Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1993 Dec;143(6):1713–1723.

Protection from tumor necrosis factor-mediated cytolysis by platelets.

C Philippe 1, B Philippe 1, B Fouqueray 1, J Perez 1, M Lebret 1, L Baud 1
PMCID: PMC1887257  PMID: 8256858

Abstract

Infiltrating macrophages elicit tumor-destructive reactions by releasing cytolytic factors including tumor necrosis factor alpha (TNF-alpha). Because platelets represent another major component of the cell infiltrate in tumors, we examined whether they could affect TNF alpha-induced cell death. Exposure of L-929 fibrosarcoma cells to human platelets reduced TNF alpha-induced cytotoxicity and cytolysis, as determined by 51Cr release assay and DNA fragmentation assay. This inhibitory effect, which depended on the concentration of platelets (0.1 to 10 x 10(6)/0.1 ml), was as high as 50%. The decrease in responsiveness to TNF-alpha reflected neither a degradation of TNF-alpha nor an inability of L-929 cells to bind TNF-alpha. Indeed, even though Scatchard analysis indicated the presence of 100 to 150 125I-TNF-alpha binding sites/platelet with a kd of 3.8 to 6.4 nM, addition of platelets up to 5 x 10(6)/0.1 ml did not compete with 125I-TNF-alpha binding to L-929 cells. Furthermore, addition of platelets 1 or 2 hours after that of TNF-alpha was still protective suggesting that platelets rather promoted hyporesponsiveness of L-929 cells to a postbinding effect of TNF-alpha Platelet-induced reduction of TNF-alpha response could be reproduced with supernatant fluids from platelets incubated at 37 degrees C for 24 hours. The platelet-derived factor responsible for this effect was found to be a lipid of low molecular weight with high affinity for albumin and charcoal. A role for 12(S) hydroxyeicosatetraenoic acid is proposed because this metabolite reduced TNF-alpha-induced cytolysis in a dose-dependent manner, whereas other platelet-derived lipids including thromboxane A2 and platelet activating factor were inactive. These observations indicate that the role of associated platelets has to be considered when analyzing the cytotoxic and cytolytic activity of macrophage-derived TNF-alpha on tumor cells.

