10-31 2003-05-08 Universite Libre De Bruxelles Generation and use of new types of dendritic cells
AR039067A1 (en) 2001-11-09 2005-02-09 Pfizer Prod Inc ANTIBODIES FOR CD40

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5316920A (en) * 1992-04-17 1994-05-31 Dana-Faber Cancer Institute, Inc. Lymphocyte activation antigen HB15, a member of the immunoglobulin superfamily
US5766570A (en) * 1992-04-17 1998-06-16 Dana-Farber Cancer Institute, Inc. Lymphocyte activation antigens and antibodies thereto
US6068984A (en) * 1992-04-17 2000-05-30 Dana-Farber Cancer Institute, Inc. Antibodies to lymphocyte activation antigens and uses therefor
US6238664B1 (en) * 1997-11-22 2001-05-29 Boehringer Mannheim Gmbh Process for stabilizing proteins
US20060083740A1 (en) * 2001-11-21 2006-04-20 Celltech R&D, Inc. Manipulation of cytokine levels using cd83 gene products

Cited By (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7893200B2 (en) 2002-11-19 2011-02-22 Argos Therapeutics, Inc. Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US20070167607A1 (en) * 2002-11-19 2007-07-19 Alexander Steinkasserer Use of soluble forms of cd83 and nucleic acids encoding them for the treatment or prevention of diseases
US9732140B2 (en) 2002-11-19 2017-08-15 Argos Therapeutics, Inc. Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US8759505B2 (en) 2002-11-19 2014-06-24 Argos Therapeutics, Inc. Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US20040116338A1 (en) * 2002-12-04 2004-06-17 Alexander Steinkasserer Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US7169898B2 (en) 2002-12-04 2007-01-30 Alexander Steinkasserer Soluble CD83 proteins and use thereof for the treatment or prevention of a disease or medical condition caused by dysfunction or undesired function of a cellular immune response involving T cells
US20040110673A1 (en) * 2002-12-04 2004-06-10 Alexander Steinkasserer Use of soluble forms of CD83 and nucleic acids encoding them for the treatment or prevention of diseases
US9102726B2 (en) 2002-12-04 2015-08-11 Argos Therapeutics, Inc. Nucleic acid of recombination expression vector encoding soluble forms of CD83, host cells transformed/transfected therewith and pharmaceutical compositions containing same
US8124099B2 (en) 2002-12-04 2012-02-28 Argos Therapeutics, Inc. Soluble CD83 proteins and use thereof for the treatment and prevention of a disease or medical condition caused by dysfunction or undesired function of a cellular immune response involving T cells
US20070269868A1 (en) * 2006-05-19 2007-11-22 Carvalho Jensen Anne E Culture method for obtaining a clonal population of antigen-specific B cells
US10233239B2 (en) 2007-05-21 2019-03-19 Alderbio Holdings Llc Isolated host cells expressing anti-IL-6 antibodies
US10344086B2 (en) 2007-05-21 2019-07-09 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US11827700B2 (en) 2007-05-21 2023-11-28 Vitaeris Inc. Treatment or prevention of diseases and disorders associated with cells that express IL-6 with Anti-IL-6 antibodies
US9725509B2 (en) 2007-05-21 2017-08-08 Alderbio Holdings Llc Expression vectors containing isolated nucleic acids encoding anti-human IL-6 antibody
US20090291089A1 (en) * 2007-05-21 2009-11-26 Smith Jeffrey T L Antagonists of IL-6 to prevent or treat Thrombosis
US10913794B2 (en) 2007-05-21 2021-02-09 Vitaeris Inc. Antibodies to IL-6 and use thereof
US10800841B2 (en) 2007-05-21 2020-10-13 Vitaeris, Inc. Methods of treating autoimmunity using specific anti-IL-6 antibodies
US20100290993A1 (en) * 2007-05-21 2010-11-18 Leon Garcia-Martinez Antibodies to IL-6 and use thereof
US10787507B2 (en) 2007-05-21 2020-09-29 Vitaeris Inc. Antagonists of IL-6 to prevent or treat thrombosis
US10759853B2 (en) 2007-05-21 2020-09-01 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US7935340B2 (en) 2007-05-21 2011-05-03 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
WO2008144753A2 (en) * 2007-05-21 2008-11-27 Alder Biopharmaceuticals, Inc. Antibodies to tnf alpha and use thereof
US10160804B2 (en) 2007-05-21 2018-12-25 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat cachexia, weakness, fatigue, and/or fever
US20110217303A1 (en) * 2007-05-21 2011-09-08 Smith Jeffrey T L Antagonists of il-6 to prevent or treat cachexia, weakness, fatigue, and/or fever
US10040851B2 (en) 2007-05-21 2018-08-07 Alderbio Holdings Llc Antagonists to IL-6 to raise albumin and/or lower CRP
