Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1988 Sep;56(9):2330–2335. doi: 10.1128/iai.56.9.2330-2335.1988

Detection of pilus subunits (pilins) and filaments by using anti-P pilin antisera.

I E Salit 1, J Hanley 1, L Clubb 1, S Fanning 1
PMCID: PMC259568  PMID: 2900813

Abstract

P pilus filaments are important in binding to globoside through an adhesin located at the tip of the pilus. There is considerable antigenic variation among P pili, and the immunologic response is usually serotype specific. We purified denatured pilin subunits and used them as immunogens to prepare more broadly cross-reactive antisera. Although antifilament antisera (AFA) detected predominantly the homologous strain, antisubunit antisera (ASA) prepared from two different strains detected P pili in 16 of 16 and 14 of 16 P-piliated strains by Western blotting (immunoblotting). The binding of ASA to the homologous pilus filament was inhibited by only 3 of 17 strains. ASA agglutinated only two of nine heterologous strains and immunoprecipitated pili from one of three heterologous strains. By immunoelectron microscopy ASA was seen to bind to pilus filaments but not as strongly as AFA. Antiserum raised to the denatured pilin subunit was not substantially more reactive with pilus filaments derived from heterologous strains than was AFA. ASA was, however, a very useful probe for detecting most P pilins.

Full text

PDF
2330

Images in this article

2332Image
on p.2332
2332Image
on p.2332
2332Image
on p.2332
2333Image
on p.2333
2333Image
on p.2333
2333Image
on p.2333
2334Image
on p.2334

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berzofsky J. A. Intrinsic and extrinsic factors in protein antigenic structure. Science. 1985 Sep 6;229(4717):932–940. doi: 10.1126/science.2410982. [DOI] [PubMed] [Google Scholar]
  2. Clegg S., Evans D. J., Jr, Evans D. G. Antigenic heterogeneity of haemagglutination type VI fimbriae produced by Escherichia coli isolated from patients with bacteremia. J Clin Microbiol. 1982 Jul;16(1):174–180. doi: 10.1128/jcm.16.1.174-180.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clegg S. Serological heterogeneity among fimbrial antigens causing mannose-resistant hemagglutination by uropathogenic Escherichia coli. Infect Immun. 1982 Feb;35(2):745–748. doi: 10.1128/iai.35.2.745-748.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans D. G., Evans D. J., Jr, Tjoa W. Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect Immun. 1977 Nov;18(2):330–337. doi: 10.1128/iai.18.2.330-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hanley J., Salit I. E., Hofmann T. Immunochemical characterization of P pili from invasive Escherichia coli. Infect Immun. 1985 Sep;49(3):581–586. doi: 10.1128/iai.49.3.581-586.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Jacobsen S. H., Lins L. E., Svenson S. B., Källenius G. P fimbriated Escherichia coli in adults with acute pyelonephritis. J Infect Dis. 1985 Aug;152(2):426–427. doi: 10.1093/infdis/152.2.426. [DOI] [PubMed] [Google Scholar]
  7. Karch H., Leying H., Goroncy-Bermes P., Kroll H. P., Opferkuch W. Three-dimensional structure of fimbriae determines specificity of immune response. Infect Immun. 1985 Nov;50(2):517–522. doi: 10.1128/iai.50.2.517-522.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Korhonen T. K., Väisänen V., Saxén H., Hultberg H., Svenson S. B. P-antigen-recognizing fimbriae from human uropathogenic Escherichia coli strains. Infect Immun. 1982 Jul;37(1):286–291. doi: 10.1128/iai.37.1.286-291.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Lindberg F., Lund B., Johansson L., Normark S. Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus. Nature. 1987 Jul 2;328(6125):84–87. doi: 10.1038/328084a0. [DOI] [PubMed] [Google Scholar]
  11. Lund B., Lindberg F., Marklund B. I., Normark S. The PapG protein is the alpha-D-galactopyranosyl-(1----4)-beta-D-galactopyranose-binding adhesin of uropathogenic Escherichia coli. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5898–5902. doi: 10.1073/pnas.84.16.5898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. O'Hanley P., Lark D., Falkow S., Schoolnik G. Molecular basis of Escherichia coli colonization of the upper urinary tract in BALB/c mice. Gal-Gal pili immunization prevents Escherichia coli pyelonephritis in the BALB/c mouse model of human pyelonephritis. J Clin Invest. 1985 Feb;75(2):347–360. doi: 10.1172/JCI111707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Pere A., Väisänen-Rhen V., Rhen M., Tenhunen J., Korhonen T. K. Analysis of P fimbriae on Escherichia coli O2, O4, and O6 strains by immunoprecipitation. Infect Immun. 1986 Feb;51(2):618–625. doi: 10.1128/iai.51.2.618-625.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roberts J. A., Hardaway K., Kaack B., Fussell E. N., Baskin G. Prevention of pyelonephritis by immunization with P-fimbriae. J Urol. 1984 Mar;131(3):602–607. doi: 10.1016/s0022-5347(17)50513-3. [DOI] [PubMed] [Google Scholar]
  16. Salit I. E., Gotschlich E. C. Hemagglutination by purified type I Escherichia coli pili. J Exp Med. 1977 Nov 1;146(5):1169–1181. doi: 10.1084/jem.146.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Salit I. E., Hanley J., Clubb L., Fanning S. The human antibody response to uropathogenic Escherichia coli: a review. Can J Microbiol. 1988 Mar;34(3):312–318. doi: 10.1139/m88-057. [DOI] [PubMed] [Google Scholar]
  18. Salit I. E., Vavougios J., Hofmann T. Isolation and characterization of Escherichia coli pili from diverse clinical sources. Infect Immun. 1983 Nov;42(2):755–762. doi: 10.1128/iai.42.2.755-762.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schmidt M. A., O'Hanley P., Schoolnik G. K. Gal-Gal pyelonephritis Escherichia coli pili linear immunogenic and antigenic epitopes. J Exp Med. 1985 Apr 1;161(4):705–717. doi: 10.1084/jem.161.4.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Svanborg-Edén C., Mårild S., Korhonen T. K. Adhesion inhibition by antibodies. Scand J Infect Dis Suppl. 1982;33:72–78. [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. de Ree J. M., Schwillens P., van den Bosch J. F. Monoclonal antibodies that recognize the P fimbriae F71, F72, F9, and F11 from uropathogenic Escherichia coli. Infect Immun. 1985 Dec;50(3):900–904. doi: 10.1128/iai.50.3.900-904.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES