Human polypeptides causing or leading to the killing of cells including lymphoid tumor cells Number:7,521,047 from the United States Patent and Trademark Office (PTO) owispatent The present invention relates to polypeptide compositions which bind to cell surface epitopes and, in multivalent forms, cause or lead to the killing of cells including lymphoid tumor cells, and in the case of monovalent forms, cause immunosuppression or otherwise inhibit activation of lymphocytes. The invention further relates to nucleic acids encoding the polypeptides, methods for the production of the polypeptides, methods for killing cells, methods for immunosuppressing a patient, pharmaceutical, diagnostic and multivalent compositions and kits comprising the polypeptides and uses of the polypeptides.<H polypeptides, including, Human, polypepti, 7521047.html, Patent, pto, 7521047

Title: Human polypeptides causing or leading to the killing of cells including lymphoid tumor cells

Abstract: The present invention relates to polypeptide compositions which bind to cell surface epitopes and, in multivalent forms, cause or lead to the killing of cells including lymphoid tumor cells, and in the case of monovalent forms, cause immunosuppression or otherwise inhibit activation of lymphocytes. The invention further relates to nucleic acids encoding the polypeptides, methods for the production of the polypeptides, methods for killing cells, methods for immunosuppressing a patient, pharmaceutical, diagnostic and multivalent compositions and kits comprising the polypeptides and uses of the polypeptides.

Patent Number: 7,521,047 Issued on 04/21/2009 to Nagy, et al.

Inventors: Nagy; Zoltan (Wolfratshausen, DE), Brunner; Christoph (Bad Heilbrunn, DE), Tesar; Michael (Weilheim, DE), Thomassen-Wolf; Elisabeth (Munchen, DE), Rauchenberger; Robert (Farchant, DE)
Assignee: GPC Biotech AG (Munich, DE)
Morphosys AG (Munich, DE)

Appl. No.: 10/001,934

Filed: November 15, 2001

Related U.S. Patent Documents


Application Number	Filing Date	Patent Number	Issue Date
PCT/US01/15625	May., 2001

Current U.S. Class: 424/133.1 ; 424/141.1; 424/143.1; 424/155.1; 424/156.1; 424/173.1; 424/174.1; 530/387.9; 530/388.1; 530/388.15; 530/388.7; 530/388.73; 530/388.8; 530/388.85

Current International Class: A61K 39/395 (20060101); C07K 16/00 (20060101); C07K 16/28 (20060101); C07K 16/30 (20060101); C07K 16/46 (20060101)

Field of Search: 424/130.1,133.1,135.1,141.1,142.1,152.1,153.1,143.1,178.1 530/378.1,387.3,388.1,388.15,388.22

References Cited [Referenced By]

U.S. Patent Documents


6180377	January 2001	Morgan et al.
6300064	October 2001	Knappik et al.

Foreign Patent Documents


WO-94/29451	Dec., 1994	WO
WO-96/17874	Jun., 1996	WO
WO 96/17874	Jun., 1996	WO
WO-97/08320	Mar., 1997	WO
WO 98/37200	Aug., 1998	WO
WO-99/45031	Sep., 1999	WO
WO 99/45031	Sep., 1999	WO
WO-99/53953	Oct., 1999	WO
WO-00/12560	Mar., 2000	WO

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Primary Examiner: Canella; Karen A
Attorney, Agent or Firm: Yankwich; Leon R. O'Brien; David G. Yankwich & Associates, P.C.

Parent Case Text

APPLICATION INFORMATION

This application is a continuation-in-part of International Application No. PCT/US01/15625, filed on May 14, 2001, which designated the U.S. and will be published under PCT Article 21(2) in English, and which claims priority to EP 00110065.0, filed on May 12, 2000, the entire contents of which are incorporated by reference herein.

Claims

We claim:

1. A composition including a polypeptide comprising an antibody-based antigen-binding domain of human composition with binding specificity for a HLA-DR antigen expressed on the surface of a human cell, wherein treating cells expressing said antigen with a multivalent polypeptide having two or more of said antigen-binding domains causes or leads to killing of said cells, wherein said antigen-binding domain includes a combination of a VH domain and a VL domain, wherein said combination is found in one of the clones selected from the group consisting of MS-GPC-1 (SEQ ID NOs. 37 and 38, respectively), MS-GPC-6 (SEQ ID NOs. 39 and 40, respectively), MS-GPC-8 (SEQ ID NOs. 41 and 42, respectively), MS-GPC-10 (SEQ ID NOs. 43 and 44, respectively), MS-GPC-8-1 (SEQ ID NOs. 41 and 28, respectively), MS-GPC-8-6 (SEQ ID NOs. 41 and 46, respectively), MS-GPC-8-9 (SEQ ID NOs. 41 and 31, respectively), MS- GPC-8-10 (SEQ ID NOs. 41 and 48, respectively), MS-GPC-8-17 (SEQ ID NOs. 41 and 50, respectively), MS-GPC-8-18 (SEQ ID NOs. 41 and 32, respectively), MS-GPC-8-27 (SEQ ID NOs. 41 and 52, respectively), MS-GPC-8-6-2 (SEQ ID NOs. 41 and 45, respectively), MS-GPC-8-6-19 (SEQ ID NOs. 41 and 47, respectively), MS-GPC-8-6-27 (SEQ ID NOs. 41 and 49, respectively), MS-GPC-8-6-45 (SEQ ID NOs. 41 and 51, respectively), MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-6-47 (SEQ ID NOs. 41 and 53, respectively), MS-GPC-8-10-57 (SEQ ID NOs. 41 and 56, respectively), MS-GPC-8-27-7 (SEQ ID NOs. 41 and 55, respectively), MS-GPC-8-27-10 (SEQ ID NOs. 41 and 57, respectively) and MS-GPC-8-27-41 (SEQ ID NOs. 41 and 58, respectively).

