Car expression vector and car-expressing T cells
An object of the present invention is to provide CAR-expressing T cells that coexpress a chimeric antigen receptor (CAR) and a T cell immune function-enhancing factor and have a high immunity-inducing effect and antitumor activity, and to provide a CAR expression vector for the preparation of the CAR-expressing T cells. A CAR expression vector comprises a nucleic acid encoding a chimeric antigen receptor (CAR) and a nucleic acid encoding a T cell immune function-enhancing factor, wherein the nucleic acid encoding an immune function-enhancing factor is a nucleic acid encoding interleukin-7 and a nucleic acid encoding CCL19, a nucleic acid encoding a dominant negative mutant of SHP-1, or a nucleic acid encoding a dominant negative mutant of SHP-2, or a CAR-expressing T cell introduced with the CAR expression vector are prepared.
1. A CAR expression vector comprising a nucleic acid encoding a chimeric antigen receptor (CAR) and a nucleic acid encoding a T cell immune function-enhancing factor, wherein the nucleic acid encoding the T cell immune function-enhancing factor consists of a nucleic acid encoding interleukin-7 and a nucleic acid encoding CCL19.
2. The CAR expression vector according to claim 1 , wherein the nucleic acid encoding the CAR and the nucleic acid encoding the T cell immune function-enhancing factor are linked via a sequence encoding a self-cleaving peptide.
3. The CAR expression vector according to claim 1 , wherein the nucleic acid encoding interleukin-7 and the nucleic acid encoding CCL19 are linked via a sequence encoding a self-cleaving peptide.
4. The CAR expression vector according to claim 1 , wherein the nucleic acid encoding the CAR contains a nucleic acid encoding a polypeptide of a single chain antibody that recognizes FITC or CD20.
5. The CAR expression vector according to claim 1 , wherein the nucleic acid encoding the CAR contains a nucleic acid encoding a polypeptide of a CD8 transmembrane region.
6. The CAR expression vector according to claim 1 , wherein the nucleic acid encoding the CAR contains nucleic acids encoding polypeptides of a CD28 intracellular region, a 4-1BB intracellular region, and a CD3 intracellular region.
7. An isolated CAR-expressing T cell, or a culture comprising the isolated CAR-expressing T cell, said isolated T cell prepared by introducing the CAR expression vector according to claim 1 , and expressing the CAR, interleukin-7, and CCL19.
8. An isolated CAR-expressing T cell, or a culture comprising the isolated CAR-expressing T cell, said isolated T cell prepared by introducing the CAR expression vector according to claim 2 , and expressing the CAR, interleukin-7, and CCL19.
9. An isolated CAR-expressing T cell, or a culture comprising the isolated CAR-expressing T cell, said isolated T cell prepared by introducing the CAR expression vector according to claim 3 , and expressing the CAR, interleukin-7, and CCL19.
10. An isolated CAR-expressing T cell, or a culture comprising the isolated CAR-expressing T cell, said isolated T cell prepared by introducing the CAR expression vector according to claim 4 , and expressing the CAR, interleukin-7, and CCL19.
11. An isolated CAR-expressing T cell, or a culture comprising the isolated CAR-expressing T cell, said isolated T cell prepared by introducing a CAR expression vector that expresses a nucleic acid encoding a CAR and a nucleic acid encoding a first T cell immune function-enhancing factor, said first T cell immune function-enhancing factor consisting of interleukin-7, and a CAR expression vector that expresses a nucleic acid encoding a CAR and a nucleic acid encoding a second T cell immune function-enhancing factor, said second T cell immune function-enhancing factor consisting of CCL19; and expressing the CAR, interleukin-7, and CCL19.
12. An anticancer agent comprising the isolated CAR-expressing T cell according to claim 7 and a pharmaceutically acceptable additive.
13. An anticancer agent comprising the isolated CAR-expressing T cell according to claim 8 and a pharmaceutically acceptable additive.
14. An anticancer agent comprising the isolated CAR-expressing T cell according to claim 9 and a pharmaceutically acceptable additive.
15. An anticancer agent comprising the isolated CAR-expressing T cell according to claim 10 and a pharmaceutically acceptable additive.
16. An anticancer agent comprising the isolated CAR-expressing T cell according to claim 11 and a pharmaceutically acceptable additive.
17. The CAR expression vector of claim 1 wherein said interleukin-7 is human interleukin-7 and said CCL19 is human CCL19.
18. The isolated CAR-expressing T cell or cell culture of claim 7 , wherein said interleukin-7 is human interleukin-7 and said CCL19 is human CCL19.
19. The isolated CAR-expressing T cell or cell culture of claim 11 , wherein said interleukin-7 is human interleukin-7 and said CCL19 is human CCL19.