LDH analysis of human thymocytes and thymocyte subsets.

The lactate dehydrogenase (LDH) isoenzyme pattern (expressed as the B:A subunit ratio) and two enzymes of the purine metabolism adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) (expressed as the ADA/PNP ratio) were studied in human prenatal thymocytes, in subsets of human infant thymocytes, and in peripheral T lymphocytes. Prothymocytes were enriched by E rosette depletion, cortical thymocytes were enriched with a monoclonal antibody rosette technique using OKT6, and medullary thymocytes were enriched either with a monoclonal antibody rosette technique using OKT3 or with complement-mediated cytolysis using normal fresh rabbit serum. Peripheral T lymphocytes were isolated from normal adult peripheral blood by E rosette sedimentation. Prenatal thymocytes had the lowest B:A ratio. In the infant thymuses, prothymocytes had a lower B:A ratio (0.99 +/- 0.10) than the cortical thymocytes (1.04 +/- 0.08). The medullary thymocytes obtained either by OKT3 selection or normal rabbit serum cytotoxic treatment had higher B:A ratios (1.30 +/- 0.15 and 1.42 +/- 0.17, respectively). The highest B:A ratio is found in peripheral T lymphocytes (2.07 +/- 0.28) together with the lowest ADA/PNP ratio (0.50 +/- 0.07). The B:A ratios are paralleled by a progressive decrease in the ADA/PNP ratio. These findings indicate that during intrathymic T cell development, important changes occur not only in the activities of the enzymes of the purine metabolism but also in the distribution of the LDH isoenzymes.