Rapid detection, enrichment and propagation of specific T cell subsets based on cytokine secretion.
T cell lines with defined cytokine profiles are an invaluable tool for assessing the control of immune responses both in vitro and in vivo. Production of such cell lines can be complex and time-consuming. Here we present a powerful technique to assay the cytokines produced by T cells activated polyclonally or with specific antigens. This paper presents a detailed methodology for the identification and isolation of cytokine-producing T cells activated with the artificial superantigen, CytoStim, or viral and fungal antigens. These cells can be analysed for different cytokines simultaneously, or cultured further to rapidly establish T cell lines making known cytokine types. We highlight the enumeration, isolation and phenotype of interleukin-17-producing T cells, and the rapid generation of virus-specific Th1 T cell lines.
Campbell
J D M
JD
Research and Development, Miltenyi Biotec GmbH, Friedrich Ebert Street 68, Bergisch Gladbach, Germany. johnc@miltenyibiotec.co.uk
Foerster
A
A
Lasmanowicz
V
V
Niemöller
M
M
Scheffold
A
A
Fahrendorff
M
M
Rauser
G
G
Assenmacher
M
M
Richter
A
A
D000818:Animals; D000946:Antigens, Fungal / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D016207:Cytokines / Q000032:analysis*; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D020381:Interleukin-17 / Q000032:analysis / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D009000:Monocytes / Q000276:immunology; D018089:Superantigens / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000276:immunology; D018417:Th1 Cells / Q000166:cytology* / Q000276:immunology
D000946:Antigens, Fungal; D000956:Antigens, Viral; D016207:Cytokines; D020381:Interleukin-17; D018089:Superantigens; D007371:Interferon-gamma