US20090104187A1 (en) * 2007-05-21 2009-04-23 Alder Biopharmaceuticals, Inc. Novel Rabbit Antibody Humanization Methods and Humanized Rabbit Antibodies
US8252286B2 (en) 2007-05-21 2012-08-28 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US9926370B2 (en) 2007-05-21 2018-03-27 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US9890214B2 (en) 2007-05-21 2018-02-13 Alderbio Holdings Llc Antibodies to TNF α and use thereof
US9884912B2 (en) 2007-05-21 2018-02-06 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US9834603B2 (en) 2007-05-21 2017-12-05 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US9771421B2 (en) 2007-05-21 2017-09-26 Alderbio Holdings Llc Treating anemia in chronic IL-6 associated diseases using anti-IL-6 antibodies
US9758579B2 (en) 2007-05-21 2017-09-12 Alder Bioholdings, Llc Nucleic acids encoding anti-IL-6 antibodies of defined epitopic specificity
US9056905B2 (en) 2007-05-21 2015-06-16 Alderbio Holdings Llc Antibodies to TNF-α and use thereof
US20090028784A1 (en) * 2007-05-21 2009-01-29 Alder Biopharmaceuticals, Inc. Antibodies to IL-6 and use thereof
US8999330B2 (en) 2007-05-21 2015-04-07 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US9701747B2 (en) 2007-05-21 2017-07-11 Alderbio Holdings Llc Method of improving patient survivability and quality of life by anti-IL-6 antibody administration
US20090022659A1 (en) * 2007-05-21 2009-01-22 Katie Olson Antibodies to TNF alpha and use thereof
US9546213B2 (en) 2007-05-21 2017-01-17 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat cachexia, weakness, fatigue, and/or fever
WO2008144753A3 (en) * 2007-05-21 2009-01-29 Alder Biopharmaceuticals Inc Antibodies to tnf alpha and use thereof
US9241990B2 (en) 2007-05-21 2016-01-26 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRIP
US9259440B2 (en) 2008-01-22 2016-02-16 Multimerics Aps Methods for passive immunization
US8475789B2 (en) 2008-01-22 2013-07-02 Multimerics Aps Products and methods to prevent infections
US20110200591A1 (en) * 2008-01-22 2011-08-18 Multimerics Aps Products and methods to prevent infections
WO2009092383A3 (en) * 2008-01-22 2009-10-01 Multimerics Aps Products and methods to prevent infection
WO2009092383A2 (en) * 2008-01-22 2009-07-30 Multimerics Aps Products and methods to prevent infection
US20110008367A1 (en) * 2008-03-26 2011-01-13 Epitomics, Inc. Anti-vegf antibody
US8088375B2 (en) 2008-03-26 2012-01-03 Epitomics, Inc. Anti-VEGF antibody
WO2009120178A1 (en) * 2008-03-26 2009-10-01 Epitomics, Inc. Anti-vegf antibody
US20090246190A1 (en) * 2008-03-26 2009-10-01 Yaohuang Ke Anti-vegf antibody
US7803371B2 (en) 2008-03-26 2010-09-28 Epitomics, Inc. Anti-VEGF antibody
CN103992405A (en) * 2008-03-26 2014-08-20 宜康公司 Anti-vegf antibody
US20110182903A1 (en) * 2008-05-23 2011-07-28 Stephen Brand Novel soluble cd83 polypeptides, formulations and methods of use
US9212223B2 (en) 2008-11-25 2015-12-15 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US10858424B2 (en) 2008-11-25 2020-12-08 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of arthritis
US10053506B2 (en) 2008-11-25 2018-08-21 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat cachexia, weakness, fatigue, and/or fever
US10787511B2 (en) 2008-11-25 2020-09-29 Vitaeris Inc. Antagonists of IL-6 to raise albumin and/or lower CRP
US10117955B2 (en) 2008-11-25 2018-11-06 Alderbio Holdings Llc Methods of aiding in the diagnosis of diseases using anti-IL-6 antibodies
US8992920B2 (en) 2008-11-25 2015-03-31 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of arthritis
US10640560B2 (en) 2008-11-25 2020-05-05 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat cachexia, weakness, fatigue, and /or fever
US9765138B2 (en) 2008-11-25 2017-09-19 Alderbio Holdings Llc Isolated anti-IL-6 antibodies
US9187560B2 (en) 2008-11-25 2015-11-17 Alderbio Holdings Llc Antagonists of IL-6 to treat cachexia, weakness, fatigue, and/or fever
US9452227B2 (en) 2008-11-25 2016-09-27 Alderbio Holdings Llc Methods of treating or diagnosing conditions associated with elevated IL-6 using anti-IL-6 antibodies or fragments
US9265825B2 (en) 2008-11-25 2016-02-23 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRP
US20100150829A1 (en) * 2008-11-25 2010-06-17 Leon Garcia-Martinez Antibodies to IL-6 and use thereof
US9085615B2 (en) 2008-11-25 2015-07-21 Alderbio Holdings Llc Antibodies to IL-6 to inhibit or treat inflammation
US9994635B2 (en) 2008-11-25 2018-06-12 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRP
