Differential Expression of IL-1β,TNF-α,IL-6,and IL-8 in Human Monocytes in Response to Lipopolysaccharides from Different Microbes

Abstract

Macrophages respond to bacterial lipopolysaccharides (LPS) and activate several host defense functions through production of mediators. However, it is not clear whether the degree of macrophage responsiveness to different sources of LPS is equivalent to or varies with the source of LPS. Therefore, in this report, we examined the extent of the human monocyte response to LPS derived from two oral pathogens, Actinobacillus actinomycetemcomitans (Aa) and Porphyromonas gingivalis (Pg). Additionally, due to its well-established ability to activate monocytes, we used LPS from Escherichia coli (Ec). Human monocytes, when activated with a specific source of LPS, exhibited rapid expression of mRNA for IL-1β, TNF-a, and IL-8, which was followed by IL-6, as measured by RNA-PCR. Moreover, the expression of mRNA for these cytokines was followed by cytokine synthesis. Monocytes from the same subject, when activated with LPS from Pg, Aa, or Ec expressed quantitatively different levels of mRNA and proteins for all four cytokines. A given LPS induced either high or low expression of the battery of cytokines tested, indicating that the expression of these pro-inflammatory cytokines may be regulated by a single or a cluster of gene(s). However, no apparent differences in the time course of mRNA expression for these cytokines were observed in response to any of the LPS tested. Furthermore, the relative ability of the different sources of LPS to induce mRNA for cytokines varied throughout a wide range of LPS concentrations. This suggests that differences exist in the sensitivity of monocytes to a specific LPS, rather than in the kinetics of the secretory process itself. The ability of LPS to induce cytokine-specific mRNA also depended on the source of monocytes. Our results demonstrate that monocyte activation and cytokine release depend on the physicochemical form of LPS as well as the source of monocytes. These critical determinants may be significant in the pathogenesis of periodontal infections.

Keywords

lipopolysaccharide human monocytes pro-inflammatory cytokines

Get full access to this article

View all access options for this article.

References

Adams DO , Koener TJ (1988). Gene regulation in macrophage development and activation. In: The year in immunology. Vol. 4. Cellular, molecular and clinical aspects. Cruse JM , Lewis RE Jr , eds. Basel: Karger, p. 159.

Beutler B. , Cerami A. (1989). The biology of cachectin/TNF: a primary mediator of the host response. Ann Rev Immunol 7:625-655.

Beutler B, Krochin N, Milsark IW, Leudke C, Cerami A (1986). Control of cachectin (tumor necrosis factor) synthesis: Mechanism of endotoxin resistance. Science 232:977-980. Cavaillon JM, Haeffner-Cavaillon N (1990). Signals involved in interleukin-1 synthesis and release by lipopolysaccharide-stimulated monocytes/macrophages. Cytokine 2:1-11. Chomczynski P, Sacchi N (1987). Single step method of RNA isolation by acid guanidium thiocyanate-phenolchloroform extraction. Anal Chem 162:156-159.

Couturier C. , Haeffner-Cavaillon N. , Caroff M. , Kazatchkine MD (1991). Binding sites for endotoxins (lipopolysaccharides) on human monocytes. J Immunol 147:1899-1904.

Cramer EB (1992). Development and distribution of mononuclear phagocytes In: Inflammation. Gallin JI , Goldstein IM , Snyderman R , eds. New York: Raven Press , p. 325.

Dinarello CA , Cannon JG , Mier JW , Bernheim HA , Lo Preste G. , Lynn DL , et al. (1986). Multiple biological activities of human recombinant interleukin-1. J Clin Invest 77 :1734-1742.

Ernst TJ , Ritchie AR , Griffin JD (1989). Regulation of G- and M-CSF in monocytes. J Biol Chem 264:5700-5705.

Garrison SW , Holt SC , Nichols FC (1987). Lipopolysaccharide-stimulated PGE2 release from human monocytes. LPSelicited secretory responses in monocytes. Comparison of LPS prepared from suspected periodontal pathogens. J Periodontol 59:684-687.

Garrison SW , Holt SC , Nichols FC (1988). Lipopolysaccharide-stimulated PGE2 release from human monocytes. J Periodontol 59:684-691.

10.

Gemmel E. , Seymour GJ (1993). Interleukin 1, interleukin 6 and transforming growth factor-ß production by human gingival mononuclear cells following stimulation with Porphyromonas gingivalis and Fusobacterium nucleatum. J Periodont Res 28:122-129.

11.

Han J. , Brown T. , Beutler B. (1990). Endotoxin-responsive sequences control cachectin/tumor necrosis factor biosynthesis at the translational level. J Exp Med 171 :465-

12.

475.

13.

Hanazawa S, Nakada K, Ohmori Y, Miyoshi T, Amano S, Kitano S (1985). Functional role of interleukin-1 in periodontal disease: Induction of IL-1 production by Bacteroides gingivalis lipopolysaccharide in macrophages from C3H/Hen and C3NH/HeJ mice. Infect Immun 50:262-270. Heasman PA, Collins JG, Offenbacher S (1993). Changes in crevicular fluid levels of interleukin-1 beta, leukotriene B4, prostaglandin E2, thromboxane B2, and tumour necrosis factor alpha in experimental gingivitis in humans. J Periodont Res 28:241-247.

14.

Henricson BE, Benjamin WR, Vogel SN (1990). Differential cytokine induction by doses of lipopolysaccharide and monophosphoryl lipid A that result in equivalent early endotoxin tolerance. J Am Soc Microbiol 58:2429-2437. Keller R, Fischer W, Keist R, Bassetti S (1992). Macrophage response to bacteria: Induction of marked secretory and cellular activities by lipoteichoic acids. Infect Immun 60:3664-3672.

