Application of intracellular ATP determination in lymphocytes for HLA-typing.
The amount of adenosine triphosphate (ATP) in human lymphocytes was determined using a technique based on light emission from a bioluminescent reaction with luciferin-luciferase. The amount of ATP changed when cells were incubated in the presence of specific HLA antisera and complement. For determination of intracellular ATP a modified method was applied, which was based on reduction of extracellular ATP by the addition of ATPase. The results of titration of an anti-human lymphocyte serum using the bioluminescence assay were in agreement with the results of fluorescence vitality staining. Bioluminescent HLA-determination in 57 cell samples each tested with 5 different antisera also gave good agreement (95.8%) with the conventional method. From these experimental data the calculated ATP content per lymphocyte was 0.135 +/- 0.058 pg ATP.
D000255:Adenosine Triphosphate / Q000032:analysis*; D049454:Firefly Luciferin; D006680:HLA Antigens / Q000032:analysis*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008156:Luciferases; D008163:Luminescent Measurements; D008214:Lymphocytes / Q000032:analysis / Q000276:immunology*
D006680:HLA Antigens; D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D008156:Luciferases