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Immunisation With Immunodominant Linear B Cell Epitopes Vaccine of Manganese Transport Protein C Confers Protection against Staphylococcus aureus Infection.

Yang HJ, Zhang JY, Wei C, Yang LY, Zuo QF, Zhuang Y, Feng YJ, Srinivas S, Zeng H, Zou QM - PLoS ONE (2016)

Bottom Line: On the basis of the results, linear B cell epitopes within MntC were finely mapped using a series of overlapping synthetic peptides.Further studies indicate that MntC113-136, MntC209-232, and MntC263-286 might be the original linear B-cell immune dominant epitope of MntC, furthermore, three-dimensional (3-d) crystal structure results indicate that the three immunodominant epitopes were displayed on the surface of the MntC antigen.On the basis of immunodominant MntC113-136, MntC209-232, and MntC263-286 peptides, the epitope vaccine for S. aureus induces a high antibody level which is biased to TH2 and provides effective immune protection and strong opsonophagocytic killing activity in vitro against MRSA infection.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Research Centre for Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University, Chongqing 400038, PR China.

ABSTRACT
Vaccination strategies for Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA) infections have attracted much research attention. Recent efforts have been made to select manganese transport protein C, or manganese binding surface lipoprotein C (MntC), which is a metal ion associated with pathogen nutrition uptake, as potential candidates for an S. aureus vaccine. Although protective humoral immune responses to MntC are well-characterised, much less is known about detailed MntC-specific B cell epitope mapping and particularly epitope vaccines, which are less-time consuming and more convenient. In this study, we generated a recombinant protein rMntC which induced strong antibody response when used for immunisation with CFA/IFA adjuvant. On the basis of the results, linear B cell epitopes within MntC were finely mapped using a series of overlapping synthetic peptides. Further studies indicate that MntC113-136, MntC209-232, and MntC263-286 might be the original linear B-cell immune dominant epitope of MntC, furthermore, three-dimensional (3-d) crystal structure results indicate that the three immunodominant epitopes were displayed on the surface of the MntC antigen. On the basis of immunodominant MntC113-136, MntC209-232, and MntC263-286 peptides, the epitope vaccine for S. aureus induces a high antibody level which is biased to TH2 and provides effective immune protection and strong opsonophagocytic killing activity in vitro against MRSA infection. In summary, the study provides strong proof of the optimisation of MRSA B cell epitope vaccine designs and their use, which was based on the MntC antigen in the development of an MRSA vaccine.

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Related in: MedlinePlus

ELISA detection of B cell epitope mapping of MntC with antiserum of immunised and infected BALB/c mice.To determine the immunodominant peptides of MntC, microtitre plates were coated with synthetic overlapping peptides that spanned the entire length of the MntC of MRSA252 or GST141-158 (negative control peptides) or MntC protein and BSA. B cell epitope mapping of MntC using an overlapping 18-mer peptide ELISA with antiserum samples from BALB/c mice that were immunised with rMntC plus CFA adjuvant (A) and antiserum samples from BALB/c mice that were infected after immunisation with rMntC plus CFA adjuvant (B) were detected. The absorbances read at 450 nm for peptides MntC113-136, MntC209-232, and MntC263-286 were significantly higher than BSA (P < 0.05) and higher than GST141-158 (P < 0.01).
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pone.0149638.g002: ELISA detection of B cell epitope mapping of MntC with antiserum of immunised and infected BALB/c mice.To determine the immunodominant peptides of MntC, microtitre plates were coated with synthetic overlapping peptides that spanned the entire length of the MntC of MRSA252 or GST141-158 (negative control peptides) or MntC protein and BSA. B cell epitope mapping of MntC using an overlapping 18-mer peptide ELISA with antiserum samples from BALB/c mice that were immunised with rMntC plus CFA adjuvant (A) and antiserum samples from BALB/c mice that were infected after immunisation with rMntC plus CFA adjuvant (B) were detected. The absorbances read at 450 nm for peptides MntC113-136, MntC209-232, and MntC263-286 were significantly higher than BSA (P < 0.05) and higher than GST141-158 (P < 0.01).

Mentions: Crossing the entire length of the MntC of MRSA252, we synthesised forty six 18-mer overlapping peptides to identify the linear B-cell epitope mapping of MntC by ELISA, and the antiserum samples form rMntC-immunised mice and MRSA-infected mice post rMntC vaccinations were tested respectively. As shown in Fig 2, the strongest IgG antibody reactivity results of immunised mice antiserum(Fig 2A) were consistent with infected mice antiserum with immunised before infection (Fig 2B), which concentrated on the three major immunodominant peptides: MntC113-136 (KKVIAVSKDVKPIYLNGEEGNKDK), MntC209-232 (FKYFSKQYGITPGYIWEINTEKQG), and MntC263-286 (MESLSEETKKDIFGEVYTDSIGKE) of MntC antigen and indicated a homologous immunodominant antibody response. Therefore, peptides of MntC113-136, MntC209-232, and MntC263-286 may contain novel linear B-cell immunodominant epitopes that induced major immunodominant huromal responses in MntC vaccination. This is the first study to map the B-cell epitopes of MntC to date, which nearly contained all of the linear B-cell epitopes of MntC from MRSA252. It may be helpful for understanding the immune protective efficacy of MntC antigen against S. aureus infection.


