T cells for expression of chimeric antigen receptors and other receptors

Patent metadata

Field	Value
Publication number	US-12115189-B2
Application number	US-201615746212-A
Country	US
Kind code	B2
Filing date	Jul 21, 2016
Priority date	Jul 21, 2015
Publication date	Oct 15, 2024
Grant date	Oct 15, 2024

Abstract

Methods for preparing T cell populations useful for a variety of purposes requiring a highly active, long-lived T cell population. The T cell populations are enriched for: naive T cells (TN), memory stem cells (TSCM) and central memory T cells (TCM). These cell populations can be derived from peripheral blood mononuclear cells (PBMC) by both: 1) depleting unwanted cell populations such as CD14 expressing myeloid cells and CD25 expressing cells; and 2) enriching for CD62L expressing memory and naive T cells.

First claim

What is claimed is:

1. A method for preparing a population of human cells comprising T cells, wherein the T cells comprise central memory T cells, memory stem T cells, and naïve T cells, wherein greater than 40% of the cells are CD45RA+ and greater than 70% are CD62L+, comprising;
(a) providing sample of human PBMC;
(b) treating the sample of human PBMC to deplete cells expressing CD25 and cells expressing CD14 to prepare a depleted cell population; and
(c) treating the depleted cell population to enrich for cells expressing CD62L, thereby preparing a population of human cells comprising T cells, wherein the T cells comprise central memory T cells; memory stem T cells, and naïve T cells, wherein greater than 40% of the cells are CD45RA+ and greater than 70% are CD62L+,
wherein the method does not comprise a step of depleting cells expressing CD45RA.

2. The method of claim 1 further comprising activating the population of human cells comprising T cells, wherein the T cells comprise central memory T cells; memory stem T cells, and naïve T cells, wherein greater than 40% of the cells are CD45RA+ and greater than 70% are CD62L+ and transducing or transfecting the activated cells with a recombinant nucleic acid molecule to provide a population of T cells comprising T cells harboring a recombinant nucleic acid molecule.

3. The method of claim 2 further comprising culturing the population of human cells comprising T cells harboring a recombinant nucleic acid molecule.

4. The method of claim 3 , wherein the culturing step comprises culturing in the presence of exogenous IL-2 and exogenous IL-15.

5. The method of claim 2 , wherein the activating step comprises exposing the cells to an anti-CD3 antibody and an anti-CD28 antibody.

6. The method of claim 2 , wherein the recombinant nucleic acid molecule is a viral vector.

7. The method of claim 6 , wherein the viral vector comprises a nucleotide sequence encoding a chimeric antigen receptor or a T cell receptor.

Assignees

• Hope City

Inventors

• Brown Christine E
• Forman Stephen J

Classifications

• A61K40/4217

  Receptors for interleukins [IL] · CPC title

• A61K40/4211

  CD19 or B4 · CPC title

• A61K40/4204

  Epidermal growth factor receptors [EGFR] · CPC title

• A61K40/46

  Viral antigens · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title