Full text

PDF
1713

Images in this article

1717Figure 2
on p.1717
1717Figure 3
on p.1717

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams D. O. Effector mechanisms of cytolytically activated macrophages. I. Secretion of neutral proteases and effect of protease inhibitors. J Immunol. 1980 Jan;124(1):286–292. [PubMed] [Google Scholar]
  2. Baud L., Affres H., Perez J., Ardaillou R. Reduction in tumor necrosis factor binding and cytotoxicity by hydrogen peroxide. J Immunol. 1990 Jul 15;145(2):556–560. [PubMed] [Google Scholar]
  3. Bauvois B., Sancéau J., Wietzerbin J. Human U937 cell surface peptidase activities: characterization and degradative effect on tumor necrosis factor-alpha. Eur J Immunol. 1992 Apr;22(4):923–930. doi: 10.1002/eji.1830220407. [DOI] [PubMed] [Google Scholar]
  4. Chang J., Blazek E., Kreft A. F., Lewis A. J. Inhibition of platelet and neutrophil phospholipase A2 by hydroxyeicosatetraenoic acids (HETES). A novel pharmacological mechanism for regulating free fatty acid release. Biochem Pharmacol. 1985 May 1;34(9):1571–1575. doi: 10.1016/0006-2952(85)90701-4. [DOI] [PubMed] [Google Scholar]
  5. Damonneville M., Wietzerbin J., Pancré V., Joseph M., Delanoye A., Capron A., Auriault C. Recombinant tumor necrosis factors mediate platelet cytotoxicity to Schistosoma mansoni larvae. J Immunol. 1988 Jun 1;140(11):3962–3965. [PubMed] [Google Scholar]
  6. Fehsel K., Kolb-Bachofen V., Kolb H. Analysis of TNF alpha-induced DNA strand breaks at the single cell level. Am J Pathol. 1991 Aug;139(2):251–254. [PMC free article] [PubMed] [Google Scholar]
  7. Ferluga J., Schorlemmer H. U., Baptista L. C., Allison A. C. Production of the complement cleavage product, C3a, by activated macrophages and its tumorolytic effects. Clin Exp Immunol. 1978 Mar;31(3):512–517. [PMC free article] [PubMed] [Google Scholar]
  8. Gralnick H. R. von Willebrand factor, integrins, and platelets: their role in cancer. J Lab Clin Med. 1992 May;119(5):444–447. [PubMed] [Google Scholar]
  9. Hasday J. D., Shah E. M., Lieberman A. P. Macrophage tumor necrosis factor-alpha release is induced by contact with some tumors. J Immunol. 1990 Jul 1;145(1):371–379. [PubMed] [Google Scholar]
  10. Hawrylowicz C. M., Santoro S. A., Platt F. M., Unanue E. R. Activated platelets express IL-1 activity. J Immunol. 1989 Dec 15;143(12):4015–4018. [PubMed] [Google Scholar]
  11. Honn K. V., Nelson K. K., Renaud C., Bazaz R., Diglio C. A., Timar J. Fatty acid modulation of tumor cell adhesion to microvessel endothelium and experimental metastasis. Prostaglandins. 1992 Nov;44(5):413–429. doi: 10.1016/0090-6980(92)90137-i. [DOI] [PubMed] [Google Scholar]
  12. Honn K. V., Steinert B. W., Moin K., Onoda J. M., Taylor J. D., Sloane B. F. The role of platelet cyclooxygenase and lipoxygenase pathways in tumor cell induced platelet aggregation. Biochem Biophys Res Commun. 1987 May 29;145(1):384–389. doi: 10.1016/0006-291x(87)91333-7. [DOI] [PubMed] [Google Scholar]
  13. Ibele G. M., Kay N. E., Johnson G. J., Jacob H. S. Human platelets exert cytotoxic effects on tumor cells. Blood. 1985 May;65(5):1252–1255. [PubMed] [Google Scholar]
  14. Jättelä M. Biologic activities and mechanisms of action of tumor necrosis factor-alpha/cachectin. Lab Invest. 1991 Jun;64(6):724–742. [PubMed] [Google Scholar]
  15. Kamijo R., Takeda K., Nagumo M., Konno K. Suppression of TNF-stimulated proliferation of diploid fibroblasts and TNF-induced cytotoxicity against transformed fibroblasts by TGF-beta. Biochem Biophys Res Commun. 1989 Jan 16;158(1):155–162. doi: 10.1016/s0006-291x(89)80191-3. [DOI] [PubMed] [Google Scholar]
  16. Kinzer D., Lehmann V. Extracellular ATP and adenosine modulate tumor necrosis factor-induced lysis of L929 cells in the presence of actinomycin D. J Immunol. 1991 Apr 15;146(8):2708–2711. [PubMed] [Google Scholar]
  17. Kroll M. H., Schafer A. I. Biochemical mechanisms of platelet activation. Blood. 1989 Sep;74(4):1181–1195. [PubMed] [Google Scholar]
  18. Kumar S., Baglioni C. Protection from tumor necrosis factor-mediated cytolysis by overexpression of plasminogen activator inhibitor type-2. J Biol Chem. 1991 Nov 5;266(31):20960–20964. [PubMed] [Google Scholar]
