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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

2-D gel electrophoresis profiles of whole cell lysates of biofilm-grownS. equispp.zooepidemicuswith the immunoreactive proteins indicated.A Protein staining with Coomassie B-250. B Western blot analysis of the immunoreactive proteins using pig convalescent serum. Immunoreactive spots are indicated by the abbreviation of biofilm (BF) followed by an arbitrary number.
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Fig1: 2-D gel electrophoresis profiles of whole cell lysates of biofilm-grownS. equispp.zooepidemicuswith the immunoreactive proteins indicated.A Protein staining with Coomassie B-250. B Western blot analysis of the immunoreactive proteins using pig convalescent serum. Immunoreactive spots are indicated by the abbreviation of biofilm (BF) followed by an arbitrary number.

Mentions: A 2-DE covering a pH range of 4–7 (IPG linear gradient) was performed to separate whole cell proteins prepared from biofilm-grown S. equi spp. zooepidemicus. The spots were detected by Coomassie Brilliant Blue G-250 staining (Figure 1A). Western blotting with pig convalescent sera revealed the presence of seventeen immunoreactive proteins in the biofilm-grown bacterial samples (Figure 1B), which was consistent with our observations of the duplicated 2-D gel (Figure 1A). The seventeen spots were excised and were characterized by MALDI-TOF–MS and MALDI-TOF-TOF–MS, and the data were compared to those in the NCBI sequence database. The probability score for the match, MW, pI, and number of peptide matches were used to identify the spots. The seventeen immunoreactive spots, listed in Table 1, corresponded to seventeen different proteins. Nine of these immunoreactive proteins were also identified in planktonic-grown bacteria using the same protocol [4]: a DNA polymerase III delta subunit, elongation factor G, pyruvate kinase, transketolase, the GroEL molecular chaperone, glyceraldehyde-3-phosphate dehydrogenase, pyridine nucleotide-disulfide oxidoreductase, oligopeptide ABC transporter periplasmic oligopeptide-binding protein OppA, and elongation factor Ts. Since the GroEL molecular chaperone is essential for biofilm formation in other bacteria [18], it was selected for further analysis.Figure 1


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)

2-D gel electrophoresis profiles of whole cell lysates of biofilm-grownS. equispp.zooepidemicuswith the immunoreactive proteins indicated.A Protein staining with Coomassie B-250. B Western blot analysis of the immunoreactive proteins using pig convalescent serum. Immunoreactive spots are indicated by the abbreviation of biofilm (BF) followed by an arbitrary number.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig1: 2-D gel electrophoresis profiles of whole cell lysates of biofilm-grownS. equispp.zooepidemicuswith the immunoreactive proteins indicated.A Protein staining with Coomassie B-250. B Western blot analysis of the immunoreactive proteins using pig convalescent serum. Immunoreactive spots are indicated by the abbreviation of biofilm (BF) followed by an arbitrary number.
Mentions: A 2-DE covering a pH range of 4–7 (IPG linear gradient) was performed to separate whole cell proteins prepared from biofilm-grown S. equi spp. zooepidemicus. The spots were detected by Coomassie Brilliant Blue G-250 staining (Figure 1A). Western blotting with pig convalescent sera revealed the presence of seventeen immunoreactive proteins in the biofilm-grown bacterial samples (Figure 1B), which was consistent with our observations of the duplicated 2-D gel (Figure 1A). The seventeen spots were excised and were characterized by MALDI-TOF–MS and MALDI-TOF-TOF–MS, and the data were compared to those in the NCBI sequence database. The probability score for the match, MW, pI, and number of peptide matches were used to identify the spots. The seventeen immunoreactive spots, listed in Table 1, corresponded to seventeen different proteins. Nine of these immunoreactive proteins were also identified in planktonic-grown bacteria using the same protocol [4]: a DNA polymerase III delta subunit, elongation factor G, pyruvate kinase, transketolase, the GroEL molecular chaperone, glyceraldehyde-3-phosphate dehydrogenase, pyridine nucleotide-disulfide oxidoreductase, oligopeptide ABC transporter periplasmic oligopeptide-binding protein OppA, and elongation factor Ts. Since the GroEL molecular chaperone is essential for biofilm formation in other bacteria [18], it was selected for further analysis.Figure 1

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