ABSTRACT
Objective To isolate and identify the putative Japanese encephalitis virus (JEV) re-ceptors from C6/36 and Vero cells. Methods Molecules binding with JEV were isolated from C6/36 and Vero cells by co-immunoprecipitation (Co-IP) approach, identified by mass spectrometry, and detected by Western blot. The location of putative JEV receptor on cells membrane and the binding with JEV were ob-served by laser scanning confocal microscopy (LCM). Results Several molecules binding with JEV were isolated from C6/36 and Vero cells by Co-IP, and only one molecule was identified as heat shock cognate 70 (HSC70) by mass spectrometry. Antibody against HSC'70 was able to detect a 74 ×103 protein isolated by Co-IP from C6/36 and Vero cells membrane in Western blot assays. It was observed by LSCM that when JEV attached on the surface of C6/36 cells, JEV and HSCT0 protein were co-localization. Conclusion 74 x 103 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.
ABSTRACT
Aim To study the mimicry peptide of JEV E protein by screening a phage 15-mer peptide library with anti-JEV E protein mAb 2H4. Methods After three rounds biopanning, the enriched positive phage clones were identified by ELISA. 10 positive phage clones were sequenced and compared homologously with JEV E protein. Results The short peptide displayed on screened positive phage could bind specifically to mAb 2H4, and the binding could be inhibited by natural JEV Ag. Amino acid sequences of the 10 positive phage clones were consensus, that is, RQDPQWPYANSTIAR. By homology analysis, two higher homologous sequences STXAR and WXXAXST were found in different regions of JEV E protein. The peptide displayed on positive phage could react specifically with the mouse antiserum against natural JEV Ag . Conclusion This peptide displayed on positive phage may mimic partial antigenicity of JEV E protein.