  19. Köller M., König W. 12-Hydroxyeicosatetraenoic acid (12-HETE) induces heat shock proteins in human leukocytes. Biochem Biophys Res Commun. 1991 Mar 29;175(3):804–809. doi: 10.1016/0006-291x(91)91636-q. [DOI] [PubMed] [Google Scholar]
  20. König M., Wallach D., Resch K., Holtmann H. Induction of hyporesponsiveness to an early post-binding effect of tumor necrosis factor by tumor necrosis factor itself and interleukin 1. Eur J Immunol. 1991 Jul;21(7):1741–1745. doi: 10.1002/eji.1830210723. [DOI] [PubMed] [Google Scholar]
  21. Larrick J. W., Wright S. C. Cytotoxic mechanism of tumor necrosis factor-alpha. FASEB J. 1990 Nov;4(14):3215–3223. doi: 10.1096/fasebj.4.14.2172061. [DOI] [PubMed] [Google Scholar]
  22. Loetscher H., Gentz R., Zulauf M., Lustig A., Tabuchi H., Schlaeger E. J., Brockhaus M., Gallati H., Manneberg M., Lesslauer W. Recombinant 55-kDa tumor necrosis factor (TNF) receptor. Stoichiometry of binding to TNF alpha and TNF beta and inhibition of TNF activity. J Biol Chem. 1991 Sep 25;266(27):18324–18329. [PubMed] [Google Scholar]
  23. Matsuda M., Masutani H., Nakamura H., Miyajima S., Yamauchi A., Yonehara S., Uchida A., Irimajiri K., Horiuchi A., Yodoi J. Protective activity of adult T cell leukemia-derived factor (ADF) against tumor necrosis factor-dependent cytotoxicity on U937 cells. J Immunol. 1991 Dec 1;147(11):3837–3841. [PubMed] [Google Scholar]
  24. Molinas F. C., Wietzerbin J., Falcoff E. Human platelets possess receptors for a lymphokine: demonstration of high specific receptors for HuIFN-gamma. J Immunol. 1987 Feb 1;138(3):802–806. [PubMed] [Google Scholar]
  25. Nathan C. F., Brukner L. H., Silverstein S. C., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. I. Pharmacologic triggering of effector cells and the release of hydrogen peroxide. J Exp Med. 1979 Jan 1;149(1):84–99. doi: 10.1084/jem.149.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Perussia B. Tumor infiltrating cells. Lab Invest. 1992 Aug;67(2):155–157. [PubMed] [Google Scholar]
  27. Reiss M., Roos D. Differences in oxygen metabolism of phagocytosing monocytes and neutrophils. J Clin Invest. 1978 Feb;61(2):480–488. doi: 10.1172/JCI108959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Robaye B., Mosselmans R., Fiers W., Dumont J. E., Galand P. Tumor necrosis factor induces apoptosis (programmed cell death) in normal endothelial cells in vitro. Am J Pathol. 1991 Feb;138(2):447–453. [PMC free article] [PubMed] [Google Scholar]
  29. Ruggiero V., Johnson S. E., Baglioni C. Protection from tumor necrosis factor cytotoxicity by protease inhibitors. Cell Immunol. 1987 Jul;107(2):317–325. doi: 10.1016/0008-8749(87)90240-1. [DOI] [PubMed] [Google Scholar]
  30. Russell F. A., Deykin D. The use of high pressure liquid chromatography (HPLC) for the separation of radiolabeled arachidonic acid and its metabolities produced by thrombin-treated human platelets. I. The validation of the technique. Prostaglandins. 1979 Jul;18(1):11–18. doi: 10.1016/s0090-6980(79)80018-0. [DOI] [PubMed] [Google Scholar]
  31. Setty B. N., Graeber J. E., Stuart M. J. The mitogenic effect of 15- and 12-hydroxyeicosatetraenoic acid on endothelial cells may be mediated via diacylglycerol kinase inhibition. J Biol Chem. 1987 Dec 25;262(36):17613–17622. [PubMed] [Google Scholar]
  32. Todoroki N., Watanabe Y., Akaike T., Katagiri Y., Tanoue K., Yamazaki H., Tsuji T., Toyoshima S., Osawa T. Enhancement by IL-1 beta and IFN-gamma of platelet activation: adhesion to leukocytes via GMP-140/PADGEM protein (CD62). Biochem Biophys Res Commun. 1991 Sep 16;179(2):756–761. doi: 10.1016/0006-291x(91)91881-c. [DOI] [PubMed] [Google Scholar]
  33. Zhang X. M., Weber I., Chen M. J. Site-directed mutational analysis of human tumor necrosis factor-alpha receptor binding site and structure-functional relationship. J Biol Chem. 1992 Nov 25;267(33):24069–24075. [PubMed] [Google Scholar]
  34. van Kessel K. P., van Strijp J. A., Verhoef J. Inactivation of recombinant human tumor necrosis factor-alpha by proteolytic enzymes released from stimulated human neutrophils. J Immunol. 1991 Dec 1;147(11):3862–3868. [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES