1887

Abstract

To explore an epitope-based vaccine against , we screened the epitopes in the N2N3 subdomain of fibronectin-binding protein A (FnBPA) as a surface component of .

We expressed N2N3 proteins and prepared monoclonal antibodies (mAbs) against N2N3 by the hybridoma technique, before screening the B-cell epitopes in N2N3 using a phage-displayed random 12-mer peptide library with these mAbs against N2N3. Finally, we analysed the characters of the screened epitopes using immunofluorescence and an infection assay.

In this paper, we identified a linear B-cell epitope in N2N3 through screening a phage-displayed peptide library with a 3C3 mAb against the N2N3. The 3C3 mAb recognized the IETFNKANNRFSH sequence of the N2N3 subdomain. Subsequently, site-directed mutagenic analysis demonstrated that residues F162, K164, N167, R168 and F169 formed the core of IETFNKANNRFSH, and this core motif was the minimal determinant of the B-cell epitope recognized by the 3C3 mAb. The epitope IETFNKANNRFSH showed high homology among different strains. Moreover, this epitope was exposed on the surface of the by using an enzyme-linked immunosorbent assay (ELISA) assay and an indirect immunofluorescence assay. As expected, the epitope peptide evoked a protective immune response against infection in immunized mice.

We identified a novel linear B-cell epitope, IETFNKANNRFSH, in the N2N3 subdomain of fibronectin-binding protein A that is recognized by 3C3 mAb, which will contribute to the further study of an epitope-based vaccine candidate against .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000633
2018-03-01
2024-10-05
Loading full text...

Full text loading...

/deliver/fulltext/jmm/67/3/423.html?itemId=/content/journal/jmm/10.1099/jmm.0.000633&mimeType=html&fmt=ahah

