# US 12,180,458 B2 · Method for detecting CD4+CD25+glycoprotein A33(GPA33)high and/or CD4+CD127-GPA33high T cells

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- Number: US 12,180,458 B2
- Document ID: US12180458B2
- Status: Granted
- Kind: B2
- Application number: 16/499,599
- Filed: 2018-03-29
- Published: 2020-04-02
- Granted: 2024-12-31
- Earliest claimed: 2017-03-29
- Assignees: Stichting Sanquin Bloedvoorziening; Stichting Het Nederlands Kanker Institmit-Antoni van Leeuwenhoek Ziekenhuis
- Applicants: Stichting Sanquin Bloedvoorziening; Stichting Het Nederlands Kanker Instituut-Antoni van Leeuwenhoek Ziekenhuis
- Inventors: Derk Amsen; Eloy Cuadrado Godia; Jannetje Geertruida Borst
- Web page: https://patentgrep.com/patent/US12180458B2

## CPC

- C12N 5/0637: {Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg}
- A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
- A61K 40/22: Immunosuppressive or immunotolerising
- A61K 40/416: {Antigens related to auto-immune diseases; Preparations to induce self-tolerance}
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- G01N 33/505: {involving T-cells}
- A61K 2035/122: {for inducing tolerance or supression of immune responses}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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
- 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)

## Abstract

The invention relates to isolated populations of CD4+GPA33high regulatory T cells, to methods for detecting and/or isolating CD4+GPA33high cells, methods for enriching for CD4+GPA33high cells and to uses of CD4+GPA33high cells and populations of such cells. The invention further relates to methods for classifying an individual suffering from cancer and typing a tumor sample as well as for determining the etiology of autoimmune diseases.

## Claims

### Claim 1

A method for detecting CD4+CD25+ glycoprotein A33(GPA33)high and/or CD4+CD127−GPA33high T cells in a sample comprising:
obtaining a cell sample; and
detecting CD4+CD25+GPA33high and/or CD4+CD127−GPA33high T cells by contacting the cell sample with an anti-CD4 antibody or antigen-binding fragment thereof and an anti-GPA33 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD4 antibody or antigen-binding fragment thereof and the anti-GPA33 antibody or antigen-binding fragment thereof, and contacting the cell sample with an anti-CD25 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD25 antibody or antigen-binding fragment thereof to detect CD4+CD25+GPA33high T cells, and/or contacting the cell sample with an anti-CD127 antibody or antigen-binding fragment thereof and detecting binding or the lack thereof between cells in the sample and the anti-CD127 antibody or antigen-binding fragment thereof to detect CD4+CD127−GPA33high T cells,
wherein GPA33high is defined as the expression level of GPA33 that is higher than the average level of expression of GPA33 on CD4+ T cells in the cell sample.

### Claim 2 (depends on 1)

A method according to claim 1, wherein said cells are obtained from a tumor or a tumor sample.

### Claim 3 (depends on 1)

The method according to claim 1 further comprising quantifying the level of CD4+CD25+GPA33high and/or CD4+CD127−GPA33high cells in said sample.

### Claim 4 (depends on 3)

The method according to claim 3, comprising quantifying the level of CD4+CD25+GPA33high cells.

### Claim 5 (depends on 1)

The method according to claim 1, further comprising isolating said CD4+CD25+GPA33high and/or CD4+CD127−GPA33high T cells.

### Claim 6 (depends on 5)

The method according to claim 5, further comprising culturing the isolated cells in the presence of one or more factors promoting proliferation, activation and/or growth of said cells.

### Claim 7 (depends on 1)

The method according to claim 1 wherein CD4+CD25+GPA33high T cells are detected.

### Claim 8 (depends on 1)

The method according to claim 1 wherein CD4+CD25+CD127−GPA33high T cells are detected.

### Claim 9 (depends on 1)

The method according to claim 1 wherein GPA33high refers to a level of expression that is at least 2-fold the average level of expression of GPA33 on CD4+ T cells in the cell sample.

### Claim 10

A method for detecting CD4+CD25+ glycoprotein A33(GPA33)high and/or CD4+CD127−GPA33high T cells in a sample, comprising:
obtaining a cell sample; and
detecting CD4+CD25+GPA33high and/or CD4+CD127−GPA33high T cells by contacting the cell sample with an anti-CD4 antibody or antigen-binding fragment thereof and an anti-GPA33 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD4 antibody or antigen-binding fragment thereof and the anti-GPA33 antibody or antigen-binding fragment thereof, and contacting the cell sample with an anti-CD25 antibody or antigen-binding fragment thereof and detecting binding between cells in the sample and the anti-CD25 antibody or antigen-binding fragment thereof to detect CD4+CD25+GPA33high T cells, and/or contacting the cell sample with an anti-CD127 antibody or antigen-binding fragment thereof and detecting binding or the lack thereof between cells in the sample and the anti-CD127 antibody or antigen-binding fragment thereof to detect CD4+CD127−GPA33high T cells, wherein GPA33high is defined as the expression level of GPA33 at which 75% of CD4+CD25+CD45RA+ T cells are GPA33+.