Carbohydrate metabolism of lymphocytes: Modified methodology and comparisons of diabetics with non-diabetics

Abstract

Changes in the hexose monophosphate shunt (HMPS) and Krebs cycle activity during lymphocyte blast transformation are reported in 50 patients with diabetes mellitus and 50 non-diabetics. A modified technique using 12 × 75 mm sterile tubes and micropipette tips stuffed with filter paper was used. The filter paper, soaked with hyamine hydroxide, absorbed radioactively labeled CO 2 produced from [ 14C]-labeled glucose incorporated by cells. [ 14C]CO 2 from glucose labeled at the C-1 position measured the activity of the HMPS. [ 14C]CO 2 from glucose labeled at the C-2 position measured the HMPS activity associated with the feedback of pentose sugars. [ 14C]-labeled CO 2 from glucose labeled at the C-6 position was used to measure Krebs cycle activity. A statistically significant decrease in HMPS activity was found in diabetic cells exposed to the mitogens phytohemagglutinin-P (PHA-P), concanavalin-A (CON-A) and pokeweed mitogen (PWM) ( P < 0.01). This decrease in HMPS activity and its relation to lymphocyte blast transformation may be related to the increased incidence of infection known to occur in patients with diabetes mellitus.