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Identification and characterization of a Streptococcus equi ssp. zooepidemicus immunogenic GroEL protein involved in biofilm formation.

Yi L, Wang Y, Ma Z, Lin HX, Xu B, Grenier D, Fan HJ, Lu CP - Vet. Res. (2016)

Bottom Line: Biofilm formation by this bacterium has been previously reported.In this study, we used an immunoproteomic approach to search for immunogenic proteins expressed by biofilm-grown S. equi spp. zooepidemicus.Seventeen immunoreactive proteins were found, of which nine common immunoreactive proteins were identified in planktonic and biofilm-grown bacteria.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Streptococcus equi ssp. zooepidemicus (S. equi spp. zooepidemicus) is an opportunistic pathogen that causes major economic losses in the swine industry in China and is also a threat for human health. Biofilm formation by this bacterium has been previously reported. In this study, we used an immunoproteomic approach to search for immunogenic proteins expressed by biofilm-grown S. equi spp. zooepidemicus. Seventeen immunoreactive proteins were found, of which nine common immunoreactive proteins were identified in planktonic and biofilm-grown bacteria. The immunogenicity and protective efficacy of the S. equi spp. zooepidemicus immunoreactive GroEL chaperone protein was further investigated in mice. The protein was expressed in vivo and elicited high antibody titers following S. equi spp. zooepidemicus infections of mice. An animal challenge experiment with S. equi spp. zooepidemicus showed that 75% of mice immunized with the GroEL protein were protected. Using in vitro biofilm inhibition assays, evidence was obtained that the chaperonin GroEL may represent a promising target for the prevention and treatment of persistent S. equi spp. zooepidemicus biofilm infections. In summary, our results suggest that the recombinant GroEL protein, which is involved in biofilm formation, may efficiently stimulate an immune response, which protects against S. equi spp. zooepidemicus infections. It may therefore be a candidate of interest to be included in vaccines against S. equi spp. zooepidemicus infections.

No MeSH data available.


Related in: MedlinePlus

SDS-PAGE and Western immunoblotting analysis of the rGroEL protein expressed inE. coliBL21. Lane M, protein molecular mass marker; lane 1, Western blot analysis of puried rGroEL protein using pre-immune mini-pig sera as the negative control; lane 2, Western blot analysis of puried rGroEL protein using mini-pig convalescent sera; lane 3, the elution of the purified rGroEL were separated on an SDS-PAGE and stained with coomassie; lane 4, rGroEL protein in E. coli BL21induced with 1 mM IPTG for 4 h; lane 5, rGroEL protein without IPTG; lane 6, empty expression vector pET28a (+).
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Fig2: SDS-PAGE and Western immunoblotting analysis of the rGroEL protein expressed inE. coliBL21. Lane M, protein molecular mass marker; lane 1, Western blot analysis of puried rGroEL protein using pre-immune mini-pig sera as the negative control; lane 2, Western blot analysis of puried rGroEL protein using mini-pig convalescent sera; lane 3, the elution of the purified rGroEL were separated on an SDS-PAGE and stained with coomassie; lane 4, rGroEL protein in E. coli BL21induced with 1 mM IPTG for 4 h; lane 5, rGroEL protein without IPTG; lane 6, empty expression vector pET28a (+).

Mentions: The GroEL protein of S. equi spp. zooepidemicus was cloned in E. coli. The immunogenicity of the rGroEL protein was confirmed by Western blotting using mini-pig convalescent sera (Figure 2).Figure 2


Identification and characterization of a Streptococcus equi ssp. zooepidemicus immunogenic GroEL protein involved in biofilm formation.

Yi L, Wang Y, Ma Z, Lin HX, Xu B, Grenier D, Fan HJ, Lu CP - Vet. Res. (2016)

SDS-PAGE and Western immunoblotting analysis of the rGroEL protein expressed inE. coliBL21. Lane M, protein molecular mass marker; lane 1, Western blot analysis of puried rGroEL protein using pre-immune mini-pig sera as the negative control; lane 2, Western blot analysis of puried rGroEL protein using mini-pig convalescent sera; lane 3, the elution of the purified rGroEL were separated on an SDS-PAGE and stained with coomassie; lane 4, rGroEL protein in E. coli BL21induced with 1 mM IPTG for 4 h; lane 5, rGroEL protein without IPTG; lane 6, empty expression vector pET28a (+).
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4834820&req=5

Fig2: SDS-PAGE and Western immunoblotting analysis of the rGroEL protein expressed inE. coliBL21. Lane M, protein molecular mass marker; lane 1, Western blot analysis of puried rGroEL protein using pre-immune mini-pig sera as the negative control; lane 2, Western blot analysis of puried rGroEL protein using mini-pig convalescent sera; lane 3, the elution of the purified rGroEL were separated on an SDS-PAGE and stained with coomassie; lane 4, rGroEL protein in E. coli BL21induced with 1 mM IPTG for 4 h; lane 5, rGroEL protein without IPTG; lane 6, empty expression vector pET28a (+).
Mentions: The GroEL protein of S. equi spp. zooepidemicus was cloned in E. coli. The immunogenicity of the rGroEL protein was confirmed by Western blotting using mini-pig convalescent sera (Figure 2).Figure 2

Bottom Line: Biofilm formation by this bacterium has been previously reported.In this study, we used an immunoproteomic approach to search for immunogenic proteins expressed by biofilm-grown S. equi spp. zooepidemicus.Seventeen immunoreactive proteins were found, of which nine common immunoreactive proteins were identified in planktonic and biofilm-grown bacteria.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Streptococcus equi ssp. zooepidemicus (S. equi spp. zooepidemicus) is an opportunistic pathogen that causes major economic losses in the swine industry in China and is also a threat for human health. Biofilm formation by this bacterium has been previously reported. In this study, we used an immunoproteomic approach to search for immunogenic proteins expressed by biofilm-grown S. equi spp. zooepidemicus. Seventeen immunoreactive proteins were found, of which nine common immunoreactive proteins were identified in planktonic and biofilm-grown bacteria. The immunogenicity and protective efficacy of the S. equi spp. zooepidemicus immunoreactive GroEL chaperone protein was further investigated in mice. The protein was expressed in vivo and elicited high antibody titers following S. equi spp. zooepidemicus infections of mice. An animal challenge experiment with S. equi spp. zooepidemicus showed that 75% of mice immunized with the GroEL protein were protected. Using in vitro biofilm inhibition assays, evidence was obtained that the chaperonin GroEL may represent a promising target for the prevention and treatment of persistent S. equi spp. zooepidemicus biofilm infections. In summary, our results suggest that the recombinant GroEL protein, which is involved in biofilm formation, may efficiently stimulate an immune response, which protects against S. equi spp. zooepidemicus infections. It may therefore be a candidate of interest to be included in vaccines against S. equi spp. zooepidemicus infections.

No MeSH data available.


Related in: MedlinePlus