Quantitative assessment of interleukin-2-producing alloreactive human T cells by limiting dilution analysis.
A limiting dilution (LD) culture system was developed to estimate the frequency of IL-2 producing alloantigen-reactive human helper T lymphocytes (HTL). E rosette-purified T cells or cell sorter-separated CD4+ and CD8+ subsets were co-cultured under LD conditions with irradiated allogeneic Epstein-Barr virus-transformed lymphoblastoid cell lines (LCL) in the absence of additional growth factors. IL-2 in microculture supernatants was detected in a bioassay combined with rapid colorimetric visualization (cleavage of MTT). Under these conditions, one out of 250-800 E+, 180-280 CD4+, and 440-1000 CD8+ T cells produced IL-2 after a culture period of 3 days. This quantitative analysis of alloantigen-specific human HTL is complete within 4 days and thus provides a rapid method with potential applications in various clinical situations, e.g., transplantation medicine.
Jooss
J
J
Department of Medical Microbiology and Immunology, University of Ulm, F.R.G.
Zanker
B
B
Wagner
H
H
Kabelitz
D
D
D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology*; D002461:Cell Line, Transformed; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007376:Interleukin-2 / Q000096:biosynthesis*; D007700:Kinetics; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D013601:T-Lymphocytes / Q000145:classification / Q000276:immunology / Q000378:metabolism*; D006377:T-Lymphocytes, Helper-Inducer
D000945:Antigens, Differentiation, T-Lymphocyte; D007376:Interleukin-2