Application of intracellular ATP determination in lymphocytes for HLA-typing

✍ Scribed by Beckers, B. ;Lang, H. R. M. ;Weber, W. ;Pranada, F. ;Schweizer, K.

• Publisher: John Wiley and Sons
• Year: 1986
• Weight: 506 KB
• Volume: 1
• Category: Article
• ISSN: 0884-3996
• DOI: 10.1002/bio.1170010202

✦ Synopsis

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.