A Fluorospot assay to detect single T lymphocytes simultaneously producing multiple cytokines.
Various subpopulations of T lymphocytes-i.e. Type 1, Type 2, Tr1 T cells-play a major role in the homeostasis of the immune system and in the pathogenesis of many inflammatory and auto-immune diseases. At present, in the absence of specific surface markers, these T cells can only be reliably distinguished on the basis of their cytokine production profile. The Elispot assay detects cytokine-producing cells, but in most cases can detect only one secreted cytokine, which represents a major limitation of this technique. We have developed a Fluorospot assay to detect single cells that simultaneously produce multiple cytokines. The Fluorospot assay permits the detection of regulatory T cells with an immunosuppressive activity, identified by their coexpression of IL-10 and IFNgamma. Polarized type 1 and type 2 specific tetanus toxoid T cells are also directly detected using a dual color Fluorospot. This technique will therefore be useful for detailed analysis of T lymphocytes in various disease states in which an imbalance of T cell subpopulations is suspected, but will also provide a better characterization of polarized specific immune responses.
Gazagne
Agnès
A
Unité d'Immunologie, Hopital Européen Georges Pompidou, AP-HP, INSERM U255, Université Pierre et Marie Curie, 20 Rue Leblanc, 75908 Cedex 15 Paris, France.
Claret
Emmanuel
E
Wijdenes
John
J
Yssel
Hans
H
Bousquet
François
F
Levy
Eric
E
Vielh
Philippe
P
Scotte
Florian
F
Goupil
Thierry Le
TL
Fridman
Wolf H
WH
Tartour
Eric
E
D016764:Cell Polarity; D016207:Cytokines / Q000032:analysis* / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis; D016753:Interleukin-10 / Q000032:analysis; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000378:metabolism*
D016207:Cytokines; D016753:Interleukin-10; D007371:Interferon-gamma