Analysis of alloreactive helper T lymphocyte precursor frequencies. Influence of interleukin-2 produced by the stimulating cells.
In the determination of alloreactive helper T lymphocyte precursor frequencies (HTLpf) by limiting dilution analysis (LDA), peripheral blood mononuclear cells (PBMC) are used as stimulating cells. Interleukin-2 (IL-2) production by stimulating T cells constitute a source of error in these assays. We found that 100-150 Gy of gamma irradiation was required to abrogate IL-2 production by stimulating PBMC. This dose of irradiation, however, greatly reduced their allostimulatory capability. Here we describe how non-irradiated PBMC, immunomagnetically depleted of T cells and thus of IL-2 producing cells, can be used as stimulators in assays to determine alloresponsive HTLpf.
Reisaeter
A V
AV
Institute of Transplantation Immunology, National Hospital, Oslo, Norway.
Thorsby
E
E
Brinchmann
J E
JE
D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004307:Dose-Response Relationship, Radiation; D005720:Gamma Rays; D006801:Humans; D018189:Immunomagnetic Separation; D007376:Interleukin-2 / Q000096:biosynthesis / Q000502:physiology* / Q000528:radiation effects; D008213:Lymphocyte Activation / Q000528:radiation effects; D018655:Lymphocyte Count / Q000528:radiation effects; D007959:Lymphocyte Culture Test, Mixed; D013234:Stem Cells / Q000276:immunology* / Q000378:metabolism / Q000528:radiation effects; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology* / Q000378:metabolism / Q000528:radiation effects
D007376:Interleukin-2