Cell Biology and Toxicology

, Volume 9, Issue 3, pp 259–267 | Cite as

Biochemical manipulation of intracellular glutathione levels influences cytotoxicity to isolated human lymphocytes by sulfur mustard

  • Clark L. Gross
  • Joy K. Innace
  • Renee C. Hovatter
  • Henry L. Meier
  • William J. Smith
Reviews in Cell Toxicology


Glutathione (GSH) is the major nonprotein thiol that can protect cells from damage due to electrophilic alkylating agents by forming conjugates with the agent. Sulfur mustard (HD) is an electrophilic alkylating agent that has potent mutagenic, carcinogenic, cytotoxic, and vesicant properties. Compounds that elevate or reduce intracellular levels of GSH may produce changes in cytotoxicity induced by sulfur mustard. Pretreatment of human peripheral blood lymphocytes (PBL) for 72 hr with 1 mM buthionine sulfoximine (BSO), which reduces intracellular GSH content to approximately 26% of control, appears to sensitize these in vitro cells to the cytotoxic effects of 10 μM HD but not to higher HD concentrations. Pretreatment of PBL for 48 hr with 10 mM N-acetyl cysteine (NAC), which elevates intracellular glutathione levels to 122% of control, appears to partially protect these in vitro cells from the cytotoxic effects of 10 μM HD but not to higher HD concentrations. Augmentation of intracellular levels of glutathione may provide partial protection against cytotoxicity of sulfur mustard.

