Study On Linear Epitopes Of HPV58 Virus E1 ^ E4 Protein B Cells

Human papillomavirus (HPV) plays a necessary, though insufficient, role in the development of cervical cancer, which is the third most common cancer in women worldwide, just following breast and colorectal cancers. HPV16contribute to most cervical cancers. While HPV58, as a "high-risk" type, has lower prevalence than HPV16and HPV18among cervical cancers across the world, there is a strong body of evidence to show that the contribution of HPV58varies geographically and HPV58attributes to a substantial proportion of cervical cancers in the eastern and southern parts of China, also in other parts of East Asia.E1ï¼¾E4, as one of an early nonstructural protein of HPV, has a relationship with the viral DNA replication and invasion, but its function is still not fully understood, with little research reports of HPV58E1ï¼¾E4protein. It is beneficial to the vaccine and antibody drug development, once we obtain its accurate epitope information.First, we analyzed E1ï¼¾E4protein through bioinformatics. E1ï¼¾E4is a hydrophilic protein, with several higher hydrophilic areas, and is not stable in solution. We predicted the B cell linear epitope of E1ï¼¾E4protein according to several prediction algorithms such as hydrophilicity, accessibility, flexibility, secondary structure, ABCpred and BepiPred, founding that these results have certain consistency. We analyzed the area that may contain B cell linear epitope, and had a roughly understanding to the E1ï¼¾E4protein.We expressed and purified the E1ï¼¾E4protein of HPV58, succeeded to obtain its polyclonal antibodies by inoculating rabbits. After that, we expressed13polypeptides covering HPV58E1ï¼¾E4protein by pXXGST-1, a short peptide expression vector, and two adjacent polypeptides overlap9amino acids. In the end, we proceeded western blot with the antiserum prepared. The results suggested that there are9B cell linear epitopes in HPV58E1ï¼¾E4protein, all located on the N terminal. Also, we succeeded to identify a dominant B cell linear epitope QSDDDSIYQTPETTP, which may contribute to more immune response.In this study, we focused on HPV58E1ï¼¾E4protein, identified its full B cells linear epitopes for the first time in the world. Our results will contribute to the development of HPV related diagnostic reagent, vaccines and antibody drug.