2. A composition including a polypeptide comprising an antibody-based antigen-binding domain of human composition with binding specificity for a HLA-DR antigen expressed on the surface of a human cell, wherein treating cells expressing said antigen with a multivalent polypeptide having two or more of said antigen-binding domains causes or leads to killing of said cells, wherein said antigen-binding domain includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3, VL CDR1 and VL CDR3 is found in one of the clones selected from the group consisting of MS-GPC-1 (SEQ ID NOs. 37 and 38, respectively), MS-GPC-6 (SEQ ID NOs. 39 and 40, respectively), MS-GPC-8 (SEQ ID NOs. 41 and 42, respectively), MS-GPC-10 (SEQ ID NOs. 43 and 44, respectively), MS-GPC-8-1 (SEQ ID NOs. 41 and 28, respectively), MS-GPC-8-6 (SEQ ID NOs. 41 and 46, respectively), MS-GPC-8-9 (SEQ ID NOs. 41 and 31, respectively), MS-GPC-8-10 (SEQ ID NOs. 41 and 48, respectively), MS-GPC-8-17 (SEQ ID NOs. 41 and 50, respectively), MS-GPC-8-18 (SEQ ID NOs. 41 and 32, respectively), MS-GPC-8-27 (SEQ ID NOs. 41 and 52, respectively), MS-GPC-8-6-2 (SEQ ID NOs. 41 and 45, respectively), MS-GPC-8-6-19 (SEQ ID NOs. 41 and 47, respectively), MS-GPC-8-6-27 (SEQ ID NOs. 41 and 49, respectively), MS-GPC-8-6-45 (SEQ ID NOs. 41 and 51, respectively), MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-6-47 (SEQ ID NOs. 41 and 53, respectively), MS-GPC-8-10-57 (SEQ ID NOs. 41 and 56, respectively), MS-GPC-8-27-7 (SEQ ID NOs. 41 and 55, respectively), MS-GPC-8-27-10 (SEQ ID NOs. 41 and 57, respectively) and MS-GPC-8-27-41 (SEQ ID NOs. 41 and 58, respectively).

3. A composition including a polypeptide comprising an antibody-based antigen-binding domain of human composition with binding specificity for a HLA-DR antigen expressed on the surface of a human cell, wherein treating cells expressing said antigen with a multivalent polypeptide having two or more of said antigen-binding domains causes or leads to killing of said cells, wherein said antigen-binding domain includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3 sequence is taken from the consensus CDR3 sequence XXXXRGXFDX (SEQ ID NO: 1) wherein each X independently represents any amino acid residue; and/or wherein the VL CDR3 sequence is taken from the consensus CDR3 sequence QSYDXXXX (SEQ ID NO: 2) wherein each X independently represents any amino acid residue.

4. The composition of claim 3, wherein the VH CDR3 sequence of said antigen-binding domain is SPRYRGAFDY (SEQ ID NO: 3) and/or the VL CDR3 sequence of said antigen-binding domain is QSYDLIRH (SEQ ID NO: 4) or QSYDMNVH (SEQ ID NO: 5).

5. A composition including a polypeptide comprising an antibody-based antigen-binding domain of human composition with binding specificity for a HLA-DR antigen expressed on the surface of a human cell, wherein treating cells expressing said antigen with a multivalent polypeptide having two or more of said antigen-binding domains causes or leads to killing of said cells, wherein said antigen-binding domain includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the V.lamda.1 CDR1 sequence is represented in the general formula SGSXXNIGXNYVX (SEQ ID NO: 6) wherein each X independently represents any amino acid residue.

6. The composition of claim 5, wherein the CDR1 sequence is SGSESNIGNNYVQ (SEQ ID NO: 7).

7. The composition of any one of claims 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 for killing transformed cells at least 5-fold lower than the EC.sub.50 for killing normal cells.

8. The composition of any one of claims and 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 for killing activated cells at least 5-fold lower than the EC.sub.50 for killing unactivated cells.

9. The composition of any of claims and 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 of 50 nM or less for killing transformed cells.

10. The composition of any of claims and 1-4, 5, and 6, wherein the multivalent polypeptide has an EC50 for killing lymphoid tumor cells of 10 nM or less.

11. The composition of any of claims 1-4, 5, and 6, wherein the multivalent polypeptide kills activated lymphoid cells.

12. The composition of claim 11, wherein said activated lymphoid cells are lymphoid tumor cells representing a disease selected from the group consisting of B cell non-Hodgkin lymphoma, B cell lymphoma, B cell acute lymphoid leukemia, Burkitt lymphoma, Hodgkin lymphoma, hairy cell leukemia, acute myeloid leukemia, T cell lymphoma, T cell non-Hodgkin lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, and multiple myeloma.

13. The composition of claim 11, wherein said activated lymphoid cells are from a cell line selected from the group consisting of PRIESS (ECACC Accession No: 86052111), GRANTA-519 (DSMZ Accession No: ACC 342), and KARPAS-422 (DSMZ Accession No: ACC 32) cell lines.

14. The composition of any of claims 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 of 100 nM or less for killing KARPAS-422 (DSMZ Accession No: ACC 32) cells.

15. The composition of any of claims and 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 of 50 nM or less for killing KARPAS-422 (ACC 32 from DSMZ) cells.

16. The composition of any of claims and 1-4, 5, and 6, wherein the multivalent polypeptide has an EC.sub.50 of 10 nM or less for killing PRIESS (ECACC Accession No: 86052111) cells.

17. The composition of any of claims and 1-4, 5, and 6, wherein said cells are non-lymphoid cells that express HLA-DR molecules.

18. The composition of any of claims and 1-4, 5, and 6, wherein said antigen-binding domain binds to the .beta.-chain of HLA-DR.

19. The composition of claim 18, wherein said antigen-binding domain binds to the first domain of the .beta.-chain of HLA-DR.

20. The composition of any of claims 1-4, 5, and 6, wherein said antigen-binding domain binds to one or more HLA-DR types selected from the group consisting of DR1-0101, DR2-15021, DR3-0301, DR4Dw4-0401, DR4Dw10-0402, DR4Dw14-0404, DR6-1302, DR6-1401, DR8-8031, DR9-9012, DRw53-B4*0101 and DRw52-B3*0101.

21. The composition of claim 20, wherein said antigen-binding domain binds to at least 5 different of said HLA-DR types.

22. The composition of any one of claims 1-4, 5, and 6, wherein said antibody-based antigen-binding domain is part of a multivalent polypeptide including at least a F(ab').sub.2 antibody fragment or a mini-antibody fragment.

23. The composition of any one of claims 1-4, 5, and 6, wherein said antibody-based antigen-binding domain is part of a multivalent polypeptide comprising at least two monovalent antibody fragments selected from Fv, scFv, dsFv and Fab fragments, and further comprises a cross-linking moiety or moieties.

24. The composition of any one of claims 1-4, 5, and 6, wherein said antibody-based antigen-binding domain is part of a multivalent polypeptide comprising at least one full antibody selected from the antibodies of classes IgG.sub.1, 2a, 2b, 3, 4, IgA, and IgM.

25. The composition of any one of claims 1-4, 5, and 6, wherein said antibody-based antigen-binding domain is part of a multivalent polypeptide that is formed prior to binding to a cell.

26. The composition of any one of claims and 1-4, 5, and 6, wherein said antibody-based antigen-binding domain is part of a multivalent polypeptide that is formed after binding to a cell.

27. The composition of any one of claims 1-4, 5, and 6, formulated in a pharmaceutically acceptable carrier and/or diluent.

