Quantification of lymphocyte activation by measurement of DNA polymerase alpha activity.
Measurement of the activity of the enzyme DNA polymerase alpha has been investigated with regard to its potential usefulness as a method for the detection and quantification of lymphocyte activation in vivo. A modified enzyme assay was developed in order to optimise measurement of activity in crude homogenates of cells or tissues, thus allowing the convenient handling of multiple samples. Specificity of the assay for polymerase alpha was ensured by the inclusion in the assay mixture of dideoxythymidine triphosphate, an inhibitor of the other eukaryotic DNA polymerases. The activity of DNA polymerase alpha was found to be closely correlated with [3H]thymidine incorporation in a mitogen-stimulated in vitro system. The usefulness of the polymerase alpha method for the quantification of lymphocyte activation was validated in 3 different in vivo systems of either immune-mediated or drug-induced lymphoid cell response.
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003520:Cyclophosphamide / Q000494:pharmacology; D004257:DNA Polymerase II / Q000032:analysis*; D054306:Dideoxynucleotides; D005260:Female; D016027:Heart Transplantation; D006801:Humans; D007111:Immunity, Cellular; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000201:enzymology; D008297:Male; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000276:immunology; D009206:Myocardium / Q000201:enzymology; D019384:Nucleic Acid Synthesis Inhibitors; D009929:Organ Size; D010835:Phytohemagglutinins / Q000494:pharmacology; D051381:Rats; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013942:Thymine Nucleotides / Q000494:pharmacology; D014181:Transplantation Immunology
D054306:Dideoxynucleotides; D019384:Nucleic Acid Synthesis Inhibitors; D010835:Phytohemagglutinins; D013942:Thymine Nucleotides; D003520:Cyclophosphamide; D004257:DNA Polymerase II; C017216:2',3'-dideoxythymidine triphosphate