Search tips
Search criteria 


Logo of molmedLink to Publisher's site
Mol Med. 1997 December; 3(12): 864–875.
PMCID: PMC2230288

Lipopolysaccharide-induced cytokine cascade and lethality in LT alpha/TNF alpha-deficient mice.


BACKGROUND: Tumor necrosis factor alpha (TNF-alpha) is often considered the main proinflammatory cytokine induced by lipopolysaccharide (LPS) and consequently the critical mediator of the lethality associated with septic shock. MATERIALS AND METHODS: We used mice carrying a deletion of both the lymphotoxin alpha (LT-alpha) and TNF-alpha genes to assess the role of TNF in the cytokine cascade and lethality induced by LPS. RESULTS: Initial production of IL-1 alpha, IL-1 beta, IL-6, and IL-10 is comparable in wild-type and mutant mice. However, at later times, expression of IL-1 alpha, IL-1 beta, and IL-10 is prolonged, whereas that of IL-6 decreases in mutant mice. Expression of IFN-gamma is almost completely abrogated in mutants, which is in agreement with a more significant alteration of the late phase of the cytokine cascade. We measured similar LD50 (600 micrograms) for the intravenous injection of LPS in mice of the three genotypes (+/+, +/-, -/-), demonstrating that the absence of TNF does not confer long-term protection from lethality. However, death occurred much more slowly in mutant mice, who were protected more efficiently from death by CNI 1493, an inhibitor of proinflammatory cytokine production, than were wild-type mice. DISCUSSION: Thus, while TNF-alpha is not required for the induction of these cytokines by LPS, it modulates the kinetics of their expression. The lethality studies simultaneously confirm a role for TNF as a mediator of early lethality and establish that, in the absence of these cytokines, other mediators take over, resulting in the absence of long-term protection from LPS toxicity.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Smith DM, Lackides GA, Epstein LB. Coordinated induction of autocrine tumor necrosis factor and interleukin 1 in normal human monocytes and the implications for monocyte-mediated cytotoxicity. Cancer Res. 1990 Jun 1;50(11):3146–3153. [PubMed]
  • Van Damme J, Opdenakker G, Simpson RJ, Rubira MR, Cayphas S, Vink A, Billiau A, Van Snick J. Identification of the human 26-kD protein, interferon beta 2 (IFN-beta 2), as a B cell hybridoma/plasmacytoma growth factor induced by interleukin 1 and tumor necrosis factor. J Exp Med. 1987 Mar 1;165(3):914–919. [PMC free article] [PubMed]
  • Cassatella MA, Guasparri I, Ceska M, Bazzoni F, Rossi F. Interferon-gamma inhibits interleukin-8 production by human polymorphonuclear leucocytes. Immunology. 1993 Feb;78(2):177–184. [PubMed]
  • Wanidworanun C, Strober W. Predominant role of tumor necrosis factor-alpha in human monocyte IL-10 synthesis. J Immunol. 1993 Dec 15;151(12):6853–6861. [PubMed]
  • Yamada H, Iwase S, Mohri M, Kufe D. Involvement of a nuclear factor-kappa B-like protein in induction of the macrophage colony-stimulating factor gene by tumor necrosis factor. Blood. 1991 Oct 15;78(8):1988–1995. [PubMed]
  • Debs RJ, Fuchs HJ, Philip R, Montgomery AB, Brunette EN, Liggitt D, Patton JS, Shellito JE. Lung-specific delivery of cytokines induces sustained pulmonary and systemic immunomodulation in rats. J Immunol. 1988 May 15;140(10):3482–3488. [PubMed]
  • Brouckaert P, Spriggs DR, Demetri G, Kufe DW, Fiers W. Circulating interleukin 6 during a continuous infusion of tumor necrosis factor and interferon gamma. J Exp Med. 1989 Jun 1;169(6):2257–2262. [PMC free article] [PubMed]
  • Sheron N, Williams R. IL-8 as a circulating cytokine: induction by recombinant tumour necrosis factor-alpha. Clin Exp Immunol. 1992 Jul;89(1):100–103. [PubMed]
