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Effect of Ascorbic Acid in Vitro on Lymphocyte Reactivity to Mitogens^1,2,

Department of Pediatrics, The University of Texas System Cancer Center, M. D. Anderson Hospital and Tumor Institute, Texas Medical Center, Houston, TX 77030

The direct effects of ascorbic acid and dehydroascorbic acid in vitro on human lymphocyte proliferation to phytohemagglutinin (PHA) and concanavalin A (Con A) stimulation were determined. Cells exposed to physiologic and high concentrations of ascorbic acid and dehydroascorbic acid showed poorer tritiated thymidine ([³H]TdR) incorporation than controls without the vitamin. The inhibitory effect was dose-dependent, with the greatest inhibition occurring as the concentration of ascorbic acid and dehydroascorbic acid was increased. At supraoptimal concentrations of PHA and Con A, there was no recovery of the mitogen response, indicating that ascorbic acid did not inhibit the response by competition. Viability studies of cells in culture showed that concentrations of ascorbic acid and dehydroascorbic acid, which consistently inhibited mitogenic stimulation of lymphocytes, were noncytotoxic throughout the culture period. Timed addition of ascorbic acid to PHA-stimulated lymphocytes in culture demonstrated inhibition of [³H]TdR incorporation when ascorbic acid was added as late as 96 hours after initiation of culture. However, the greatest inhibitory effect was observed when ascorbic acid was added at initiation or early in culture. Inhibition was also evident when RNA and protein synthesis were determined. The results suggest that physiologic and high concentrations of ascorbic and dehydroascorbic acid affect early metabolic events in the process of mitogen-stimulated lymphocyte activation.

KEY WORDS: • ascorbic acid • dehydroascorbic acid • mitogen • lymphocyte • transformation • inhibition

¹ Supported by NIH Grant CA09070.

² Part of the work reported in this paper was presented at the April 1979 FASEB meeting. Fed Proc. 38, 763, (abs. 2825).

Manuscript received 2 April 1980.