# US 11,306,289 B2 · Methods of using IL-21 for adoptive immunotherapy and identification of tumor antigens

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- Number: US 11,306,289 B2
- Document ID: US11306289B2
- Status: Granted
- Kind: B2
- Application number: 15/926,874
- Filed: 2018-03-20
- Published: 2019-10-17
- Granted: 2022-04-19
- Earliest claimed: 2004-11-24
- Assignees: Fred Hutchinson Cancer Research Center
- Applicants: Fred Hutchinson Cancer Research Center
- Inventors: Cassian Yee
- Web page: https://patentgrep.com/patent/US11306289B2

## CPC

- G01N 33/505: {involving T-cells}
- 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)
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/4269: {NY-ESO}
- A61K 40/4272: {Melan-A/MART}
- A61K 40/4273: {Glycoprotein 100 [Gp100]}
- A61P 35/00: Antineoplastic agents
- A61P 37/00: Drugs for immunological or allergic disorders
- C12N 5/0636: {T lymphocytes}
- C12N 5/0638: {Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]}
- G01N 33/5047: {Cells of the immune system}
- G01N 33/575: for cancer
- G01N 33/5758: {involving compounds serving as markers for tumours, cancers or neoplasias, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides or metabolites}
- G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells}
- 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}
- A61K 2239/38: characterised by the dose, timing or administration schedule
- A61K 2239/57: Skin; melanoma
- C12N 2501/23: Interleukins [IL]
- C12N 2501/2321: Interleukin-21 (IL-21)
- C12N 2502/1121: Dendritic cells
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## 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

### Claim 1

A method of preparing a T cell population comprising:
obtaining the T cell population from PBMCs;
activating the obtained T cell population,
transducing the activated T cell population with a viral vector comprising a nucleic acid encoding a protein,
expanding the transduced T cell population, and
wherein the activating, transducing, and expanding are performed in the presence of IL-21.

### Claim 2 (depends on 1)

The method of claim 1, wherein the PBMCs are obtained from a patient or a healthy donor.

### Claim 3 (depends on 1)

The method of claim 1, wherein the activating, transducing, and expanding are further performed in the presence of at least one cytokine selected from IL2, IL-12, and IL15.

### Claim 4 (depends on 1)

The method of claim 1, wherein the protein is a T cell receptor.

### Claim 5 (depends on 1)

The method of claim 1, further comprising restimulating the expanded transduced T cell population.

### Claim 6 (depends on 5)

The method of claim 5, wherein the restimulating is performed in the presence of a feeder cell and an anti-CD3 antibody.

### Claim 7 (depends on 6)

The method of claim 6, wherein the feeder cell is an irradiated allogenic feeder cell.

### Claim 8 (depends on 7)

The method of claim 7, wherein the feeder cell comprises a PBL and a LCL.

### Claim 9 (depends on 8)

The method of claim 8, further comprising culturing the restimulated cells in the presence of IL-2.