Restor Dent Endod.  2015 Nov;40(4):306-311. 10.5395/rde.2015.40.4.306.

Identification of Enterococcus faecalis antigens specifically expressed in vivo

• Lee SW
• Shet UK
• Park SW
• Lim HP
• Yun KD
• Kang SS
• Kim SE

• KMID: 2316951
• DOI: http://doi.org/10.5395/rde.2015.40.4.306

Abstract

OBJECTIVES
Molecular mechanism of the pathogenicity of Enterococcus faecalis (E. faecalis), a suspected endodontic pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here we report the identification of in vivo expressed antigens of E. faecalis by using a novel immunoscreening technique called change-mediated antigen technology (CMAT) and an experimental animal model of endodontic infection.
MATERIALS AND METHODS
Among 4,500 E. coli recombinant clones screened, 19 positive clones reacted reproducibly with hyperimmune sera obtained from rabbits immunized with E. faecalis cells isolated from an experimental endodontic infection. DNA sequences from 16 of these in vivo-induced (IVI) genes were determined.
RESULTS
Identified protein antigens of E. faecalis included enzymes involved in housekeeping functions, copper resistance protein, putative outer membrane proteins, and proteins of unknown function.
CONCLUSIONS
In vivo expressed antigens of E. faecalis could be identified by using a novel immune-screening technique CMAT and an experimental animal model of endodontic infection. Detailed analysis of these IVI genes will lead to a better understanding of the molecular mechanisms involved in the endodontic infection of E. faecalis.

Keyword

Antigen; Enterococcus faecalis; In vivo; Pulpitis; Virulence factor

MeSH Terms

Base Sequence
Clone Cells
Copper
Enterococcus faecalis*
Enterococcus*
Housekeeping
Membrane Proteins
Models, Animal
Pulpitis
Rabbits
Virulence
Virulence Factors
Copper
Membrane Proteins
Virulence Factors

Figure

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  Figure 1 Confirmation of bacterial antigens by SDS-PAGE. In lanes 4 - 6, different amount of bacterial cell pellets were loaded. SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis; BSA, bovine serum albumin.

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  Figure 2 Testing an efficacy of the adsorption procedure for hyperimmune rabbit antisera against E. faecalis whole cell pellets by ELISA analysis. Adsorbed serum exhibited a significant decrease of the reactivity. ELISA, enzyme-linked immunosorbent assay.

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