28. A diagnostic composition including the composition of any of claims 1-4, 5, and 6.

29. The diagnostic composition of claim 28, further comprising a cross-linking moiety or moieties.

30. A kit to identify patients that can be treated with a composition of any of claims 1-4, 5, and 6, formulated in a pharmaceutically acceptable carrier and/or diluent comprising: a. a composition of any of claims and 1-4, 5, and 6; and b. means to measure the degree of killing or immunosuppression of said cells.

31. A kit comprising: a. a composition according to any one of claims 1-4, 5, and 6, and b. a cross-linking moiety.

32. A kit comprising: a. a composition according to any one of claims 1-4, 5, and 6, and b. a detectable moiety or moieties, and c. reagents and/or solutions to effect and/or detect binding of (a) to an antigen.

33. The composition of any one of claims 1-4, 5, and 6 operably linked to a cytotoxic agent.

34. The composition of any one of claims 1-4, 5, and 6 operably linked to an immunogenic agent.

35. The composition of claim 3, wherein said antigen-binding domain further comprises a VL CDR1 sequence represented in the general formula SGSXXNIGXNYVX (SEQ ID NO: 6) wherein each X independently represents any amino acid residue.

36. The composition of claim 35, wherein the VL CDR1 sequence is SGSESNIGNNYVQ (SEQ ID NO: 7).

37. The composition of any of claims 1-4, 5, and 6, wherein said antigen-binding domain binds to human HLA-DR with a K.sub.d of 1 .mu.M or less.

38. The composition of any of claims 1-4, 5, and 6, wherein said antigen-binding domain binds to the .alpha.-chain of HLA-DR.

39. The composition of any of claims 1-4, 5, and 6, wherein said multivalent polypeptide has an EC.sub.50 of 100 nM or less for killing activated lymphoid cells.

40. A composition including a polypeptide comprising at least one antibody-based antigen-binding domain with a binding specificity for human HLA-DR antigen, wherein treating cells expressing HLA-DR with said polypeptide causes or leads to suppression of an immune response, and wherein said antigen-binding domain includes a combination of a VH domain and a VL domain, wherein said combination is found in one of the clones taken from the group consisting of MS-GPC-1 (SEQ ID NOs. 37 and 38, respectively), MS-GPC-6 (SEQ ID NOs. 39 and 40, respectively), MS-GPC-8 (SEQ ID NOs. 41 and 42, respectively), MS-GPC-10 (SEQ ID NOs. 43 and 44, respectively), MS-GPC-8-1 (SEQ ID NOs. 41 and 28, respectively), MS-GPC-8-6 (SEQ ID NOs. 41 and 46, respectively), MS-GPC-8-9 (SEQ ID NOs. 41 and 31, respectively), MS-GPC-8-10 (SEQ ID NOs. 41 and 48, respectively), MS-GPC-8-17 (SEQ ID NOs. 41 and 50, respectively), MS-GPC-8-18 (SEQ ID NOs. 41 and 32, respectively), MS-GPC-8-27 (SEQ ID NOs. 41 and 52, respectively), MS-GPC-8-6-2 (SEQ ID NOs. 41 and 45, respectively), MS-GPC-8-6-19 (SEQ ID NOs. 41 and 47, respectively), MS-GPC-8-6-27 (SEQ ID NOs. 41 and 49, respectively), MS-GPC-8-6-45 (SEQ ID NOs. 41 and 51, respectively), MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-6-47 (SEQ ID NOs. 41 and 53, respectively), MS-GPC-8-10-57 (SEQ ID NOs. 41 and 56, respectively), MS-GPC-8-27-7 (SEQ ID NOs. 41 and 55, respectively), MS-GPC-8-27-10 (SEQ ID NOs. 41 and 57, respectively) and MS-GPC-8-27-41 (SEQ ID NOs. 41 and 58, respectively).

41. A composition including a polypeptide comprising at least one antibody-based antigen-binding domain with a binding specificity for a human HLA-DR antigen with a K.sub.d of 1 .mu.M or less, wherein treating cells expressing said antigen with said polypeptide causes or leads to suppression of an immune response, wherein said antigen-binding domain includes of a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3, VL CDR1 and VL CDR3 is found in one of the clones selected from the group consisting of MS-GPC-1 (SEQ ID NOs. 37 and 38, respectively), MS-GPC-6 (SEQ ID NOs. 39 and 40, respectively), MS-GPC-8 (SEQ ID NOs. 41 and 42, respectively), MS-GPC-10 (SEQ ID NOs. 43 and 44, respectively), MS-GPC-8-1 (SEQ ID NOs. 41 and 28, respectively), MS-GPC-8-6 (SEQ ID NOs. 41 and 46, respectively), MS-GPC-8-9 (SEQ ID NOs. 41 and 31, respectively), MS-GPC-8-10 (SEQ ID NOs. 41 and 48, respectively), MS-GPC-8-17 (SEQ ID NOs. 41 and 50, respectively), MS-GPC-8-18 (SEQ ID NOs. 41 and 32, respectively), MS-GPC-8-27 (SEQ ID NOs. 41 and 52, respectively), MS-GPC-8-6-2 (SEQ ID NOs. 41 and 45, respectively), MS-GPC-8-6-19 (SEQ ID NOs. 41 and 47, respectively), MS-GPC-8-6-27 (SEQ ID NOs. 41 and 49, respectively), MS-GPC-8-6-45 (SEQ ID NOs. 41 and 51, respectively), MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-6-47 (SEQ ID NOs. 41 and 53, respectively), MS-GPC-8-lO-57 (SEQ ID NOs. 41 and 56, respectively), MS-GPC-8-27-7 (SEQ ID NOs. 41 and 55, respectively), MS-GPC-8-27-10 (SEQ ID NOs. 41 and 57, respectively) and MS-GPC-8-27-41 (SEQ ID NOs. 41 and 58, respectively).

42. A composition including a polypeptide comprising at least one antibody-based antigen-binding domain with a binding specificity for a human HLA-DR antigen with a K.sub.d of 1 .mu.M or less, wherein treating cells expressing said antigen with said polypeptide causes or leads to suppression of an immune response, wherein said antigen-binding domain includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3 sequence is taken from the consensus CDR3 sequence XXXXRGXFDX (SEQ ID NO: 1) wherein each X independently represents any amino acid residue; and/or wherein the VL CDR3 sequence is taken from the consensus CDR3 sequence QSYDXXXX (SEQ ID NO: 2) wherein each X independently represents any amino acid residue.

43. The composition of claim 42, wherein the VH CDR3 sequence of said antigen-binding domain is SPRYRGAFDY (SEQ ID NO: 3) and/or the VL CDR3 sequence of said antigen-binding domain is QSYDLIRH (SEQ ID NO: 4) or QSYDMNVH (SEQ ID NO: 5).

