Platelet Activation Induced by lnterleukin-6: Evidence for a Mechanism Involving Arachidonic Acid Metabolism

Summary

The effect of IL-6 on in vitro platelet function was investigated. Platelet-rich plasma (PRP) incubated with IL-6 showed a dose dependent enhancement of agonist induced maximum aggregation (AIMA) and secretion of thromboxane B₂ (TXB₂) as measured by RIA, in short term incubations. Dazoxiben (0.2 to 160 (µM) pretreated PRP incubated with IL-6 and aggregated with ionophore A23187, showed a dose dependent inhibition of TXB₂ secretion concomitant with a dose dependent abrogation of IL-6’s enhancement of AIMA. A similar abrogation of AIMA was observed when these experiments were repeated using indomethacin. Further, PRP incubated with IL-6 showed a dose dependent increase in TXB₂ and BTG secretion as measured by RIA and an increased incorporation of actin binding protein, talin, and myosin into the cytoskeletal core (triton insoluble residue) as shown by SDS-PAGE. The integrin glycoprotein Ilia (GPIIIa) was also observed to be retained into the cytoskeleton by immunoblot. These results suggest that IL-6 activates platelets in vitro and enhances AIMA via a mechanism involving arachidonic acid metabolism.

• References

• 1 Kishimoto T. The biology of interleukin-6. Blood 1989; 74: 1-10
• 2 Smyth MJ, Ortaldo RJ. Comparison of the effect of interleukin-2 and interleukin-6 on lytic activity of purified human peripheral blood large granular lymphocytes. J of Immunology 1990; 146: 1380-1384
• 3 Van Snick J. Interleukin-6: an overview. Annu Rev Immunol 1990; 8: 253-278
• 4 Cheson B, Carter CL, Sznol M. Clinical Trials Referral Resource: NCI-sponsored Trials of Interleukin-6. Oncology 1993; 7: 43-44
• 5 Ishibashi T, Kimura H, Shikama Y, Uchida T, Kariyone S, Hirano T, Kishimoto T, Takatsuki F, Akiyana Y. Interleukin-6 is a potent thrombo-poietic factor in vivo in mice. Blood 1989; 74: 1241-1244
• 6 Mayer P, Geissler K, Valent P, Ceska M, Bettelheim P, Liehl E. Recombinant human interleukin-6 is a potent inducer of acute phase response and elevates the blood platelets in nonhuman primates. Exp-Hematol 1991; 7: 688-696
• 7 Mei RL, Burstein SA. Megakaryocytic maturation in murine long term bone marrow culture; role of interleukin-6. Blood 1991; 78: 1438-1447
• 8 Stahl DP, Zucker-Franklin D, Evatt BL, Winton EF. Effects of human interleukin-6 on megakaryocyte development and thrombocytopoiesis in primates. Blood 1991; 78: 1467-1475
• 9 Burstein SA, Peng J, Dale GL, Hamilton KK, Comp PC, George JN. Pro-hemostatic effects of Interleukin-6: augmentation of platelet activation and plasma von Willebrand factor and reduction in free protein S in dogs. Thromb Haemost 1993; 69: 749 (Abstract)
• 10 Waring P, Wall Dominic, Dauer R, Parkin D, Metcalf D. The effects of leukaemia inhibitory factor on platelet function. British Journal of Hematology 1993; 83: 80-87
• 11 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-683
• 12 Gunn HC, Weber J, Myers LA, Mullarkey MA, Beggs JL, Habucky K, Levitt D. Pharmacokinetic profile of rIL-6 in four patients. Blood 1992; (Suppl. 01) 80: 414 (abstract)
• 13 Spinas GA, Bloesch D, Keller U, Zimmerli W, Cammisuli S. Pretreatment with ibuprofen augments circulating tumor necrosis factor-a, interleukin-6, and elastase during acute endotoxinemia. J Infect Dis 1991; 163: 89-95
• 14 Waage A, Brandtzaeg P, Halstensen A, Kierulf P, Espevik T. The complex pattern of cytokines in serum from patients with meningococcal septic shock. J Exp Med 1989; 168: 333-338
• 15 Symer DE, Paznekas A, Shin HS. A requirement for membrane-associated phospholipase A2 in platelet cytotoxicity activated by receptors for immunoglobulin G and complement. J Exp Med 1993; 177: 937-947
• 16 Oleksowicz L, Zuckerman D, Mrowiec Z, Puszkin E, Dutcher J. Effects of Interleukin-2 on platelet function in cancer patients. Am J Hematol 1994; 45: 224-231
• 17 Avenarius HJ, Freund M, Deinhaardt J, Poliwoda H. Effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on circulating platelets. Ann Hematol 1992; 65: 6-9
• 18 Lapetina EG, Chandrabose KA, Cuatrecasas P. Ionophore A23187- and thrombin-induced platelet aggregation: Independence from cyclooxygenase products. Proc Natl Acad Sci USA 1978; 75: 818-822
