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Iran Biomed J. 2012;16(3):133-9. doi: 10.6091/ibj.1059.2012.

A DNA vaccine against Escherichia coli O157:H7.

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  • 1Dept. of Biology, Shahed University, Tehran, Iran.

Abstract

BACKGROUND:

Infection with Escherichia coli O157:H7 rarely leads to bloody diarrhea and causes hemolytic uremic syndrome with renal failure that can be deadly dangerous. Intimin, translocated Intimin receptor (Tir), and enterohemorrhagic E. coli (EHEC) secreted protein A (EspA) proteins are the virulence factors expressed by locus of enterocyte effacement locus of EHEC. This bacterium needs EspA as a conduit for Tir delivery into the host cell and the surface arrayed Intimin, which docks the bacterium to the translocated Tir.

METHODS:

Here we used triplet synthetic gene (eit) which was designed from three genes: espA coding EspA 120 lacking 36 amino acids from the N-terminal of the protein, eae coding Intimin constructed of 282 amino acids from the C-terminal and tir coding Tir 103, residues 258-361 which interacts with Intimin. The multimeric gene was cloned in two eukaryotic vectors pAAV-multiple cloning site-green fluorescent protein and pCI-neo. The pAAV was used for gene expression assay in cell line 293T and pCI-neo-EIT (EspA, Intimin, Tir) was used as DNA vaccine in mice. Test groups were injected intramuscularly with pCI-neo-EIT four times and mice control group was injected under the same conditions with PBS or pCI-neo vector.

RESULTS:

The titration of serums showed that BALB/c mice were successfully immunized with DNA vaccine compared to control groups and also they were protected against challenges of live oral using E. coli O157:H7.

CONCLUSION:

The results suggest that the DNA vaccine could induce protective immunity either alone or in combination with purified antigens to reduce EHEC infection.

PMID:
23023214
PMCID:
PMC3629931
[PubMed - indexed for MEDLINE]
Free PMC Article
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