Please use this identifier to cite or link to this item:
http://hdl.handle.net/1942/4088
Title: | A CFSE based assay for measuring CD4(+)CD25(+) regulatory T cell mediated suppression of auto-antigen specific and polyclonal T cell responses | Authors: | VENKEN, Koen THEWISSEN, Marielle HELLINGS, Niels SOMERS, Veerle HENSEN, Karen STINISSEN, Piet RUMMENS, Jean-Luc |
Issue Date: | 2007 | Publisher: | ELSEVIER SCIENCE BV | Source: | JOURNAL OF IMMUNOLOGICAL METHODS, 322(1-2). p. 1-11 | Abstract: | CD4(+)CD25(+) regulatory T cells (Tregs) are considered to play a key role as suppressors of immune inediated reactions. The analysis of Treg function in patients with autoimmune, allergic or oncogenic diseases has emerged over the past years. In the present study we describe a CFSE based protocol to measure Treg mediated suppression of CD4(+) T cells. Measuring Treg suppressive capacity towards proliferation of anti-CD3 Ab stimulated CD4(+)CD25(-) T cells in coculture experiments by means of a CFSE based and a classical [H-3]thymidine incorporation assay gave similar results, provided that CD4(+)CD25(+) T cells were anergic. However, when CD4(+)CD25(+) T cells proliferated upon mitogenic stimulation, data obtained by the CFSE assay allowed the detection of a significant Treg suppression whereas this was clearly underestimated using the [H-3]thymidine assay. In addition, an indirect CFSE based method was developed to analyze antigen specific responses of total CD4(+) T cells and Treg depleted CD4(+) T cells (i.e. CD4(+)CD25(-) T cells). Our results indicate that, in healthy individuals, CD4(+) T cell responses against the multiple sclerosis (MS) auto-antigens, myelin basic protein (MBP) and myelin oligodendrocyte glycoprotein (MOG), were increased in Treg depleted CD4(+) T cells as compared to total CD4(+) T cells. Our initial data suggest that Tregs in MS patients show an impaired suppression of myelin reactive T cells when compared to healthy controls. Moreover, this experimental setup permits the measurement of cytokine production of the antigen proliferated CFSElow T cells by additional flow cytometric analyses. In conclusion, the described CFSE based Treg suppression assay is a valuable tool to study suppressor T cells in (auto)immune disorders. (C) 2007 Elsevier B.V. All rights reserved. | Notes: | Hasselt Univ, Biomed Onderzoeksinst, B-3590 Diepenbeek, Belgium. Transnatl Univ Limburg, Sch Life Sci, Diepenbeek, Belgium. Virga Jesse Hosp, Clin Lab Expt Hematol, Hasselt, Belgium.STINISSEN, P, Hasselt Univ, Biomed Onderzoeksinst, Agoralaan Bldg A, B-3590 Diepenbeek, Belgium.piet.stinissen@uhasselt.be | Keywords: | CD4(+)CD25(+) regulatory T cells; CFSE; Myelin responses | Document URI: | http://hdl.handle.net/1942/4088 | ISSN: | 0022-1759 | e-ISSN: | 1872-7905 | DOI: | 10.1016/j.jim.2007.01.025 | ISI #: | 000246422100001 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2008 |
Appears in Collections: | Research publications |
Show full item record
SCOPUSTM
Citations
110
checked on Sep 2, 2020
WEB OF SCIENCETM
Citations
114
checked on Apr 22, 2024
Page view(s)
86
checked on Sep 7, 2022
Google ScholarTM
Check
Altmetric
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.