• 19 Mobely A, Tai H. Synergistic stimulation of thromboxane biosynthesis by calcium ionophore and phorbol ester or thrombin in human platelets. Biochemical and Biophysical Research Communications 1985; 130: 717-723
• 20 Rittenhouse-Simmons S. Differential Activation of platelet phospholipases by thrombin and ionophore A23187. J of Biol Chem 1981; 256: 4153-4155
• 21 Sano K, Nakamura H, Matsuo T, Kawahara Y, Fukuzaki H, Kaibuchi K, Takai Y. Comparison of the modes of action of Ca2+ ionophore A23187 and thrombin in protein kinase C activation in human platelets. FEBS lett 1985; 192: 4-8
• 22 Feinman RD, Detwiler TC. Platelet secretion induced by divalent cation ionophores. Nature 1974; 249: 172-175
• 23 Vavarra P, Tsagarakis S, Faria MS, Rees LH, Besser GM, Grossman AB. Interleukins-1 and -6 stimulate the release of corticotropin-releasing hormone-41 from rat hypothalamus in vitro via the eicosanoid cyclooxygenase pathway. Endocrinology 1991; 128: 37-44
• 24 Crowl RM, Stoller TJ, Conroy RR, Stoner CR. Induction of phospholipase A2 gene expression in human hepatoma cells by mediators of the acute phase reponse. J of Biol Chem 1991; 266: 2647-2651
• 25 Stasi M, Gresele P, Porcellati S, Quero E, Nenci GG, Coracci G. Activation of phospholipase A2 and beta-thromboglobulin release in human platelets: comparative effects of thrombin and fluoroaluminate stimulation. Bio-chimica et Biophysica Acta 1992; 1124: 279-287
• 26 Shimoda K, Okamura S, Harada N, Okamura T, Niho Y. Identification of a functional receptor for granulocytes colony-stimulating factor on platelets. J Clin Invest 1993; 91: 1310-1313
• 27 Molinas FC, Wietzwebin J, Falcoff E. Human platelets possess receptors for a lymphokine: demonstration of high specific receptors for HuIFN-gam-ma. J Immunol 1987; 138: 802-806
• 28 Todoroki N, Watanabe Y, Akaike T, Katagiri Y, Tanoue K, Yamazaki H, Tsuji T, Toyoshima S, Osawa T. Enhancement by IL-1 and IFN-gamma of platelet activation: Adhesion to leukocytes via GMP-140/PADEM protein (CD62). Biochem Biophys Res Commun 1991; 179: 756-761
• 29 Navarro S, Debili N, Couedic JP, Klein B, Breton-Gorius J, Doly J, Vainchenker W. Interleukin-6 and its receptor are expressed by human megakaryocytes: in vitro effects on proliferation and endoreplication. Blood 1991; 77: 461-471
• 30 Pancre V, Monte D, Delanoye A, Captron A, Auriault C. Interleukin-6 is the main mediator of the interaction between monocytes and platelets in the killing of Schistosoma mansoni. Eur-Cytokine-Netw 1990; 1: 15-19
• 31 Bauer J, Bauer TM, Kalb T, Taga T, Lengyel G, Hirano T, Kishimoto T, Acs G, Mayer L, Gerok W. Regulation of interleukin 6 receptor expression in human monocytes and monocyte-derived macrophages. Comparison with the expression in human hepatocytes. J Exp Med 1989; 170: 1537-1549
• 32 Oleksowicz L, Paciucci AP, Zuckerman D, Colerito A, Rand JH, Holland JF. Alterations of platelet function induced by interleukin-2. J Immunotherapy 1991; 10: 369-370
• 33 Fox JEB. The platelet cytoskeleton. In: Thrombosis and Haemostasis Ver-traete M, Vermylen J, Lijnen HR, Arnout J. eds Leuven University Press; Leuven: 1987: 175-185
• 34 Phillips DR, Jenings LK, Edwards HH. Identification of membrane proteins mediating the interaction of human platelets. J Cell Biol 1980; 86: 77-86
• 35 Puszkin EG, Mauss EA, Milot DC, Zucker MB. Effect of the thiol group inhibitor monobromobimane and other inhibitors on the composition of the platelet cytoskeletal core and its association with glycoprotein Ilia. J Cell Biochem 1989; 39: 339-354
• 36 Wheeler ME, Cox AC, Carroll RC. Retention of the glycoprotein Ilb-IIIa complex in the isolated platelet cytoskeleton. Effect of separable assembly of platelet pseudopodal and contractile cytoskeletons. J Clin Invest 1984; 74: 1080-1089
• 37 Collier NC, Wang K. Human platelet P235: A high Mr. protein which restricts the length of actin filaments. FEBS lett 1982; 143: 205-209
• 38 Puszkin EG, Mauss EA, Zucker MB. Assembly and GPIIIa content of cytoskeletal core in platelets agglutinated with bovine von Willebrand Factor. Blood 1990; 76: 1572-1579