  • van der Poll T, Jansen J, Levi M, ten Cate H, ten Cate JW, van Deventer SJ. Regulation of interleukin 10 release by tumor necrosis factor in humans and chimpanzees. J Exp Med. 1994 Nov 1;180(5):1985–1988. [PMC free article] [PubMed]
  • Doherty GM, Lange JR, Langstein HN, Alexander HR, Buresh CM, Norton JA. Evidence for IFN-gamma as a mediator of the lethality of endotoxin and tumor necrosis factor-alpha. J Immunol. 1992 Sep 1;149(5):1666–1670. [PubMed]
  • Michie HR, Manogue KR, Spriggs DR, Revhaug A, O'Dwyer S, Dinarello CA, Cerami A, Wolff SM, Wilmore DW. Detection of circulating tumor necrosis factor after endotoxin administration. N Engl J Med. 1988 Jun 9;318(23):1481–1486. [PubMed]
  • Cannon JG, Tompkins RG, Gelfand JA, Michie HR, Stanford GG, van der Meer JW, Endres S, Lonnemann G, Corsetti J, Chernow B, et al. Circulating interleukin-1 and tumor necrosis factor in septic shock and experimental endotoxin fever. J Infect Dis. 1990 Jan;161(1):79–84. [PubMed]
  • Redl H, Schlag G, Bahrami S, Schade U, Ceska M, Stütz P. Plasma neutrophil-activating peptide-1/interleukin-8 and neutrophil elastase in a primate bacteremia model. J Infect Dis. 1991 Aug;164(2):383–388. [PubMed]
  • Martich GD, Danner RL, Ceska M, Suffredini AF. Detection of interleukin 8 and tumor necrosis factor in normal humans after intravenous endotoxin: the effect of antiinflammatory agents. J Exp Med. 1991 Apr 1;173(4):1021–1024. [PMC free article] [PubMed]
  • Wollenberg GK, DeForge LE, Bolgos G, Remick DG. Differential expression of tumor necrosis factor and interleukin-6 by peritoneal macrophages in vivo and in culture. Am J Pathol. 1993 Oct;143(4):1121–1130. [PubMed]
  • Fong Y, Tracey KJ, Moldawer LL, Hesse DG, Manogue KB, Kenney JS, Lee AT, Kuo GC, Allison AC, Lowry SF, et al. Antibodies to cachectin/tumor necrosis factor reduce interleukin 1 beta and interleukin 6 appearance during lethal bacteremia. J Exp Med. 1989 Nov 1;170(5):1627–1633. [PMC free article] [PubMed]
  • Emerson TE, Jr, Lindsey DC, Jesmok GJ, Duerr ML, Fournel MA. Efficacy of monoclonal antibody against tumor necrosis factor alpha in an endotoxemic baboon model. Circ Shock. 1992 Oct;38(2):75–84. [PubMed]
  • Teti G, Mancuso G, Tomasello F, Chiofalo MS. Production of tumor necrosis factor-alpha and interleukin-6 in mice infected with group B streptococci. Circ Shock. 1992 Oct;38(2):138–144. [PubMed]
  • Hinshaw LB, Emerson TE, Jr, Taylor FB, Jr, Chang AC, Duerr M, Peer GT, Flournoy DJ, White GL, Kosanke SD, Murray CK, et al. Lethal Staphylococcus aureus-induced shock in primates: prevention of death with anti-TNF antibody. J Trauma. 1992 Oct;33(4):568–573. [PubMed]
  • van der Poll T, Levi M, van Deventer SJ, ten Cate H, Haagmans BL, Biemond BJ, Büller HR, Hack CE, ten Cate JW. Differential effects of anti-tumor necrosis factor monoclonal antibodies on systemic inflammatory responses in experimental endotoxemia in chimpanzees. Blood. 1994 Jan 15;83(2):446–451. [PubMed]
  • Van Zee KJ, Kohno T, Fischer E, Rock CS, Moldawer LL, Lowry SF. Tumor necrosis factor soluble receptors circulate during experimental and clinical inflammation and can protect against excessive tumor necrosis factor alpha in vitro and in vivo. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4845–4849. [PubMed]
  • Suffredini AF, Reda D, Banks SM, Tropea M, Agosti JM, Miller R. Effects of recombinant dimeric TNF receptor on human inflammatory responses following intravenous endotoxin administration. J Immunol. 1995 Nov 15;155(10):5038–5045. [PubMed]
  • Espat NJ, Cendan JC, Beierle EA, Auffenberg TA, Rosenberg J, Russell D, Kenney JS, Fischer E, Montegut W, Lowry SF, et al. PEG-BP-30 monotherapy attenuates the cytokine-mediated inflammatory cascade in baboon Escherichia coli septic shock. J Surg Res. 1995 Jul;59(1):153–158. [PubMed]