US9879074B2 (en) 2008-11-25 2018-01-30 Alderbio Holdings Llc Antibodies to IL-6 and use thereof
US10391169B2 (en) 2009-07-28 2019-08-27 Alderbio Holdings Llc Method of treating allergic asthma with antibodies to IL-6
US9724410B2 (en) 2009-11-24 2017-08-08 Alderbio Holdings Llc Anti-IL-6 antibodies or fragments thereof to treat or inhibit cachexia, associated with chemotherapy toxicity
US10471143B2 (en) 2009-11-24 2019-11-12 Alderbio Holdings Llc Antagonists of IL-6 to raise albumin and/or lower CRP
US9821057B2 (en) 2009-11-24 2017-11-21 Alderbio Holdings Llc Anti-IL-6 antibody for use in the treatment of cachexia
US9775921B2 (en) 2009-11-24 2017-10-03 Alderbio Holdings Llc Subcutaneously administrable composition containing anti-IL-6 antibody
US9468676B2 (en) 2009-11-24 2016-10-18 Alderbio Holdings Llc Antagonists of IL-6 to prevent or treat thrombosis
US9717793B2 (en) 2009-11-24 2017-08-01 Alderbio Holdings Llc Method of improving patient survivability and quality of life by administering an anti-IL-6 antibody
US8992908B2 (en) 2010-11-23 2015-03-31 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of oral mucositis
US9957321B2 (en) 2010-11-23 2018-05-01 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of oral mucositis
US9304134B2 (en) 2010-11-23 2016-04-05 Alderbio Holdings Llc Anti-IL-6 antibodies for the treatment of anemia
EP2726509A4 (en) * 2011-07-01 2015-01-28 Genentech Inc Use of anti-cd83 agonist antibodies for treating autoimmune diseases
EP2726509A1 (en) * 2011-07-01 2014-05-07 Genentech, Inc. Use of anti-cd83 agonist antibodies for treating autoimmune diseases
CN103906766A (en) * 2011-07-01 2014-07-02 弗·哈夫曼-拉罗切有限公司 Use of anti-cd83 agonist antibodies for treating autoimmune diseases
US11725054B2 (en) 2013-02-01 2023-08-15 Kira Biotech Pty Limited Anti-CD83 antibodies and use thereof
US10781255B2 (en) 2013-02-01 2020-09-22 Dendrocyte Biotech Pty Ltd Anti-CD83 antibodies and use thereof
US9840559B2 (en) 2013-02-01 2017-12-12 The Regents Of The University Of California Anti-CD83 antibodies and use thereof
WO2015061209A1 (en) * 2013-10-21 2015-04-30 Genentech, Inc. ANTI-Ly6E ANTIBODIES AND METHODS OF USE
CN105658237A (en) * 2013-10-21 2016-06-08 基因泰克公司 Anti-ly6e antibodies and methods of use
US9290578B2 (en) 2013-10-21 2016-03-22 Genentech, Inc. Anti-Ly6E antibodies and methods of use
US10066022B2 (en) 2013-10-21 2018-09-04 Genentech, Inc. Anti-Ly6E antibodies and methods of use
US10889651B2 (en) 2013-10-21 2021-01-12 Genentech, Inc. Anti-Ly6E antibodies and methods of use
US10351897B2 (en) 2014-02-14 2019-07-16 Bioventures, Llc Acetaminophen adducts and methods of use thereof
US11987834B2 (en) 2014-02-14 2024-05-21 Bioventures, Llc Acetaminophen adducts and methods of use thereof
WO2016018562A1 (en) * 2014-07-31 2016-02-04 Salk Institute For Biological Studies Generation and use of polyclonal and monoclonal antibodies specific for 3-phosphohistidine
US9850319B2 (en) 2014-07-31 2017-12-26 Salk Institute For Biological Studies Generation and use of polyclonal and monoclonal antibodies specific for 3-phosphohistidine
US10870704B2 (en) 2014-10-23 2020-12-22 Kira Biotech Pty Limited CD83 binding proteins and uses thereof
US10570216B2 (en) 2014-12-03 2020-02-25 Bioventures, Llc Anti-acetaminophen antibodies and acetaminophen protein adducts
US11566081B2 (en) 2014-12-03 2023-01-31 Bioventures, Llc Anti-acetaminophen antibodies and acetaminophen protein adducts
WO2016090163A3 (en) * 2014-12-03 2016-08-25 Board Of Trustees Of The University Of Arkansas Anti-acetaminophen antibodies and acetaminophen protein adducts
US10112997B2 (en) 2015-05-28 2018-10-30 Oncomed Pharmaceuticals, Inc. Tight-binding agents and uses thereof
US10544219B2 (en) 2015-05-28 2020-01-28 Oncomed Pharmaceuticals, Inc. TIGIT-binding agents and uses thereof
WO2016191643A3 (en) * 2015-05-28 2017-01-12 Oncomed Pharmaceuticals, Inc. Tigit-binding agents and uses thereof
EA039540B1 (en) * 2016-03-25 2022-02-08 Онкомед Фармасьютикалс, Инк. Tigit-binding agents and uses thereof
US11136384B2 (en) 2016-11-30 2021-10-05 Mereo Biopharma 5, Inc. Methods for treatment of cancer comprising TIGIT-binding agents
US11230596B2 (en) 2016-11-30 2022-01-25 Mereo Biopharma 5, Inc. Methods for treatment of cancer comprising TIGIT-binding agents
US10736976B2 (en) 2016-12-01 2020-08-11 Regeneron Pharmaceuticals, Inc. Radiolabeled anti-PD-L1 antibodies for immuno-PET imaging

