The Effects of Carcinogenic Methylcholanthrene on Carbohydrate Residues of Nk Cells

Abstract

The present study examines the effect of methylcholanthrene (MCA), a carcinogenic polycylic hydrocarbon, on the carbohydrate receptor determinants (RD) on natural killer (NK) cell surface using the bead-coupled lectin assay. Murine NK cells exhibited different degrees of preferential binding to the specific lectins tested. Of the ten lectins tested, five exhibited a positive binding affinity while the remaining five exhibited no or insignificant binding. NK cells bind to beads derivatized with mannose specific lectins: Concanavalin A (Con A), Lens culinaris, and Pisum sativum. NK cells also bind to other lectin beads such as Triticum vulgaris (GalNac) and Vicia villosa (D-G1cNAc). All these lectin beads exhibited greater than 90% adhesion. The underivatized control beads exhibited no NK binding. The NK cells that were exposed to MCA for 2 h demonstrated a significant decrease in lectin bead-cell coupling in a dose dependent manner. MCA (10μg/mL) caused a 17.8%, 40% and 4.7% decrease in binding affinity when introduced to the mannose specific lectins; Con A, L. culinaris and P. sativum beads, respectively. The binding of T. vulgaris and V. villosa to NK cells was inhibited (23.4% and 28%) by MCA treatment. An increase in the dose to 20μg/mL resulted in a greater inhibition in binding affinity towards lectin beads, Con A, 35.3%, L. culinaris, 62.6%, P. sativum, 30.9%, T. vulgaris, 44.2% and V. villosa, 46.2%. The effect of MCA on the synthesis of NK polyphosphoinositides such as phosphatidylinositol 4-phosphate (PIP), phosphotidylinositol 4, 5- biphosphate (PIP₂) was examined. The results showed lower phosphate 32 (³²P)-labeling of PIP and PIP₂ of MCA-treated NK cells as compared to untreated cells. Both of these MCA-induced cellular events (carbohydrate RD, PIP, and PIP₂) may be interrelated at the level of transmembrane signal transduction, and may represent important steps in the inhibition of NK mediated cytotoxicity by this carcinogen.

Get full access to this article

View all access options for this article.

References

Chambers, W.H. , Forbes, J.T. , and Olteman, T.N. (1985). "Role of soluble lytic factors in NK lytic activity ." In: Mechanisms of Cytotoxicity by NK Cells (R.B. Herberman and D. Callewaert, eds.). Academic Press, New York, NY. pp. 351-354.

Chambers, W.H. and Oeltman, T.N. (1986). "The effects of Hexose 6-0-sulfate esters on human natural killer cell lytic function." J. Immunol. 137:1469-1474.

Fletcher, M.A. , Baron, G.C. , Ashman, M.R. , Fischl, M.A. , and Klimas, N.G. (1987). "Use of whole blood methods in assessment of immune parameters in immunodeficiency states." Diag. Clin. Immunol. 5:69-81.

Forbes, J.T. , Bretthauer, R.K. , and Oeltmann, T.N. (1981). "Mannose 6-fructose 1-, and fructose 6-phosphates inhibit human natural cell-mediated cytotoxicity." Proc. Natl. Acad. Sci. 78:5797-5801.

Ghoneum, M. , Gill, G. , Wojdani, A. , Payne, C. , and Alfred, L. (1987a). "Suppression of basal and corynebacterium parvum-augmented NK activity during chemically induced tumor development." Intl. J. Immunopharmacol. 9:71-79.

Ghoneum, M. , Jackson, K. , Gill, G. , and Salem, F. (1991). "Murine intersex difference in tumor development and its correlation with NK activity." Intl. J. Immunopath Pharmac. 4:9-18.

Ghoneum, M. , Salem, F. , Gill, G. , Jackson, K. , Betru, Y. , and Allen, H. (1989). "NK cell activity and tumor development of age-related variation to 3-methylcholantrhene." In: Natural Killer and Host Defense (E.W. Ades and G. Lopes, eds.). Basel Karger , New York, NY. pp. 278-284.

Ghoneum, M. , Wojdani, A. , and Alfred, L. (1987b). "Effect of methylcholanthrene on human natural killer cell cytotoxicity and lymphokine production in vitro." Immunolpharmacology. 14:27-33.

Herberman, RB (1983) "Natural killer cells and tumor immunity." Surv. Immunol. Res. 2:306-8.

10.

Hiserodt, J.C. , Britvan, L.J. , and Targan, S.R. (1982). "Characterization of the cytolytic reaction mechanism of the human natural killer (NK) lymphocyte: resolution into binding, programming, and killer cell-independent steps." J. Immunol. 129(4):1782-7.