References

  1. Deleo FR, Otto M, Kreiswirth BN, Chambers HF. Community-associated meticillin-resistant Staphylococcus aureus. Lancet 2010; 375:1557–1568 [View Article][PubMed]
    [Google Scholar]
  2. Durai R, Ng PC, Hoque H. Methicillin-resistant Staphylococcus aureus: an update. Aorn J 2010; 91:599–609 [View Article][PubMed]
    [Google Scholar]
  3. Krishna S, Miller LS. Innate and adaptive immune responses against Staphylococcus aureus skin infections. Semin Immunopathol 2012; 34:261–280 [View Article][PubMed]
    [Google Scholar]
  4. Otto M. MRSA virulence and spread. Cell Microbiol 2012; 14:1513–1521 [View Article][PubMed]
    [Google Scholar]
  5. Ma X, Wu Y, Li L, Xu Q, Hu B et al. First multicenter study on multidrug resistant bacteria carriage in Chinese ICUs. BMC Infect Dis 2015; 15:358 [View Article][PubMed]
    [Google Scholar]
  6. Mehraj J, Akmatov MK, Strömpl J, Gatzemeier A, Layer F et al. Methicillin-sensitive and methicillin-resistant Staphylococcus aureus nasal carriage in a random sample of non-hospitalized adult population in northern Germany. PLoS One 2014; 9:e107937 [View Article][PubMed]
    [Google Scholar]
  7. Scali F, Camussone C, Calvinho LF, Cipolla M, Zecconi A. Which are important targets in development of S. aureus mastitis vaccine?. Res Vet Sci 2015; 100:88–99 [View Article][PubMed]
    [Google Scholar]
  8. Lee JW, O'Brien CN, Guidry AJ, Paape MJ, Shafer-Weaver KA et al. Effect of a trivalent vaccine against Staphylococcus aureus mastitis lymphocyte subpopulations, antibody production, and neutrophil phagocytosis. Can J Vet Res 2005; 69:11–18[PubMed]
    [Google Scholar]
  9. Leitner G, Yadlin N, Lubashevsy E, Ezra E, Glickman A et al. Development of a Staphylococcus aureus vaccine against mastitis in dairy cows. II. Field trial. Vet Immunol Immunopathol 2003; 93:153–158 [View Article][PubMed]
    [Google Scholar]
  10. Malito E, Faleri A, Lo Surdo P, Veggi D, Maruggi G et al. Defining a protective epitope on factor H binding protein, a key meningococcal virulence factor and vaccine antigen. Proc Natl Acad Sci USA 2013; 110:3304–3309 [View Article][PubMed]
    [Google Scholar]
  11. Correia BE, Bates JT, Loomis RJ, Baneyx G, Carrico C et al. Proof of principle for epitope-focused vaccine design. Nature 2014; 507:201–206 [View Article][PubMed]
    [Google Scholar]
  12. Gershoni JM, Roitburd-Berman A, Siman-Tov DD, Tarnovitski Freund N, Weiss Y. Epitope mapping: the first step in developing epitope-based vaccines. BioDrugs 2007; 21:145–156 [View Article][PubMed]
    [Google Scholar]
  13. Reason DC, Ullal A, Liberato J, Sun J, Keitel W et al. Domain specificity of the human antibody response to Bacillus anthracis protective antigen. Vaccine 2008; 26:4041–4047 [View Article][PubMed]
    [Google Scholar]
  14. Reason D, Liberato J, Sun J, Keitel W, Zhou J. Frequency and domain specificity of toxin-neutralizing paratopes in the human antibody response to anthrax vaccine adsorbed. Infect Immun 2009; 77:2030–2035 [View Article][PubMed]
    [Google Scholar]
  15. Moyle PM, Toth I. Modern subunit vaccines: development, components, and research opportunities. ChemMedChem 2013; 8:360–376 [View Article][PubMed]
    [Google Scholar]
  16. de Groot AS, Mcmurry J, Marcon L, Franco J, Rivera D et al. Developing an epitope-driven tuberculosis (TB) vaccine. Vaccine 2005; 23:2121–2131 [View Article][PubMed]
    [Google Scholar]
  17. Zhu D, Williams JN, Rice J, Stevenson FK, Heckels JE et al. A DNA fusion vaccine induces bactericidal antibodies to a peptide epitope from the PorA porin of Neisseria meningitidis. Infect Immun 2008; 76:334–338 [View Article][PubMed]
    [Google Scholar]
  18. Broughan J, Anderson R, Anderson AS. Strategies for and advances in the development of Staphylococcus aureus prophylactic vaccines. Expert Rev Vaccines 2011; 10:695–708 [View Article][PubMed]
    [Google Scholar]
  19. Verkaik NJ, van Wamel WJ, van Belkum A. Immunotherapeutic approaches against Staphylococcus aureus. Immunotherapy 2011; 3:1063–1073 [View Article][PubMed]
    [Google Scholar]
  20. Que YA, Haefliger JA, Piroth L, François P, Widmer E et al. Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis. J Exp Med 2005; 201:1627–1635 [View Article][PubMed]
    [Google Scholar]
  21. Deivanayagam CC, Wann ER, Chen W, Carson M, Rajashankar KR et al. A novel variant of the immunoglobulin fold in surface adhesins of Staphylococcus aureus: crystal structure of the fibrinogen-binding MSCRAMM, clumping factor A. Embo J 2002; 21:6660–6672 [View Article][PubMed]
    [Google Scholar]
  22. Keane FM, Loughman A, Valtulina V, Brennan M, Speziale P et al. Fibrinogen and elastin bind to the same region within the A domain of fibronectin binding protein A, an MSCRAMM of Staphylococcus aureus. Mol Microbiol 2007; 63:711–723 [View Article][PubMed]
    [Google Scholar]
  23. Roche FM, Downer R, Keane F, Speziale P, Park PW et al. The N-terminal A domain of fibronectin-binding proteins A and B promotes adhesion of Staphylococcus aureus to elastin. J Biol Chem 2004; 279:38433–38440 [View Article][PubMed]
    [Google Scholar]
  24. O'Neill E, Pozzi C, Houston P, Humphreys H, Robinson DA et al. A novel Staphylococcus aureus biofilm phenotype mediated by the fibronectin-binding proteins, FnBPA and FnBPB. J Bacteriol 2008; 190:3835–3850 [View Article][PubMed]
    [Google Scholar]
  25. Geoghegan JA, Monk IR, O'Gara JP, Foster TJ. Subdomains N2N3 of fibronectin binding protein A mediate Staphylococcus aureus biofilm formation and adherence to fibrinogen using distinct mechanisms. J Bacteriol 2013; 195:2675–2683 [View Article][PubMed]
    [Google Scholar]
  26. Huang X, Xu J, Wang Y, Guo C, Chen L et al. GP50 as a promising early diagnostic antigen for Taenia multiceps infection in goats by indirect ELISA. Parasit Vectors 2016; 9:618 [View Article][PubMed]
    [Google Scholar]
  27. Li Y, Ning YS, Wang YD, Luo J, Wang W et al. Production of mouse monoclonal antibodies against Helicobacter pylori catalase and mapping the antigenic epitope by phage display library. Vaccine 2008; 26:1263–1269 [View Article][PubMed]
    [Google Scholar]
  28. Wu X, Li X, Zhang Q, Wulin S, Bai X et al. Identification of a conserved B-cell epitope on duck hepatitis A type 1 virus VP1 protein. PLoS One 2015; 10:e0118041 [View Article][PubMed]
    [Google Scholar]
  29. Yu L, Fan Z, Ma J, Tong C, Song B et al. Cross-protective effect of a novel multi-antigen-chimeric vaccine against Streptococcus and Staphylococcus aureus infection in mice. J Med Microbiol 2014; 63:1732–1740 [View Article][PubMed]
    [Google Scholar]
  30. Su Y, Wang S, Shao J, Zhang B, Wei H. Effect of Kozak sequence on mice DNA vaccine immunization of Staphylococcus aureus adhesion fibronectin-binding protein FnBPA-A. Sheng Wu Gong Cheng Xue Bao 2013; 29:458–465[PubMed]
    [Google Scholar]
  31. Zuo QF, Cai CZ, Ding HL, Wu Y, Yang LY et al. Identification of the immunodominant regions of Staphylococcus aureus fibronectin-binding protein A. PLoS One 2014; 9:e95338 [View Article][PubMed]
    [Google Scholar]
  32. Aghebati-Maleki L, Bakhshinejad B, Baradaran B, Motallebnezhad M, Aghebati-Maleki A et al. Phage display as a promising approach for vaccine development. J Biomed Sci 2016; 23:66 [View Article][PubMed]
    [Google Scholar]
  33. Kronqvist N, Malm M, Rockberg J, Hjelm B, Uhlén M et al. Staphylococcal surface display in combinatorial protein engineering and epitope mapping of antibodies. Recent Pat Biotechnol 2010; 4:171–182 [View Article][PubMed]
    [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.000633
Loading
/content/journal/jmm/10.1099/jmm.0.000633
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error