Flow cytometric analysis of intracellular interferon-gamma synthesis in rat CD4 T cells. NLM AIDSLINE Important note: Information in this article was accurate in 1999. The state of the art may have changed since the publication date.

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Flow cytometric analysis of intracellular interferon-gamma synthesis in rat CD4 T cells.

Eur Cytokine Netw. 1998 Dec;9(4):613-8. Unique Identifier : AIDSLINE MED/99107682
Bernard I; Foucras G; Cautain B; van der Meide PH; Druet P; Guery JC; Saoudi JC; INSERM U. 28, place du Dr.-Baylac, 31059 Toulouse Cedex, France.

Abstract: To date the techniques used to analyse cytokine expression by rat T cells do not give information about the simultaneous production of different cytokines from individual cells. Recently, a method for analysing the intracellular production of cytokines at the single cell level using flow cytometry has been developed. It is well established that the most critical requirement for successful intracellular cytokine staining is the availability of appropriate antibodies. In rat, it is possible to stain for intracellular IL-4 and IL-10 (Th2 cytokines) using the commercially available antibodies but not for Th1 cytokines. In the present work, we show that DB1, a mouse anti-rat IFN-gamma monoclonal antibody, could be used for intracytoplasmic staining of IFN-gamma producing rat CD4 T cells. The specificity of the staining was confirmed using a molar excess of unlabelled antibodies or recombinant cytokine. Finally, intracellular staining for IFN-gamma correlates with cytokine production in culture supernatant as evaluated by ELISA analysis.
Keywords: JOURNAL ARTICLE Animal Antibodies, Monoclonal Antibody Specificity Cell Line CD4-Positive T-Lymphocytes/*IMMUNOLOGY Enzyme-Linked Immunosorbent Assay Flow Cytometry/*METHODS Interferon Type II/*BIOSYNTHESIS/IMMUNOLOGY Intracellular Fluid/IMMUNOLOGY Kinetics Mice Rats Staining/METHODS Support, Non-U.S. Gov't Th1 Cells/IMMUNOLOGYKWDjournalarticleanimalantibodies,monoclonalantibodyspecificitycelllinecd4-positivet-lymphocytes/KWDimmunologyenzyme-linkedimmunosorbentassayflowcytometry/KWDmethodsinterferontypeii/KWDbiosynthesis/immunologyintracellularfluid/immunologykineticsmiceratsstaining/methodssupport,non-uKWDsKWDgov'tth1cells/immunology
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A9950962

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