Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/5652
Title: Expression of a pilin subunit BfpA of the bundle-forming pilus of enteropathogenic Escherichia coli in an aroA live salmonella vaccine strain.
Authors: Schriefer, Albert
Maltez, J. R
Silva, N
Stoeckle, M.Y
Barral Netto, Manoel
Riley, Lee Woodland
Affilliation: Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Cornell University Medical College. Division of International Medicine and Infectious Diseases. New York, New York
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Cornell University Medical College. Division of International Medicine and Infectious Diseases. New York, New York
Abstract: Enteropathogenic Escherichia coli (EPEC) is a major cause of childhood diarrhea in developing countries and is a leading cause of severe diarrheal illness among Brazilian infants. As one approach to constructing a vaccine candidate against diarrhea caused by EPEC, we evaluated whether the pilin subunit (BfpA) of the bundle-forming pilus (BFP) could be expressed by a live Salmonella vaccine strain. Several copies of the coding region of BfpA (bfpA) were amplified by PCR from a preparation of the EAF plasmid of EPEC strain B171 and cloned into plasmid vectors. An intact copy of bfpA was subcloned into the heat inducible prokaryotic expression vector pCYTEXP1, and the resulting pBfpA was used to transform the aroA S. typhimurium strain SL3261, generating SL3261(pBfpA). The recombinant vaccine strain was able to express, but not to process, rBfpA as evidenced by a prominent 21 kDa protein that crossreacted with anti-BFP antiserum found only in extracts of heat-treated SL3261(pBfpA), but not in strains of untreated SL3261(pBfpA) or SL3261 not carrying the plasmid. Furthermore, rBfpA accumulation was not toxic to the Salmonella host, as evidenced by similar plating efficiencies between induced and uninduced strains of SL3261(pBfpA). Finally, SL3261(pBfpA) orally administered to BALB/c mice was capable of eliciting a sustained and vigorous humoral immune response to BfpA, achievable even with a single oral dose of approximately 10(9) organisms. Therefore, this pilin product may serve as a potential immunogen as part of a live combined vaccine strategy to prevent two of the major public health problems in Brazil--salmonellosis and EPEC childhood diahrrea.
Keywords: EPEC
BFP
Vaccine
Oral
DeCS: Proteínas da Membrana Bacteriana Externa/genética
Vacinas Bacterianas/imunologia
Proteínas de Escherichia coli
Escherichia coli/genética
Escherichia coli/imunologia
Fímbrias Bacterianas/imunologia
Proteínas de Membrana/genética
Vacinas contra Salmonella
Salmonella typhimurium/imunologia
Vacinas Tíficas-Paratíficas
Administração Oral
Animais
Anticorpos Antibacterianos/biossíntese
Especificidade de Anticorpos
Proteínas da Membrana Bacteriana
Externa/biossíntese
Vacinas Bacterianas/genética
Clonagem Molecular
Escherichia coli/patogenicidade
Feminino
Proteínas de Fímbrias
Fímbrias Bacterianas/genética
Regulação Bacteriana da Expressão
Gênica/imunologia
Vetores Genéticos/imunologia
Proteínas de Membrana/imunologia
Camundongos Endogâmicos BALB C
Salmonella typhimurium/genética
Transformação Bacteriana
Vacinas Atenuadas/genética
Vacinas Atenuadas/imunologia
Vacinas Sintéticas/imunologia
Issue Date: 1999
Citation: SCHRIEFER, A. et al. Expression of a pilin subunit BfpA of the bundle-forming pilus of enteropathogenic Escherichia coli in an aroA live salmonella vaccine strain. Vaccine, v. 17, n. 7-8, p. 770-8, feb. 1999.
ISSN: 0264-410X
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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