44. A composition including a polypeptide comprising at least one antibody-based antigen-binding domain with a binding specificity for a human HLA-DR antigen with a K.sub.d of 1 .mu.M or less, wherein treating cells expressing said antigen with said polypeptide causes or leads to suppression of an immune response, wherein said antigen-binding domain includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the V.lamda.1 CDR1 sequence is represented in the general formula SGSXXNIGXNYVX (SEQ ID NO: 6) wherein each X independently represents any amino acid residue.

45. The composition of claim 44, wherein the CDR1 sequence is SGSESNIGNNYVQ (SEQ ID NO: 7).

46. The composition of any of claims 40 or 41-43, 44 and 45, wherein said antigen-binding domain binds to the .beta.-chain of HLA-DR.

47. The composition of claim 46, wherein said antigen-binding domain binds to an epitope of the first domain of the .beta.-chain of HLA-DR.

48. The composition of any of claims 40 or 41-43, 44 and 45, wherein said cells are lymphoids cells.

49. The composition of any of claims 40 or 41-43, 44 and 45, wherein said cells are non-lymphoid cells and express HLA-DR antigens.

50. The composition of any of claims 40 or 41-43, 44 and 45, having an IC.sub.50 for suppressing an immune response of 1 .mu.M or less.

51. The composition of any of claims 40 or 41-43, 44 and 45, having an IC.sub.50 for inhibition of IL-2 secretion of 1 .mu.M or less.

52. The composition of any of claims 40 or 41-43, 44 and 45, having an IC.sub.50 for inhibiting T cell proliferation of 1 .mu.M or less.

53. The composition of any of claims 40 or 41-43, 44 and 45, wherein said antigen-binding domain binds to one or more HLA-DR types selected from the group consisting of DR1-0101, DR2-15021, DR3-0301, DR4Dw4-0401, DR4Dw10-0402, DR4Dw14-0404, DR6-1302, DR6-1401, DR8-8031, DR9-9012, DRw53-B4*0101 and DRw52-B3*0101.

54. The composition of claim 53, wherein said antigen-binding domain binds to at least 5 different of said HLA-DR types.

55. The composition of any of claims 40 or 41-43, 44 and 45, formulated in a pharmaceutically acceptable carrier and/or diluent.

56. A pharmaceutical preparation comprising the composition of claim 50 in an amount sufficient to suppress an immune response in an animal.

57. A pharmaceutical preparation comprising the composition of claim 51 in an amount sufficient to inhibit IL-2 secretion in an animal.

58. A pharmaceutical preparation comprising the composition of claim 52 in an amount sufficient to inhibit T cell proliferation in an animal.

59. The composition of any of claims 40 and 41-43, 44 and 45, wherein said antigen-binding domain binds to the .alpha.-chain of HLA-DR.

60. The composition of claim 42, wherein said antigen-binding domain further comprises a VL CDR1 sequence represented in the general formula SGSXXNIGXNYVX (SEQ ID NO: 6) wherein each X independently represents any amino acid residue.

61. The composition of claim 60, wherein the VL CDR1 sequence is SGSESNIGNNYVQ (SEQ ID NO: 7).

62. A human IgG antibody generated by cloning into an immunoglobulin expression system an antigen-binding domain of human composition with binding specificity for human HLA-DR antigen, wherein: (a) treating cells expressing said antigen with said IgG causes or leads to killing of said cells; and (b) said antigen-binding domain includes a combination of a VH and a VL domain, wherein said combination is found in one of the clones selected from the group consisting of: MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-10-57 (SEQ ID NOs. 41 and 56, respectively) and MS-GPC-8-27-41 (SEQ ID NOs. 41 and 58, respectively).

63. The human IgG antibody of claim 62, wherein the IgG antibody is an IgG.sub.4 antibody.

64. A human IgG antibody generated by cloning into an immunoglobulin expression system an antigen-binding domain of human composition with a binding specificity for human HLA-DR antigen, wherein: (a) treating cells expressing HLA-DR with said IgG causes or leads to suppression of an immune response; and, (b) said antigen-binding domain includes a combination of a VH and a VL domain, wherein said combination is found in one of the clones selected from the group consisting of: MS-GPC-8-6-13 (SEQ ID NOs. 41 and 54, respectively), MS-GPC-8-10-57 (SEQ ID NOs. 41 and 56, respectively) and MS-GPC-8-27-41(SEQ ID NOs. 41 and 58, respectively).

65. The human IgG antibody of claim 64, wherein the IgG antibody is an IgG.sub.4 antibody.

Description

BACKGROUND OF THE INVENTION

Every mammalian species, which has been studied to date, carries a cluster of genes coding for the so-called major histocompatibility complex (MHC). This tightly linked cluster of genes code for surface antigens, which play a central role in the development of both humoral and cell-mediated immune responses. In humans the products coded for by the MHC are referred to as Human Leukocyte Antigens or HLA. The MHC-genes are organized into regions encoding three classes of molecules, class I to III.

Class I MHC molecules are 45 kD transmembrane glycoproteins, noncovalently associated with another glycoprotein, the 12 kD beta-2 microglobulin (Brown et al., 1993). The latter is not inserted into the cell membrane, and is encoded outside the MHC. Human class I molecules are of three different isotypes, termed HLA-A, -B, and -C, encoded in separate loci. The tissue expression of class I molecules is ubiquitous and codominant. MHC class I molecules present peptide antigens necessary for the activation of cytotoxic T-cells.

Class II MHC molecules are noncovalently associated heterodimers of two transmembrane glycoproteins, the 35 kD .alpha. chain and the 28 kD .beta. chain (Brown et al., 1993). In humans, class II molecules occur as three different isotypes, termed human leukocyte antigen DR (HLA-DR), HLA-DP and HLA-DQ. Polymorphism in DR is restricted to the .beta. chain, whereas both chains are polymorphic in the DP and DQ isotypes. Class II molecules are expressed codominantly, but in contrast to class I, exhibit a restricted tissue distribution: they are present only on the surface of cells of the immune system, for example dendritic cells, macrophages, B lymphocytes, and activated T lymphocytes. They are also expressed on human adrenocortical cells in the zona reticularis of normal adrenal glands and on granulosa-lutein cells in corpora lutea of normal ovaries (Kahoury et al., 1990). Their major biological role is to bind antigenic peptides and present them on the surface of antigen presenting cells (APC) for recognition by CD4 helper T (Th) lymphocytes (Babbitt et al., 1985). MHC class II molecules can also be expressed on the surface of non-immune system cells, for example, cells that express MHC class II molecules during a pathological inflammatory response. These cells may include synovial cells, endothelial cells, thyroid stromal cells and glial cells.

Class III MHC molecules are also associated with immune responses, but encode somewhat different products. These include a number of soluble serum proteins, enzymes and proteins like tumor necrosis factor or steroid 21-hydroxylase enzymes. In humans, class III molecules occur as three different isotypes, termed Ca, C2 and Bf (Kuby, 1994).

