In silico analysis as a strategy to identify candidate epitopes with human IgG reactivity to study Porphyromonas gingivalis virulence factors
2019
Ellen Karla Nobre dos Santos-Lima | Kizzes Araújo Paiva Andrade Cardoso | Patrícia Mares de Miranda | Ana Carla Montino Pimentel | Paulo Cirino de Carvalho-Filho | Yuri Andrade de Oliveira | Lília Ferreira de Moura-Costa | Teresa Olczak | Isaac Suzart Gomes-Filho | Roberto José Meyer | Márcia Tosta Xavier | Soraya Castro Trindade
Abstract Porphyromonas gingivalis (Pg) is one of the main pathogens in chronic periodontitis (CP). Studies on the immunogenicity of its virulence factors may contribute to understanding the host response to infection. The present study aimed to use in silico analysis as a tool to identify epitopes from Lys-gingipain (Kgp) and neuraminidase virulence factors of the Pg ATCC 33277 strain. Protein sequences were obtained from the NCBI Protein Database and they were scanned for amino acid patterns indicative of MHC II binding using the MHC-II Binding Predictions tool from the Immune Epitope Database (IEDB). Peptides from different regions of the proteins were chemically synthesized and tested by the indirect ELISA method to verify IgG immunoreactivity in serum of subjects with CP and without periodontitis (WP). T cell epitope prediction resulted in 16 peptide sequences from Kgp and 18 peptide sequences from neuraminidase. All tested Kgp peptides exhibited IgG immunoreactivity whereas tested neuraminidase peptides presented low IgG immunoreactivity. Thus, the IgG reactivity to Kgp protein could be reaffirmed and the low IgG reactivity to Pg neuraminidase could be suggested. The novel peptide epitopes from Pg were useful to evaluate its immunoreactivity based on the IgG-mediated host response. In silico analysis was useful for preselecting epitopes for immune response studies in CP.
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