Characterization of rat lymphocyte primary culture for the development of an in-vitro mutagenesis assay: effect of interleukin-2 and 2-mercaptoethanol on the activities of intermediary metabolism enzymes and cell proliferation.
Efficient energy utilization is essential for cell growth; in an attempt to improve the growth conditions of the rat T-lymphocyte culture model for potential use in studying the mutagenic activity of carcinogens in vitro, we have investigated the effects of phytohemagglutinin (PHA), interleukin-2 (IL-2) and 2-mercaptoethanol (2-ME) on the activities of intermediary metabolism enzymes and cell proliferation. Isolated lymphocytes were cultured in the presence and absence of PHA, IL-2, or 2-ME. The intermediary metabolism enzymes investigated were glutamate dehydrogenase, glutamate-pyruvate transaminase, malate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase, pyruvate kinase, and fatty acid synthetase (FAS). Measurable activity of all enzymes investigated, except for FAS, was detected in PHA-stimulated cells cultured with IL-2 or 2-ME. The unstimulated lymphocytes had significantly lower enzyme activity than stimulated cells. The combination of all three agents showed increased enzyme activity. This increase in activity brought about by the combination of the three agents was not reproduced by either agent acting alone. In general, the increase in enzyme activity correlated with cell proliferation as measured by [3H]thymidine uptake in PHA-stimulated cultures containing IL-2 and/or 2-ME. The results suggest that the addition of exogenous IL-2 and 2-ME enhances metabolic function and may be beneficial in in vitro culture of rat lymphocytes.
Aidoo
A
A
Food and Drug Administration, National Center for Toxicological Research, Jefferson, AR 72079, USA.
Feuers
R J
RJ
Lyn-Cook
L E
LE
Bishop
M E
ME
Casciano
D A
DA
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology / Q000502:physiology; D002952:Citric Acid Cycle; D064429:Fatty Acid Synthases / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D007376:Interleukin-2 / Q000494:pharmacology*; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D008291:Malate Dehydrogenase / Q000378:metabolism; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology*; D009152:Mutagenicity Tests; D010835:Phytohemagglutinins; D011770:Pyruvate Kinase / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism; D019289:Pyruvic Acid; D051381:Rats; D011916:Rats, Inbred F344; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D000637:Transaminases / Q000378:metabolism
D000596:Amino Acids; D007376:Interleukin-2; D010835:Phytohemagglutinins; D011773:Pyruvates; D008623:Mercaptoethanol; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; D005969:Glutamate Dehydrogenase; D064429:Fatty Acid Synthases; D000637:Transaminases; C036402:glutamate aminotransferase; D011770:Pyruvate Kinase