Since Th cell activation is a crucial event of the initiation of virtually all immune responses and is mediated through class II molecules, class II MHC offers itself as a target for immunomodulation (Baxevanis et al., 1980; Rosenbaum et al., 1981; Adorini et al., 1988). Besides peptide presentation, class II molecules can transduce various signals that influence the physiology of APC. Such signals arise by the interaction of multiple class II molecules with an antibody or with the antigen receptor of Th cells (Vidovic et al., 1995a; Vidovic et al., 1995b), and can induce B cell activation and immunoglobulin secretion (Cambier et al., 1991; Palacios et al., 1983), cytokine production by monocytes (Palacios, 1985) as well as the up-regulation of co-stimulatory (Nabavi et al., 1992) and cell adhesion molecules (Mourad et al., 1990).

There is also a set of observations suggesting that class II ligation, under certain conditions, can lead to cell growth arrest or be cytotoxic. Ligation under these conditions is the interaction of a polypeptide with a class II MHC molecule. There is substantial contradiction about the latter effects and their possible mechanisms. Certain authors claim that formation of a complex of class II molecules on B cells leads to growth inhibition (Vaickus et al., 1989; Kabelitz et al., 1989), whereas according to others class II complex formation results in cell death (Vidovic et al., 1995a; Newell et al., 1993; Truman et al., 1994; Truman et al., 1997; Drenou et al., 1999). In certain experimental systems, the phenomenon was observed with resting B cells only (Newell et al., 1993), or in other systems with activated B cells only (Vidovic et al., 1995a; Truman et al., 1994).

Based on these observations, anti-class II monoclonal antibodies (mAbs) have been envisaged for a number of years as therapeutic candidates. Indeed, this proposal has been supported by the beneficial effect of mouse-derived anti-class II mAbs in a series of animal disease models (Waldor et al., 1983; Jonker et al., 1988; Stevens et al., 1990; Smith et al., 1994; Vidovic & Torral, 1998; Vidovic & Laus, 2000).

Despite these early supporting data, to date no anti-MHC class II mAb of human composition has been described that displays the desired cytotoxic and other biological properties which may include affinity, efficiency of killing and selectivity. Indeed, despite the relative ease by which mouse-derived mAbs may be derived, work using mouse-derived mAbs has demonstrated the difficulty of obtaining an antibody with the desired biological properties. For example, significant and not fully understood differences were observed in the T cell inhibitory capacity of different murine anti-class II mAbs (Naquet et al., 1983). Furthermore, the application of certain mouse-derived mAbs in vivo was associated with unexpected side effects, sometimes resulting in death of laboratory primates (Billing et al., 1983; Jonker et al., 1991).

It is generally accepted that mouse-derived mAbs (including chimeric and so-called "humanized" mAbs) carry an increased risk of generating an adverse immune response (Human anti-murine antibody--HAMA) in patients compared to treatment with a human mAb (for example, Vose et al, 2000; Kashmiri et al., 2001). This risk is potentially increased when treating chronic diseases such as rheumatoid arthritis or multiple sclerosis with any mouse-derived mAb or where regular treatment may be required, for example in the treatment of certain cancers; prolonged exposure of the human immune system to a non-human molecule often leads to the development of an adverse immune reaction. Furthermore, it has proven very difficult to obtain mouse-derived antibodies with the desired specificity or affinity to the desired antigen (Pichla et al. 1997). Such observation may significantly reduce the overall therapeutic effect or advantage provided by mouse-derived mAbs. Examples of disadvantages for mouse-derived mAbs may include the following. First, mouse-derived mAbs may be limited in the medical conditions or length of treatment for a condition for which they are appropriate. Second, the dose rate for mouse-derived mAbs may need to be relatively high in order to compensate for a relatively low affinity or therapeutic effect, hence making the dose not only more severe but potentially more immunogenic and perhaps dangerous. Third, such restrictions in suitable treatment regimes and high-dose rates requiring high production amounts may significantly add to the cost of treatment and could mean that such a mouse-derived mAb be uneconomical to develop as a commercial therapeutic. Finally, even if a mouse mAb could be identified that displayed the desired specificity or affinity, often these desired features are detrimentally affected during the "humanization" or "chimerization" procedures necessary to reduce immunogenic potential (Slavin-Chiorini et al., 1997). Once a mouse-derived mAb has been "humanized" or chimerized, then it is very difficult to optimize its specificity or affinity.

The art has sought over a number of years for anti-MHC class II mAbs of human composition that show biological properties suitable for use in a pharmaceutical composition for the treatment of humans. Workers in the field have practiced the process steps of first identifying a mouse-derived mAb, and then modifying the structure of this mAb with the aim of improving immunotolerance of this non-human molecule for human patients (for further details, see Jones et al., 1986; Riechmann et al., 1988; Presta, 1992). This modification is typically made using so-called "humanization" procedures or by fabricating a human-mouse chimeric mAb. Other workers have attempted to identify human antibodies that bind to human antigens having desired properties within natural repertoires of human antibody diversity. For example, by exploring the fetal-tolerance mechanism in pregnant women (Bonagura et al., 1987) or by panning libraries of natural diversities of antibodies (Stausbol-Gron et al., 1996; Winter et al., 1994). However, to date no anti-MHC class II mAb of human composition has been described that displays the desired biological properties of cytotoxicity, selectivity, specificity, low immunogenicity and affinity.

For therapeutic purposes a polypeptide reacting with most or at least many of the common allelic forms of a human class II MHC molecule would be desirable--e.g., to enable its use in diverse patient populations. Moreover, the candidate polypeptide should be cytotoxic to a wide range of lymphoid tumors, and preferably is cytotoxic by way of a mechanism common to such a range of tumor cells. To allow for a wide range of possible applications, the polypeptide desired should mediate its cytotoxic effect without the dependence on further components of the immune system. For therapeutic purposes, most patients receive for the treatment of, e.g. cancer, standard chemo- or radiotherapy. Most of these treatments leave the patient immunocompromised. Any additional treatment that relies on an intact immune system is therefore likely to fail. The underlying problem is further demonstrated in humans who suffer from a disease that destroys the immune system, e.g. HIV. Opportunistic infections and malignant transformations are able to escape the immune-surveillance and cause further complications.

SUMMARY OF THE INVENTION

One aspect of the present invention relates to a composition including a polypeptide comprising at least one antibody-based antigen-binding domain of human composition with binding specificity for an antigen expressed on the surface of a human cell, wherein treating cells expressing the antigen with a multivalent polypeptide having two or more of said antigen binding domains causes or leads to killing of the cells in a manner where neither cytotoxic entities nor immunological mechanisms are needed for killing. In certain preferred embodiments the antigen is an MHC antigen, preferably an MHC class II antigen, such as DR/DP/DQ or DR. For instance, in certain preferred embodiments, the subject compositions include a polypeptide comprising at least one antibody-based antigen-binding domain which binds to human HLA-DR with a K.sub.d of 1 .mu.M, 100 nM, 10 nM or even 1 nM or less.

