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. 1987 May;55(5):1063–1069. doi: 10.1128/iai.55.5.1063-1069.1987

Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli O159:H4.

C O Tacket, D R Maneval, M M Levine
PMCID: PMC260469  PMID: 2883122

Abstract

The ability to colonize the small intestine is essential for the pathogenesis of diarrhea caused by enterotoxigenic Escherichia coli (ETEC). Colonization is mediated by fimbriae (pili), of which there are several antigenically distinct types, including colonization factor antigen I, colonization factor antigen II (CS1, CS2, and CS3), and PCF8775 (CS4, CS5, and CS6). These fimbriae are associated with certain ETEC O serogroups. Serogroup O159 has had no known colonization factor. We found a distinct plasmid-encoded fimbria composed of 19-kilodalton protein subunits associated with ETEC serotype O159:H4. Rabbit antibody against this purified fimbria reacted with a single 19-kilodalton protein band as seen by Western immunoblot of sheared-cell preparations. The rabbit antibody, treated with colloidal-gold-labeled goat anti-rabbit immunoglobulin G, bound specifically to fimbriae when cells were examined with an electron microscope. Of 10 available ETEC O159:H4 strains from Europe, Bangladesh, and Kenya, 6 expressed this type of fimbria; its true prevalence among ETEC strains is unknown. This putative colonization factor of O159:H4 joins other ETEC fimbriae as potentially useful immunogens against human diarrhea.

