METHODS OF USING IL-21 FOR ADOPTIVE IMMUNOTHERAPY AND IDENTIFICATION OF TUMOR ANTIGENS

• US 20190316088A1
• Filed: 03/20/2018
• Published: 10/17/2019
• Est. Priority Date: 11/24/2004
• Status: Active Grant

• Current Assignee: Fred Hutchinson Cancer Center
• Original Assignee: Fred Hutchinson Cancer Research Center (Fred Hutchinson Cancer Research Center|Argus Genetics|Mars)
• Inventors: YEE, Cassian
• Granted Patent: US 11,306,289 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2239/57 Skin; melanoma
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/464488 NY-ESO
• CPC Class Codes: A61K 39/464491 Melan-A/MART
• CPC Class Codes: A61K 39/464492 Glycoprotein 100 [Gp100]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/2321 Interleukin-21 (IL-21)
• CPC Class Codes: C12N 2502/1121 Dendritic cells
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12N 5/0638 Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]
• CPC Class Codes: G01N 33/5047 Cells of the immune system
• CPC Class Codes: G01N 33/505 involving T-cells
• CPC Class Codes: G01N 33/574 for cancer
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors

Abstract

Methods for preparing ex vivo T cell cultures using IL-21 compositions for use in adoptive immunotherapy are described. Addition of IL-21 to cultures of non-terminally differentiated T cells population, either isolated or present in peripheral blood mononuclear cells are exposed to one or more tumor antigens, and in the presence of IL-21 compositions and antigen presenting cells (APCs), the resulting T cell population has an enhanced antigen-specificity, and can be reintroduced into the patient. Methods are also disclosed for identifying tumor antigens by culturing T cell populations exposed to IL-21 compositions and APCs in the presence of tumor material.

Claims

1. A method for identifying a tumor antigen comprising:

co-culturing tumor material isolated from a subject with peripheral blood mononuclear cells (PBMCs) in the presence of an IL-21 composition and antigen presenting cells (APCs);

isolating a T cell population from the culture;

cloning individual T cells from the T cell population; and

characterizing T cell clones for antigen-specificity.

2. The method of claim 1, wherein the T cell population is a T cell population that is non-terminally differentiated.

3. The method of claims 1, wherein the PBMCs or T cell population is an autologous cell population.

4. The method of claim 1, wherein the tumor material comprises total RNA, lysed tumor cells or apoptotic bodies.

5. The method of claim 2, wherein the isolated T cell population does not include CD4+ cells.

6. The method of claim 1, wherein co-culturing is in the presence of the IL-21 composition and one or more additional cytokines.

7. A method of preparing a naive CD8+ human T cell population for in use adoptive immunotherapy comprising:

isolating peripheral blood mononuclear cells (PBMCs) from a biological sample;

identifying PBMCs having a histocompatible phenotype to a subject having a tumor;

isolating a naï

ve CD8+ human T cell population;

co-culturing tumor material isolated from the subject with the naï

ve CD8+ human T cell population in the presence of an IL-21 composition and antigen presenting cells (APCs); and

expanding the cells in culture.

8. The method of claim 7, wherein the PBMCs are autologous.

9. The method of claim 7, wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched.

10. A method of preparing a naï

ve CD8+ human T cell population for use in adoptive immunotherapy comprising;

isolating a biological sample containing T cells;

identifying a T cell population having a histocompatible phenotype to a subject having a tumor;

isolating a naï

ve CD8+ human T cell population;

co-culturing tumor material from the subject with the naï

ve CD8+ human T cell population in the presence of an IL-21 composition and APCs; and

expanding these cells in culture.

11. The method of claim 10, wherein the T cell population is autologous.

12. The method of claims 7, 10, or 16 wherein the tumor material comprises total RNA, lysed tumor cells or apoptotic bodies.

13. The method of claim 10, wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched.

14. A method for ex vivo expansion of antigen-specific cytotoxic T cells comprising:

isolating a biological sample containing T cells;

identifying a T cell population having a histocompatible phenotype to a subject;

co-culturing antigenic material from the subject with the T cell population in the presence of an IL-21 composition;

expanding the cells in culture; and

and reintroducing the cells back into the subject.

15. The method of claim 14, wherein after the step of co-culturing the tumor material and with the T cell population, the T cells are enriched.

16. A method of preparing a non-terminally differentiated CD8+ human T cell population for use in adoptive immunotherapy comprising:

isolating a biological sample containing T cells;

identifying a T cell population having a histocompatible phenotype to a subject having a tumor;

isolating a non-terminally differentiated CD8+ human T cell population;

co-culturing tumor material from the subject with the non-terminally differentiated CD8+ human T cell population in the presence of an IL-21 composition and APCs; and

expanding these cells in culture.

17. The method of claim 16, wherein the T cell population is autologous.