Rapid assessment of antigen induced cytokine expression in memory T cells by flow cytometry

Veterinary Immunology and Immunopathology
V C Maino

Abstract

Methods for analysis of T cell responses to specific antigen have traditionally relied on measurements of proliferation or cytokine expression in bulk cultures of PBMC in long term incubations with putative antigen. These techniques suffer from the drawback that they do not enable analysis of single cell responses in the context of unselected cellular backgrounds. It is increasingly important to not only identify cells on the basis of expression of unique surface antigens but also to determine functional and molecular parameters of individual cells in response to a variety of stimuli. We have recently developed methodologies to rapidly assess T-cell subset responses to polyclonal activators and specific antigen in whole blood. These procedures determine the percentages of activated cells and the identification of leucocyte subsets capable of expressing various cytokines and cell surface antigens. Multiparameter analysis of CD69+ /cytokine + expression in T cells in response to specific antigen (e.g. CMV, mumps) demonstrated a range of frequencies from 0.05% to 5.0% within 6 h. Frequencies of responding T cells were consistent but varied depending upon the antigen. Antigen specific T cell responses were host specific and both po...Continue Reading

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Citations

Sep 12, 2006·Journal of Clinical Immunology·Susan M MooreDeborah J Briggs
Mar 3, 2009·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·J J E M KitzenJ Verweij
Jul 7, 2000·Clinical and Diagnostic Laboratory Immunology·M C CheeranJ R Lokensgard
Apr 9, 2005·Clinical and Diagnostic Laboratory Immunology·Leonor RodríguezRocío Ortiz
Feb 23, 2002·The Journal of Immunology : Official Journal of the American Association of Immunologists·Mohammed L GarbaJeffrey A Frelinger

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