The relative radioresistance of interleukin-2 production by human peripheral blood lymphocytes: consequences for the development of a new limiting dilution assay for the enumeration of helper T lymphocyte precursor frequencies.
We describe a limiting dilution assay for the enumeration of alloreactive helper T lymphocyte precursor frequencies in human peripheral blood. The proliferation rate of the murine indicator cell line, cytotoxic T lymphoblastic line 2 (CTLL-2) induced by interleukin-2 (IL-2) culture supernatants was determined by staining with the fluorescent DNA dye propidium-iodide. Lymphocytes from healthy individuals as well as from patients with end stage kidney disease and no previous allosensitization exhibited a relative radioresistance of their IL-2 production up to gamma irradiation doses of 40-60 Gy. This differs from previous findings in the literature, showing a total inhibition of the IL-2 production in unsensitized individuals using a gamma irradiation dose of 20 Gy. The consequences of this relative radioresistance are that for a reliable stimulator cell inactivation in assays for the enumeration of helper T lymphocyte precursors gamma irradiation doses of at least 50 (-60) Gy are needed. Increasing the gamma irradiation dose for the inactivation of the stimulator cells can result in a decrease of the antigen presenting capacity of these cells.
Schanz
U
U
Department of Immunohematology and Blood Bank, University Hospital, Leiden, Netherlands.
Roelen
D L
DL
Bruning
J W
JW
Kardol
M J
MJ
van Rood
J J
JJ
Claas
F H
FH
D000818:Animals; D017951:Antigen Presentation / Q000276:immunology; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D005312:Fetal Blood / Q000276:immunology; D006684:HLA-DR Antigens / Q000276:immunology; D006801:Humans; D007158:Immunologic Techniques; D007376:Interleukin-2 / Q000096:biosynthesis*; D007676:Kidney Failure, Chronic / Q000276:immunology / Q000473:pathology; D007958:Leukocyte Count; D051379:Mice; D011836:Radiation Tolerance; D013234:Stem Cells / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000528:radiation effects; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000528:radiation effects*
D006684:HLA-DR Antigens; D007376:Interleukin-2; D004247:DNA