Abstract
The antigenic relationships between heat-labile enterotoxin (LT) produced by a human strain of enterotoxigenic Escherichia coli (strain 286C2) and cholera toxin (CT) were examined by using antisera raised against LT and CT and specific antisera prepared against each subunit of both enterotoxins. Double immunodiffusion analysis revealed reactions of partial identity between the A subunits of LT and CT, as well as between the B subunits. Rabbit antisera raised against LT subunit A reacted with only subunit A, whereas rabbits immunized with LT subunit B produced antibodies which reacted with only subunit B. A high degree of CT neutralization was observed with antisera raised against LT. Data from neutralization assays with specific antisera to each enterotoxin showed that LT was more effectively neutralized by homologous anti-LT than CT (3.7-fold); however, anti-CT was only slightly more effective in neutralization of homologous CT compared with LT (1.9-fold). In contrast, antisera raised against the B subunit of CT (choleragenoid) exhibited significantly higher neutralization activity against CT than LT (5.8-fold); however, the amount of CT neutralized by anticholeragenoid was less (4.1-fold) than anti-CT. These results suggested that anti-CT serum contained neutralizing antibodies reactive with a shared determinant formed by interaction of the A and B subunits, whereas anti-LT and anti-choleragenoid sera did not. Sensitive solid-phase radioimmunoassays were developed to examine the affinity and degree of specificity involved in homologous and heterologous antigen-antibody interactions between LT, CT, their subunits, and specific antibodies. Only unlabeled LT competed with radiolabeled LT in polystyrene tubes coated with anti-LT, and only unlabeled CT competed with radiolabeled CT in tubes coated with anti-CT. However, when radiolabeled CT was incubated in tubes coated with anti-LT, competitive inhibition responses were observed with both unlabeled toxins. When radiolabeled LT was incubated with tubes coated with anti-CT, competitive inhibition responses were observed with both unlabeled toxins. Similar competitive inhibition responses were observed with the A subunits of LT and CT and with the B subunits using antisera specific for the subunits of each enterotoxin. Double immunodiffusion analysis and radioimmunoassay data supported the presence of unique and shared immunodeterminants in each subunit.
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Selected References
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- Boesman-Finkelstein M., Finkelstein R. A. Protection in rabbits induced by the Texas Star-SR attenuated A-B+ mutant candidate live oral cholera vaccine. Infect Immun. 1982 Apr;36(1):221–226. doi: 10.1128/iai.36.1.221-226.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clements J. D., Finkelstein R. A. Demonstration of shared and unique immunological determinants in enterotoxins from Vibrio cholerae and Escherichia coli. Infect Immun. 1978 Dec;22(3):709–713. doi: 10.1128/iai.22.3.709-713.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clements J. D., Finkelstein R. A. Immunological cross-reactivity between a heat-labile enterotoxin(s) of Escherichia coli and subunits of Vibrio cholerae enterotoxin. Infect Immun. 1978 Sep;21(3):1036–1039. doi: 10.1128/iai.21.3.1036-1039.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Clements J. D., Yancey R. J., Finkelstein R. A. Properties of homogeneous heat-labile enterotoxin from Escherichia coli. Infect Immun. 1980 Jul;29(1):91–97. doi: 10.1128/iai.29.1.91-97.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dallas W. S., Falkow S. The molecular nature of heat-labile enterotoxin (LT) of escherichia coli. Nature. 1979 Feb 1;277(5695):406–407. doi: 10.1038/277406a0. [DOI] [PubMed] [Google Scholar]
- Donta S. T., Moon H. W., Whipp S. C. Detection of heat-labile Escherichia coli enterotoxin with the use of adrenal cells in tissue culture. Science. 1974 Jan 25;183(4122):334–336. doi: 10.1126/science.183.4122.334. [DOI] [PubMed] [Google Scholar]
- Donta S. T. Neutralization of cholera enterotoxin-induced steroidogenesis by specific antibody. J Infect Dis. 1974 Mar;129(3):284–288. doi: 10.1093/infdis/129.3.284. [DOI] [PubMed] [Google Scholar]
