Method of expanding double negative t cells

Details Method of expanding double negative t cells Method of expanding double negative t cells

C - Chemistry, Metallurgy

12

N

C12N 5/0783 (2010.01) A61K 35/12 (2006.01) A61K 39/00 (2006.01) A61P 31/00 (2006.01) A61P 35/00 (2006.01) A61P 37/02 (2006.01) C12N 5/02 (2006.01)

Patent

CA 2629532

A method of expanding double negative T cells in culture is described. The method comprises (a) providing a starting sample comprising DN T cells or precursors thereof; (b) substantially depleting CD8+ and CD4+ T cells from the starting sample; (c) culturing the sample from step (b) with an immobilized T cell mitogen in a culture medium comprising an agent that can stimulate DN T cell growth; (d) washing the cells obtained in step (c) and resuspending in a culture medium comprising the agent without the T cell mitogen; and (e) washing the cells obtained in step (d) and resuspending in a culture medium comprising the agent and a soluble T cell mitogen. The DN T cells obtained by the method are useful in a variety of applications including the treatment of cancer, infectious diseases, graft versus host disease and autoimmune disease.

L'invention concerne un procédé d'expansion de lymphocytes T doubles négatifs (DN) en culture. Ce procédé consiste à (a) préparer un échantillon de départ comprenant des lymphocytes T DN ou des précurseurs de ceux-ci ; (b) éliminer sensiblement les lymphocytes T CD8+ et CD4+ de l'échantillon de départ ; (c) cultiver l'échantillon issu de l'étape (b) avec un mitogène des lymphocytes T immobilisé dans un milieu de culture comprenant un agent qui peut stimuler la croissance des lymphocytes T DN ; (d) laver les lymphocytes obtenus à l'étape (c) et les remettre en suspension dans un milieu de culture comprenant l'agent susmentionné sans le mitogène des lymphocytes T ; et (e) laver les lymphocytes obtenus à l'étape (d) et les remettre en suspension dans un milieu de culture comprenant l'agent susmentionné et un mitogène des lymphocytes T soluble. Les lymphocytes T DN obtenus par ce procédé sont utiles dans diverses applications, notamment en traitement du cancer, de maladies infectieuses, de la maladie du greffon contre l'hôte et d'une maladie auto-immune.

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