Chimeric receptor genes and cells transformed therewith

• US 7,741,465 B1
• Filed: 07/02/1993
• Issued: 06/22/2010
• Est. Priority Date: 03/18/1992
• Status: Active Grant

• Current Assignee: Cabaret Biotech, Ltd., The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: Gideon Gross, Tova Waks, Zelig Eshhar
• Inventors: Schindler, Daniel, Gross, Gideon, Eshhar, Zelig, Waks, Tova
• Primary Examiner(s): WEHBE, ANNE MARIE SABRINA
• Application Number:
  US08/084,994
• Time in Patent Office: 6,199 Days
• Field of Search: 424/93.2, 536/23.53, 536/23.1, 536/23.5, 536/23.4, 435/320.1, 435/325
• US Class Current: 536/23.4
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464406 Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 16/4291 against IgE
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment

Abstract

Chimeric receptor genes suitable for endowing lymphocytes with antibody-type specificity include a first gene segment encoding a single-chain Fv domain of a specific antibody and a second gene segment encoding all or part of the transmembrane and cytoplasmic domains, and optionally the extracellular domain, of an immune cell-triggering molecule. The chimeric receptor gene, when transfected to immune cells, expresses the antibody-recognition site and the immune cell-triggering moiety into one continuous chain. The transformed lymphocytes are useful in therapeutic treatment methods.

Claims

1. A chimeric DNA comprising:

a first DNA segment encoding a single-chain Fv domain (scFv) comprising a V_L linked to a V_H of a specific antibody by a flexible linker, and

a second DNA segment encoding partially or entirely the transmembrane and cytoplasmic, and optionally the extracellular, domains of an endogenous protein, wherein said endogenous protein is expressed on the surface of lymphocytes and triggers the activation and/or proliferation of said lymphocytes,

which chimeric DNA, upon transfection to lymphocytes, expresses both said scFv domain and said domains of said endogenous protein in one single, continuous chain on the surface of the transfected lymphocytes such that the transfected lymphocytes are triggered to activate and/or proliferate and have MHC non-restricted antibody-type specificity when said expressed scFv domain binds to its antigen.

2. A chimeric DNA according to claim 1, wherein the second DNA segment further comprises partially or entirely the extracellular domain of said endogenous protein.

3. A chimeric DNA according to claim 1, wherein the first DNA segment encodes the scFv domain of an antibody against tumor cells.

4. A chimeric DNA according to claim 1, wherein the first DNA segment encodes the scFv domain of an antibody against virus infected cells.

5. A chimeric DNA according to claim 4, wherein the virus is HIV.

6. A chimeric DNA according to claim 1, wherein said endogenous protein is a lymphocyte receptor chain, a polypeptide of the TCR/CD3 complex, or a subunit of the Fc or IL-2 receptor.

7. A chimeric DNA according to claim 1, wherein the second DNA segment encodes a lymphocyte receptor chain.

8. A chimeric DNA according to claim 7, wherein the second DNA segment encodes a chain of the T cell receptor.

9. A chimeric DNA according to claim 8, wherein the second DNA segment encodes the α

, β

, γ

or δ

chain of the antigen-specific T cell receptor.

10. A chimeric DNA according to claim 1, wherein the second DNA segment encodes a polypeptide of the TOR/CD3 complex.

11. A chimeric DNA according to claim 10, wherein the second DNA segment encodes the zeta or eta isoform chain.

12. A chimeric DNA according to claim 1, wherein the second DNA segment encodes a subunit of the Fc receptor or IL-2 receptor.

13. A chimeric DNA according to claim 12, wherein the second DNA segment encodes a common subunit of IgE and IgG binding Fc receptors.

14. A chimeric DNA according to claim 13, wherein said subunit is the gamma subunit.

15. A chimeric DNA according to claim 13, wherein the second DNA segment encodes the CD16α

chain of the Fcγ

RIII or Fcγ

RII.

16. A chimeric DNA according to claim 12, wherein the second DNA segment encodes the α

or β

subunit of the IL-2 receptor.

17. An expression vector comprising a chimeric DNA according to claim 1.

18. A lymphocyte, isolated from an individual, transformed with an expression vector according to claim 17, wherein expression of said chimeric DNA endows said lymphocyte with antibody specificity.

19. A lymphocyte, isolated from an individual, transformed with a chimeric DNA according to claim 1, wherein expression of said chimeric DNA endows said lymphocyte with antibody specificity.

20. A lymphocyte according to claim 19. selected from the group consisting of a natural killer cell, a lymphokine activated killer cell, a cytotoxic T cell, a helper T cell and a subtype thereof.