Chimeric antigen receptors targeting CD-19
The invention is directed to a chimeric antigen receptor (CAR) directed against CD19, which comprises an amino acid sequence of any one of SEQ ID NO: 1-SEQ ID NO: 13. The invention also provides T-cells expressing the CAR and methods for destroying malignant B-cells.
1. A chimeric antigen receptor (CAR) directed against CD19, which comprises an amino acid sequence of SEQ ID) NO: 1, SEQ ID) NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13.
2. A nucleic acid encoding a chimeric antigen receptor(CAR) directed against CD19, the CAR comprises an amino acid sequence of SEQ ID) NO: 1, SEQ ID) NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, or SEQ ID NO: 13.
3. The nucleic acid of claim 2 which is present within a vector.
4. The vector of claim 3 which is present within a T-cell or NK cell.
5. A method of destroying malignant B-cells, which method comprises contacting one or more vector-containing T-cells or NK cells of claim 4 with a population of malignant B-cells that express CD19, whereby the CAR binds to CD19 on the malignant B-cells and the malignant B-cells are destroyed.
6. The method of claim 5 , wherein the malignant B-cells are lymphoma cells.
7. The method of claim 5 , wherein the malignant B-cells are leukemia cells.
8. The method of claim 5 , wherein the malignant B-cells are in a human.
9. The method of claim 5 , wherein the malignant B-cells are in vitro.
10. A chimeric antigen receptor (CAR) comprising the amino acid sequence of each of the following that are present in SEQ ID NO: 4 or SEQ ID NO: 9;
(i) the extracellular spacer,
(ii) the transmembrane domain derived from a human CD8α molecule, and
(iii) the intracellular T-cell signaling domains derived from
(a) one or both of a human CD28 molecule and a human CD27 molecule, and
(b) a human CD3 ζmolecule.
11. A chimeric antigen receptor (CAR) comprising the amino acid sequence of each of the following that are present in SEQ ID NO: 10 or SEQ ID NO: 11;
(i) the extracellular spacer,
(ii) the transmembrane domain derived from a human CD8α , molecule, and
(iii) the intracellular T-cell signaling domains derived from
(a) one or both of a human CD28 molecule and a human CD27 molecule, and
(b) the gamma chain of FcεRI.
12. A chimeric antigen receptor (CAR) comprising the amino acid sequence of each of the following that are present in SEQ ID NO: 12 or SEQ ID NO: 13;
(i) the extracellular spacer,
(ii) the transmembrane domain derived from a human CD8α molecule, and
(iii) the intracellular T-cell signaling domains derived from a human CD28molecule and the gamma chain of FcεRI.
13. A method of preventing or treating a B-cell malignancy in a subject, the method comprising administering to the subject an effective amount of one or more vector-containing T-cell or NK cell of claim 4 .
14. (A method of making a cell expressing a chimeric antigen receptor (CAR), the method comprising introducing a vector encoding the CAR of claim 1 into a T-cell or NK cell.
15. A method of making a cell expressing a chimeric antigen receptor (CAR), the method comprising introducing a vector encoding the CAR of claim 10 into a T-cell or NK cell.
16. A method of making a cell expressing a chimeric antigen receptor (CAR), the method comprising introducing a vector encoding the CAR of claim 11 into a T-cell or NK cell.
17. A method of making a cell expressing a chimeric antigen receptor (CAR), the method comprising introducing a vector encoding the CAR of chum 12 into a T-cell or NK cell.
18. The CAR of claim 1 , wherein the CAR comprises SEQ ID NO: 10.