11.

Hubbard, S.C. and Ivatt, R.J. (1981). "Synthesis and processing of asparagine-lined oligosaccharidase." Ann. Rev. Biochem. 50:555-9.

12.

Huhti E. (1981). "Smoking and lung cancer - now." Euro. J. Respir. Dis. 62:147-9.

13.

Kalland, T. and Forsberg, J.G. (1983). "3-methylcholanthrene: transient inhibition of the lytic step of mouse natural killer cells." J. Natl. Cancer Inst. 71:385-90.

14.

Kiyohara, T. , Dennis, J.W. , Boegman, R.J. , and Roder, J.C. (1985). "An exoglycosidase-sensitive triggering site on NK cells which is coupled to transmethylation of membrane phospholipids ." J. Immunol. 135:659-64.

15.

Kumagai., K. , Itoh, K. , Suzuki, R. , Hinuma, S. , and Saitoh, F. (1982). "Murine large granular lymphocytes identification as effector cells in NK and K cytotoxicity." J. Immunol. 129:388.

16.

Lagos, N. and Vergara, J. (1990). "Phosphoinositides in frog skeletal muscle: a quantitative analysis." Biochem. Biophys. Acta. 1043:235-44.

17.

Macdermott, R.P. , Kienker, L.J. , Bertovich, M.J. , and Muchmore, A.V. (1981). "Inhibition of spontaneous but not antibody-dependent cell-mediated cytotoxicity by simple sugars: Evidence that endogenous lectins may mediate spontaneous cell-mediated cytotoxicity." Immunol . 44:143-52.

18.

McCoy, J.P. , Hanley-Yanez, K. , Brumfield, A. , Herberman, R.B. , and Chambers, W. (1990). "Alteration in cell surface carbohydrates of rat large granular lymphocytes associated with interleukin-2 activation." Cell Immunol. 127:275-83.

19.

Ortaldo, J.R. , Timonen, T.T. , and Herberman, R.B. (1984). "Inhibition of activity of human NK and K cells by simple sugars: discrimination between binding and postbinding events." Clin. Immunol. Immunopath. 31:439-43.

20.

Roder, J.C. , Haliotis, T. , Klein, M. , Korec, S. , Jett, J.R. , Ortaldo, J. , Heberman, R.B. , Katz, P. , and Fauci, A.S. (1980). "A new immunodeficiency disorder in humans involving NK cells." Nature 10, 284(5756):553-5.

21.

Sadek, I.A. , Ghoneum, M. , and Cooper, E. (1987). "Effect of methylcholanthrene on amphibian NK activity." Dis. Aquat. Organ. 3:155-8.

22.

Silverberg, E. (1985). Cancer statistics Ca: a Cancer Journal for Clinicians . 35:19-35.

23.

Stutman, O. , Dein, P. , Wisun, R.E. , and Lattime, E.C. (1980). "Natural cytotxic cells against solid tumors in mice: Blocking of cytotoxicity by D-mannose." Proc. Natl. Acad. Sci. 77:2895-8.

24.

Thorgeirsson, S.S. and Nebert, D.W. (1977). "The Ah locus and the metabolism of chemical carcinogens and other foreign compounds." Adv. Can. Res. 25:149-93.

25.

Timonen, T. and Saksela, E. (1980). "Isolation of human NK cells by density gradient centrifugation." J. Immunol. Meth. 36:285-91.

26.

Vose, B.M. , Harding, M. , White, W. , Moore, M. , and Gallagher, J. (1983). "Effect of simple sugars on natural killing: evidence against the involvement of a lectin like mechanism in target recognition ." Clin. Exp. Immunol. 51:517-24.

27.

Werkmeister, J.A. , Roder, J.C. , Curry, C. , and Pross, H.F. (1983). "The effect of unphosphorylated and phosphorylated sugar moieties on human and mouse natural killer cell activity: Is there selective inhibition at the level of target recognition and lytic acceptor site?" Cell. Immunol. 80:172-7.

28.

Wright, S.C. and Bonavida, B. (1982). "Studies on the mechanism of natural killer (NK) cell-mediated cytotoxicity (CMC). I. Release of cytotoxic factors specific for NK-sensitive target cells (NKCF) during co-culture of NK effector cells with NK target cells." J. Immunol. 129:433-9.

29.

Yamamoto, H. , Fuyama, S. , Arai, S. , and Sendo, F. (1985). "Inhibition of mouse natural killer cytotoxicity by heparin." Cell. Immunol. 96:409-17.