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. 1985 Nov;50(2):459–466. doi: 10.1128/iai.50.2.459-466.1985

Monoclonal antibodies to enterobacterial common antigen and to Escherichia coli lipopolysaccharide outer core: demonstration of an antigenic determinant shared by enterobacterial common antigen and E. coli K5 capsular polysaccharide.

H Peters, M Jürs, B Jann, K Jann, K N Timmis, D Bitter-Suermann
PMCID: PMC261975  PMID: 2414223

Abstract

We established hybridoma cell lines producing monoclonal antibodies against enterobacterial common antigen (ECA) and a substructure of the outer core of different Escherichia coli lipopolysaccharides (LPSs). Anti-ECA antibodies 865 and 898 reacted with ECA in extracts of heated E. coli and with ECA-bound R1 and R4 core-containing LPS preparations, as well as with a purified sample of ECA from Salmonella montevideo. Antibody 865, but not antibody 898, cross-reacted with K5 capsular polysaccharide, suggesting that 4-linked alpha-N-acetylglucosamine is part of an antigenic determinant shared by both K5 polysaccharide and ECA. Anti-LPS antibody 786 recognized an outer core structure common to E. coli K-12, B, R2, and R4 core type LPS, but not to R1 and R3 core type LPS. Its most probable target is the trisaccharide sequence Hexp(1----2)-alpha-D -Glcp(1----3) alpha-D-Glcp----(Hepp) (where Hex is hexose, p is phosphate, Glc is glucose, and Hep is heptose), the first glucose being the immunodominant moiety. These monoclonal antibodies may be used not only for the detection of ECA, K5, and LPS core structures but also for analysis of the molecular forms resolved on polyacrylamide gels (banding patterns) of both ECA and LPS, independently of one another.

