Confirmation of Mycobacterium tuberculosis infection by flow cytometry after ex vivo incubation of peripheral blood T cells with an ESAT-6-derived peptide pool

Cytometry. Part B, Clinical Cytometry
Lydia TesfaFlorian Kern

Abstract

The presence of a T-cell response to the early secretory antigenic target-6 (ESAT-6) indicates previous infection with or exposure to Mycobacterium tuberculosis. Measuring this response is useful for identifying individuals infected with M. tuberculosis. It was also reported that the frequencies of ESAT-6-specific T cells correlate with disease state. Established procedures measure secreted T-cell cytokines following whole blood stimulation with recombinant ESAT-6 protein or use Elispot as a read-out. A single ESAT-6- spanning pool of overlapping peptides (15 amino acids length with 11 overlaps) was used for overnight stimulation of peripheral blood mononuclear cells (PBMCs) from 15 patients infected with Mycobacterium tuberculosis and 11 healthy controls. T-cell responses were rated positive if interferon-gamma (IFN-gamma)-producing T cells were identified above background level, using 4-color cytokine flow cytometry. Thirteen of 15 (87%) patients, but none of the healthy controls, had a positive CD4 T-cell response to the ESAT-6 spanning peptide pool. The frequencies of IFN-gamma-producing cells varied between 1 and 167 per 10,000 CD4 T cells. The test performed as well as the tests described in the literature. Cytokine flow ...Continue Reading

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Citations

Jun 25, 2009·BMC Infectious Diseases·Valeria SargentiniRoberto Nisini
Apr 16, 2013·Methods : a Companion to Methods in Enzymology·Giulia Freer, Laura Rindi
Sep 24, 2005·Clinical and Experimental Immunology·A J HughesP D R Johnson
Aug 9, 2008·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Stephan FuhrmannFlorian Kern
Feb 14, 2006·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·M H HammerP Reinke
Oct 24, 2012·Cytometry. Part B, Clinical Cytometry·Frederic I Preffer
Nov 11, 2009·Cytometry. Part B, Clinical Cytometry·Dong Il Won, Jung Ran Park
Dec 28, 2006·International Archives of Allergy and Immunology·Lorenzo CosmiFrancesco Annunziato
Sep 16, 2009·Scandinavian Journal of Immunology·O ParkashM Pai
Jul 24, 2015·Cytometry. Part B, Clinical Cytometry·Chrysovalantis V PapageorgiouNikolaos Poulakis

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