Infect Immun 67(7): 3284-3289

Carrageenan Primes Leukocytes To Enhance Lipopolysaccharide-Induced Tumor Necrosis Factor Alpha Production

Department of Anesthesiology,^{Department of Molecular Biology,}^{and Department of Forensic Medicine,}^{School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan}

^{Corresponding author. Mailing address: Department of Anesthesiology, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishiku, Kitakyushu 807, Japan. Phone: (093) 691-7265. Fax: (093) 601-2910. E-mail:}pj.ca.u-heou.dem@atagom.

Received 1998 Dec 7; Revisions requested 1999 Feb 24; Accepted 1999 Apr 2.

Abstract

We have previously reported that pretreatment with carrageenan (CAR) enhances lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-α) production in and lethality for mice. Whole blood cultured in vitro was used to show that CAR pretreatment results in about a 200-fold increase in LPS-induced TNF-α production. CAR by itself did not induce TNF-α production. However, CAR-treated cultured medium sensitized whole blood to make more LPS-induced TNF than did saline-treated cultured medium in vitro. It was also demonstrated that CAR pretreatment increases TNF-α mRNA levels of both blood cells and peritoneal exudate cells, but not of bone marrow cells. Immunoelectron microscopic analysis revealed that polymorphonuclear leukocytes and macrophages are TNF-α-producing cells in CAR-treated mice. In CAR-treated mice, TNF-α was seen early after LPS injection in leukocytes in hepatic sinusoids and on the surfaces of endothelial cells. TNF-α was also detected late after LPS injection in hepatocytes which become edematous. These results suggest that CAR primes leukocytes to produce TNF-α in response to LPS and that they play an important role in the pathogenesis of liver injury.

Abstract

Tumor necrosis factor alpha (TNF-α) has a variety of biological activities which affect a number of cells, such as inhibition of cellular growth, production of cytokines, induction of shock, and so on (2, 4, 33). In general, TNF-α is produced in macrophages by stimulation of lipopolysaccharide (LPS) (19). It has been demonstrated that Mycobacterium bovis BCG and Propionibacterium acnes (Corynebacterium parvum) increase the sensitivity of macrophages to LPS (31, 37), and this priming effect on macrophages appears at least 4 days after the administration of macrophage activators (37). d-Galactosamine and actinomycin D have been also used as endotoxin sensitizers (9–12, 32). Unlike BCG and C. parvum, treatment with d-galactosamine increases susceptibility of mice to the lethal effects of LPS several thousand-fold immediately (10, 11). UTP depletion by d-galactosamine is considered to be responsible for the development of sensitization to LPS (11). In addition, d-galactosamine does not affect LPS-induced cytokine gene expression in Kupffer cells (7).

On the other hand, carrageenan (CAR; a high-molecular-weight sulfated polygalactose isolated from marine plants) increases LPS-induced TNF-α production at least 2 h after it is injected (22). Since CAR is known to destroy macrophages (6, 27), it is likely that TNF-α is not produced in macrophages but in other cells. Polymorphonuclear leukocytes (PMNs) are widely accepted as key effector cells in both host defense and tissue destruction. Although PMNs are viewed as terminally differentiated cells which are devoid of RNA and protein synthesis, several lines of evidence have shown that PMNs release various cytokines, including TNF-α (5, 8, 18).

In the present study, we demonstrated that the pretreatment of mice with CAR increases TNF-α mRNA levels in leukocytes. Further, primed leukocytes including PMNs produce a large amount of TNF-α in response to LPS and play a major role in the pathogenesis of liver injury.

ACKNOWLEDGMENTS

We thank Robert S. Munford, Southwestern Medical School, Dallas, Tex., for his critical review of the manuscript and helpful advice.

This work was supported in part by Grant-in-Aid for Scientific Research (C) 10671453 from the Ministry of Education, Science, Sport and Culture of Japan.