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  1. Batteiger B., Newhall W. J., 5th, Jones R. B. The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 1982 Dec 30;55(3):297–307. doi: 10.1016/0022-1759(82)90089-8. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  4. Evans D. G., Evans D. J., Jr New surface-associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups O6 and O8. Infect Immun. 1978 Aug;21(2):638–647. doi: 10.1128/iai.21.2.638-647.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Evans D. G., Evans D. J., Jr, Tjoa W. S., DuPont H. L. Detection and characterization of colonization factor of enterotoxigenic Escherichia coli isolated from adults with diarrhea. Infect Immun. 1978 Feb;19(2):727–736. doi: 10.1128/iai.19.2.727-736.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans D. G., Satterwhite T. K., Evans D. J., Jr, DuPont H. L. Differences in serological responses and excretion patterns of volunteers challenged with enterotoxigenic Escherichia coli with and without the colonization factor antigen. Infect Immun. 1978 Mar;19(3):883–888. doi: 10.1128/iai.19.3.883-888.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faulk W. P., Taylor G. M. An immunocolloid method for the electron microscope. Immunochemistry. 1971 Nov;8(11):1081–1083. doi: 10.1016/0019-2791(71)90496-4. [DOI] [PubMed] [Google Scholar]
  9. Gorbach S. L., Kean B. H., Evans D. G., Evans D. J., Jr, Bessudo D. Travelers' diarrhea and toxigenic Escherichia coli. N Engl J Med. 1975 May 1;292(18):933–936. doi: 10.1056/NEJM197505012921801. [DOI] [PubMed] [Google Scholar]
  10. Gothefors L., Ahrén C., Stoll B., Barua D. K., Orskov F., Salek M. A., Svennerholm A. M. Presence of colonization factor antigens on fresh isolates of fecal Escherichia coli: a prospective study. J Infect Dis. 1985 Dec;152(6):1128–1133. doi: 10.1093/infdis/152.6.1128. [DOI] [PubMed] [Google Scholar]
  11. Honda T., Arita M., Miwatani T. Characterization of new hydrophobic pili of human enterotoxigenic Escherichia coli: a possible new colonization factor. Infect Immun. 1984 Mar;43(3):959–965. doi: 10.1128/iai.43.3.959-965.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Levine M. M., Nalin D. R., Hoover D. L., Bergquist E. J., Hornick R. B., Young C. R. Immunity to enterotoxigenic Escherichia coli. Infect Immun. 1979 Mar;23(3):729–736. doi: 10.1128/iai.23.3.729-736.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Levine M. M., Rennels M. B., Daya V., Hughes T. P. Hemagglutination and colonization factors in enterotoxigenic and enteropathogenic Escherichia coli that cause diarrhea. J Infect Dis. 1980 Jun;141(6):733–737. doi: 10.1093/infdis/141.6.733. [DOI] [PubMed] [Google Scholar]
  15. Levine M. M., Ristaino P., Marley G., Smyth C., Knutton S., Boedeker E., Black R., Young C., Clements M. L., Cheney C. Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infect Immun. 1984 May;44(2):409–420. doi: 10.1128/iai.44.2.409-420.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Levine M. M., Ristaino P., Sack R. B., Kaper J. B., Orskov F., Orskov I. Colonization factor antigens I and II and type 1 somatic pili in enterotoxigenic Escherichia coli: relation to enterotoxin type. Infect Immun. 1983 Feb;39(2):889–897. doi: 10.1128/iai.39.2.889-897.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  18. McConnell M. M., Thomas L. V., Day N. P., Rowe B. Enzyme-linked immunosorbent assays for the detection of adhesion factor antigens of enterotoxigenic Escherichia coli. J Infect Dis. 1985 Dec;152(6):1120–1127. doi: 10.1093/infdis/152.6.1120. [DOI] [PubMed] [Google Scholar]
  19. Merson M. H., Morris G. K., Sack D. A., Wells J. G., Feeley J. C., Sack R. B., Creech W. B., Kapikian A. Z., Gangarosa E. J. Travelers' diarrhea in Mexico. A prospective study of physicians and family members attending a congress. N Engl J Med. 1976 Jun 10;294(24):1299–1305. doi: 10.1056/NEJM197606102942401. [DOI] [PubMed] [Google Scholar]
  20. Morgan R. L., Isaacson R. E., Moon H. W., Brinton C. C., To C. C. Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified 987 or K99 pili: protection correlates with pilus homology of vaccine and challenge. Infect Immun. 1978 Dec;22(3):771–777. doi: 10.1128/iai.22.3.771-777.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Moseley S. L., Huq I., Alim A. R., So M., Samadpour-Motalebi M., Falkow S. Detection of enterotoxigenic Escherichia coli by DNA colony hybridization. J Infect Dis. 1980 Dec;142(6):892–898. doi: 10.1093/infdis/142.6.892. [DOI] [PubMed] [Google Scholar]
  22. Nagy B., Moon H. W., Isaacson R. E., To C. C., Brinton C. C. Immunization of suckling pigs against enteric enterotoxigenic Escherichia coli infection by vaccinating dams with purified pili. Infect Immun. 1978 Jul;21(1):269–274. doi: 10.1128/iai.21.1.269-274.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. ORNSTEIN L. DISC ELECTROPHORESIS. I. BACKGROUND AND THEORY. Ann N Y Acad Sci. 1964 Dec 28;121:321–349. doi: 10.1111/j.1749-6632.1964.tb14207.x. [DOI] [PubMed] [Google Scholar]
  24. Orskov I., Orskov F. Special O:K:H serotypes among enterotoxigenic E. coli strains from diarrhea in adults and children. Occurrence of the CF (colonization factor) antigen and of hemagglutinating abilities. Med Microbiol Immunol. 1977 Jul 18;163(2):99–110. doi: 10.1007/BF02121825. [DOI] [PubMed] [Google Scholar]
  25. Ristaino P. A., Levine M. M., Young C. R. Improved GM1-enzyme-linked immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin. J Clin Microbiol. 1983 Oct;18(4):808–815. doi: 10.1128/jcm.18.4.808-815.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosenberg M. L., Koplan J. P., Wachsmuth I. K., Wells J. G., Gangarosa E. J., Guerrant R. L., Sack D. A. Epidemic diarrhea at Crater Lake from enterotoxigenic Escherichia coli. A large waterborne outbreak. Ann Intern Med. 1977 Jun;86(6):714–718. doi: 10.7326/0003-4819-86-6-714. [DOI] [PubMed] [Google Scholar]
  27. Sack R. B. Human diarrheal disease caused by enterotoxigenic Escherichia coli. Annu Rev Microbiol. 1975;29:333–353. doi: 10.1146/annurev.mi.29.100175.002001. [DOI] [PubMed] [Google Scholar]
  28. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  29. Taylor W. R., Schell W. L., Wells J. G., Choi K., Kinnunen D. E., Heiser P. T., Helstad A. G. A foodborne outbreak of enterotoxigenic Escherichia coli diarrhea. N Engl J Med. 1982 May 6;306(18):1093–1095. doi: 10.1056/NEJM198205063061807. [DOI] [PubMed] [Google Scholar]
  30. Thomas L. V., Cravioto A., Scotland S. M., Rowe B. New fimbrial antigenic type (E8775) that may represent a colonization factor in enterotoxigenic Escherichia coli in humans. Infect Immun. 1982 Mar;35(3):1119–1124. doi: 10.1128/iai.35.3.1119-1124.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Thomas L. V., McConnell M. M., Rowe B., Field A. M. The possession of three novel coli surface antigens by enterotoxigenic Escherichia coli strains positive for the putative colonization factor PCF8775. J Gen Microbiol. 1985 Sep;131(9):2319–2326. doi: 10.1099/00221287-131-9-2319. [DOI] [PubMed] [Google Scholar]
  32. Thomas L. V., Rowe B. The occurrence of colonisation factors (CFA/I, CFA/II and E8775) in enterotoxigenic Escherichia coli from various countries in South East Asia. Med Microbiol Immunol. 1982;171(2):85–90. doi: 10.1007/BF02124915. [DOI] [PubMed] [Google Scholar]
  33. Thompson M. R., Brandwein H., LaBine-Racke M., Giannella R. A. Simple and reliable enzyme-linked immunosorbent assay with monoclonal antibodies for detection of Escherichia coli heat-stable enterotoxins. J Clin Microbiol. 1984 Jul;20(1):59–64. doi: 10.1128/jcm.20.1.59-64.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Yamamoto T., Tamura Y., Yokota T. Enteroadhesion fimbriae and enterotoxin of Escherichia coli: genetic transfer to a streptomycin-resistant mutant of the galE oral-route live-vaccine Salmonella typhi Ty21a. Infect Immun. 1985 Dec;50(3):925–928. doi: 10.1128/iai.50.3.925-928.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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