Key Words

cytotoxicity glutathione lymphocytes sulfur mustard 



L-buthionine (S,R)-sulfoximine




sulfur mustard




peripheral blood lymphocytes


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  1. AHMAD, S., OKINE, L., LE, B., NALARIAN, P., and VISTICA, D.T. (1987). “Elevation of glutathione in phenylalanine mustard-resistant murine L1210 leukemia cells” J. Biol. Chem. 262: 15048–15053.Google Scholar
  2. AUERBACH, C. (1949). “Chemical mutagenesis”. Biol. Rev. 24: 355–391.Google Scholar
  3. BELLOMO, G., VAITETTI, M., STIVALA, L., MIRABELLA, F., RICHELEMI, P., and ORRENIUS, S. (1992). “Demonstration of nuclear compartmentalization of glutathione in hepatocytes”. Proc. Natl. Acad. Sci. USA 89: 4412–4416.Google Scholar
  4. BLACK, R.M., BREWSTER, L., CLARKE, R.J., HAMBROOKE, J.L., HARRISON, J.M., and HOWELLS, D.J., (1992). “Biological fate of sulphur mustard 1,1′-thiobis (2-chloethane): isolation and identification of urinary metabolites following intraperitoneal administration to rat”. Xenobiotica 22: 405–418.Google Scholar
  5. DAVISON, C., ROZMAN, R.S., and SMITH, P.K. (1961). “Metabolism of bis-β-chloroethyl sulfide (sulfur mustard gas)”. Biochem. Pharmacol. 7: 64–74.Google Scholar
  6. DETHMERS, J.K., and MEISTER, A. (1981). “Glutathione export by human lymphoid cells: Depletion of glutathione by inhibition of its synthesis decreases export and increases sensitivity to irradiation. Proc. Natl. Acad. Sci. USA 78: 7492–7495.Google Scholar
  7. ELSAYED, N.B., OMAYE, S.T., KLAIN, G.J., INASE, J.L., DAHLBERG, E.T., WHEELER, C.R. KORTE, D.W. (1989) “Response of mouse brain to a single subcutaneous injection of the monofunctional sulfur mustard, butyl 2-chloroethyl sulfide (BCS)”. Toxicology. 58: 11–20.Google Scholar
  8. GENTILHOMME, E., NEVEUX, Y., TUA, A., THIRIOT, M., FAURE, M., and THIVOLET, J. (1992). “Action of bis (betachloroethyl) sulfide (BCES) on human epidermis reconstituted in culture: Morphological alterations and biochemical depletion of glutathione”. Toxicol. in Vitro 6: 139–147.Google Scholar
  9. GRIFFITH, O.W. (1980). “Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine”. Anal. Biochem. 106: 207–212.Google Scholar
  10. GRIFFITH, O. W., and MEISTER, A. (1979). “Potent and specific inhibition of glutathione synthesis by buthionine sulfoximine (S-n-butyl homocysteine sulfoximine)”. J. Biol. Chem. 254: 7558–7560.Google Scholar
  11. HESTON, W.E. (1953). “Occurrence of tumors in mice injected subcutaneously with sulfur mustard and nitrogen mustard”. J. Natl. Cancer Inst. 14: 131–140.Google Scholar
  12. KINSEY, V.E., and GRANT, W.M. (1947). “Action of mustard gas and other poisons on yeast cells III. Distribution of fixed mustard gas in yeast.” J. Cell. Comp. Physiol. 29: 75–84.Google Scholar
  13. KRAMER, R.A., GREENE, K., AHMAD, S., and VISTICA, D.T. (1987). “Chemosensitization of L-phenylalanine mustard by the thiol-modulating agent, buthionine sulfoximine.” Cancer Res. 47: 1593–1597.Google Scholar
  14. MEIER, H.L., and JOHNSON, B.J. (1992). “The determination and prevention of cytotoxic effects induced in human lymphocytes by the alkylating agent 2,2′-dichlorodiethyl sulfide (sulfur mustard, HD).” Toxicol. Appl. Pharmacol. 113: 234–239.Google Scholar
  15. MEIER, H.L., GROSS, C.L., and PAPIRMEISTER, B. (1987). “2,2′-Dichlorodiethyl sulfide decreases NAD+ levels in human leukocytes.” Toxicol. Lett. 39: 109–122.Google Scholar
  16. MEISTER, A., and ANDERSON, M.E. (1983). “Glutathione.” Annu. Rev. Biochem. 52: 711–760.Google Scholar
  17. ONO, K., SHRIEVE, D.C. (1987). “Effect of glutathione depletion byl-buthionine sulfoximine on the cytotoxicity of cyclophosphamide in single and fractionated doses to EMT6/SF mouse tumors and bone marrow.” J. Natl. Cancer Inst. 79: 811–815.Google Scholar
  18. PAPIRMEISTER, B., and DAVISON, C.L. (1965). “Unbalanced growth and latent killing ofEscherichia coli following exposure to sulfur mustard.” Biochim. Biophys. Acta 103: 70–92.Google Scholar
  19. PAPIRMEISTER, B., GROSS, C.L., MEIER, H.L., PETRALI, J.P., and JOHNSON, J.B. (1985). “Molecular basis for mustard-induced vesication.” Fund. Appl. Toxicol. 5: S134-S149.Google Scholar
  20. PURI, R.N., MEISTER, A. (1983). “Transport of glutathione, as γ-glutamylcysteinylglycl ester, into liver and kidney.” Proc. Natl. Acad. Sci. 80: 5258–5260.Google Scholar
  21. SMITH, W.J., GROSS, C.L., CHAN, P., MEIER, H.L. (1990). “The use of human epidermal keratinocytes in culture as a model for studying the biochemical mechanisms of sulfur mustard toxicity.” Cell Biol. Toxicol. 6: 285–291.Google Scholar
  22. SMITH, W.J., SANDERS, K.M., GALES, Y.A., and GROSS, C.L. (1991). “Flow cytometric analysis of toxicity by vesicating agents in human cellsin vitro.” J. Toxicol.-Cutan. Ocular Toxicol. 10: 33–42.Google Scholar
  23. VIJAYARAGHAVAN, R., SUGENDRAN, K., PANT, S.C., HUSAIN, K., and MALHOTRA, R.C. (1991). “Dermal intoxication of mice with bis(2-chloroethyl) sulfide and the protective effect of flavonoids.” Toxicology 69: 35–42.Google Scholar
  24. VOJVODIC, V., MILOSAVLJEVIC, Z., BOSKOVIC, B., and BOJANIC, N. (1985). “The protective effect of different drugs in rats poisoned by sulfur and nitrogen mustards.” Fund. Appl. Toxicol. 5: S160–168.Google Scholar
  25. WALKER, I.G., and SMITH, J.F. (1969). “Protection of L-cells by thiols against the toxicity of sulfur mustard.” Can. J. Physiol. Pharmacol. 47: 143–151.Google Scholar
  26. WARTHIN, S.W., and WELLER, C.V. (1919) The Medical Aspects of Mustard Gas Poisoning. C. V. Mosby, St. Louis, MO.Google Scholar
  27. WONG, B.K., CHAN, H.C., and CORCORAN, G.B. (1986). “Selective effects ofN-acetyl cysteine stereoisomers on hepatic glutathione and plasma sulfate in mice.” Toxicol. Appl. Pharmacol. 86: 421–429.Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Clark L. Gross
    • 1
  • Joy K. Innace
    • 1
  • Renee C. Hovatter
    • 1
  • Henry L. Meier
    • 1
  • William J. Smith
    • 1
  1. 1.Biochemical Pharmacology BranchUS Army Medical Research Institute of Chemical DefenseAberdeen Proving GroundUSA

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