| Summary: | Objective To screen, identify and verify the immunoreactive epitopes on Hantaan virus glycoprotein C-terminal (HTNV Gc). Methods On the basis of predicting the major histocompatibility complex (MHC) gene in more than 98% of the global population, the affinity of 9-peptide and 15-peptide epitopes on HTNV Gc was systematically analyzed by bioinformatics cutting-edge algorithms such as Immune Epitope Database (IEDB) and Technical University of Denmark (DTU). Their immunogenicity was analyzed by using the Vaxijen tool, and the interspecies intraspecific conservation was analyzed with the Blastp tool. Bidirectional hierarchical clustering was employed to compare the cross-immunoreactivity of MHC-class Ⅰ and class Ⅱ restricted epitopes. Possible results of dominant epitopes docking with MHC were simulated by using molecular docking. Enzyme-linked immunospot (ELISpot) assay was performed to validate the results of computer analyses. Results After integrating the results of various bioinformatics algorithms, we obtained 8 9-peptide epitopes and 80 15-peptide epitopes with high affinity, intraspecific conservation and strong immunogenicity, including 8 human-mouse common dominant epitopes. Cluster analysis showed that H-2 genotype had certain similarity as HLA genotype. The results of molecular docking revealed that dominant epitopes had good docking with various MHC genotypes, which were consistent with the results of affinity analysis. The amount of IFN-γ secreted from the spleen cells was higher in the mice with dominant epitopes than those with non-dominant epitopes. Conclusion With the aids of various bioinformatics algorithms, we successfully screen out the dominant epitopes on HTNV Gc, and validate the cellular immunoreactivity of these epitopes through ELISpot assay.
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