Compositions and methods for immunotherapy
The present invention provides immunoresponsive cells, including T cells, cytotoxic T cells, regulatory T cells, and Natural Killer (NK) cells, expressing an antigen recognizing receptor and an inhibitory chimeric antigen receptor (iCAR). Methods of using the immunoresponsive cell include those for the treatment of neoplasia and other pathologies where an increase in an antigen-specific immune response is desired.
1. A T cell comprising:
a) a chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a first antigen that is expressed at the surface of a tumor cell, and an intracellular signaling domain that is capable of activating the T cell and comprises a signaling domain of CD28, and
b) an inhibitory chimeric antigen receptor (iCAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a second antigen that is not expressed on the tumor cell surface, and a signaling domain of an immunoinhibitory receptor selected from the group consisting of CTLA-4, PD-1, LAG-3, 2B4, and BTLA,
wherein binding of the CAR to the first antigen induces cytotoxicity of the T cell, an binding of the iCAR to the second antigen reduces the cytotoxicity of the T cell induced by the CAR.
2. The T cell of claim 1 , wherein said iCAR is recombinantly expressed.
3. The T cell of claim 1 , wherein the iCAR is expressed from a vector.
4. The T cell of claim 1 , wherein said CAR is recombinantly expressed.
5. The T cell of claim 1 , wherein the CAR is expressed from a vector.
6. The T cell of claim 1 , wherein the T cell is selected from the group consisting of effector T cells, and memory T cells..
7. The T cell of claim 1 , wherein said T cell is autologous.
8. The T cell of claim 1 , wherein said T cell is non-autologous..
9. The T cell of claim 1 , wherein said first antigen selected from the group consisting of CD19, CD7, CD10, CD20, CD22, CD30, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD123, CD133, CD138, CAIX, CEA, CD5, EGP-2, EGP-40, EpCAM, Erb-B2, Erb-B3, Erb-B4, FBP, Fetal acetylcholine receptor, folate receptor-a, GD2, GD3, HER-2, IL-13R-α2, κ-light chain, LeY, L1 cell adhesion molecule, Mesothelin, Muc-1, Muc-16, oncofetal antigen (h5T4), PSCA, PSMA, ROR1, TAG-72, and VEGF-R2.
10. The T cell of claim 1 , wherein said iCAR further comprises a transmembrane domain selected from the group consisting of a CD4 polypeptide, a CD8 polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, and a BTLA polypeptide.
11. The T cell of claim 6 , wherein said effector T cells are selected from the group consisting of helper T cells (CD4 + T cells), and cytotoxic T cells (CD8 + T cells).
12. The T cell of claim 1 , wherein said first antigen is selected from the group consisting of CD19, PSMA, mesothelin, and CD56.
13. The T cell of claim 1 , wherein the second antigen is selected from the group consisting of an Epithelial-mesenchymal transition (EMT) antigen, cytokeratin, human leukocyte antigens (HLAs), Opioid-binding protein/cell adhesion molecule (OPCML), HYAL2, Deleted in Colorectal Carcinoma (DCC), Scaffold/Matrix attachment region-binding protein 1 (SMAR1), CD33, CD38, and E-cadherin.
14. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and an effective amount of T cells, wherein said T cell comprises:
(a) a chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a first antigen that is expressed at the surface of a tumor cell, and an intracellular signaling domain that is capable of activating the T cell and comprises a signaling domain of CD28, and
(b) an inhibitory chimeric antigen receptor (iCAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a second antigen that is not expressed on the tumor cell surface, and a signaling domain of an immunoinhibitory receptor selected from the group consisting of CTLA-4, PD-1, LAG-3, 2B4, and BTLA, wherein binding of the CAR to the first antigen induces cytotoxicity of the T cell, and binding of the iCAR to the second antigen reduces the cytotoxicity of the T cell induced by the CAR.
15. The pharmaceutical composition of claim 14 , wherein the second antigen is selected from the group consisting of an Epithelial-mesenchymal transition (EMT) antigen, cytokeratin, human leukocyte antigens (HLAs), Opioid-binding protein/cell adhesion molecule (OPCML), HYAL2, Deleted in Colorectal Carcinoma (DCC), Scaffold/Matrix attachment region-binding protein 1 (SMAR1), CD33, CD38, and E-cadherin.
16. A kit comprising a T cell, wherein said T cells comprises:
(a) a chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a first antigen that is expressed at the surface of a tumor cell, and an intracellular signaling domain that is capable of activating the T cell and comprises a signaling domain of CD28, and
(b) an inhibitory chimeric antigen receptor (iCAR) comprising an extracellular antigen-binding domain comprising a single chain variable fragment (scFv) that binds to a second antigen that is not expressed on the tumor cell surface, and a signaling domain of an immunoinhibitory receptor selected from the group consisting of CTLA-4, PD-1, LAG-3, 2B4, and BTLA,
wherein binding of the CAR to the first antigen induces cytotoxicity of the T cell, and binding of the iCAR to the second antigen reduces the cytotoxicity of the T cell induced by the CAR.
17. The kit of claim 16 , wherein the kit further comprises written instructions for using said cell for the treatment of a subject having a neoplasm.
18. The kit of claim 16 , wherein the second antigen is selected from the group consisting of an Epithelial-mesenchymal transition (EMT) antigen, cytokeratin, human leukocyte antigens (HLAs), Opioid-binding protein/cell adhesion molecule (OPCML), HYAL2, Deleted in Colorectal Carcinoma (DCC), Scaffold/Matrix attachment region-binding protein 1 (SMAR1), CD33, CD38, and E-cadherin.