Methods for isolating, culturing, and genetically engineering immune cell populations for adoptive therapy

Patent metadata

Field	Value
Publication number	US-12296010-B2
Application number	US-202418983137-A
Country	US
Kind code	B2
Filing date	Dec 16, 2024
Priority date	Apr 23, 2014
Publication date	May 13, 2025
Grant date	May 13, 2025

Abstract

The present disclosure relates in some aspects to methods, cells, and compositions for preparing cells and compositions for genetic engineering and cell therapy. Provided in some embodiments are streamlined cell preparation methods, e.g., for isolation, processing, incubation, and genetic engineering of cells and populations of cells. Also provided are cells and compositions produced by the methods and methods of their use. The cells can include immune cells, such as T cells, and generally include a plurality of isolated T cell populations or types. In some aspects, the methods are capable of preparing of a plurality of different cell populations for adoptive therapy using fewer steps and/or resources and/or reduced handling compared with other methods.

First claim

The invention claimed is:

1. A method of treating systemic lupus erythematosus (SLE) in a human subject, the method comprising intravenously administering a dose of genetically engineered CD4+ and CD8+ T cells to a human subject with SLE, wherein:
the CD4+ and CD8+ T cells of the dose are at a ratio, wherein the ratio is between at or about 5:1 and at or about 1:3;
the dose of genetically engineered CD4+ and CD8+ T cells is between at or about 5×10 6 to 500×10 6 cells; and
the CD4+ and CD8+ T cells are genetically engineered with a chimeric antigen receptor (CAR), wherein the CAR comprises an scFv that binds CD19, a transmembrane domain, and an intracellular signaling domain comprising a 4-1BB costimulatory signaling domain and a CD3zeta signaling domain.

2. A method of treating systemic lupus erythematosus (SLE) in a human subject, the method comprising intravenously administering a dose of genetically engineered CD4+ and CD8+ T cells to a human subject with SLE, wherein:
the CD4+ and CD8+ T cells of the dose are at a ratio, wherein the ratio is between at or about 5:1 and at or about 1:3;
the dose of genetically engineered CD4+ and CD8+ T cells is between at or about 5×10 6 to 500×10 6 cells; and
the CD4+ and CD8+ T cells are genetically engineered with a chimeric antigen receptor (CAR), wherein the CAR comprises an scFv that binds CD19, a transmembrane domain, and an intracellular signaling domain comprising a CD28 costimulatory signaling domain and a CD3zeta signaling domain.

3. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is between at or about 5×10 6 to 25×10 6 cells.

4. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is at or about 5×10 6 cells.

5. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is at or about 10×10 6 cells.

6. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 60×10 6 cells.

7. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 90×10 6 cells.

8. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 250×10 6 cells.

9. The method of claim 1 , wherein the dose of CD4+ and CD8+ T cells is between 30×10 6 cells and 90×10 6 cells.

10. The method of claim 1 , wherein the percentage of CD4+ and CD8+ T cells in the dose is at least about 90%.

11. The method of claim 8 , wherein the percentage of CD4+ and CD8+ T cells in the dose is at least about 90%.

12. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is between at or about 5×10 6 to 25×10 6 cells.

13. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is at or about 5×10 6 cells.

14. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is at or about 10×10 6 cells.

15. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 60×10 6 cells.

16. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 90×10 6 cells.

17. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is between 20×10 6 cells and 250×10 6 cells.

18. The method of claim 2 , wherein the dose of CD4+ and CD8+ T cells is between 30×10 6 cells and 90×10 6 cells.

19. The method of claim 2 , wherein the percentage of CD4+ and CD8+ T cells in the dose is at least about 90%.

20. The method of claim 17 , wherein the percentage of CD4+ and CD8+ T cells in the dose is at least about 90%.

Assignees

• Juno Therapeutics Inc

Inventors

• Ramsborg Chris
• Bonyhadi Mark L
• Chan Calvin
• Beauchesne Pascal

Classifications

• G01N2333/70517

  CD8 · CPC title

• G01N2333/70514

  CD4 · CPC title

• G01N33/56972

  White blood cells · CPC title

• C12M47/04 Primary

  Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04) · CPC title

• A61K2035/124

  the cells being hematopoietic, bone marrow derived or blood cells · CPC title