Another aspect of the present invention provides a composition including a multivalent polypeptide comprising a plurality of antibody-based antigen-binding domains of human composition with binding specificity for human HLA-DR. Treating cells expressing HLA-DR with the multivalent polypetide causes or leads to killing of the cell in a manner where neither cytotoxic entities nor immunological mechanisms are needed for killing. In certain preferred embodiments, the said antigen-binding domains individually bind to the human HLA-DR with a K.sub.d of 1 .mu.M, 100 nM, 10 nM or even 1 nM or less. In certain preferred embodiments, the multivalent polypeptide has an EC.sub.50 of 100 nM, 10 nM or even 1 nM or less for killing activated lymphoid cells, transformed cells and/or lymphoid tumor cells.

Still another aspect of the present invention provides a composition including a polypeptide comprising at least one antibody-based antigen-binding domain that binds to human HLA-DR with a K.sub.d of 1 .mu.M, 100 nM, 10 nM or even 1 nM or less, the antigen-binding domain being isolated by a method which includes isolation of human VL and VH domains from a recombinant antibody library by ability to bind to at least one epitope of human HLA-DR. Treating a cell expressing HLA-DR with a multivalent polypeptide having two or more of the antigen binding domains causes or leads to killing of the cells in a manner where neither cytotoxic entities nor immunological mechanisms are needed for killing. In certain embodiments, the method for isolating the antigen-binding domain includes the further steps of: a) generating a library of variants of at least one of the CDR1, CDR2 and CDR3 sequences of one or both of the VL and VH domains, and, b) isolation of VL and VH domains from the library of variants by ability to bind to human HLA-DR with a K.sub.d of 1 .mu.M or less.

In certain preferred embodiments, the composition of the present invention can be characterized as including multivalent polypeptides having an EC.sub.50 for killing transformed cells at least 5-fold lower than the EC.sub.50 for killing normal cells, and even more preferably at least 10-fold, 100-fold and even 1000-fold less than for killing normal cells.

In certain preferred embodiments, the composition of the present invention are characterized as including multivalent polypeptides having an EC.sub.50 for killing activated cells at least 5-fold lower than the EC.sub.50 for killing unactivated cells, and even more preferably at least 10-folded, 100-fold and even 1000-fold less than for killing unactivated cells.

In certain preferred embodiments, the composition of the present invention are characterized as including multivalent polypeptides having an EC.sub.50 of 50 nM or less for killing transformed cells, and even more preferably an EC.sub.50 of less than 10 nM, 1 nM and even 0.1 nM. In certain embodiments, the subject multivalent polypeptides have an EC.sub.50 for killing activated lymphoid cells, transformed cells and/or lymphoid tumor cells of 100 nM, 10 nM or even 1 nM or less.

In certain embodiments, the subject compositions including multivalent polypeptides that selectively kill activated lymphoid cells. For example, such multivalent forms of the subject compositions can be used to kill activated lymphoid cells are lymphoid tumor cells representing a disease selected from B cell non-Hodgkin lymphoma, B cell lymphoma, B cell acute lymphoid leukemia, Burkitt lymphoma, Hodgkin lymphoma, hairy cell leukemia, acute myeloid leukemia, T cell lymphoma, T cell non-Hodgkin lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, and multiple myeloid leukemia. Exemplary activated lymphoid tumor cells which can be killed include PRIESS (ECACC Accession No: 86052111), GRANTA-519 (DSMZ Accession No: ACC 342), KARPAS-422 (DSMZ Accession No: ACC 32), KARPAS-299, DOHH-2, SR-786, MHH-CALL-4, MN-60, BJAB, RAJI, L-428, HDLM-2, HD-MY-Z, KM-H2, L1236, BONNA-12, HC-1, NALM-1, L-363, EOL-1, LP-1, RPMI-8226, and MHH-PREB-1 cell lines. In certain preferred embodiments, the subject compositions have an EC.sub.50 of 100 nM or less, and preferably less than 10 uM or even 1 nM, for killing at least one of B cell lymphoma cells and T cell lymphoma cells selected from the list of KARPAS-422, DOHH-2, SR-786 MHH-CALL-4, MN-60, HD-MY-Z, NALM-1 and LP-1. In certain instances, to effect cell killing, the target cells may require further activation or pre-activation, such as by by incubation with Lipopolysaccharide (LPS, 10 .mu.g/ml), Interferon-gamma (IFN-.gamma., Roche, 40 ng/ml) and/or phyto-hemagglutinin (PHA, 5 .mu.g/ml) to name but a few.

In certain embodiments, the multivalent forms of the subject compositions can be used to kill non-lymphoid cells that express MHC class II molecules.

In certain embodiments, one or more of the antigen binding domains of the subject compositions bind to the .beta.-chain of HLA-DR, e.g., the antigen-binding domain binds to the first domain of the .beta.-chain of HLA-DR.

In certain other embodiments, one or more of the antigen binding domains of the subject compositions bind to the .alpha.-chain of HLA-DR, e.g., the antigen-binding domain binds to the first domain of the .alpha.-chain of HLA-DR.

In certain preferred embodiments, the antigen binding domain(s) of the subject compositions bind to one or more HLA-DR types selected from the group consisting of DR1-0101, DR2-15021, DR3-0301, DR4Dw4-0401, DR4Dw10-0402, DR4Dw14-0404, DR6-1302, DR6-1401, DR8-8031, DR9-9012, DRW53-B4*0101 and DRW52-B3*0101. In preferred embodiments, the antigen binding domains of the subject compositions provide broad-DR reactivity, that is, the antigen-binding domain(s) of a given composition binds to epitopes on at least 5 different of said HLA-DR types. In certain embodiments, the antigen binding domain(s) of a polypeptide(s) of the subject compositions binds to a plurality of HLA-DR types as to bind to HLA-DR expressing cells for at least 60 percent of the human population, more preferably at least 75 percent, and even more preferably 85 percent of the human population.

In certain embodiments, the antigen-binding domains of the subject compositions include a combination of a VH domain and a VL domain, wherein said combination is found in one of the clones taken from the list of MS-GPC-1, MS-GPC-6, MS-GPC-8, MS-GPC-10, MS-GPC-8-1, MS-GPC-8-6, MS-GPC-8-9, MS-GPC-8-10, MS-GPC-8-17, MS-GPC-8-18, MS-GPC-8-27, MS-GPC-8-6-2, MS-GPC-8-6-19, MS-GPC-8-6-27, MS-GPC-8-6-45, MS-GPC-8-6-13, MS-GPC-8-6-47, MS-GPC-8-10-57, MS-GPC-8-27-7, MS-GPC-8-27-10 and MS-GPC-8-27-41.

