Sulfated Polysaccharide Anticoagulants Suppress Natural Killer Cell Activity In Vitro

Summary

In contrast to the well studied anticoagulant activity of sulfated polysaccharides (SPS), little is known about their influence on immune competent cells. Using two naturally derived SPSs (unfractionated heparin and low molecular weight heparin), one semi-synthetic SPS (pentosanpolysulfate), and one synthetic SPS (lactobionic acid) as well as the polypeptide hirudin we investigated the effect of these drugs on natural killer cell activity in vitro. We demonstrate that all SPSs tested significantly suppress the activity of natural killer cells at clinically relevant dosages. At all concentrations and at all effector:target ratios tested pentosanpolysulfate was the most potent natural killer cell inhibitor. In contrast, hirudin had no effect on natural killer cell function. Furthermore, scanning electron microscopy revealed that reduced natural killer cell activity is paralleled by decreased lymphocyte cell size and altered cell surface structures. Our results indicate that defined therapeutically applied SPSs can interfere with the tumor cell killing process.

• References

• 1 Fareed J, Hoppensteadt D, Walenga JM, Bick RL. Current trends in the development of anticoagulant and antithrombotic drugs. Med Clin North Am 1994; 78: 713-731
• 2 Laghi-Pasini F, Pasqui AL, Ceccatelli L, DiPerri T. Heparin inhibits fMLP and ConA dependent activation of human polymorphonuclear leukocytes in vitro. Int J Tiss Reac 1983; 2: 144-51
• 3 Lazarowski ER, Santome JA, Behrens NH, Sanches Avalos JC. Aggregation of human neutrophils by heparin. Thrombosis Research 1986; 41: 437-446
• 4 Labrouche S, Freyburger G, Belloc F, Boisseau MR. Influence of selected heparins on human neutrophil functions in vitro. Thromb Haemost 1992; 68: 556-562
• 5 Johann S, Zoller C, Haas S, Blümel G, Förster R. Anticoagulant glycosaminoglycans activate respiratory burst in neutrophils and monocytes. Clin Appl Throm Haemost. 1995 in press
• 6 Sugawara I, Ishizaka S. Polysaccharides with sulfate groups are human T-cell mitogens and murine polyclonal B-cell activators (PBAs). Cell Immunol 1982; 74: 162-171
• 7 Frieri M, Metcalf DD. Analysis of effect of mast cell granules on lymphocyte blaszogenesis in the absence and presence of mitogens: identification of heparin as a granule-associated suppressor factor. J Immunol 1983; 131: 1942-1948
• 8 Eskinazi DP, Perna JJ, Ershow AG, Sharrow SO. Effects of heparin on in vitro immune parameters. J Biol Response Mod 1988; 7: 173-184
• 9 Ferrarini M, Grossi CE. Definition of the cell types within the so called ”null lymphocytes” population of human peripheral blood. An analysis of phenotypes and functions Sem Hematol 1984; 21: 270-286
• 10 Herberman RB. Natural killer cells. Anu Rev Med 1986; 37: 347-352
• 11 Trinchieri G. Biology of natural killer cells. Adv Immunol 1989; 47: 187-376
• 12 Giorda R, Rudert WA, Vavassori C, Chambers WH, Hiserodt JC, Trucco M. NKR-P1, a signal transduction molecule on natural killer cells. Science 1990; 249: 1298-1300
• 13 Chambers WH, Vujanovic NL, De LA, Olszowy MW, Herberman RB, Hiserodt JC. Monoclonal antibody to a triggering structure expressed on rat natural killer cells and adherent lymphokine-activated killer cells. J Exp Med 1989; 169: 1373-1389
• 14 Ryan JC, Niemi EC, Goldfien RD, Hiserodt JC, Deaman ME. NKR-P1, an activating molecule on rat natural killer cells, stimulates phosphoinositide turnover and rise in intracellular calcium. J Immunol 1991; 147: 3244-3250
• 15 Bezouska K, Yuen CT, O’Brirn J, Childs RA, Chai E, Lawson AM, Drbal K, Fiserova A, Pospisil M, Feizi T. Oligosaccharide ligands for NKR-P1 protein activate NK cells and cytotoxicity. Nature 1994; 372: 150-157
• 16 Böjum A. Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Invest 1968; 21: 77-89
• 17 Johann S, Bliimel G, Lipp M, Förster R. A versatile flow cytometry based assay for the determination of short- and long-term natural killer cell activity. J Immunol Meth. 1995 in press
• 18 Ludwig H, Metzger H. In: The human reproductive tract. A scanning microscopic atlas Ludwig H, Metzger H. eds. Heidelberg: Springer; 1976: 3-6
• 19 Aren C. Heparin and protamine therapy. Semin Thorac Cardiovasc Surg 1990; 2: 364-372
• 20 Mebius RE, Watson SR. L- and E-selectin can recognize the same naturally occurring ligands on high endothelial venules. J Immunol 1993; 151: 3252-3260
• 21 Nelson RM, Cecconi O, Roberts WG, Aruffo A, Linhardt RJ, Bevilacqua MP. Heparin oligosaccharides bind L- and P-selectin and inhibit acute inflammation. Blood 1993; 82: 3253-3258
• 22 Norgard SK, Varki NM, Varki A. Calcium-dependent heparin-like ligands for L-selectin in nonlymphoid endothelial cells. Science 1993; 261: 480-483
• 23 Bradley LM, Watson SR, Swain SL. Entry of native CD4 T cells into peripheral lymphnodes requires L-selectin. J Exp Med 1994; 180: 2401-2406
• 24 Barrowcliffe TW, Johnson EA, Thomas D. Antithrombin III and heparin. Brit Med Bull 1978; 34: 143-154
• 25 Forbes JT, Bretthauer RK, Oeltmann TN. Mannose 6-, fructose 1-, and fructose 6-phosphate inhibit natural cell-mediated cytotoxicity. Proc Nat Acad Sci 1981; 78: 5797-5802
• 26 Chambers WH, Oeltmann RN. The effects of hexose 6-O-sulfate esters on human natural killer lytic function. J Immunol 1986; 137: 1469-1475