Genetically-engineered MHC molecules

• US 7,319,143 B2
• Filed: 05/31/2001
• Issued: 01/15/2008
• Est. Priority Date: 06/01/2000
• Status: Expired due to Fees

• Current Assignee: Gavish-Galilee Bio Applications Ltd.
• Original Assignee: Gavish-Galilee Bio Applications Ltd.
• Inventors: Gross, Gideon, Margalit, Alon
• Primary Examiner(s): Chan; Christina
• Assistant Examiner(s): VANDERVEGT, FRANCOIS P
• Application Number:
  US10/297,060
• Publication Number:
  US 20040137562A1
• Time in Patent Office: 2,420 Days
• Field of Search: None
• US Class Current: 536/23.4
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules

Abstract

The invention provides DNA molecules encoding a chimeric polypeptide comprising (a) a component of a MHC molecule capable of association on a cell surface with an endogenous MHC molecule component of the same class, and (b) an intracellular region of a signal transduction element capable of activating T cells. Component (a) may be a monomorphic component and is preferably beta 2-microglobulin, or a polymorphic class I or class II component. The signal transduction element (b) capable of activating T cells may be a component of T-cell receptor CD3, preferably the CD3 zeta (zeta) polypeptide, a B cell receptor polypeptide or an Fc receptor polypeptide. Immune cells such as a CTLs expressing said chimeric MHC molecules specifically eliminate or inactivate harmful T cells and are useful for treating graft rejection and autoimmune diseases.

Claims

1. A DNA molecule encoding a chimeric polypeptide comprising (a) a component of a MHC molecule capable of association on a cell surface with an endogenous MHC molecule component of the same class, and (b) an intracellular region of a signal transduction element capable of activating T cells, wherein said signal transduction element is a component of T cell receptor CD3 and said component of T-cell receptor CD3 comprises the transmembranal and cytoplasmic regions of the human zeta (ζ

) polypeptide.

2. The DNA molecule of claim 1, wherein said MHC component (a) is a monomorphic component selected from the group consisting of β

₂-microglobulin and the monomorphic α

chain of a HLA-DR molecule.

3. The DNA molecule of claim 2, wherein said MHC component (a) is human β

₂-microglobulin capable of association on a cell surface with an endogenous class I heavy chain HLA molecule, and said β

₂-microglobulin is linked to component (b) by a bridge peptide having about 10-15 amino acid residues.

4. The DNA molecule of claim 3, wherein said bridge peptide has a sequence comprised within the membrane-proximal sequence of a class I heavy chain HLA molecule.

5. The DNA molecule of claim 4, wherein said bridge peptide has 13 amino acid residues comprised within the extracellular membrane-proximal sequence of the class I heavy chain HLA-A2 molecule, and has the sequence:

Leu Arg Trp Glu Pro Ser Ser Gln Pro Thr Ile Pro Ile (SEQ ID NO;

9).

6. The DNA molecule of claim 2, wherein said MHC component (a) is a monomorphic α

chain of a HLA-DR molecule.

7. The DNA molecule of claim 1, wherein said MHC component (a) is a polymorphic component selected from the group consisting of:

(i) an α

chain of class I HLA-A, HLA-B or HLA-C molecule, which is capable of association on a cell surface with endogenous β

₂m;

(ii) an α

chain of class II HLA-DP or HLA-DQ molecule, which is capable of association on a cell surface with endogenous HLA-DPβ

or HLA-DQβ

chain;

or (iii) a β

chain of class II HLA-DP, HLA-DR or HLA-DQ molecule, which is capable of association on a cell surface with an endogenous HLA-DPα

, HLA-DRα

or HLA-DQα

chain.

8. The DNA molecule of claim 1, wherein said chimeric polypeptide further comprises an antigenic peptide related to an autoimmune disease, said antigenic peptide being linked to said chimeric polypeptide by a peptide linker.

9. The DNA molecule of claim 8, wherein said antigenic peptide has 8-10 amino acid residues and binds to a product of a certain HLA allele.

10. An isolated immune cell which expresses a chimeric polypeptide as defined in claim 9 and binds to an lyses autoreactive cells causing an autoimmune disease.

11. An isolated immune cell which expresses a chimeric polypeptide as defined in claim 8 and binds to and lyses autoreactive cells causing an autoimmune disease.

12. A vector comprising the DNA molecule of claim 1.

13. An isolated immune cell which expresses a chimeric polypeptide as defined in claim 1 and binds to, and eliminates, alloreactive cells causing transplant rejection.

14. The immune cell of claim 13 which is a cytotoxic T lymphocyte (CTL).

15. The cell which expresses a chimeric polypeptide comprising (a) a component of a MHC molecule capable of association on a cell surface with an endogenous MHC molecule component of the same class, and (b) an intracellular region of a signal transduction element capable of activating T cells, wherein said signal transduction element is a component of T-Cell receptor CD3 and said component of T-cell receptor CD3 comprises the transmembranal and cytoplasmic regions of the human zeta (ζ

) polypeptide.

16. The cell of claim 15, wherein said cell is an immune cell selected from the group consisting of T helper cells (CD4⁺), cytotoxic T lymphocytes (CD8⁺) and natural killer (NK) cells, capable of recognizing and binding to harmful T cells and causing their lysis or inactivation.