In certain embodiments, the antigen-binding domains of the subject compositions include a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3, VL CDR1 And VL CDR3 is found in one of the clones taken from the list of MS-GPC-1, MS-GPC-8, MS-GPC-10, MS-GPC-8-1, MS-GPC-8-6, MS-GPC-8-9, MS-GPC-8-10, MS-GPC-8-17, MS-GPC-8-18, MS-GPC-8-27, MS-GPC-8-6-2, MS-GPC -8-6-19, MS-GPC-8-6-27, MS-GPC-8-6-45, MS-GPC-8-6-13, MS-GPC-8-6-47, MS-GPC-8-10-57, MS-GPC-8-27-7, MS-GPC-8-27-10 and MS-GPC-8-27-41.

In a further preferred embodiment, the antigen-binding domain is modified compared to a parental antigen-binding domain of the present invention by addition, deletion and/or substitution of amino acid residues, while maintaining the properties according to the present invention, or improving one or more of said properties, of said parental antigen-binding domain. This may include, but is not limited to, the modification of a nucleic acid sequence encoding a parental antigen-binding domain for cloning purposes, the modification of CDR regions in order to improve or modify antigen-binding affinity and/or specificity, including the exchange of one or more CDR sequences of a parental antigen-binding domain by corresponding CDR sequences from one or more different antigen-binding domains, and the addition of peptide sequences for detection and/or purification purposes. It is well within the scope of one of ordinary skill in the art to identify positions in a given parental antigen-binding domain where an addition, deletion and/or substitution should occur, to design and pursue the approach to achieve said addition, deletion and/or substitution, and to test or assay whether the modified antigen-binding domain has maintained the properties of, or exhibits one or more improved properties compared to, the parental antigen-binding domain. Furthermore, one of ordinary skill would be able to design approaches where collections or libraries of modified antigen-binding domains are designed, constructed and screened to identify one or more modified antigen-binding domain which have maintained the properties, or exhibit one or more improved properties compared to the parental antigen-binding domain. In one example, the first amino acid residue of a HuCAL VH domain comprised in any antigen-binding domain or the present invention, which is either E or Q depending on the expression construct, may be exchanged by Q or E, respectively. Preferred regions to optimize an antigen-binding domain by designing, constructing and screening collections or libraries of modified antigen-binding domains according to the present invention comprise the CDR regions, and most preferably CDR3 of VH and VL, CDR1 of VL and CDR2 of VH domains.

In certain embodiments, the antigen-binding domains includes a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3 sequence is taken from the consensus CDR3 sequence: XXXXRGXFDX (SEQ ID No. 1)

wherein each X independently represents any amino acid residue; and/or,

wherein the VL CDR3 sequence is taken from the consensus CDR3 sequence: QSYDXXXX (SEQ ID No. 2)

wherein each X independently represents any amino acid residue. For instance, the VH CDR3 sequence can be SPRYRGAFDY (SEQ ID No. 3) and/or the VL CDR3 sequence can be QSYDLIRH (SEQ ID No. 4) or QSYDMNVH (SEQ ID No. 5).

In certain embodiments, the antigen-binding domains of the subject antigen-binding domain competes for antigen binding with an antibody including a combination of HuCAL VH2 and HuCAL V.lamda.1, wherein the VH CDR3 sequence is taken from the consensus CDR3 sequence: XXXXRGXFDX (SEQ ID No. 1)

each X independently represents any amino acid residue; and/or,

the VL CDR3 sequence is taken from the consensus CDR3 sequence: QSYDXXXX (SEQ ID No. 2)

each X independently represents any amino acid residue. For instance, the VH CDR3 sequence can be SPRYRGAFDY (SEQ ID No. 3) and/or the VL CDR3 sequence can be QSYDLIRH (SEQ ID No. 4) or QSYDMNVH (SEQ ID No. 5).

In certain preferred embodiments, the antigen-binding domain includes a VL CDR1 sequence represented in the general formula: SGSXXNIGXNYVX (SEQ ID No. 6)

wherein each X independently represents any amino acid residue. For instance, the CDR1 sequence is SGSESNIGNNYVQ (SEQ ID No. 7).

In preferred embodiments, the mechanism of killing by multivalent forms of the subject compositions involves an innate pre-programmed process of said cell. For instance, the killing is non-apoptotic. Killing by the subject compositions can be dependent on the action of non-caspase proteases, and/or killing which cannot be inhibited by zVAD-fmk or zDEVD-fmk.

In certain preferred embodiments, the antibody-based antigen-binding domain is part of a multivalent polypeptide including at least a F(ab').sub.2 antibody fragment or a mini-antibody fragment.

In certain preferred embodiments, the antibody-based antigen-binding domain is part of a multivalent polypeptide comprising at least two monovalent antibody fragments selected from Fv, scFv, dsFv and Fab fragments, and further comprises a cross-linking moiety or moieties.

In certain preferred embodiments, the antibody-based antigen-binding domain is part of a multivalent polypeptide comprising at least one full antibody selected from the antibodies of classes IgG.sub.1, 2a, 2b, 3, 4, IgA, and IgM.

In certain preferred embodiments, the antibody-based antigen-binding domain is part of a multivalent polypeptide is formed prior to binding to said cell.

In certain preferred embodiments, the antibody-based antigen-binding domain is part of a multivalent polypeptide is formed after binding to said cell.

In certain preferred embodiments, the antigen binding sites are cross-linked to a polymer.

Another aspect of the present invention provides a nucleic acid comprising a coding sequence for an antigen-binding domain, such as those antigen binding domains described above, or a multivalent polypeptide thereof. For example, in certain embodiments, the nucleic acid includes a coding sequence for a polypeptide comprising at least one antibody-based antigen-binding domain of human composition with binding specificity for an antigen expressed on the surface of a human cell, wherein treating cells expressing the antigen with a multivalent form of the polypeptide causes or leads to killing of said cell in a manner where neither cytotoxic entities nor immunological mechanisms are needed for killing. In certain embodiments, the nucleic acid includes a coding sequence for a polypeptide comprising at least one antibody-based antigen-binding domain which binds to at least one epitope of human HLA-DR with a K.sub.d of 1 .mu.M, 100 nM, 10 nM or even 1 nM or less.

In certain embodiments, the nucleic acid includes a coding sequence for a polypeptide comprising a plurality of antibody-based antigen-binding domains of human composition with binding specificity for human HLA-DR, wherein treating a cell expressing HLA-DR with the multivalent polypeptide causes or leads to killing of the cell in a manner where neither cytotoxic entities nor immunological mechanisms are needed for killing. In preferred embodiments, the antigen-binding domains individually bind to epitopes on the human HLA-DR with a K.sub.d of 1 .mu.M, 100 nM, 10 nM or even 1 nM or less.