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Selected References

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  1. Bitter-Suermann D., Peters H., Jürs M., Nehrbass R., Montenegro M., Timmis K. N. Monoclonal antibody detection of IncF group plasmid-encoded TraT protein in clinical isolates of Escherichia coli. Infect Immun. 1984 Nov;46(2):308–313. doi: 10.1128/iai.46.2.308-313.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burger R., Deubel U., Hadding U., Bitter-Suermann D. Identification of functionally relevant determinants on the complement component C3 with monoclonal antibodies. J Immunol. 1982 Nov;129(5):2042–2050. [PubMed] [Google Scholar]
  3. Dell A., Oates J., Lugowski C., Romanowska E., Kenne L., Lindberg B. The enterobacterial common-antigen, a cyclic polysaccharide. Carbohydr Res. 1984 Oct 1;133(1):95–104. doi: 10.1016/0008-6215(84)85186-1. [DOI] [PubMed] [Google Scholar]
  4. Feige U., Jann B., Jann K., Schmidt G., Stirm S. On the primary structure of the Escherichia coli R4 cell wall lipopolysaccharide core. Biochem Biophys Res Commun. 1977 Nov 7;79(1):88–95. doi: 10.1016/0006-291x(77)90064-x. [DOI] [PubMed] [Google Scholar]
  5. Frosch M., Görgen I., Boulnois G. J., Timmis K. N., Bitter-Suermann D. NZB mouse system for production of monoclonal antibodies to weak bacterial antigens: isolation of an IgG antibody to the polysaccharide capsules of Escherichia coli K1 and group B meningococci. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1194–1198. doi: 10.1073/pnas.82.4.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  7. Gotschlich E. C., Fraser B. A., Nishimura O., Robbins J. B., Liu T. Y. Lipid on capsular polysaccharides of gram-negative bacteria. J Biol Chem. 1981 Sep 10;256(17):8915–8921. [PubMed] [Google Scholar]
  8. Gotschlich E. C., Rey M., Etienne J., Sanborn W. R., Triau R., Cvjetanović B. The immunological responses observed in field studies in Africa with group A meningococcal vaccines. Prog Immunobiol Stand. 1971;5:485–491. [PubMed] [Google Scholar]
  9. Gross R. J., Cheasty T., Rowe B. Isolation of bacteriophages specific for the K1 polysaccharide antigen of Escherichia coli. J Clin Microbiol. 1977 Dec;6(6):548–550. doi: 10.1128/jcm.6.6.548-550.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hase S., Rietschel E. T. Isolation and analysis of the lipid A backbone. Lipid A structure of lipopolysaccharides from various bacterial groups. Eur J Biochem. 1976 Mar 16;63(1):101–107. doi: 10.1111/j.1432-1033.1976.tb10212.x. [DOI] [PubMed] [Google Scholar]
  11. Hämmerling G., Lüderitz O., Westphal O., Mäkelä P. H. Structural investigations on the core polysaccharide of Escherichia coli 0100. Eur J Biochem. 1971 Oct 14;22(3):331–344. doi: 10.1111/j.1432-1033.1971.tb01549.x. [DOI] [PubMed] [Google Scholar]
  12. Jansson P. E., Lindberg A. A., Lindberg B., Wollin R. Structural studies on the hexose region of the core in lipopolysaccharides from Enterobacteriaceae. Eur J Biochem. 1981 Apr;115(3):571–577. doi: 10.1111/j.1432-1033.1981.tb06241.x. [DOI] [PubMed] [Google Scholar]
  13. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  14. Manning P. A., Timmis J. K., Moll A., Timmis K. N. Mutants that overproduce TraTp, a plasmid-specified major outer membrane protein of Escherichia coli. Mol Gen Genet. 1982;187(3):426–431. doi: 10.1007/BF00332623. [DOI] [PubMed] [Google Scholar]
  15. Mayer H., Schmidt G. Chemistry and biology of the enterobacterial common antigen (ECA). Curr Top Microbiol Immunol. 1979;85:99–153. doi: 10.1007/978-3-642-67322-1_3. [DOI] [PubMed] [Google Scholar]
  16. Mäkelä P. H., Mayer H. Enterobacterial common antigen. Bacteriol Rev. 1976 Sep;40(3):591–632. doi: 10.1128/br.40.3.591-632.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Männel D., Mayer H. Isolation and chemical characterization of the enterobacterial common antigen. Eur J Biochem. 1978 May 16;86(2):361–370. doi: 10.1111/j.1432-1033.1978.tb12318.x. [DOI] [PubMed] [Google Scholar]
  18. Orskov F., Orskov I., Sutton A., Schneerson R., Lin W., Egan W., Hoff G. E., Robbins J. B. Form variation in Escherichia coli K1: determined by O-acetylation of the capsular polysaccharide. J Exp Med. 1979 Mar 1;149(3):669–685. doi: 10.1084/jem.149.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Orskov I., Orskov F., Jann B., Jann K. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriol Rev. 1977 Sep;41(3):667–710. doi: 10.1128/br.41.3.667-710.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Palva E. T., Mäkelä P. H. Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Eur J Biochem. 1980;107(1):137–143. doi: 10.1111/j.1432-1033.1980.tb04634.x. [DOI] [PubMed] [Google Scholar]
  21. Prehm P., Schmidt G., Jann B., Jann K. The cell-wall lipopolysaccharide of Escherichia coli K-12. Structure and acceptor site for O-antigen and other substituents. Eur J Biochem. 1976 Nov 1;70(1):171–177. doi: 10.1111/j.1432-1033.1976.tb10967.x. [DOI] [PubMed] [Google Scholar]
  22. Prehm P., Stirm S., Jann B., Jann K., Boman H. G. Cell-wall lipopolysaccharides of ampicillin-resistant mutants of Escherichia coli K-12. Eur J Biochem. 1976 Jul 1;66(2):369–377. doi: 10.1111/j.1432-1033.1976.tb10526.x. [DOI] [PubMed] [Google Scholar]
  23. Prehm P., Stirm S., Jann B., Jann K. Cell-wall lipopolysaccharide from Escherichia coli B. Eur J Biochem. 1975 Aug 1;56(1):41–55. doi: 10.1111/j.1432-1033.1975.tb02205.x. [DOI] [PubMed] [Google Scholar]
  24. Tsui F. P., Boykins R. A., Egan W. Structural and immunological studies of the Escherichia coli K7 (K56) capsular polysaccharide. Carbohydr Res. 1982 Apr 16;102:263–271. doi: 10.1016/s0008-6215(00)88068-4. [DOI] [PubMed] [Google Scholar]
  25. Vann W. F., Jann K. Structure and serological specificity of the K13-antigenic polysaccharide (K13 antigen) of urinary tract-infective Escherichia coli. Infect Immun. 1979 Jul;25(1):85–92. doi: 10.1128/iai.25.1.85-92.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Weisgerber C., Jann B., Jann K. Biosynthesis of the 09 antigen of Escherichia coli. Core structure of rfe mutant as indication of assembly mechanism. Eur J Biochem. 1984 May 2;140(3):553–556. doi: 10.1111/j.1432-1033.1984.tb08137.x. [DOI] [PubMed] [Google Scholar]
  27. Ziegler E. J., McCutchan J. A., Fierer J., Glauser M. P., Sadoff J. C., Douglas H., Braude A. I. Treatment of gram-negative bacteremia and shock with human antiserum to a mutant Escherichia coli. N Engl J Med. 1982 Nov 11;307(20):1225–1230. doi: 10.1056/NEJM198211113072001. [DOI] [PubMed] [Google Scholar]

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