  • Teti G, Mancuso G, Tomasello F. Cytokine appearance and effects of anti-tumor necrosis factor alpha antibodies in a neonatal rat model of group B streptococcal infection. Infect Immun. 1993 Jan;61(1):227–235. [PMC free article] [PubMed]
  • Junger WG, Hoyt DB, Redl H, Liu FC, Loomis WH, Davies J, Schlag G. Tumor necrosis factor antibody treatment of septic baboons reduces the production of sustained T-cell suppressive factors. Shock. 1995 Mar;3(3):173–178. [PubMed]
  • Ferran C, Dautry F, Mérite S, Sheehan K, Schreiber R, Grau G, Bach JF, Chatenoud L. Anti-tumor necrosis factor modulates anti-CD3-triggered T cell cytokine gene expression in vivo. J Clin Invest. 1994 May;93(5):2189–2196. [PMC free article] [PubMed]
  • Cavaillon JM, Marie C, Pitton C, Fitting C. The production of TNF alpha and IL-8 in whole blood assays are differently regulated upon activation by LPS. Prog Clin Biol Res. 1995;392:433–439. [PubMed]
  • Vasilescu C, Berger D, Buttenschön K, Seidelmann M, Beger HG. Endotoxin-induced release of interleukin 6 and interleukin 1 beta in human blood is independent of tumor necrosis factor alpha. Eur Surg Res. 1996;28(1):55–62. [PubMed]
  • Schandené L, Vandenbussche P, Crusiaux A, Alègre ML, Abramowicz D, Dupont E, Content J, Goldman M. Differential effects of pentoxifylline on the production of tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) by monocytes and T cells. Immunology. 1992 May;76(1):30–34. [PubMed]
  • Mándi Y, Farkas G, Ocsovszky I, Nagy Z. Inhibition of tumor necrosis factor production and ICAM-1 expression by pentoxifylline: beneficial effects in sepsis syndrome. Res Exp Med (Berl) 1995;195(5):297–307. [PubMed]
  • Netea MG, Blok WL, Kullberg BJ, Bemelmans M, Vogels MT, Buurman WA, van der Meer JW. Pharmacologic inhibitors of tumor necrosis factor production exert differential effects in lethal endotoxemia and in infection with live microorganisms in mice. J Infect Dis. 1995 Feb;171(2):393–399. [PubMed]
  • Amiot F, Bellkaid Y, Lebastard M, Ave P, Dautry F, Milon G. Abnormal organisation of the splenic marginal zone and the correlated leukocytosis in lymphotoxin-alpha and tumor necrosis factor alpha double deficient mice. Eur Cytokine Netw. 1996 Dec;7(4):733–739. [PubMed]
  • Amiot F, Boussadia O, Cases S, Fitting C, Lebastard M, Cavaillon JM, Milon G, Dautry F. Mice heterozygous for a deletion of the tumor necrosis factor-alpha and lymphotoxin-alpha genes: biological importance of a nonlinear response of tumor necrosis factor-alpha to gene dosage. Eur J Immunol. 1997 Apr;27(4):1035–1042. [PubMed]
  • Beutler B, Milsark IW, Cerami AC. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985 Aug 30;229(4716):869–871. [PubMed]
  • Tracey KJ, Fong Y, Hesse DG, Manogue KR, Lee AT, Kuo GC, Lowry SF, Cerami A. Anti-cachectin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia. Nature. 1987 Dec 17;330(6149):662–664. [PubMed]
  • Fiedler VB, Loof I, Sander E, Voehringer V, Galanos C, Fournel MA. Monoclonal antibody to tumor necrosis factor--alpha prevents lethal endotoxin sepsis in adult rhesus monkeys. J Lab Clin Med. 1992 Oct;120(4):574–588. [PubMed]
  • Jesmok G, Lindsey C, Duerr M, Fournel M, Emerson T., Jr Efficacy of monoclonal antibody against human recombinant tumor necrosis factor in E. coli-challenged swine. Am J Pathol. 1992 Nov;141(5):1197–1207. [PubMed]
  • Freudenberg MA, Galanos C. Tumor necrosis factor alpha mediates lethal activity of killed gram-negative and gram-positive bacteria in D-galactosamine-treated mice. Infect Immun. 1991 Jun;59(6):2110–2115. [PMC free article] [PubMed]
  • Franks AK, Kujawa KI, Yaffe LJ. Experimental elimination of tumor necrosis factor in low-dose endotoxin models has variable effects on survival. Infect Immun. 1991 Aug;59(8):2609–2614. [PMC free article] [PubMed]
  • Rothe J, Lesslauer W, Lötscher H, Lang Y, Koebel P, Köntgen F, Althage A, Zinkernagel R, Steinmetz M, Bluethmann H. Mice lacking the tumour necrosis factor receptor 1 are resistant to TNF-mediated toxicity but highly susceptible to infection by Listeria monocytogenes. Nature. 1993 Aug 26;364(6440):798–802. [PubMed]
  • Abraham E, Wunderink R, Silverman H, Perl TM, Nasraway S, Levy H, Bone R, Wenzel RP, Balk R, Allred R, et al. Efficacy and safety of monoclonal antibody to human tumor necrosis factor alpha in patients with sepsis syndrome. A randomized, controlled, double-blind, multicenter clinical trial. TNF-alpha MAb Sepsis Study Group. JAMA. 1995 Mar 22;273(12):934–941. [PubMed]
  • Fisher CJ, Jr, Agosti JM, Opal SM, Lowry SF, Balk RA, Sadoff JC, Abraham E, Schein RM, Benjamin E. Treatment of septic shock with the tumor necrosis factor receptor:Fc fusion protein. The Soluble TNF Receptor Sepsis Study Group. N Engl J Med. 1996 Jun 27;334(26):1697–1702. [PubMed]
  • Sansonetti PJ, Arondel J, Cavaillon JM, Huerre M. Role of interleukin-1 in the pathogenesis of experimental shigellosis. J Clin Invest. 1995 Aug;96(2):884–892. [PMC free article] [PubMed]
  • Bianchi M, Bloom O, Raabe T, Cohen PS, Chesney J, Sherry B, Schmidtmayerova H, Calandra T, Zhang X, Bukrinsky M, et al. Suppression of proinflammatory cytokines in monocytes by a tetravalent guanylhydrazone. J Exp Med. 1996 Mar 1;183(3):927–936. [PMC free article] [PubMed]
  • Firestein GS, Boyle D, Bullough DA, Gruber HE, Sajjadi FG, Montag A, Sambol B, Mullane KM. Protective effect of an adenosine kinase inhibitor in septic shock. J Immunol. 1994 Jun 15;152(12):5853–5859. [PubMed]
  • Marino MW, Dunn A, Grail D, Inglese M, Noguchi Y, Richards E, Jungbluth A, Wada H, Moore M, Williamson B, et al. Characterization of tumor necrosis factor-deficient mice. Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8093–8098. [PubMed]
  • Hinson RM, Williams JA, Shacter E. Elevated interleukin 6 is induced by prostaglandin E2 in a murine model of inflammation: possible role of cyclooxygenase-2. Proc Natl Acad Sci U S A. 1996 May 14;93(10):4885–4890. [PubMed]
  • Sower LE, Froelich CJ, Carney DH, Fenton JW, 2nd, Klimpel GR. Thrombin induces IL-6 production in fibroblasts and epithelial cells. Evidence for the involvement of the seven-transmembrane domain (STD) receptor for alpha-thrombin. J Immunol. 1995 Jul 15;155(2):895–901. [PubMed]
  • Thivierge M, Rola-Pleszczynski M. Platelet-activating factor enhances interleukin-6 production by alveolar macrophages. J Allergy Clin Immunol. 1992 Nov;90(5):796–802. [PubMed]
  • Brach MA, de Vos S, Arnold C, Gruss HJ, Mertelsmann R, Herrmann F. Leukotriene B4 transcriptionally activates interleukin-6 expression involving NK-chi B and NF-IL6. Eur J Immunol. 1992 Oct;22(10):2705–2711. [PubMed]
  • Musso T, Espinoza-Delgado I, Pulkki K, Gusella GL, Longo DL, Varesio L. IL-2 induces IL-6 production in human monocytes. J Immunol. 1992 Feb 1;148(3):795–800. [PubMed]
  • Schmidt JA, Abdulla E. Down-regulation of IL-1 beta biosynthesis by inducers of the heat-shock response. J Immunol. 1988 Sep 15;141(6):2027–2034. [PubMed]
  • Snyder DS, Unanue ER. Corticosteroids inhibit murine macrophage Ia expression and interleukin 1 production. J Immunol. 1982 Nov;129(5):1803–1805. [PubMed]
  • Fushimi T, Okayama H, Seki T, Shimura S, Shirato K. Dexamethasone suppressed gene expression and production of interleukin-10 by human peripheral blood mononuclear cells and monocytes. Int Arch Allergy Immunol. 1997 Jan;112(1):13–18. [PubMed]
  • Barsig J, Küsters S, Vogt K, Volk HD, Tiegs G, Wendel A. Lipopolysaccharide-induced interleukin-10 in mice: role of endogenous tumor necrosis factor-alpha. Eur J Immunol. 1995 Oct;25(10):2888–2893. [PubMed]