Immunisation With Immunodominant Linear B Cell Epitopes Vaccine of Manganese Transport Protein C Confers Protection against Staphylococcus aureus Infection.

Yang HJ, Zhang JY, Wei C, Yang LY, Zuo QF, Zhuang Y, Feng YJ, Srinivas S, Zeng H, Zou QM - PLoS ONE (2016)

ELISA detection of B cell epitope mapping of MntC with antiserum of immunised and infected BALB/c mice.To determine the immunodominant peptides of MntC, microtitre plates were coated with synthetic overlapping peptides that spanned the entire length of the MntC of MRSA252 or GST141-158 (negative control peptides) or MntC protein and BSA. B cell epitope mapping of MntC using an overlapping 18-mer peptide ELISA with antiserum samples from BALB/c mice that were immunised with rMntC plus CFA adjuvant (A) and antiserum samples from BALB/c mice that were infected after immunisation with rMntC plus CFA adjuvant (B) were detected. The absorbances read at 450 nm for peptides MntC113-136, MntC209-232, and MntC263-286 were significantly higher than BSA (P < 0.05) and higher than GST141-158 (P < 0.01).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4764517&req=5

pone.0149638.g002: ELISA detection of B cell epitope mapping of MntC with antiserum of immunised and infected BALB/c mice.To determine the immunodominant peptides of MntC, microtitre plates were coated with synthetic overlapping peptides that spanned the entire length of the MntC of MRSA252 or GST141-158 (negative control peptides) or MntC protein and BSA. B cell epitope mapping of MntC using an overlapping 18-mer peptide ELISA with antiserum samples from BALB/c mice that were immunised with rMntC plus CFA adjuvant (A) and antiserum samples from BALB/c mice that were infected after immunisation with rMntC plus CFA adjuvant (B) were detected. The absorbances read at 450 nm for peptides MntC113-136, MntC209-232, and MntC263-286 were significantly higher than BSA (P < 0.05) and higher than GST141-158 (P < 0.01).
Mentions: Crossing the entire length of the MntC of MRSA252, we synthesised forty six 18-mer overlapping peptides to identify the linear B-cell epitope mapping of MntC by ELISA, and the antiserum samples form rMntC-immunised mice and MRSA-infected mice post rMntC vaccinations were tested respectively. As shown in Fig 2, the strongest IgG antibody reactivity results of immunised mice antiserum(Fig 2A) were consistent with infected mice antiserum with immunised before infection (Fig 2B), which concentrated on the three major immunodominant peptides: MntC113-136 (KKVIAVSKDVKPIYLNGEEGNKDK), MntC209-232 (FKYFSKQYGITPGYIWEINTEKQG), and MntC263-286 (MESLSEETKKDIFGEVYTDSIGKE) of MntC antigen and indicated a homologous immunodominant antibody response. Therefore, peptides of MntC113-136, MntC209-232, and MntC263-286 may contain novel linear B-cell immunodominant epitopes that induced major immunodominant huromal responses in MntC vaccination. This is the first study to map the B-cell epitopes of MntC to date, which nearly contained all of the linear B-cell epitopes of MntC from MRSA252. It may be helpful for understanding the immune protective efficacy of MntC antigen against S. aureus infection.

Bottom Line: On the basis of the results, linear B cell epitopes within MntC were finely mapped using a series of overlapping synthetic peptides.Further studies indicate that MntC113-136, MntC209-232, and MntC263-286 might be the original linear B-cell immune dominant epitope of MntC, furthermore, three-dimensional (3-d) crystal structure results indicate that the three immunodominant epitopes were displayed on the surface of the MntC antigen.On the basis of immunodominant MntC113-136, MntC209-232, and MntC263-286 peptides, the epitope vaccine for S. aureus induces a high antibody level which is biased to TH2 and provides effective immune protection and strong opsonophagocytic killing activity in vitro against MRSA infection.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Research Centre for Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University, Chongqing 400038, PR China.

ABSTRACT
Vaccination strategies for Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA) infections have attracted much research attention. Recent efforts have been made to select manganese transport protein C, or manganese binding surface lipoprotein C (MntC), which is a metal ion associated with pathogen nutrition uptake, as potential candidates for an S. aureus vaccine. Although protective humoral immune responses to MntC are well-characterised, much less is known about detailed MntC-specific B cell epitope mapping and particularly epitope vaccines, which are less-time consuming and more convenient. In this study, we generated a recombinant protein rMntC which induced strong antibody response when used for immunisation with CFA/IFA adjuvant. On the basis of the results, linear B cell epitopes within MntC were finely mapped using a series of overlapping synthetic peptides. Further studies indicate that MntC113-136, MntC209-232, and MntC263-286 might be the original linear B-cell immune dominant epitope of MntC, furthermore, three-dimensional (3-d) crystal structure results indicate that the three immunodominant epitopes were displayed on the surface of the MntC antigen. On the basis of immunodominant MntC113-136, MntC209-232, and MntC263-286 peptides, the epitope vaccine for S. aureus induces a high antibody level which is biased to TH2 and provides effective immune protection and strong opsonophagocytic killing activity in vitro against MRSA infection. In summary, the study provides strong proof of the optimisation of MRSA B cell epitope vaccine designs and their use, which was based on the MntC antigen in the development of an MRSA vaccine.

Show MeSH
Related in: MedlinePlus