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. 1993 Sep;61(9):3976–3980. doi: 10.1128/iai.61.9.3976-3980.1993

Tumor necrosis factor alpha (TNF-alpha) production in mice immunized with Escherichia coli: correlation with mortality after lethal challenge and production of a native inhibitor of TNF-alpha activity.

G Raponi 1, N Keller 1, M Rozenberg-Arska 1, M T Lun 1, C Mancini 1, J Verhoef 1
PMCID: PMC281102  PMID: 8359919

Abstract

The kinetics of tumor necrosis factor alpha (TNF-alpha) production, the correlation between cytokine levels and mortality rates after lethal challenge, and the production of a native inhibitor of TNF-alpha activity was investigated in mice immunized with formalin-killed Escherichia coli. Groups of mice were injected for 8 weeks with either untreated bacteria or bacteria treated with 0.5 MIC of aztreonam and subsequently challenged with 100 50% lethal doses of viable E. coli. Mice receiving saline only (controls) died within 24 h. The mortality of mice immunized with aztreonam-treated E. coli was significantly lower than that of mice immunized with untreated E. coli. There were no measurable levels of TNF-alpha in sera obtained from control mice during the entire period of immunization. TNF-alpha levels ranging from 90 to 306 U/ml were measured 90 min after each vaccination in sera obtained from mice immunized with untreated E. coli. Sera from mice immunized with antibiotic-treated E. coli showed lower TNF-alpha levels, ranging from 40 to 128 U/ml. TNF-alpha levels measured 90 min after lethal challenge correlated with the mortality rate observed in each group (r = 0.95). The cytotoxic activity of recombinant murine TNF-alpha was inhibited by the sera from immunized mice but not by the sera from controls. The inhibition was drastically reduced by preincubation of the sera with staphylococcal protein A. In immunoblot experiments, class G immunoglobulins reacting with recombinant murine TNF-alpha were observed in the sera of immunized mice. These data confirm the correlation existing between TNF-alpha levels and mortality and suggest that native inhibitors may play a role in the regulation of the biological function of the cytokine.