Also Published As

Publication number Publication date
US20100249380A1 (en) 2010-09-30
US20080299122A1 (en) 2008-12-04
US7872103B2 (en) 2011-01-18
US7700740B2 (en) 2010-04-20

Similar Documents

Publication Publication Date Title
US7700740B2 (en) Antibodies to CD83
US7850968B2 (en) Manipulation of cytokine levels using CD83 gene products
WO2004048552A2 (en) Modulating immune responses
JP7277047B2 (en) ANTI-HUMAN VISTA ANTIBODY AND USES THEREOF
KR102046666B1 (en) Anti-kir antibodies for the treatment of inflammatory disorders
AU2012296613B2 (en) Anti-B7-H4 antibodies and their uses
ES2645026T3 (en) NKG2D antibodies for use in the treatment of rheumatoid arthritis or Crohn&#39;s disease
AU2008350535B2 (en) Fc receptor-binding polypeptides with modified effector functions
KR20090114388A (en) Method of providing disease-specific binding molecules and targets
JP2009530423A (en) Methods for preventing and treating amyloidogenic diseases
CA2480404A1 (en) Fc receptor homolog, reagents, and uses thereof
KR20090094848A (en) Cd44 antibodies
EA036660B1 (en) Sp35 ANTIBODY OR ANTIGEN-BINDING FRAGMENT THEREOF AND USE THEREOF IN TREATING DISORDERS OF THE CENTRAL NERVOUS SYSTEM
KR20180087430A (en) Antibodies specifically binding to HLA-DR and uses thereof
CA2629715A1 (en) Anti-alpha2 integrin antibodies and their uses
WO2007098283A2 (en) Use of antagonists of maif receptor complex molecules and neurotrophic factors for treatment of neurological diseases and disorders
KR20090078339A (en) Prophylactic or therapeutic agent for cancer
RU2724742C2 (en) Anti-orai1 antibody
KR20210142638A (en) CD3 antigen-binding fragment and applications thereof
JP2021527395A (en) CD79-specific chimeric antigen receptor
CN114174536A (en) anti-TREM-1 antibodies and uses thereof

Legal Events

Date Code Title Description
AS Assignment

Owner name: CELLTECH R & D LIMITED, UNITED KINGDOM

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:GARCIA-MARTINEZ, LEON FERNANDO;CHEN, YUCHING;ANDREWS, DAWN;REEL/FRAME:014521/0231;SIGNING DATES FROM 20040408 TO 20040412

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION