# US 2010/0167399 A1 · Method of generating regulatory t cells using cord blood and adult blood cd14+ monocyte cells

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- Number: US 2010/0167399 A1
- Document ID: US20100167399A1
- Status: Published application
- Kind: A1
- Application number: 12/644,140
- Filed: 2009-12-22
- Published: 2010-07-01
- Earliest claimed: 2008-12-29
- Assignees: Loyola University of Chicago
- Inventors: Makio Iwashima
- Web page: https://patentgrep.com/patent/US20100167399A1

## CPC

- C12N 5/0636: {T lymphocytes}
- A61K 2035/122: {for inducing tolerance or supression of immune responses}
- C12N 2502/11: blood or immune system cells
- 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)

## Abstract

A method of inducing stable and functional Tregs from human umbilical cord blood and adult blood without requiring the expansion of pre-existing Tregs. The method utilizes CD14+ monocyte cells present in or isolated from cord blood or adult blood to induce functional Tregs from T cells, and particularly T cells that express CD4+, which may also be obtained from cord blood or adult blood. The developed Tregs are long-lasting and maintain their suppressive functions.

## Claims

### Claim 1

A method of inducing functional regulatory T cells by stimulating T cells with CD14+ monocyte cells obtained from cord blood or adult blood.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the T cells are at least one of CD4+ T cells and CD8+ T cells.

### Claim 3 (depends on 2)

The method according to claim 2, wherein the T cells are CD4+CD25− T cells.

### Claim 4 (depends on 2)

The method according to claim 2, wherein the T cells are CD8+ T cells.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the T cells are present in whole lymphocyte/monocyte fraction during stimulation with the CD14+ monocyte cells.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the whole lymphocyte/monocyte fraction is obtained from cord blood or adult blood.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the T cells are purified from cord blood or adult blood prior to being stimulated with the CD14+ monocyte cells.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the CD14+ monocyte cells are cord blood CD14+ monocyte cells.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the CD14+ monocyte cells are adult peripheral blood CD14+ monocyte cells.

### Claim 10 (depends on 9)

The method according to claim 9, further comprising enriching the CD14+ monocyte cells prior to stimulation of the T cells to induce the functional regulatory T cells.

### Claim 11 (depends on 1)

The method according to claim 1, wherein the CD14+ monocyte cells are isolated from cord blood or adult blood prior to stimulation of the T cells to induce the functional regulatory T cells.

### Claim 12 (depends on 1)

The method according to claim 1, further comprising treating the CD14+ monocyte cells with at least one stimulator prior to or during the induction of the functional regulatory T cells.

### Claim 13 (depends on 12)

The method according to claim 12, wherein the stimulator is at least one chosen from the group consisting of allogenic MHC molecules, antigen peptides (both self and non-self), and antigenic proteins.

### Claim 14 (depends on 12)

The method according to claim 12, wherein the stimulator comprises anti-CD3 antibody.

### Claim 15 (depends on 1)

The method according to claim 1, wherein the functional regulatory T cells comprise Foxp3+ regulatory T cells.

### Claim 16 (depends on 1)

The method according to claim 1, wherein the regulatory T cells are induced by direct contact with the CD14+ monocyte cells.

### Claim 17 (depends on 1)

The method according to claim 1, wherein the regulatory T cells are induced by non-direct interaction with the CD14+ monocyte cells.

### Claim 18 (depends on 17)

The method according to claim 17, wherein the regulatory T cells are induced by a soluble factor or factors.

### Claim 19 (depends on 1)

The method according to claim 1, wherein the functional regulatory T cells are stable for at least three weeks.

### Claim 20 (depends on 1)

The method according to claim 1, wherein the induction of the functional regulatory T cells comprises activin C TGF-β signaling.