title: Differentiation of human alloreactive CD8(+) T cells in vitro.
abstract: Expansion and differentiation of alloantigen-reactive CD8(+) T cells in mixed lymphocyte cultures was followed by measurement of the loss of carboxyfluorescein diacetate succinimidyl ester (CFSE) fluorescence of responder cells. Proliferation of CD8(+) T cells became detectable on day 4 of culture and, 2 days later, > 60% of the CD8(+) T cells in culture were dividing alloreactive lymphocytes. In parallel with expansion, CD8(+) T-cell differentiation was initiated, as evidenced by an increase in the number of CD45RA(-) and CD27(-) T cells and acquisition of the ability to produce interferon-gamma after restimulation with the specific alloantigen. Finally, although short-term stimulation and measurement of intracellular cytokine production allowed visualization of alloreactive CD8(+) T cells expanded in vitro, this procedure did not detect circulating alloreactive CD8(+) T cells activated in vivo in recipients of allogeneic kidney grafts.
authors.affiliation: Renal Transplant Unit, Department of Internal Medicine, Academic Medical Center, Amsterdam, The Netherlands. rrentenaar@yahoo.com
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Rentenaar
authors.forename: Jelle L G
authors.initials: JL
authors.lastname: Vosters
authors.forename: Frank N J
authors.initials: FN
authors.lastname: van Diepen
authors.forename: Ester B M
authors.initials: EB
authors.lastname: Remmerswaal
authors.forename: René A W
authors.initials: RA
authors.lastname: van Lier
authors.forename: Ineke J M
authors.initials: IJ
authors.lastname: ten Berge
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D007519:Isoantigens / Q000276:immunology*; D016030:Kidney Transplantation / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000032:analysis; D007959:Lymphocyte Culture Test, Mixed; D016176:T-Lymphocyte Subsets / Q000276:immunology; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7 / Q000032:analysis
chemical_list: D007519:Isoantigens; D018127:Tumor Necrosis Factor Receptor Superfamily, Member 7; D007371:Interferon-gamma; D017493:Leukocyte Common Antigens