# US 2022/0275331 A1 · Selective expansion of different subpopulations of t cells by the alteration of cell surfacing signals and signal ratio

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- Number: US 2022/0275331 A1
- Document ID: US20220275331A1
- Status: Published application
- Kind: A1
- Application number: 17/578,172
- Filed: 2022-01-18
- Published: 2022-09-01
- Earliest claimed: 2015-10-28
- Assignees: Life Technologies Corporation
- Applicants: Life Technologies AS
- Inventors: Hilde ALMAASBAK; Tanja AARVAK; Kerstin BERNSTROEM; Oystein AAMELLEM
- Web page: https://patentgrep.com/patent/US20220275331A1

## CPC

- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- C12N 5/0637: {Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg}
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 1/18: for pancreatic disorders, e.g. pancreatic enzymes
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 17/00: Drugs for dermatological disorders
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 35/00: Antineoplastic agents
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- A61P 37/08: Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14)
- C12N 5/0636: {T lymphocytes}
- C12N 2501/51: B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand)
- C12N 2501/515: CD3, T-cell receptor complex
- C12N 2501/599: with CD designations not provided for elsewhere
- 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

This invention relates, inter alia, to compositions of expanded T cell populations, methods for the expansion of T cell populations and methods for using such populations of cells. In some aspects, the invention relates to compositions and methods for the selective expansion of T cell subpopulations present in mixed T cell populations, as well as T cell subpopulations produced by methods for the invention.

## Claims

### Claim at position 1

1-106. (canceled)

### Claim 107

A method for expanding regulatory T cells, the method comprising contacting the regulatory T cells with one more cytokines and stimulating the regulatory T cells with an anti-CD3 antibody and an anti-CD28 antibody, and
wherein the regulatory T cells expand from 500 fold to 1,000 fold after 15 days post-activation.

### Claim 108 (depends on 107)

The method of claim 107, wherein at least one of the one or more cytokines is Interleukin-2.

### Claim 109 (depends on 107)

The method of claim 107, wherein the anti-CD3 antibody and the anti-CD28 antibody are bound to a solid support.

### Claim 110 (depends on 109)

The method of claim 109, wherein the solid support is a bead.

### Claim 111 (depends on 109)

The method of claim 109, wherein the anti-CD3 antibody and the anti-CD28 antibody are bound to the same solid support.

### Claim 112 (depends on 107)

The method of claim 107, wherein the regulatory T cells are separated from T cells of other subtypes prior to contact with the one more cytokines and stimulation with the anti-CD3 antibody and the anti-CD28 antibody.

### Claim 113

A method for expanding antigen experienced T cells, the method comprising activating the antigen experienced T cells by contacting the antigen experienced T cells with at one or more cytokines and with an antibody combination selected from the group consisting of:
(a) an anti-CD3 antibody and an anti-CD28 antibody,
(b) an anti-CD3 antibody and an anti-CD137 antibody, and
(c) an anti-CD3 antibody, an anti-CD28 antibody and an anti-CD137 antibody, and
wherein the antigen experienced T cells expand at least 50 fold after 10 days post-activation.

### Claim 114 (depends on 113)

The method of claim 113, wherein at least one of the one or more cytokines is Interleukin-2.

### Claim 115 (depends on 113)

The method of claim 113, wherein the anti-CD3 antibody, the anti-CD28 antibody, and/or the anti-CD137 antibody are bound to a solid support.

### Claim 116 (depends on 115)

The method of claim 115, wherein the solid support is a bead.

### Claim 117 (depends on 115)

The method of claim 115, wherein the anti-CD3 antibody, the anti-CD28 antibody, and/or the anti-CD137 antibody are bound to the same solid support.

### Claim 118

A method for increasing the percentage of Th17 cells present in a T cell population, the method comprising contacting a population of T cells with at least one cytokine and with an antibody combination selected from the group consisting of:
(a) an anti-CD3 antibody and an anti-CD5 antibody,
(b) an anti-CD3 and an anti-CD28 antibody,
(c) an anti-CD3 and an anti-CD238 antibody,
(d) an anti-CD3 antibody, an anti-CD28 antibody and an anti-CD238 antibody,
(e) an anti-CD3 antibody, an anti-CD5 antibody and an anti-CD238 antibody,
(f) an anti-CD3 antibody, an anti-CD5 antibody and an anti-CD28 antibody, and
(g) an anti-CD3 antibody, an anti-CD5 antibody, an anti-CD28 antibody and an anti-CD238 antibody, and
wherein the T cell surface receptor agonists remain in contact with the population of T cells for a period of 1 to 5 days.

### Claim 119 (depends on 118)

The method of claim 118, wherein the Th17 cells expand up to 200 fold after 13 days in culture.

### Claim 120 (depends on 118)

The method of claim 118, wherein at least one of the one or more cytokines is Interleukin-1 beta.

### Claim 121 (depends on 118)

The method of claim 118, wherein the anti-CD3 antibody, the anti-CD5 antibody, the anti-CD28 antibody, and/or the anti-CD238 antibody are bound to a solid support.

### Claim 122 (depends on 121)

The method of claim 121, wherein the solid support is a bead.

### Claim 123 (depends on 121)

The method of claim 121, wherein the anti-CD3 antibody, the anti-CD5 antibody, the anti-CD28 antibody, and/or the anti-CD238 antibody are bound to the same solid support.

### Claim 124 (depends on 118)

The method of claim 118, wherein the Th17 cells are separated from T cells of other subtypes prior to contact with the at least one cytokine and antibody combination.

### Claim 125 (depends on 124)

The method of claim 124, wherein the Th17 cells are separated from T cells of other subtypes by flow cytometry.

### Claim 126 (depends on 118)

The method of claim 118, wherein the population of T cells is contacted with an aryl hydrocarbon receptor agonists.