Chimeric receptor genes and cells transformed therewith
View Patent ↗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.
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 FcγRIII or FcγRII.
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.