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Screening and Identification of Efficient T and B Combined Epitopes of Vibrio vulnificus Hemolysin Based on Phage Display Technique
Bo Wang, Hui Ding, Dan-Li Xie, Yong-Liang Lou*, Mei-Ying Xiao, Bi-Fang Pang, Jie Yan1
Zhejiang Provincial Key Laboratory of Medical Genetics, School of Medical Lab Science, Wenzhou Medical College,Wenzhou 325035, China; 1Department of Medical Microbiology and Parasitology, College of Medicine,Zhejiang University, Hangzhou 310031
Abstract: The epitopes of T and B cells in Vibrio vulnificus hemolysin (VvhA) were predicted and analyzed by bioinformatics technique. We discovered four major T and B combined epitopes， named Vvha1， Vvha2， Vvha3 and Vvha4， respectively. The four recombinant epitopes were successfully expressed by phase display system， named M13KE-Vvha1， M13KE-Vvha2， M13KE-Vvah3， M13KE-Vvha4， and the antigenicities of M13KE-Vvah2 and M13KE-Vvha4 were better than that of M13KE-Vvha1 and M13KE-Vvha3 through Western blot. The immunogenicities of combined epitopes were further confirmed by flow cytometry and ELISA according to the changes of CD4+， CD8+ T cells and cytokines， such as IL-2， IL-4， IL-6， IFN-g. The subgroup of CD4+ T cells of ICR mice which were injected intraperitoneally with M13KE-Vvha1， M13KE-Vvha2 and M13KE-Vvha4 changed apparently， and the cytokines also had a significant change compared with cytokines of control groups. But when mice were intraperitoneally injected with M13KE-Vvha3， only IL-4 changed significantly. In conclusion， we had discovered and screened two efficient combined epitopes successfully， Vvha2 and Vvha4， and they could be as candidates of multipleantigenic peptide vaccine.
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