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Microbiology 147 (2001), 861-867
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Microbiology (2001), 147, 861-867.
© 2001 Society for General Microbiology

Antigens and Immunity

Antibody response against Escherichia coli heat-stable enterotoxin expressed as fusions to flagellin

Cátia M. Pereira1, Beatriz E. Cabilio Guth1, Maria Elisabete Sbrogio-Almeida2 and Beatriz A. Castilho1

Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Botucatu, 862, CEP 04023-062, São Paulo, Brazil1
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Avenida Vital Brasil 1500, CEP 05503-900, São Paulo, Brazil2

Author for correspondence: Cátia M. Pereira. Tel: +55 11 5084 3213. Fax: +55 11 571 6504. e-mail: pereiracm{at}hotmail.com

The heat-stable toxin (ST) produced by enterotoxigenic Escherichia coli strains causes diarrhoea by altering the fluid secretion in intestinal epithelial cells. Here, the effectiveness of a flagellin fusion protein of Salmonella containing a 19-amino-acid sequence derived from the ST sequence (FLA–ST) in generating antibodies capable of neutralizing the toxic activity of ST was evaluated. This fusion protein, and an alternative construction where two cysteine residues in the ST sequence were substituted by alanines (STmt), were delivered to the immune system by three distinct strategies: (i) orally, using an attenuated Salmonella strain expressing FLA–ST; (ii) intraperitoneally, by injection of purified FLA–ST; (iii) orally, using attenuated Salmonella carrying a eukaryotic expression plasmid (pCDNA3) with the gene encoding FLA–ST. The results showed that the flagellin system can be used as a carrier to generateST-neutralizing antibodies. However, it should be mentioned that humoral immune response against ST was only obtained when the mutated ST sequence was employed. FLA–ST was found to be non-immunogenic when delivered via the oral route with attenuated Salmonella strains. However, a flagellin antibody response was obtained by immunizing mice with Salmonella carrying pCDNA3/FLA-STmt. Oral immunization with Salmonella carrying the eukaryotic expression plasmid (pCDNA3/FLA–STmt) seems to be a promising method to elicit an appropriate response against fusions to flagellin.

Keywords: ETEC, ST, attenuated Salmonella, flagellin fusions, mucosal vaccines

Abbreviations: ETEC; enterotoxigenic Escherichia coli; OPD, o-phenylenediamine dihydrochloride




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