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. 1984 Mar;43(3):1027–1032. doi: 10.1128/iai.43.3.1027-1032.1984

Characterization of monoclonal antibodies to heat-labile enterotoxin encoded by a plasmid from a clinical isolate of Escherichia coli.

B W Belisle, E M Twiddy, R K Holmes
PMCID: PMC264288  PMID: 6365779

Abstract

Eight selected hybridoma cell lines that produced monoclonal antibodies against heat-labile enterotoxin from an Escherichia coli strain of human origin (LTh) were characterized. Antibodies produced by these cell lines were tested for binding specificity in a series of solid-phase radioimmunoassays and Western blots by using as test antigens LTh, the A, A1, A2, and B polypeptides of LTh, the heat-labile enterotoxin from an E. coli strain of porcine origin, and cholera toxin. The monoclonal antibodies were also tested for isotype and ability to neutralize LTh. Two of the anti-LTh monoclonal antibodies cross-reacted with cholera toxin, and six were specific for determinants of LTh that were not present on cholera toxin. One was specific for a unique epitope of LTh that was not shared by the heat-labile enterotoxin from an E. coli strain of porcine origin or cholera toxin. Four antibodies specific for epitopes on the B subunit of LTh (LTh-B) reacted with pentameric LTh-B but did not react in Western blots with monomeric LTh-B. The remaining four antibodies were specific for epitopes on LTh-A; two of these antibodies bound to A1, one reacted with A2, and one recognized only intact LTh-A. Only one monoclonal antibody had detectable neutralizing activity, and it was specific for LTh-A.

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Selected References

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  1. Bohn W. A fixation method for improved antibody penetration in electron microscopical immunoperoxidase studies. J Histochem Cytochem. 1978 Apr;26(4):293–297. doi: 10.1177/26.4.77869. [DOI] [PubMed] [Google Scholar]
  2. Clements J. D., Finkelstein R. A. Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun. 1979 Jun;24(3):760–769. doi: 10.1128/iai.24.3.760-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clements J. D., Flint D. C., Klipstein F. A. Immunological and physicochemical characterization of heat-labile enterotoxins isolated from two strains of Escherichia coli. Infect Immun. 1982 Nov;38(2):806–809. doi: 10.1128/iai.38.2.806-809.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dallas W. S., Falkow S. Amino acid sequence homology between cholera toxin and Escherichia coli heat-labile toxin. Nature. 1980 Dec 4;288(5790):499–501. doi: 10.1038/288499a0. [DOI] [PubMed] [Google Scholar]
  5. Finkelstein R. A., Boesman M., Neoh S. H., LaRue M. K., Delaney R. Dissociation and recombination of the subunits of the cholera enterotoxin (choleragen). J Immunol. 1974 Jul;113(1):145–150. [PubMed] [Google Scholar]
  6. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  7. Geary S. J., Marchlewicz B. A., Finkelstein R. A. Comparison of heat-labile enterotoxins from porcine and human strains of Escherichia coli. Infect Immun. 1982 Apr;36(1):215–220. doi: 10.1128/iai.36.1.215-220.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gyles C. L. Relationships among heat-labile enterotoxins of Escherichia coli and Vibrio cholerae. J Infect Dis. 1974 Mar;129(3):277–283. doi: 10.1093/infdis/129.3.277. [DOI] [PubMed] [Google Scholar]
  9. Hejtmancik K. E., Peterson J. W., Markel D. E., Kurosky A. Radioimmunoassay for the antigenic determinants of cholera toxin and its components. Infect Immun. 1977 Sep;17(3):621–628. doi: 10.1128/iai.17.3.621-628.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Holmes R. K., Twiddy E. M. Characterization of monoclonal antibodies that react with unique and cross-reacting determinants of cholera enterotoxin and its subunits. Infect Immun. 1983 Dec;42(3):914–923. doi: 10.1128/iai.42.3.914-923.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Holmgren J., Svennerholm A. M. Mechanisms of disease and immunity in cholera: a review. J Infect Dis. 1977 Aug;136 (Suppl):S105–S112. doi: 10.1093/infdis/136.supplement.s105. [DOI] [PubMed] [Google Scholar]
  12. Honda T., Tsuji T., Takeda Y., Miwatani T. Immunological nonidentity of heat-labile enterotoxins from human and porcine enterotoxigenic Escherichia coli. Infect Immun. 1981 Nov;34(2):337–340. doi: 10.1128/iai.34.2.337-340.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Lindholm L., Holmgren J., Wikström M., Karlsson U., Andersson K., Lycke N. Monoclonal antibodies to cholera toxin with special reference to cross-reactions with Escherichia coli heat-labile enterotoxin. Infect Immun. 1983 May;40(2):570–576. doi: 10.1128/iai.40.2.570-576.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Neill R. J., Ivins B. E., Holmes R. K. Synthesis and secretion of the plasmid-coded heat-labile enterotoxin of Escherichia coli in Vibrio cholerae. Science. 1983 Jul 15;221(4607):289–291. doi: 10.1126/science.6857285. [DOI] [PubMed] [Google Scholar]
  17. Peterson J. W., Hejtmancik K. E., Markel D. E., Craig J. P., Kurosky A. Antigenic specificity of neutralizing antibody to cholera toxin. Infect Immun. 1979 Jun;24(3):774–779. doi: 10.1128/iai.24.3.774-779.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Remmers E. F., Colwell R. R., Goldsby R. A. Production and characterization of monoclonal antibodies to cholera toxin. Infect Immun. 1982 Jul;37(1):70–76. doi: 10.1128/iai.37.1.70-76.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Robb M., Nichols J. C., Whoriskey S. K., Murphy J. R. Isolation of hybridoma cell lines and characterization of monoclonal antibodies against cholera enterotoxin and its subunits. Infect Immun. 1982 Oct;38(1):267–272. doi: 10.1128/iai.38.1.267-272.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  22. Spicer E. K., Noble J. A. Escherichia coli heat-labile enterotoxin. Nucleotide sequence of the A subunit gene. J Biol Chem. 1982 May 25;257(10):5716–5721. [PubMed] [Google Scholar]
  23. Takeda Y., Honda T., Sima H., Tsuji T., Miwatani T. Analysis of antigenic determinants in cholera enterotoxin and heat-labile enterotoxins from human and porcine enterotoxigenic Escherichia coli. Infect Immun. 1983 Jul;41(1):50–53. doi: 10.1128/iai.41.1.50-53.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Yamamoto T., Yokota T. Sequence of heat-labile enterotoxin of Escherichia coli pathogenic for humans. J Bacteriol. 1983 Aug;155(2):728–733. doi: 10.1128/jb.155.2.728-733.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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