Differentiation of human alloreactive CD4+ and CD8+ T cells in vitro

Transplantation
Natalia NikolaevaIneke J M Ten Berge

Abstract

Recent studies have revealed that, during viral infection, ordered phenotypic and functional changes occur in human antigen-specific T cells. We analyzed whether a similar differentiation program is induced after alloantigen stimulation in vitro. Peripheral blood mononuclear cells and purified CD4(+)CD45RA+, CD4(+)CD45RO+, and CD8+ T cells from healthy individuals were labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE). Cells were co-cultured with allogeneic irradiated cells. Flow cytometric analysis was performed on days 3, 5, and 7 of culture using surface CD45RA, CD27, CD28, CCR7, and intracellular perforin and granzyme B markers in relation to CFSE dilution. Based on the decrease in CFSE fluorescence, both CD4+ and CD8+ T cells showed an early and vigorous response to allogeneic stimulation. Loss of CD45RA expression and up-regulation of CD27 and CD28 costimulatory molecules was an early event occurring in the first generations of dividing cells. Differentiation at later stages of proliferation was characterized by loss of CCR7 homing receptor expression, more pronounced in CD4+ than in CD8+ T cells, indicating the decreased ability of these cells to traffic to secondary lymphoid organs. Productio...Continue Reading

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Citations

Jun 14, 2006·Transplantation·Seiichiro InoueMarc H Dahlke
Sep 30, 2008·Human Immunology·Sven D KochIneke J M ten Berge
Feb 17, 2007·Clinical and Experimental Immunology·V FaivreA Haziot
Jan 6, 2009·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·I E DijkeC C Baan
Apr 5, 2012·Clinical and Experimental Immunology·P J E J van de BergI J M ten Berge
Nov 8, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Ruth D FritschPeter E Lipsky
Aug 22, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Elena UssIneke J M ten Berge

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