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. 1995 Apr;63(4):1263–1269. doi: 10.1128/iai.63.4.1263-1269.1995

Loss of the O4 antigen moiety from the lipopolysaccharide of an extraintestinal isolate of Escherichia coli has only minor effects on serum sensitivity and virulence in vivo.

T A Russo 1, G Sharma 1, C R Brown 1, A A Campagnari 1
PMCID: PMC173145  PMID: 7890383

Abstract

The O-specific antigen in extraintestinal isolates of Escherichia coli is believed to be an important virulence factor. To assess its role in the pathogenic process, proven isogenic derivatives with either a complete (CP921) or nearly complete (CP920) deficiency of the O4 antigen were obtained by TnphoA'1-mediated transposon mutagenesis of an O4/K54/H5 blood isolate (CP9). By utilizing a previously reported isogenic K54 capsule-deficient derivative (CP9.137), additional isogenic derivatives deficient in both the K54 capsular antigen and either all (CP923) or nearly all (CP922) of the O4 antigen were also constructed. These strains and their wild-type parent were evaluated in vitro for serum sensitivity and in vivo by intraperitoneal challenge of outbred mice. The complete or nearly complete loss of the O4 antigen (CP920 and CP921) resulted in only a minor increase in serum sensitivity. In contrast, CP9.137 had a significant increase in serum sensitivity, and CP922 and CP923 were extremely serum sensitive. When tested in vivo, the complete or nearly complete loss of the O4 antigen resulted in a small but significant increase (P < or = 0.05), not the expected decrease, in virulence compared with its wild-type parent. In contrast, CP9.137 and CP922 were significantly less virulent (P < or = 0.05). These studies do not exclude a role for the O4 antigen moiety of lipopolysaccharide in the pathogenesis of extraintestinal E. coli infection; however, they demonstrate that the O4 antigen plays only a minor role in serum resistance in vitro and that its loss does not diminish and perhaps enhances the virulence of CP9 in vivo after intraperitoneal challenge.

