Abstract
Gardnerella vaginalis produces vaginolysin (VLY), a cholesterol-dependent cytolysin, responsible of the cellular lysis and epithelial damage. In recently years, IgY antibodies produced in hens present several uses in diagnostic, treatment and research, however they present the disadvantage of unspecific recognition, which could be improved by the preselection of the epitope based on in silico analysis. Finally, multiantigenic peptides with eight ramification (MAP8) are groups of identical peptides synthesized on polylylisine core that increase immunogenicity, and allows its use to improve in vivo antibodies production and troubleshooting. In this work, we designed and administrated a MAP8 synthetic peptide in a hen to test its ability to induce the production of IgY antibodies. Further, we tested the reactivity of anti-VLY IgY antibody (derived of VLY-MAP8 peptide administration) in detecting the denaturing and native cognate antigen in the complete bacteria by ELISA, dot blot, western blot and inhibition of hemagglutination assay. Results showed the induction of IgY antibodies in the immunized hen, that were able to recognize the native and denatured cognate antigen in samples of culture supernatant and complete bacteria of Gardnerella vaginalis ATCC 14018 strain. Additionally, anti-VLY IgY antibody inhibit > 90% erythrocyte’s lysis. In conclusion, the MAP8 peptide and the produced anti-VLY IgY antibody could serve as a potential diagnostic tool in bacterial vaginosis. Also, our study proposed a method for the improvement of IgY antibodies production supported by the administration of MAP8 peptides selected by in silico analysis of the complete antigen.
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Acknowledgements
We are thankful to Graciella Castro-Escarpulli that kindly donated the G. vaginalis ATCC 14018 strain and Hugo Alberto Rodriguez Ruiz for its support in Fig. 1 design.
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This work was supported with 2018 PFCE founds granted to the academic body UAGRO CA-120.
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KCS, IBS, ACS, AVV conceived and designed research. KCS, IBS, ACS, AKEM, ARP conducted experiments. GFT, EFA, IPR contributed new reagents, analytical tools and analyzed the obtained data. KCS, AVV, IBS, ACS wrote the manuscript. All authors read and approved the manuscript.
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Procedures involving animal manipulation were performed in accordance with the bioethical regulations of the Universidad Autónoma de Guerrero and the NOM-062-ZOO-1999.
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Cortés-Sarabia, K., Bautista-Santos, I., Cisneros-Sarabia, A. et al. Gardnerella vaginalis Vaginolysin (VLY)-Derived MAP8 Peptide (VLY-MAP8) Induced the Production of Egg Yolk IgY Antibodies that Inhibit Erythrocytes Lysis. Int J Pept Res Ther 27, 413–420 (2021). https://doi.org/10.1007/s10989-020-10099-3
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DOI: https://doi.org/10.1007/s10989-020-10099-3