  • DeForge LE, Kenney JS, Jones ML, Warren JS, Remick DG. Biphasic production of IL-8 in lipopolysaccharide (LPS)-stimulated human whole blood. Separation of LPS- and cytokine-stimulated components using anti-tumor necrosis factor and anti-IL-1 antibodies. J Immunol. 1992 Apr 1;148(7):2133–2141. [PubMed]
  • McGeehan GM, Becherer JD, Bast RC, Jr, Boyer CM, Champion B, Connolly KM, Conway JG, Furdon P, Karp S, Kidao S, et al. Regulation of tumour necrosis factor-alpha processing by a metalloproteinase inhibitor. Nature. 1994 Aug 18;370(6490):558–561. [PubMed]
  • Lesslauer W, Tabuchi H, Gentz R, Brockhaus M, Schlaeger EJ, Grau G, Piguet PF, Pointaire P, Vassalli P, Loetscher H. Recombinant soluble tumor necrosis factor receptor proteins protect mice from lipopolysaccharide-induced lethality. Eur J Immunol. 1991 Nov;21(11):2883–2886. [PubMed]
  • Ashkenazi A, Marsters SA, Capon DJ, Chamow SM, Figari IS, Pennica D, Goeddel DV, Palladino MA, Smith DH. Protection against endotoxic shock by a tumor necrosis factor receptor immunoadhesin. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10535–10539. [PubMed]
  • Mohler KM, Torrance DS, Smith CA, Goodwin RG, Stremler KE, Fung VP, Madani H, Widmer MB. Soluble tumor necrosis factor (TNF) receptors are effective therapeutic agents in lethal endotoxemia and function simultaneously as both TNF carriers and TNF antagonists. J Immunol. 1993 Aug 1;151(3):1548–1561. [PubMed]
  • Martin RA, Silva AT, Cohen J. Effect of anti-TNF-alpha treatment in an antibiotic treated murine model of shock due to Streptococcus pyogenes. FEMS Microbiol Lett. 1993 Jun 15;110(2):175–178. [PubMed]
  • Ohlsson K, Björk P, Bergenfeldt M, Hageman R, Thompson RC. Interleukin-1 receptor antagonist reduces mortality from endotoxin shock. Nature. 1990 Dec 6;348(6301):550–552. [PubMed]
  • Block MI, Berg M, McNamara MJ, Norton JA, Fraker DL, Alexander HR. Passive immunization of mice against D factor blocks lethality and cytokine release during endotoxemia. J Exp Med. 1993 Sep 1;178(3):1085–1090. [PMC free article] [PubMed]
  • Bernhagen J, Calandra T, Mitchell RA, Martin SB, Tracey KJ, Voelter W, Manogue KR, Cerami A, Bucala R. MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia. Nature. 1993 Oct 21;365(6448):756–759. [PubMed]
  • Cohen PS, Nakshatri H, Dennis J, Caragine T, Bianchi M, Cerami A, Tracey KJ. CNI-1493 inhibits monocyte/macrophage tumor necrosis factor by suppression of translation efficiency. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3967–3971. [PubMed]
  • Bianchi M, Ulrich P, Bloom O, Meistrell M, 3rd, Zimmerman GA, Schmidtmayerova H, Bukrinsky M, Donnelley T, Bucala R, Sherry B, et al. An inhibitor of macrophage arginine transport and nitric oxide production (CNI-1493) prevents acute inflammation and endotoxin lethality. Mol Med. 1995 Mar;1(3):254–266. [PMC free article] [PubMed]
  • Erickson SL, de Sauvage FJ, Kikly K, Carver-Moore K, Pitts-Meek S, Gillett N, Sheehan KC, Schreiber RD, Goeddel DV, Moore MW. Decreased sensitivity to tumour-necrosis factor but normal T-cell development in TNF receptor-2-deficient mice. Nature. 1994 Dec 8;372(6506):560–563. [PubMed]
  • Pfeffer K, Matsuyama T, Kündig TM, Wakeham A, Kishihara K, Shahinian A, Wiegmann K, Ohashi PS, Krönke M, Mak TW. Mice deficient for the 55 kd tumor necrosis factor receptor are resistant to endotoxic shock, yet succumb to L. monocytogenes infection. Cell. 1993 May 7;73(3):457–467. [PubMed]
  • Garcia I, Miyazaki Y, Araki K, Araki M, Lucas R, Grau GE, Milon G, Belkaid Y, Montixi C, Lesslauer W, et al. Transgenic mice expressing high levels of soluble TNF-R1 fusion protein are protected from lethal septic shock and cerebral malaria, and are highly sensitive to Listeria monocytogenes and Leishmania major infections. Eur J Immunol. 1995 Aug;25(8):2401–2407. [PubMed]

Articles from Molecular Medicine are provided here courtesy of The Feinstein Institute for Medical Research at North Shore LIJ