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Selected References

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  1. Campagnari A. A., Spinola S. M., Lesse A. J., Kwaik Y. A., Mandrell R. E., Apicella M. A. Lipooligosaccharide epitopes shared among gram-negative non-enteric mucosal pathogens. Microb Pathog. 1990 May;8(5):353–362. doi: 10.1016/0882-4010(90)90094-7. [DOI] [PubMed] [Google Scholar]
  2. Cross A. S., Gemski P., Sadoff J. C., Orskov F., Orskov I. The importance of the K1 capsule in invasive infections caused by Escherichia coli. J Infect Dis. 1984 Feb;149(2):184–193. doi: 10.1093/infdis/149.2.184. [DOI] [PubMed] [Google Scholar]
  3. Goldman R. C., Joiner K., Leive L. Serum-resistant mutants of Escherichia coli O111 contain increased lipopolysaccharide, lack an O antigen-containing capsule, and cover more of their lipid A core with O antigen. J Bacteriol. 1984 Sep;159(3):877–882. doi: 10.1128/jb.159.3.877-882.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hagberg L., Hull R., Hull S., McGhee J. R., Michalek S. M., Svanborg Edén C. Difference in susceptibility to gram-negative urinary tract infection between C3H/HeJ and C3H/HeN mice. Infect Immun. 1984 Dec;46(3):839–844. doi: 10.1128/iai.46.3.839-844.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Haraguchi G. E., Hull R. A., Krallmann-Wenzel U., Hull S. I. Molecular cloning and expression of the O4 polysaccharide gene cluster from Escherichia coli. Microb Pathog. 1989 Feb;6(2):123–132. doi: 10.1016/0882-4010(89)90015-6. [DOI] [PubMed] [Google Scholar]
  6. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jann K., Jann B. Polysaccharide antigens of Escherichia coli. Rev Infect Dis. 1987 Sep-Oct;9 (Suppl 5):S517–S526. doi: 10.1093/clinids/9.supplement_5.s517. [DOI] [PubMed] [Google Scholar]
  8. Jiang X. M., Neal B., Santiago F., Lee S. J., Romana L. K., Reeves P. R. Structure and sequence of the rfb (O antigen) gene cluster of Salmonella serovar typhimurium (strain LT2). Mol Microbiol. 1991 Mar;5(3):695–713. doi: 10.1111/j.1365-2958.1991.tb00741.x. [DOI] [PubMed] [Google Scholar]
  9. Johnson J. R., Stamm W. E. Urinary tract infections in women: diagnosis and treatment. Ann Intern Med. 1989 Dec 1;111(11):906–917. doi: 10.7326/0003-4819-111-11-906. [DOI] [PubMed] [Google Scholar]
  10. Joiner K. A. Complement evasion by bacteria and parasites. Annu Rev Microbiol. 1988;42:201–230. doi: 10.1146/annurev.mi.42.100188.001221. [DOI] [PubMed] [Google Scholar]
  11. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McCabe W. R., Kaijser B., Olling S., Uwaydah M., Hanson L. A. Escherichia coli in bacteremia: K and O antigens and serum sensitivity of strains from adults and neonates. J Infect Dis. 1978 Jul;138(1):33–41. doi: 10.1093/infdis/138.1.33. [DOI] [PubMed] [Google Scholar]
  13. Medearis D. N., Jr, Camitta B. M., Heath E. C. Cell wall composition and virulence in Escherichia coli. J Exp Med. 1968 Sep 1;128(3):399–414. doi: 10.1084/jem.128.3.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moses A. M., Howanitz J., van Gemert M., Miller M. Clofibrate-induced antidiuresis. J Clin Invest. 1973 Mar;52(3):535–542. doi: 10.1172/JCI107213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Natanson C., Hoffman W. D., Suffredini A. F., Eichacker P. Q., Danner R. L. Selected treatment strategies for septic shock based on proposed mechanisms of pathogenesis. Ann Intern Med. 1994 May 1;120(9):771–783. doi: 10.7326/0003-4819-120-9-199405010-00009. [DOI] [PubMed] [Google Scholar]
  16. Orskov I., Orskov F. Escherichia coli in extra-intestinal infections. J Hyg (Lond) 1985 Dec;95(3):551–575. doi: 10.1017/s0022172400060678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Parrillo J. E. Pathogenetic mechanisms of septic shock. N Engl J Med. 1993 May 20;328(20):1471–1477. doi: 10.1056/NEJM199305203282008. [DOI] [PubMed] [Google Scholar]
  18. Patton J. P., Nash D. B., Abrutyn E. Urinary tract infection: economic considerations. Med Clin North Am. 1991 Mar;75(2):495–513. doi: 10.1016/s0025-7125(16)30466-7. [DOI] [PubMed] [Google Scholar]
  19. Pluschke G., Mayden J., Achtman M., Levine R. P. Role of the capsule and the O antigen in resistance of O18:K1 Escherichia coli to complement-mediated killing. Infect Immun. 1983 Dec;42(3):907–913. doi: 10.1128/iai.42.3.907-913.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Porat R., Johns M. A., McCabe W. R. Selective pressures and lipopolysaccharide subunits as determinants of resistance of clinical isolates of gram-negative bacilli to human serum. Infect Immun. 1987 Feb;55(2):320–328. doi: 10.1128/iai.55.2.320-328.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rajakumar K., Jost B. H., Sasakawa C., Okada N., Yoshikawa M., Adler B. Nucleotide sequence of the rhamnose biosynthetic operon of Shigella flexneri 2a and role of lipopolysaccharide in virulence. J Bacteriol. 1994 Apr;176(8):2362–2373. doi: 10.1128/jb.176.8.2362-2373.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reeves P. Evolution of Salmonella O antigen variation by interspecific gene transfer on a large scale. Trends Genet. 1993 Jan;9(1):17–22. doi: 10.1016/0168-9525(93)90067-R. [DOI] [PubMed] [Google Scholar]
