Generation of recombinant bacillus Calmette–Guérin and Mycobacterium smegmatis expressing BfpA and intimin as vaccine vectors against enteropathogenic Escherichia coli
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Data
2012
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Resumo
Enteropathogenic Escherichia coli (EPEC) is an important cause of diarrhea in children. EPEC adheres to the
intestinal epithelium and causes attaching and effacing (A/E) lesions. Recombinant Mycobacterium smegmatis
(Smeg) and Mycobacterium bovis BCG strains were constructed to express either BfpA or intimin.
The entire bfpA gene and a portion of the intimin gene were amplified by PCR from EPEC genomic DNA
and inserted into the pMIP12 vector at the BamHI/KpnI sites. The pMIP bfpA and pMIP intimin vectors
were introduced separately into Smeg and BCG. Recombinant clones were selected based on kanamycin
resistance and designated rSmeg pMIP (bfpA or intimin) and rBCG pMIP (bfpA or intimin). The expression
of bfpA and intimin was detected by Immunoblotting using polyclonal anti-BfpA and anti-intimin
antibodies. The immunogenicity of these proteins was assessed in C57BL/6 mice by assaying the feces
and serum for the presence of anti-BfpA and anti-intimin IgA and IgG antibodies. TNF- and INF- were
produced in vitro by spleen cells from mice immunized with recombinant BfpA, whereas TNF- was produced
in mice immunized with recombinant intimin. The adhesion of EPEC (E2348/69) to HEp-2 target
cells was blocked by IgA or IgG antibodies from mice immunized with recombinant BfpA or intimin but
not by antibodies from non-immunized mice. Immunogenic non-infectious vectors containing relevant
EPEC virulence genes may be promising vaccine candidates.
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Diarrhea, EPEC, BCG, M. smegmatis, Recombinant vaccine, bfpA and intimin
Citação
VASCONCELLOS, Halyka Luzorio Franzotti et al. Generation of recombinant bacillus Calmette Guérin and Mycobacterium smegmatis expressing BfpA and intimin as vaccine vectors against enteropathogenic Escherichia coli. Vaccine, Amsterdam, v. 30, n. 41, p. 5999-6005, 2012.