Title

Ex vivo expansion of memory CD8 T cells from lymph nodes or spleen through in vitro culture with interleukin-7

Authors

Authors

C. Kittipatarin;A. R. Khaled

Abbreviated Journal Title

J. Immunol. Methods

Keywords

Proliferation; Cytokine; Lymphocytes; Immunotherapy; Activation; RECEPTOR-DEFICIENT MICE; HOMEOSTATIC PROLIFERATION; PREFERENTIAL; EXPANSION; CUTTING EDGE; BONE-MARROW; IL-7; NAIVE; LYMPHOCYTES; ACTIVATION; EXPRESSION; Biochemical Research Methods; Immunology

Abstract

Interleukin-7 (IL-7) increases lymphocyte numbers, a critical feature of immune reconstitution, through mechanisms that are still poorly understood. Part of the problem is that IL-7 is produced in limited amounts by non-lymphoid cells, making in vivo studies of the cytokine's activity a challenge. To overcome this, we developed an in vitro system by which lymphocytes from secondary immune organs could be cultured to produce IL-7 responsive cells. Using this method, we showed that CD8(hi)CD44(hi)T cells accumulate in culture with IL-7 from a population of lymph node or splenic cells. These results were validated when a similar lymphocyte subset was found in mice expressing a constitutively active form of STAT5b, a key transducer of IL-7 signals. Interestingly, IL-7-expanded cells also up regulated the activation marker. CD69. The IL-7-derived CD44(hi)CD69(hi) cells were not generated from naive cells, but expanded from an existing population, since culture in IL-7 of naive lymphocytes from OT-1/Rag1(-/-) mice did not produce CD44(hi)CD69(hi) cells. Using the in vitro culture system to study lymphocytes from mice deficient in the apoptotic protein, BIM, we were able to attribute the expansion of CD8(hi)CD44(hi)CD69(hi) T cells to the proliferative and not survival activity of IL-7. The in vitro culture system provides an important new methodology to examine the activities of this essential as well as immunotherapeutic cytokine. (C) 2009 Elsevier B.V. All rights reserved

Journal Title

Journal of Immunological Methods

Volume

344

Issue/Number

1

Publication Date

1-1-2009

Document Type

Article

Language

English

First Page

45

Last Page

57

WOS Identifier

WOS:000266572900006

ISSN

0022-1759

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