In certain embodiments, the nucleic acid includes a coding sequence for a multivalent polypeptide comprising a plurality of antibody-based antigen-binding domains of human composition with binding specificity for human HLA-DR, wherein treating a cell expressing HLA-DR with said multivalent polypeptide causes or leads to killing of said cell in a manner where neither cytotoxic entities nor immunological mechanisms are needed for said cell killing. Preferably, the multivalent polypeptide has an EC.sub.50 for killing activated lymphoid cells, transformed cells and/or lymphoid tumor cells of 100 nM, 10 nM or even 1 nM or less.

Another aspect of the invention provides a vector comprising the coding sequence of any one of the subject nucleic acids, e.g., as described above, and a transcriptional regulatory sequence operably linked thereto.

Still another aspect of the present invention provides a host cell harboring at least one subject nucleic acids or the subject vector. Another aspect of the present invention provides a method for the production of a multivalent composition that causes or leads to killing of cells in a manner where neither cytotoxic entities nor immunological mechanisms are needed to cause or lead to said killing comprising culturing the host cells under conditions wherein the nucleic acid is expressed either as a polypeptide comprising a plurality of antigen binding domains or as a polypeptide comprising at least one antigen binding domains which is subsequently treated to form a multivalent composition.

Another aspect of the present invention provides forms of the subject polypeptide or nucleic acid compositions, formulated in a pharmaceutically acceptable carrier and/or diluent. The present invention specifically contemplates the use of such compositions for preparing a pharmaceutical preparation for the treatment of animals, especially humans.

Such pharmaceutical compositions can be used for the treatment of conditions involving unwanted cell proliferation, particularly the treatment of a disorder involving transformed cells expressing MHC class II antigens. For instance, the formulations can be used for the treatment of a disorder selected from B cell non-Hodgkin lymphoma, B cell lymphoma, B cell acute lymphoid leukemia, Burkitt lymphoma, Hodgkin lymphoma, hairy cell leukemia, acute myeloid leukemia, T cell lymphoma, T cell non-Hodgkin lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloid leukemia and B cell precursor leukemia.

Such pharmaceutical preparations can be used for the treatment of diseases involving unwanted activation of immune cells, such as in the treatment of a disorder selected from rheumatoid arthritis, juvenile arthritis, multiple sclerosis, Grave's disease, insulin-dependent diabetes, narcolepsy, psoriasis, systemic lupus erythematosus, ankylosing spondylitis, transplant rejection, graft vs. host disease, Hashimoto's disease, myasthenia gravis, pemphigus vulgaris, glomerulonephritis, thyroiditis, pancreatitis, insulitis, primary biliary cirrhosis, irritable bowel disease and Sjogren syndrome.

Another aspect of the present invention provides a diagnostic composition including the polypeptide or nucleic acid compositions of the present invention. In certain embodiments, the diagnostic composition includes a polypeptide composition and a cross-linking moiety or moieties.

Still another aspect of the present invention provides a method for killing a cell expressing an antigen on the surface of said cell comprising the step of contacting the cell with a multivalent polypeptide composition of the subject invention.

Another aspect of the invention provides a method to identify patients that can be treated with a multivalent polypeptide composition, formulated in a pharmaceutically acceptable carrier and/or diluent comprising the steps of: a. isolating cells from a patient; b. contacting said cells with the composition; and c. measuring the degree of killing or immunosuppression of said cells.

The present invention also provides a kit to identify patients that can be treated with a multivalent polypeptide composition of the present invention, formulated in a pharmaceutically acceptable carrier and/or diluent comprising: a. a multivalent polypeptide composition; and b. means to measure the degree of killing or immunosuppression of said cells.

In certain embodiments, the kit includes a multivalent polypeptide composition, and a cross-linking moiety. In other embodiments, the kit includes a. a multivalent polypeptide composition, b. a detectable moiety or moieties, and c. reagents and/or solutions to effect and/or detect binding of (a) to an antigen.

Another aspect of the present invention provides a cytotoxic composition comprising a multivalent polypeptide composition operably linked to a cytotoxic agent.

Stil another aspect of the invention provides an immunogenic composition comprising a multivalent polypeptide composition operablly linked to an immunogenic agent.

Another aspect of the present invention provides a method to kill a cell comprising contacting the cell with a multivalent polypeptide composition operablly linked a cytotoxic or immunogenic agent.

Another aspect of the invention provides a method for treating a human to reduce the severity of disorder involving unwanted proliferation/activation of cells expressing the human .beta.-chain of HLA-DR, comprising administering to the patient a a multivalent polypeptidepolypeptide of the present invention. In certain embodiments, the disorder involves unwanted proliferation/activation of lymphoid cells, e.g., selected from B cell non-Hodgkin lymphoma, B cell lymphoma, B cell acute lymphoid leukemia, Burkitt lymphoma, Hodgkin lymphoma, hairy cell leukemia, acute myeloid leukemia, T cell lymphoma, T cell non-Hodgkin lymphoma, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloid leukemia and B cell precursor leukemia.

Another aspect of the invention provides a use of a multivalent polypeptide composition operably linked a cytotoxic or immunogenic agent for preparing a pharmaceutical preparation for the treatment of animals

According to a preferred embodiment, the polypeptide is directed to a lymphoid cell or a non-lymphoid cell that expresses MHC class II molecules. The latter type of cells occur for example at pathological sites of inflammation and/or autoimmune diseases, e.g. synovial cells, endothelial cells, thyroid stromal cells and glial cells, or it may also comprise genetically altered cells capable of expressing MHC class II molecules.

Preferably, the polypeptide is directed to lymphoid tumor cells. More preferred are lymphoid tumor cells that represent a disease selected from B cell non-Hodgkin lymphoma, B cell lymphoma, B cell acute lymphoid leukemia, Burkitt lymphoma, Hodgkin lymphoma, hairy cell leukemia, acute myeloid leukemia and B cell precursor leukemia. Most preferred are lymphoid tumor cells from a cell line taken from the list of GRANTA-519, PRIESS, KARPAS-422, DOHH-2, MHH-CALL-4, MN-60, BJAB, L-428, BONNA-12, EOL-1, MHH-PREB-1 and MHH-CALL-2 cell lines.

In certain embodiments, the polypeptide binds to at least one epitope in the alpha-chain of an HLA-DR molecule. In such embodiments, the polypeptide preferably binds to at least one epitope in the first domain of the alpha-chain of HLA-DR, the first domain being the N-terminal domain of the chain. For instance, the polypeptide can be selected to bind to at least one epitope within the alpha-helix ranging from Glu55 to Tyr79 of the alpha-chain of HLA-DR.

In other embodiments, the polypeptide binds to at least one epitope in the beta-chain of an HLA-DR molecule