ACKNOWLEDGMENTS

REFERENCES

References

1. Bagglioni M, Dewald B, Thelen MEffects of PAF on neutrophils and mononuclear phagocytes. Prog Biochem Pharmacol. 1988;22:90.[PubMed][Google Scholar]
2. Bazzoni F, Beutler BThe tumor necrosis factor ligand and receptor families. N Engl J Med. 1996;334:1717–1725.[PubMed][Google Scholar]
3. Bazzoni F, Cassatella M A, Laudanna C, Rossi FPhagocytosis of opsonized yeast induces tumor necrosis factor-α mRNA accumulation and protein release by human polymorphonuclear leukocytes. J Leukoc Biol. 1991;50:223–228.[PubMed][Google Scholar]
4. Beutler B, Cerami ACachectin: more than a tumor necrosis factor. N Engl J Med. 1987;316:379–385.[PubMed][Google Scholar]
5. Cassatella M AThe production of cytokines by polymorphonuclear neutrophils. Immunol Today. 1995;16:21–26.[PubMed][Google Scholar]
6. Catanzaro P J, Schwartz H J, Graham R C., Jr Spectrum and possible mechanism of carrageenan cytotoxicity. Am J Pathol. 1971;64:387–404.
7. De S K, Silverstein R, Andrews G KHydrazine sulfate protection against endotoxin lethality: analysis of effects on expression of hepatic cytokine genes and as acute-phase gene. Microb Pathog. 1992;13:37–47.[PubMed][Google Scholar]
8. Djeu Y J. TNF production by neutrophils. In: Beutler B, editor. Tumor necrosis factor. New York, N.Y: Raven Press; 1992. pp. 531–537. [PubMed]
9. Freudenberg M A, Galanos CInduction of tolerance to lipopolysaccharide (LPS)-d-galactosamine lethality by pretreatment with LPS is mediated by macrophages. Infect Immun. 1988;56:1352–1357.[Google Scholar]
10. Freudenberg M A, Galanos CTumor necrosis factor alpha mediates lethal activity of killed gram-negative and gram-positive bacteria in d-galactosamine-treated mice. Infect Immun. 1991;59:2110–2115.[Google Scholar]
11. Freudenberg M A, Keppler D, Galanos CRequirement for lipopolysaccharide-responsive macrophages in galactosamine-induced sensitization to endotoxin. Infect Immun. 1986;51:891–895.[Google Scholar]
12. Gonzalez J C, Johnson D C, Morrison D C, Freudenberg M A, Galanos C, Silverstein REndogenous and exogenous glucocorticoids have different roles in modulating endotoxin lethality in d-galactosamine-sensitized mice. Infect Immun. 1993;61:970–974.[Google Scholar]
13. Han J, Huez G, Beutler BInteractive effects of the tumor necrosis factor promoter and 3′-untranslated regions. J Immunol. 1991;146:1843–1848.[PubMed][Google Scholar]
14. Hewett J A, Jean P A, Kunkel S L, Roth R ARelationship between tumor necrosis factor-α and neutrophils in endotoxin-induced liver injury. Am J Physiol. 1993;265:1011–1015.[PubMed][Google Scholar]
15. Koga K, Ogata M, Takenaka I, Matsumoto T, Shigematsu AKetamine suppresses tumor necrosis factor-α activity and mortality in carrageenan-sensitized endotoxin shock model. Circ Shock. 1995;44:160–168.[PubMed][Google Scholar]
16. Kriegler M, Perez C, DeFay K, Albert I, Lu S DA novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988;53:45–53.[PubMed][Google Scholar]
17. Lehmann V, Freudenberg M A, Galanos CLethal toxicity of lipopolysaccharide and tumor necrosis factor in normal and d-galactosamine-treated mice. J Exp Med. 1987;165:657–663.[Google Scholar]
18. Lindemann A, Riedel D, Oster W, Ziegler-Heitbrock H W, Mertelsmann R, Herrmann FGranulocyte-macrophage colony-stimulating factor induces cytokine secretion by human polymorphonuclear leukocytes. J Clin Investig. 1989;83:1308–1312.[Google Scholar]
19. Mannel D N, Moore R N, Mergenhagen S EMacrophages as a source of tumoricidal activity (tumor-necrotizing factor) Infect Immun. 1980;30:523–530.[Google Scholar]
20. Nagano T, Kita T, Tanaka NThe immunocytochemical localization of tumor necrosis factor and leukotriene in the rat liver after treatment with lipopolysaccharide. Int J Exp Pathol. 1992;73:657–683.[Google Scholar]