  23. Russo T. A., Guenther J. E., Wenderoth S., Frank M. M. Generation of isogenic K54 capsule-deficient Escherichia coli strains through TnphoA-mediated gene disruption. Mol Microbiol. 1993 Jul;9(2):357–364. doi: 10.1111/j.1365-2958.1993.tb01696.x. [DOI] [PubMed] [Google Scholar]
  24. Russo T. A., Liang Y., Cross A. S. The presence of K54 capsular polysaccharide increases the pathogenicity of Escherichia coli in vivo. J Infect Dis. 1994 Jan;169(1):112–118. doi: 10.1093/infdis/169.1.112. [DOI] [PubMed] [Google Scholar]
  25. Russo T. A., Moffitt M. C., Hammer C. H., Frank M. M. TnphoA-mediated disruption of K54 capsular polysaccharide genes in Escherichia coli confers serum sensitivity. Infect Immun. 1993 Aug;61(8):3578–3582. doi: 10.1128/iai.61.8.3578-3582.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Russo T. A., Singh G. An extraintestinal, pathogenic isolate of Escherichia coli (O4/K54/H5) can produce a group 1 capsule which is divergently regulated from its constitutively produced group 2, K54 capsular polysaccharide. J Bacteriol. 1993 Dec;175(23):7617–7623. doi: 10.1128/jb.175.23.7617-7623.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schnaitman C. A., Klena J. D. Genetics of lipopolysaccharide biosynthesis in enteric bacteria. Microbiol Rev. 1993 Sep;57(3):655–682. doi: 10.1128/mr.57.3.655-682.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Simmons D. A., Romanowska E. Structure and biology of Shigella flexneri O antigens. J Med Microbiol. 1987 Jun;23(4):289–302. doi: 10.1099/00222615-23-4-289. [DOI] [PubMed] [Google Scholar]
  30. Smith H. W., Huggins M. B. The association of the O18, K1 and H7 antigens and the Co1V plasmid of a strain of Escherichia coli with its virulence and immunogenicity. J Gen Microbiol. 1980 Dec;121(2):387–400. doi: 10.1099/00221287-121-2-387. [DOI] [PubMed] [Google Scholar]
  31. Stendahl O., Normann B., Edebo L. Influence of O and K antigens on the surface properties of Escherichia coli in relation to phagocytosis. Acta Pathol Microbiol Scand B. 1979 Apr;87B(2):85–91. doi: 10.1111/j.1699-0463.1979.tb02408.x. [DOI] [PubMed] [Google Scholar]
  32. Svanborg-Edén C., Hagberg L., Hull R., Hull S., Magnusson K. E., Ohman L. Bacterial virulence versus host resistance in the urinary tracts of mice. Infect Immun. 1987 May;55(5):1224–1232. doi: 10.1128/iai.55.5.1224-1232.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Taylor P. W. Bactericidal and bacteriolytic activity of serum against gram-negative bacteria. Microbiol Rev. 1983 Mar;47(1):46–83. doi: 10.1128/mr.47.1.46-83.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taylor P. W., Koutsaimanis K. G. Experimental Escherichia coli urinary infection in the rat. Kidney Int. 1975 Oct;8(4):233–238. doi: 10.1038/ki.1975.106. [DOI] [PubMed] [Google Scholar]
  35. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  36. VOSTI K. L., GOLDBERG L. M., MONTO A. S., RANTZ L. A. HOST-PARASITE INTERACTION IN PATIENTS WITH INFECTIONS DUE TO ESCHERICHIA COLI. I. THE SEROGROUPING OF E. COLI FROM INTESTINAL AND EXTRAINTESTINAL SOURCES. J Clin Invest. 1964 Dec;43:2377–2385. doi: 10.1172/JCI105112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiss J., Elsbach P., Shu C., Castillo J., Grinna L., Horwitz A., Theofan G. Human bactericidal/permeability-increasing protein and a recombinant NH2-terminal fragment cause killing of serum-resistant gram-negative bacteria in whole blood and inhibit tumor necrosis factor release induced by the bacteria. J Clin Invest. 1992 Sep;90(3):1122–1130. doi: 10.1172/JCI115930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss J., Victor M., Cross A. S., Elsbach P. Sensitivity of K1-encapsulated Escherichia coli to killing by the bactericidal/permeability-increasing protein of rabbit and human neutrophils. Infect Immun. 1982 Dec;38(3):1149–1153. doi: 10.1128/iai.38.3.1149-1153.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wilmes-Riesenberg M. R., Wanner B. L. TnphoA and TnphoA' elements for making and switching fusions for study of transcription, translation, and cell surface localization. J Bacteriol. 1992 Jul;174(14):4558–4575. doi: 10.1128/jb.174.14.4558-4575.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wolberg G., DeWitt C. W. Mouse virulence of K(L) antigen-containing strains of Escherichia coli. J Bacteriol. 1969 Nov;100(2):730–737. doi: 10.1128/jb.100.2.730-737.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. van den Bosch J. F., Postma P., Koopman P. A., de Graaff J., MacLaren D. M., van Brenk D. G., Guinée P. A. Virulence of urinary and faecal Escherichia coli in relation to serotype, haemolysis and haemagglutination. J Hyg (Lond) 1982 Jun;88(3):567–577. doi: 10.1017/s002217240007042x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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