# US 11,993,788 B2 · Composition comprising B regulatory cells

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- Number: US 11,993,788 B2
- Document ID: US11993788B2
- Status: Granted
- Kind: B2
- Application number: 16/610,534
- Filed: 2018-05-03
- Published: 2020-03-19
- Granted: 2024-05-28
- Earliest claimed: 2017-05-05
- Assignees: Oxford University Innovation Limited
- Applicants: Oxford University Innovation Limited
- Inventors: Kathryn J. Wood; Sushma Shankar
- Web page: https://patentgrep.com/patent/US11993788B2

## CPC

- C07K 16/2803: {against the immunoglobulin superfamily}
- A61K 40/13: B-cells
- A61K 40/22: Immunosuppressive or immunotolerising
- A61K 40/24: Antigen-presenting cells [APC]
- A61K 40/416: {Antigens related to auto-immune diseases; Preparations to induce self-tolerance}
- A61K 40/418: {Antigens related to induction of tolerance to non-self}
- A61K 40/42: Cancer antigens
- C07K 16/2866: {against receptors for cytokines, lymphokines, interferons}
- C07K 16/2875: {against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 (against NGF C07K16/22, against TNF C07K16/241)}
- C07K 16/2878: {against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95}
- C07K 16/2887: {against CD20}
- C12N 5/0635: {B lymphocytes}
- G01N 33/5091: {for testing the pathological state of an organism}
- A61K 2239/57: Skin; melanoma
- C07K 2317/31: multispecific
- C12N 2501/2302: Interleukin-2 (IL-2)
- C12N 2501/2304: Interleukin-4 (IL-4)
- C12N 2501/231: Interleukin-10 (IL-10)
- C12N 2501/2321: Interleukin-21 (IL-21)
- C12N 2501/52: CD40, CD40-ligand (CD154)
- C12N 2501/599: with CD designations not provided for elsewhere
- C12N 2502/09: epidermal cells, skin cells, oral mucosa cells
- G01N 2800/24: Immunology or allergic disorders
- G01N 2800/26: Infectious diseases, e.g. generalised sepsis
- G01N 2800/50: Determining the risk of developing a disease
- G01N 2800/52: Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
- G01N 2800/56: Staging of a disease; Further complications associated with the disease
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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 present invention relates to an expanded population of human Breg cells having the phenotype CD19+CD73−CD71+CD25+TIM-1+ and methods for producing the cell population of the invention. The invention also relates to pharmaceutical compositions comprising the cell populations of the invention and their use in the treatment of immune-mediated disorders.

## Claims

### Claim 1

An isolated population of cells, wherein the population comprises human Breg cells, having the phenotype CD19+CD73−CD71+CD25+TIM-1+, and wherein the human Breg cells additionally have the phenotype CD154+.

### Claim 2 (depends on 1)

The isolated cell population of claim 1, wherein at least 50% of the human Breg cells in the cell population express CD19 and at least 50% of the human Breg cells in the cell population do not express CD73 and at least 50% of the human Breg cells in the cell population express CD71 and at least 50% of the human Breg cells in the cell population express CD25 and at least 50% of the human Breg cells in the cell population express TIM-1.

### Claim 3 (depends on 1)

The isolated cell population of claim 1, wherein the human Breg cells additionally have a phenotype selected from the group consisting of IL-10R+ and CD39+.

### Claim 4 (depends on 1)

The isolated cell population of claim 1, wherein the human Breg cells additionally have a phenotype selected from the group consisting of CD5−, CD1d−, CD24−, CD27−, CD21− and CD38−.

### Claim 5 (depends on 1)

The isolated cell population of claim 1, wherein less than 30% of the human Breg cells in the cell population express IL-10.

### Claim 6 (depends on 1)

The isolated cell population of claim 1, wherein at least 50% of the cells in the population are human Breg cells having the phenotype CD19+CD73−CD71+CD25+TIM-1+ or CD19+CD73−CD71+CD25TIM-1+CD154+.

### Claim 7 (depends on 1)

A pharmaceutical composition comprising the isolated population of cells according to claim 1 and a pharmaceutically acceptable carrier.

### Claim 8 (depends on 7)

The pharmaceutical composition of claim 7 wherein the isolated population of cells comprises at least 50% of the composition.

### Claim 9

A method for producing a human Breg cell population comprising:
isolating human CD19+ B cells;
culturing said CD19+ B cells in the presence of CD154, or peptide fragments thereof, and/or cells expressing CD154, and at least one cytokine or growth factor for at least 3 days;
harvesting the Breg cells so produced.

### Claim 10 (depends on 9)

The method according to claim 9, wherein culturing said CD19+ B cells in the presence of a CD40 agonist comprises culturing the CD19+ B cells in the presence of cells expressing CD154 and wherein the cells expressing CD154 and the CD19+ B cells are cultured at a ratio of 1:1.

### Claim 11 (depends on 9)

The method according to claim 9, wherein the at least one cytokine is selected from the group consisting of IL-2, IL-4 and IL-10, and combinations thereof.

### Claim 12 (depends on 9)

The method according to claim 9, wherein the at least one cytokine comprises IL-21; or (ii) wherein IL-21 is not added during culturing of the human CD19+ B cells.

### Claim 13 (depends on 9)

The method according to claim 9, wherein the human CD19+ B cells are cultured for between 3-60 days, optionally 3-7 days.

### Claim 14 (depends on 9)

The method of claim 9, wherein the human Breg cells are expanded by at least 200 fold.

### Claim 15 (depends on 9)

The method according to claim 9, wherein the human CD19+ B cells are isolated from a patient sample.

### Claim 16 (depends on 9)

The method according to claim 9, wherein the human CD19+ B cells are isolated from a patient sample, and wherein the patient sample is a blood sample.

### Claim 17 (depends on 9)

The method according to claim 9, wherein the Breg cells produced have the phenotype CD19+CD73−CD71+CD25+TIM-1+ or CD19+CD73−CD71+CD25+TIM-1+CD154+.

### Claim 18 (depends on 9)

The method according to claim 9, wherein the harvested cells comprise at least 80% human Breg cells.

### Claim 19

An expanded human Breg cell population obtained by
isolating human CD19+ B cells,
culturing said CD19+ B cells in the presence of cells expressing CD154 at a ratio of 1:1, and at least one cytokine or growth factor for at least 3 days, and
harvesting the Breg cells so produced.

### Claim 20 (depends on 9)

The method according to claim 9 further comprising genetically modifying said human CD19+ B cells prior to expansion, to express a target antigen, wherein the target antigen is associated with an immune-mediated disorder.