- Field M. Mechanisms of action of cholera and Escherichia coli enterotoxins. Am J Clin Nutr. 1979 Jan;32(1):189–196. doi: 10.1093/ajcn/32.1.189. [DOI] [PubMed] [Google Scholar]
- 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]
- GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Gilligan P. H., Robertson D. C. Nutritional requirements for synthesis of heat-labile enterotoxin by enterotoxigenic strains of Escherichia coli. Infect Immun. 1979 Jan;23(1):99–107. doi: 10.1128/iai.23.1.99-107.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gyles C. L. Immunological study of the heat-labile enterotoxins of Escherichia coli and Vibrio cholerae. Infect Immun. 1974 Mar;9(3):564–570. doi: 10.1128/iai.9.3.564-570.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Holmgren J., Söderlind O., Wadström T. Cross-reactivity between heat labile enterotoxins of Vibrio cholerae and Escherichia coli in neutralization tests in rabbit ileum and skin. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Dec;81(6):757–762. doi: 10.1111/j.1699-0463.1973.tb02272.x. [DOI] [PubMed] [Google Scholar]
- Honda T., Takeda Y., Miwatani T. Isolation of special antibodies which react only with homologous enterotoxins from Vibrio cholerae and Enterotoxigenic Escherichia coli. Infect Immun. 1981 Nov;34(2):333–336. doi: 10.1128/iai.34.2.333-336.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Kantor H. S. Enterotoxins of Escherichia coli and vibriocholerae: tools for the molecular biologist. J Infect Dis. 1975 May;131 (Suppl):S22–S32. doi: 10.1093/infdis/131.supplement.s22. [DOI] [PubMed] [Google Scholar]
- Klipstein F. A., Engert R. F. Immunological interrelationships between cholera toxin and the heat-labile and heat-stable enterotoxins of coliform bacteria. Infect Immun. 1977 Oct;18(1):110–117. doi: 10.1128/iai.18.1.110-117.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kunkel S. L., Robertson D. C. Purification and chemical characterization of the heat-labile enterotoxin produced by enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):586–596. doi: 10.1128/iai.25.2.586-596.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Markel D. E., Hejtmancik K. E., Peterson J. W., Martin F. B., Kurosky A. Characterization of the antigenic determinants of cholera toxin subunits. Infect Immun. 1979 Aug;25(2):615–626. doi: 10.1128/iai.25.2.615-626.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris G. K., Merson M. H., Sack D. A., Wells J. G., Martin W. T., Dewitt W. E., Feeley J. C., Sack R. B., Bessudo D. M. Laboratory investigation of diarrhea in travelers to Mexico: evaluation of methods for detecting enterotoxigenic Echerichia coli. J Clin Microbiol. 1976 May;3(5):486–495. doi: 10.1128/jcm.3.5.486-495.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moseley S. L., Falkow S. Nucleotide sequence homology between the heat-labile enterotoxin gene of Escherichia coli and Vibrio cholerae deoxyribonucleic acid. J Bacteriol. 1980 Oct;144(1):444–446. doi: 10.1128/jb.144.1.444-446.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moss J., Osborne J. C., Jr, Fishman P. H., Nakaya S., Robertson D. C. Escherichia coli heat-labile enterotoxin. Ganglioside specificity and ADP-ribosyltransferase activity. J Biol Chem. 1981 Dec 25;256(24):12861–12865. [PubMed] [Google Scholar]
- Rappaport R. S., Bonde G. Development of a vaccine against experimental cholera and Escherichia coli diarrheal disease. Infect Immun. 1981 May;32(2):534–541. doi: 10.1128/iai.32.2.534-541.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Smith N. W., Sack R. B. Immunologic cross-reactions of enterotoxins from Escherichia coli and Vibrio cholerae. J Infect Dis. 1973 Feb;127(2):164–170. doi: 10.1093/infdis/127.2.164. [DOI] [PubMed] [Google Scholar]
- Takeda Y., Honda T., Taga S., Miwatani T. In vitro formation of hybrid toxins between subunits of Escherichia coli heat-labile enterotoxin and those of cholera enterotoxin. Infect Immun. 1981 Nov;34(2):341–346. doi: 10.1128/iai.34.2.341-346.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Heyningen S. The subunits of cholera toxin: structure, stoichiometry, and function. J Infect Dis. 1976 Mar;133 (Suppl):5–13. doi: 10.1093/infdis/133.supplement_1.s5. [DOI] [PubMed] [Google Scholar]