US 9,809,797 B2
Enhanced generation of cytotoxic T-lymphocytes by IL-21 mediated FOXP3 suppression
Cassian Yee, Seattle, WA (US); and Yongqing Li, Shoreline, WA (US)
Assigned to NATIONAL INSTITUTES OF HEALTH (NIH) U.S. DEPT. OF HEALTH AND HUMAN SERVICES (DHHS) DIVISION OF EXTRAMURAL INVENTIONS AND TECHNOLOGY, Washington, DC (US)
Appl. No. 12/677,035
Filed by Cassian Yee, Seattle, WA (US); and Yongqing Li, Shoreline, WA (US)
PCT Filed Sep. 25, 2008, PCT No. PCT/US2008/011088
§ 371(c)(1), (2), (4) Date Apr. 8, 2010,
PCT Pub. No. WO2009/045308, PCT Pub. Date Apr. 9, 2009.
Claims priority of provisional application 60/977,150, filed on Oct. 3, 2007.
Prior Publication US 2010/0330056 A1, Dec. 30, 2010
Int. Cl. C12N 5/0783 (2010.01); A61K 31/41 (2006.01); A61K 39/00 (2006.01); A61K 35/17 (2015.01); A61K 35/12 (2015.01)
CPC C12N 5/0638 (2013.01) [A61K 31/41 (2013.01); A61K 35/17 (2013.01); A61K 39/00 (2013.01); C12N 5/0636 (2013.01); A61K 35/12 (2013.01); A61K 2035/124 (2013.01); A61K 2039/51 (2013.01); A61K 2039/5154 (2013.01); C12N 2501/23 (2013.01); C12N 2501/2321 (2013.01); C12N 2501/998 (2013.01)] 7 Claims
 
1. A pharmaceutical formulation comprising a single T cell population type, wherein said T cell population consists of CD8+CD25 antigen-specific human cytotoxic T lymphocyte (CTL) cells specific for an antigen, said formulation being prepared by a process comprising:
(a) sorting a lymphocyte subpopulation depleted of CD25+ cells from a first lymphocyte population of peripheral blood mononuclear cells (PBMC) to produce a CD8+CD25 subpopulation;
(b) promoting the production of antigen-specific CTL cells in said subpopulation by in vitro culturing the sorted CD8+CD25 subpopulation with antigen in a medium containing interleukin-21, wherein the number of antigen specific CD8+ CTL are enriched in said CD8+CD25 subpopulation by at least 100-fold, as compared to that seen in the same lymphocyte population not subjected to either said separating step (a) or this promoting step (b); and
(c) formulating antigen-specific CTL cells produced therefrom into a pharmaceutical formulation, and
wherein said pharmaceutical formulation comprises at least 109 CTL cells.