Chimeric receptors with 4-1BB stimulatory signaling domain
The present invention relates to a chimeric receptor capable of signaling both a primary and a co-stimulatory pathway, thus allowing activation of the co-stimulatory pathway without binding to the natural ligand. The cytoplasmic domain of the receptor contains a portion of the 4-1BB signaling domain. Embodiments of the invention relate to polynucleotides that encode the receptor, vectors and host cells encoding a chimeric receptor, particularly including T cells and natural killer (NK) cells and methods of use.
1. A chimeric antigen receptor comprising: (a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain; (b) a transmembrane domain; and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain wherein the CD3ζ signaling domain comprises the amino acid sequence of SEQ ID No. 18.
2. The chimeric antigen receptor of claim 1 , wherein the extracellular binding domain comprises the amino acid sequence of SEQ. ID. No. 10.
3. The chimeric antigen receptor of claim 1 wherein the transmembrane domain comprises the amino acid sequence of SEQ ID No. 14.
4. The chimeric antigen receptor of claim 1 wherein the 4-1BB signaling domain comprises the amino acid sequence of SEQ ID No. 16.
5. The chimeric antigen receptor of claim 1 comprising the amino acid sequence of SEQ.ID. No. 6.
6. A method of enhancing a T lymphocyte or an NK cell activity in a mammal comprising introducing into the mammal a T lymphocyte or NK cell, which T lymphocyte or NK cell comprises a chimeric receptor comprising: (a) an extracellular ligand- binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
7. A method for treating a mammal suffering from cancer comprising introducing into the mammal a T lymphocyte or an NK cell, which T lymphocyte or NK cell comprises a chimeric receptor comprising: (a) an extracellular ligand-binding domain comprising an anti-CD19 scFv domain; (b) a hinge region and transmembrane domain of CD8a; and (c) a cytoplasmic domain comprising a signaling domain of 4-1BB and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
8. A method for stimulating a T cell-mediated immune response to a target cell population or tissue in a mammal comprising administering to a mammal an effective amount of a cell genetically modified to express a chimeric antigen receptor (CAR), said CAR comprising: a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
9. A method of providing an anti-tumor immunity in a mammal, the method comprising administering to the mammal an effective amount of a cell genetically modified to express a CAR, said CAR comprising: a) an extracellular ligand-binding domain comprising an anti-CD19 single chain variable fragment (scFv) domain, (b) a hinge and transmembrane domain, and (c) a cytoplasmic domain comprising a 4-1BB signaling domain and a CD3ζ signaling domain comprising the amino acid sequence of SEQ ID No. 18.
10. The method of any one of claims 7 to 9 , wherein the mammal is suffering from B lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, B-cell non-Hodgkin's lymphoma, leukemia and lymphoma, acute lymphoblastic leukemia, Hodgkin's lymphoma, or childhood acute lymphoblastic leukemia.