Abstract
Soluble hemagglutinin (HA) from an El Tor Vibrio cholerae strain (serotype Ogawa) was purified by means of a sequence of salt precipitation, gel filtration, and agarose electrophoresis. The purified material, which gave a single precipitation line in immunodiffusion tests with homologous antiserum, showed immunological identity reactions in double diffusion-in-gel with soluble HA produced by various classical and El Tor strains of different serotypes. Purified HA was used for development of an enzyme-linked immunosorbent assay for titration of specific antibodies against soluble HA and for quantitation of this antigen. Rabbit anti-HA serum reacted in high titer with the soluble HA coated on polystyrene microtiter plates, whereas antiserum against cholera toxin, lipopolysaccharide, or whole washed V. cholerae showed little or no reactivity. In inhibition tests as little as 2.5 ng of soluble HA could be detected with the enzyme-linked immunosorbent assay. Culture supernatants of different El Tor as well as classical V. cholerae strains all completely inhibited the binding of anti-HA antibody to solid-phase-bound homologous antigen, but the amounts of HA produced by individual strains varied at least 1,000-fold. Only 2 of 10 paired acute- and convalescent-phase sera from Bangladeshi cholera patients showed significant titer increases against soluble HA in parallel titrations.
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Selected References
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- Chaicumpa W., Atthasisiha N. Study of intestinal immunity against V. cholerae: role of antibody to V. cholerae haemagglutinin in intestinal immunity. Southeast Asian J Trop Med Public Health. 1977 Mar;8(1):13–18. [PubMed] [Google Scholar]
- Finkelstein R. A., Hanne L. F. Purification and characterization of the soluble hemagglutinin (cholera lectin)( produced by Vibrio cholerae. Infect Immun. 1982 Jun;36(3):1199–1208. doi: 10.1128/iai.36.3.1199-1208.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanne L. F., Finkelstein R. A. Characterization and distribution of the hemagglutinins produced by Vibrio cholerae. Infect Immun. 1982 Apr;36(1):209–214. doi: 10.1128/iai.36.1.209-214.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmgren J., Svennerholm A. M., Ahrén C. Nonimmunoglobulin fraction of human milk inhibits bacterial adhesion (hemagglutination) and enterotoxin binding of Escherichia coli and Vibrio cholerae. Infect Immun. 1981 Jul;33(1):136–141. doi: 10.1128/iai.33.1.136-141.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmgren J., Svennerholm A. M. Enzyme-linked immunosorbent assays for cholera serology. Infect Immun. 1973 May;7(5):759–763. doi: 10.1128/iai.7.5.759-763.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmgren J., Svennerholm A. M., Lindblad M. Receptor-like glycocompounds in human milk that inhibit classical and El Tor Vibrio cholerae cell adherence (hemagglutination). Infect Immun. 1983 Jan;39(1):147–154. doi: 10.1128/iai.39.1.147-154.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johansson B. G. Agarose gel electrophoresis. Scand J Clin Lab Invest Suppl. 1972;124:7–19. doi: 10.3109/00365517209102747. [DOI] [PubMed] [Google Scholar]
- Jones G. W., Abrams G. D., Freter R. Adhesive properties of Vibrio cholerae: adhesion to isolated rabbit brush border membranes and hemagglutinating activity. Infect Immun. 1976 Jul;14(1):232–239. doi: 10.1128/iai.14.1.232-239.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lönnroth I., Holmgren J. Subunit structure of cholera toxin. J Gen Microbiol. 1973 Jun;76(2):417–427. doi: 10.1099/00221287-76-2-417. [DOI] [PubMed] [Google Scholar]
- WADSWORTH C. A slide microtechnique for the analysis of immune precipitates in gel. Int Arch Allergy Appl Immunol. 1957;10(6):355–360. doi: 10.1159/000228394. [DOI] [PubMed] [Google Scholar]