15.

Le J. , Vilcek J. (1987). Biology of disease. Tumor necrosis factor and interleukin-1: Cytokines with multiple overlapping biological activities. Lab Invest 56:234-248.

16.

Lei M. -G, Flebbe L. , Roeder D. , Morrison DC (1990). Identification and characterization of lipopolysaccharide receptor molecules on mammalian lymphoid cells. Adv Exp Med Biol 256:445-466.

17.

Masada MP, Persson R, Kennedy JS, Lee SW, Page RC, Allison AC (1990). Measurement of interleukin-1β and-1β in gingival crevicular fluid: implications for the pathogenesis of periodontal disease. J Periodont Res 25:156-163. Matsushima K, Taguchi M, Kovacs EJ, Young HA, Oppenheim JJ (1986). Intracellular localization of human monocyte associated interleukin 1 (IL-1) activity and release of biologically active IL-1 from monocytes by trypsin and plasmin. J Immunol 136:2883-2889.

18.

McFarlane CG, Reynolds JJ, Melkie MC (1990). The release of interleukin-1β, tumor necrosis factor-a and interferon-P by cultured peripheral blood mononuclear cells from patients with periodontitis. J Periodont Res 25:207-214. Mohamed-Ali T-J, Chaby R (1990b). Specific binding of lipopolysaccharides to mouse macrophages-I. Characteristics of the interactions and inefficiency of the polysaccharide region. Molec Immunol 27:751-761. Mohamed-Ali T-J, Mondange M, LeDur A, Auzanneau F-I, Charon D, Girard R, et al. (1990a). Specific binding of lipopolysaccharides to mouse macrophages-II. Involvement of distinct lipid A substructures. Molec Immunol 27:763-770.

19.

Movat HZ , Cybulsky MI , Golditz IG , Chan MK , Dinarello CA (1987). Acute inflammation in Gram-negative infection: endotoxin, interleukin-1, tumor necrosis factor, and neutrophils. Fed Proc 46:97-104.

20.

Novotny A. (1984). Heterogeneity of endotoxin. In: Handbook of endotoxins. Vol. 1. Reitschel ET , ed. Amsterdam: Elsevier Science Publishers, p. 308.

21.

Offenbacher S. , Collins JG , Heasman PA (1993). Diagnostic potential of host response mediators . Adv Dent Res 7 :175-181.

22.

Postlethwaite EAE , Lachman LB , Mainardi CL , Kang AH ( 1983). Interleukin-1 stimulation of collagenase production by

23.

cultured fibroblasts. J Exp Med 157:801-808.

24.

Revel M. (1990). Host defense against infections in inflammations: role of the multifunctional IL-6/IFN beta2 cytokine. Experientia 45:549-557.

25.

Rietschel ET, Brade L, Schade U, Seydel U, Zahringer U, Kusumoto S, et al. (1988). Bacterial endotoxins: properties and structure of biologically active domains. In: Surface structures of microorganisms and their interactions with the mammalian host. Vol. 18. Workshop conferences. Schrinner E, Richmond MH, Seibert J, Schwarz U, editors. Weinheim: Verlag Chemie, pp. 1-42.

26.

Schifferle RE , Reddy MS , Zambone JJ , Genco RJ , Levine MJ (1989). Characterization of a polysaccharide antigen from B. gingivalis. J Immunol 143:3035-3042.

27.

Stashenko P. , Obemesser MS , Prostak L. , Haffajee AD , Socransky SS (1991). Levels of interleukin-1β from active periodontal disease. J Clin Periodontol 18:548-554.

28.

Taffet SM , Singhel KJ , Overholtzer JF , Shurtleff SA (1989). Regulation of tumor necrosis factor in a macrophage-like line by lipopolysaccharide and cylic AMP. Cell Immunol 120:291-300.

29.

Vilcek J. , Palombella VJ , Henriksen-Destefano D. , Swenson C. , Fenman R. , Hirai M. , et al. (1986). Fibroblast growth enhancing activity of tumor necrosis factor and its

30.

relationship to other polypeptide growth factors. J Exp Med 163:632-639.

31.

Vogel SN , Hogan MM (1990). The role of cytokines in endotoxin-mediated responses . In: Immunophysiology: role of cells and cytokines: Immunity and inflammation. Oppenheim JJ , Shevach E , eds. Oxford, UK: Oxford University Press, p. 238.

32.

Wilson ME , Schifferle RE (1991). Evidence that serotype B antigenic determinant of Actinobacillus actinomycetemcomitans. Y4 resides in the polysaccharide moiety of lipopolysaccharide. Infect Immun 59:1544-1551.

33.

Yoshimura T, Matsushima K, Oppenheim JJ, Leonard EJ (1987). Neutrophil chemotactic factor produced by lipopolysaccharide (LPS)-stimulated human blood mononuclear from interleukin-1 (IL-1). J Immunol 139:788-794. Yu S-H, Koerner RTJ, Adams DO (1990). Gene regulation in macrophage activation: differential regulation of genes encoding for tumor necrosis factor, interleukin-1, JE, and KC by interferon-P and lipopolysaccharide. J Leukocyte Biol 48:412-419.

34.

Zhong WW , Burke PA , Hand T. , Walsh MJ , Hughes LA , Forse RA (1993). Regulation of cytokine mRNA expression in lipopolysaccharide stimulated human macrophges. Arch Surg 128:158-164.