Methods for in vitro differentiation of TH-17+ cells

Patent metadata

Field	Value
Publication number	US-9498495-B2
Application number	US-201313850497-A
Country	US
Kind code	B2
Filing date	Mar 26, 2013
Priority date	Apr 15, 2008
Publication date	Nov 22, 2016
Grant date	Nov 22, 2016

Abstract

The present invention is directed to an in vitro method for promoting differentiation and proliferation of human T helper lymphocytes that express IL17 (Th-IL17+ cells). The instant method may be used to generate a population of human T helper lymphocytes that express IL17 (Th-IL17+ cells) in vitro. Methods for screening to identify agents capable of modulating Th-IL17+ cell differentiation are also encompassed by the present invention. Isolated, pure populations of homogeneous Th-IL17+ cells that do not express cellular markers characteristic of Th1, Th2, or Treg cells are also encompassed herein.

First claim

What is claimed is:

1. A purified, homogeneous population of antigen naive human Th-IL17+ cells, wherein the purified, homogeneous population comprises about or at least 10 9 antigen naive human Th-IL17+ cells and the purified, homogeneous population of antigen naive human Th-IL17+ cells does not include Th-IL17+ cells expressing cellular markers of Th1, Th2, or Treg cells.

2. The purified, homogeneous population of antigen naive human Th-IL 17+ cells of claim 1 , wherein the antigen naive human Th-IL17+ cells express at least one marker of human Th-IL17+ cells in addition to IL-17, wherein the at least one marker is IL-17F, IL-26, or RORC.

3. The purified, homogeneous population of antigen naive human Th-IL17+ cells of claim 1 , wherein the antigen naive human Th-IL17+ cells express at least one of the following cell surface markers or cell surface antigens comprising CCR6, IL23R, CD45RO+, CCR7, CCR5, or CXCR4.

4. The purified, homogeneous population of antigen naive human Th-IL17+ cells of claim 1 , wherein the antigen naive human Th-IL17+ cells do not express IFNγ, T-bet, IL4, IL13, GATA-3, or FoxP3.

5. A composition comprising the purified, homogeneous population of antigen naive human Th-IL17+ cells of claim 1 and a pharmaceutically acceptable carrier.

6. The composition of claim 5 , further comprising recombinant TGF-β.

7. A composition comprising a purified, homogeneous population of antigen naive human Th-IL17+ cells, wherein the purified, homogeneous population of antigen naive human Th-IL17+ cells does not include Th-IL17+ cells expressing cellular markers of Th1, Th2, or Treg cells; recombinant TGF-β; and serum-free culture medium.

8. The composition of claim 7 , wherein the composition further comprises IL-1β, and any one of IL-6, IL-21 or IL-23.

9. The A purified, homogeneous population of antigen naive human Th-IL17+ cells of claim 1 , wherein the purified, homogeneous population of antigen naive human Th-IL17+ cells is detectably labeled.

10. A composition comprising a purified homogeneous population of antigen naive human Th-IL17+ cells, wherein the purified, homogeneous population of antigen naive human Th-IL17+ cells does not include Th-IL17+ cells expressing cellular markers of Th1, Th2, or Treg cells, generated by a method comprising the steps of: isolating a population of antigen naive CD4+T cells from a human, wherein the antigen naive CD4+T cells are isolated from cord blood, buffy coats of adult humans, cell cultures comprising cells that express CD34 (CD34+ cells), or human embryonic stem cells; and incubating the population of antigen naive CD4+T cells in serum-free culture medium comprising TGF-β, IL-1β, and any one of IL-6, IL-21 or IL-23, wherein the incubating promotes differentiation of human Th-IL17+ cells.

11. A purified, homogeneous population of antigen naive human Th-IL17+ cells, wherein the purified, homogeneous population comprises about or at least 10 9 antigen naive human Th-IL17+ cells.

12. A composition comprising the purified, homogeneous population of antigen naive human Th-IL17+ cells of claim 11 and a pharmaceutically acceptable carrier.

Assignees

• Manel Nicolas
• Littman Dan R
• Univ New York

Inventors

• Manel Nicolas
• Littman Dan R

Classifications

• C12N2799/027

  where the vector is derived from a retrovirus · CPC title

• G01N33/505

  involving T-cells · CPC title

• C12N5/0636 Primary

  T lymphocytes · CPC title

• C12N2501/23

  Interleukins [IL] · CPC title

• C12N2501/15

  Transforming growth factor beta (TGF-β) · CPC title