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Selected References

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  1. Bayston K. F., Cohen J. Bacterial endotoxin and current concepts in the diagnosis and treatment of endotoxaemia. J Med Microbiol. 1990 Feb;31(2):73–83. doi: 10.1099/00222615-31-2-73. [DOI] [PubMed] [Google Scholar]
  2. Bendtzen K., Svenson M., Jønsson V., Hippe E. Autoantibodies to cytokines--friends or foes? Immunol Today. 1990 May;11(5):167–169. doi: 10.1016/0167-5699(90)90068-k. [DOI] [PubMed] [Google Scholar]
  3. Beutler B., Milsark I. W., Cerami A. C. Passive immunization against cachectin/tumor necrosis factor protects mice from lethal effect of endotoxin. Science. 1985 Aug 30;229(4716):869–871. doi: 10.1126/science.3895437. [DOI] [PubMed] [Google Scholar]
  4. Brown D. E., Morrison D. C. Possible alteration of normal mechanisms of endotoxin toxicity in vivo by actinomycin D. J Infect Dis. 1982 Dec;146(6):746–750. doi: 10.1093/infdis/146.6.746. [DOI] [PubMed] [Google Scholar]
  5. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  6. Damas P., Reuter A., Gysen P., Demonty J., Lamy M., Franchimont P. Tumor necrosis factor and interleukin-1 serum levels during severe sepsis in humans. Crit Care Med. 1989 Oct;17(10):975–978. doi: 10.1097/00003246-198910000-00001. [DOI] [PubMed] [Google Scholar]
  7. Fomsgaard A., Svenson M., Bendtzen K. Auto-antibodies to tumour necrosis factor alpha in healthy humans and patients with inflammatory diseases and gram-negative bacterial infections. Scand J Immunol. 1989 Aug;30(2):219–223. doi: 10.1111/j.1365-3083.1989.tb01204.x. [DOI] [PubMed] [Google Scholar]
  8. Georgopapadakou N. H., Smith S. A., Sykes R. B. Mode of action of azthreonam. Antimicrob Agents Chemother. 1982 Jun;21(6):950–956. doi: 10.1128/aac.21.6.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Girardin E., Grau G. E., Dayer J. M., Roux-Lombard P., Lambert P. H. Tumor necrosis factor and interleukin-1 in the serum of children with severe infectious purpura. N Engl J Med. 1988 Aug 18;319(7):397–400. doi: 10.1056/NEJM198808183190703. [DOI] [PubMed] [Google Scholar]
  10. Jackson J. J., Kropp H. beta-Lactam antibiotic-induced release of free endotoxin: in vitro comparison of penicillin-binding protein (PBP) 2-specific imipenem and PBP 3-specific ceftazidime. J Infect Dis. 1992 Jun;165(6):1033–1041. doi: 10.1093/infdis/165.6.1033. [DOI] [PubMed] [Google Scholar]
  11. Johnston R. B., Jr Current concepts: immunology. Monocytes and macrophages. N Engl J Med. 1988 Mar 24;318(12):747–752. doi: 10.1056/NEJM198803243181205. [DOI] [PubMed] [Google Scholar]
  12. Keller N., Raponi G., Hoepelman I. M., Overbeek B. P., Rozenberg-Arska M., Verhoef J. Effect of sub-minimal inhibitory concentrations of ciprofloxacin and fleroxacin on the bacterial capsular antigen and opsonophagocytosis by human polymorphonuclear leukocytes. Zentralbl Bakteriol. 1991 Jan;274(4):519–526. doi: 10.1016/s0934-8840(11)80090-3. [DOI] [PubMed] [Google Scholar]
  13. Leying H., Suerbaum S., Kroll H. P., Karch H., Opferkuch W. Influence of beta-lactam antibiotics and ciprofloxacin on composition and immunogenicity of Escherichia coli outer membrane. Antimicrob Agents Chemother. 1986 Sep;30(3):475–480. doi: 10.1128/aac.30.3.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lorian V., Atkinson B. Bactericidal effect of polymorphonuclear neutrophils on antibiotic-induced filaments of gram-negative bacilli. J Infect Dis. 1984 May;149(5):719–727. doi: 10.1093/infdis/149.5.719. [DOI] [PubMed] [Google Scholar]
  15. Mathison J. C., Wolfson E., Ulevitch R. J. Participation of tumor necrosis factor in the mediation of gram negative bacterial lipopolysaccharide-induced injury in rabbits. J Clin Invest. 1988 Jun;81(6):1925–1937. doi: 10.1172/JCI113540. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Michie H. R., Manogue K. R., Spriggs D. R., Revhaug A., O'Dwyer S., Dinarello C. A., Cerami A., Wolff S. M., Wilmore D. W. Detection of circulating tumor necrosis factor after endotoxin administration. N Engl J Med. 1988 Jun 9;318(23):1481–1486. doi: 10.1056/NEJM198806093182301. [DOI] [PubMed] [Google Scholar]