21. Ogata M, Matsumoto T, Koga K, Takenaka I, Kamochi M, Sata T, Yoshida S-I, Shigematsu AAn antagonist of platelet-activating factor suppresses endotoxin-induced tumor necrosis factor and mortality in mice pretreated with carrageenan. Infect Immun. 1993;61:699–704.[Google Scholar]
22. Ogata M, Yoshida S-I, Kamochi M, Shigematsu A, Mizuguchi YEnhancement of lipopolysaccharide-induced tumor necrosis factor production in mice by carrageenan pretreatment. Infect Immun. 1991;59:679–683.[Google Scholar]
23. Ohira H, Ueno T, Torimura T, Tanikawa K, Kasukawa RLeukocyte adhesion molecules in the liver and plasma cytokine levels in endotoxin-induced rat liver injury. Scand J Gastroenterol. 1995;30:1027–1035.[PubMed][Google Scholar]
24. Parant M, Le Contel C, Parant F, Chedid LInfluence of endogenous glucocorticoid on endotoxin-induced production of circulating TNF-α Lymphokine Cytokine Res. 1991;10:265–271.[PubMed][Google Scholar]
25. Redl H, Nikolai A, Kneidinger R, Schlag GEndothelial and leukocyte activation in experimental polytrauma and sepsis. Behring Inst Mitt. 1993;92:218–228.[PubMed][Google Scholar]
26. Robache-Gallea S, Morand V, Bruneau J M, Schoot B, Tagat E, Realo E, Chouaib S, Roman-Roman S. In vitro processing of human tumor necrosis factor-α J Biol Chem. 1995;270:23688–23692.[PubMed]
27. Sawicki J E, Catanzaro P JSelective macrophage cytotoxicity of carrageenan in vivo. Int Arch Allergy Appl Immunol. 1975;49:709–714.[PubMed][Google Scholar]
28. Schlayer H J, Laaff H, Peters T, Woort-Menker M, Estler H C, Karck U, Schaefer H E, Decker KInvolvement of tumor necrosis factor in endotoxin-triggered neutrophil adherence to sinusoidal endothelial cells of mouse liver and its modulation in acute phase. J Hepatol. 1988;7:239–249.[PubMed][Google Scholar]
29. Smedly L A, Tonnesen M G, Sandhause R A, Haslett C, Guthrie L A, Johnston R B, Jr, Henson P M, Worthen G SNeutrophil-mediated injury to endothelial cells. J Clin Investig. 1986;77:1233–1243.[Google Scholar]
30. Smith J ANeutrophils, host defense, and inflammation: a double-edged sword. J Leukoc Biol. 1994;56:672–686.[PubMed][Google Scholar]
31. Tasaka S, Ishizaka A, Urano T, Sayama K, Sakamaki F, Nakamura H, Terashita T, Waki Y, Soejima K, Oyamada Y, Fujishima S, Kanazawa MBCG priming enhances endotoxin-induced acute lung injury independent of neutrophils. Am J Respir Crit Care Med. 1995;152:1041–1049.[PubMed][Google Scholar]
32. Tiegs G, Niehorster M, Wendel ALeukocyte alterations do not account for hepatitis induced by endotoxin or TNF-α in galactosamine-sensitized mice. Biochem Pharmacol. 1990;40:1317–1322.[PubMed][Google Scholar]
33. Tracey K J, Beutler B, Lowry S F, Merryweather J, Wolpe S, Milsark I W, Hariri R J, Fahey III T J, Zentella A, Albert J D, Shires T, Cerami AShock and tissue injury induced by recombinant human cachectin. Science. 1987;234:470–474.[PubMed][Google Scholar]
34. Ullrich A, Shine J, Chirgwin J, Pictet R, Tischer E, Rutter W J, Goodman H MRat insulin genes: construction of plasmids containing the coding sequences. Science. 1977;196:1313–1319.[PubMed][Google Scholar]
35. Wang J H, Redmond H P, Watson R W, Duggan S, McCarthy J, Barry M, Bouchier-Hayes DMechanisms involved in the induction of human endothelial cell necrosis. Cell Immunol. 1995;168:91–99.[PubMed][Google Scholar]
36. Wang J H, Redmond H P, Watson R W, Bouchier-Hayes DRole of lipopolysaccharide and tumor necrosis factor-α in induction of hepatocyte necrosis. Am J Physiol. 1995;269:G297–G304.[PubMed][Google Scholar]
37. Yoshikai Y, Miake S, Mitsuyama M, Nomoto KEffects of Corynebacterium parvum on Escherichia coli infection in mice. J Gen Microbiol. 1982;128:2857–2863.[PubMed][Google Scholar]