Enzyme activity of isolated leukocyte populations. I. Cytochemical and zymographic studies of stored blood under various storage conditions

Abstract

Heparinized venous blood was stored under sterile conditions at different temperatures (4 C, 20 C, 37 C) for various intervals (0-7 days). After storage the granulocytes and lymphocytes were isolated with routine methods. Naphthol AS-D-chloroacetate esterase as a granulocyte marker and acid alpha-naphthyl acetate esterase as a T-lymphocyte marker were identified on smears of the washed cell suspension. Different enzymes were identified in the cell sediment with electrophoresis. Relatively pure lymphocyte suspensions were obtained within the first 24 h. After this time, however, the percentage of these mononuclear cells declined markedly. The percentage of isolated granulocytes varied slightly; there was a marked predominance of granulocytes (more than 70%) at all intervals investigated during the isolation. Cytochemical analysis of the granulocytes and lymphocytes indicated that the decrease in the percentage of enzyme-positive cells depends in each case on the duration of the storage interval. During the first 24 h, only PGM1 and GOTM could be identified in the lymphocyte suspension with horizontal starch gel electrophoresis. The enzymes PGM1, PGM3, PGI, MDH, GOTM, 6-PGD, ADA could always be identified in the granulocyte suspension; AK, FUCA, MEM could be occasionally identified; and GPT and GLO could never be identified.