  17. Overbeek B. P., Schellekens J. F., Lippe W., Dekker B. A., Verhoef J. Carumonam enhances reactivity of Escherichia coli with mono- and polyclonal antisera to rough Escherichia coli J5. J Clin Microbiol. 1987 Jun;25(6):1009–1013. doi: 10.1128/jcm.25.6.1009-1013.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parker M. M., Parrillo J. E. Septic shock. Hemodynamics and pathogenesis. JAMA. 1983 Dec 23;250(24):3324–3327. [PubMed] [Google Scholar]
  19. Raponi G., Keller N., Overbeek B. P., Rozenberg-Arska M., Torensma R., Verhoef J. Immunization of mice with antibiotic-treated Escherichia coli results in enhanced protection against challenge with homologous and heterologous bacteria. J Infect Dis. 1991 Jan;163(1):122–127. doi: 10.1093/infdis/163.1.122. [DOI] [PubMed] [Google Scholar]
  20. Raponi G., Keller N., Overbeek B. P., Rozenberg-Arska M., van Kessel K. P., Verhoef J. Enhanced phagocytosis of encapsulated Escherichia coli strains after exposure to sub-MICs of antibiotics is correlated to changes of the bacterial cell surface. Antimicrob Agents Chemother. 1990 Feb;34(2):332–336. doi: 10.1128/aac.34.2.332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Raponi G., Lun M. T., Lorino G., Nazzari C., Gaeta A., Mancini C., Filadoro F. Reactivity and protective capacity of a polyclonal antiserum derived from mice immunized with antibiotic exposed Escherichia coli. J Antimicrob Chemother. 1993 Jan;31(1):117–128. doi: 10.1093/jac/31.1.117. [DOI] [PubMed] [Google Scholar]
  22. Rozenberg-Arska M., Visser M. R. Infectious disease therapy in the 1990s. Where are we heading? Drugs. 1992 May;43(5):629–636. doi: 10.2165/00003495-199243050-00001. [DOI] [PubMed] [Google Scholar]
  23. Silva A. T., Bayston K. F., Cohen J. Prophylactic and therapeutic effects of a monoclonal antibody to tumor necrosis factor-alpha in experimental gram-negative shock. J Infect Dis. 1990 Aug;162(2):421–427. doi: 10.1093/infdis/162.2.421. [DOI] [PubMed] [Google Scholar]
  24. Simon D. M., Koenig G., Trenholme G. M. Differences in release of tumor necrosis factor from THP-1 cells stimulated by filtrates of antibiotic-killed Escherichia coli. J Infect Dis. 1991 Oct;164(4):800–802. doi: 10.1093/infdis/164.4.800. [DOI] [PubMed] [Google Scholar]
  25. Stephens K. E., Ishizaka A., Larrick J. W., Raffin T. A. Tumor necrosis factor causes increased pulmonary permeability and edema. Comparison to septic acute lung injury. Am Rev Respir Dis. 1988 Jun;137(6):1364–1370. doi: 10.1164/ajrccm/137.6.1364. [DOI] [PubMed] [Google Scholar]
  26. Tracey K. J., Beutler B., Lowry S. F., Merryweather J., Wolpe S., Milsark I. W., Hariri R. J., Fahey T. J., 3rd, Zentella A., Albert J. D. Shock and tissue injury induced by recombinant human cachectin. Science. 1986 Oct 24;234(4775):470–474. doi: 10.1126/science.3764421. [DOI] [PubMed] [Google Scholar]
  27. Tracey K. J., Fong Y., Hesse D. G., Manogue K. R., Lee A. T., Kuo G. C., Lowry S. F., Cerami A. Anti-cachectin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia. Nature. 1987 Dec 17;330(6149):662–664. doi: 10.1038/330662a0. [DOI] [PubMed] [Google Scholar]
  28. Tracey K. J., Lowry S. F., Fahey T. J., 3rd, Albert J. D., Fong Y., Hesse D., Beutler B., Manogue K. R., Calvano S., Wei H. Cachectin/tumor necrosis factor induces lethal shock and stress hormone responses in the dog. Surg Gynecol Obstet. 1987 May;164(5):415–422. [PubMed] [Google Scholar]
  29. Tracey K. J., Vlassara H., Cerami A. Cachectin/tumour necrosis factor. Lancet. 1989 May 20;1(8647):1122–1126. doi: 10.1016/s0140-6736(89)92394-5. [DOI] [PubMed] [Google Scholar]
  30. Waage A., Halstensen A., Espevik T. Association between tumour necrosis factor in serum and fatal outcome in patients with meningococcal disease. Lancet. 1987 Feb 14;1(8529):355–357. doi: 10.1016/s0140-6736(87)91728-4. [DOI] [PubMed] [Google Scholar]
  31. Wolff S. M. Biological effects of bacterial endotoxins in man. J Infect Dis. 1973 Jul;128(Suppl):259–264. doi: 10.1093/infdis/128.supplement_1.s259. [DOI] [PubMed] [Google Scholar]
  32. Ziegler E. J. Tumor necrosis factor in humans. N Engl J Med. 1988 Jun 9;318(23):1533–1535. doi: 10.1056/NEJM198806093182309. [DOI] [PubMed] [Google Scholar]

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