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利用生物信息学预测技术成功筛选出含部分创伤弧菌溶细胞素(Vibrio vulnificushemolysin,VvhA)基因片段的T细胞、B细胞联合表位Vvha1、Vvha2、Vvha3和Vvha4,通过噬菌体展示技术成功获得四段表位多肽M13KE-Vvha1、M13KE-Vvha2、M13KE-Vvah3、M13KE-Vvha4,且M13KE-Vvah2、M13KE-Vvha4抗原反应性强于M13KE-Vvha1、M13KE-Vvha3。将四种抗原性多肽经腹腔注射ICR小鼠后,利用流式细胞仪及ELISA方法检测小鼠CD4~+T淋巴细胞亚群及IL-2、IL-4、IL-6、IFN-γ细胞因子时发现,与M13KE-Vvha3相比,经M13KE-Vvha1、M13KE-Vvha2、M13KE-Vvha4作用的小鼠CD4~+T淋巴细胞亚群的变化更为显著。与正常组相比,该三组小鼠的IL-2、IL-4、IL-6、IFN-γ细胞因子分泌量的变化都具有统计学意义;而经M13KE-Vvha3作用的小鼠仅IL-4的改变具有明显统计学意义。因此本研究成功筛选出含部分VvhA片段的T细胞、B细胞联合表位Vvha2、Vvha4,从而为进一步研制多抗原肽(multipleantigenic peptide,MAP)疫苗打下基础。
A total of four segments of T cells, B cells combined epitopes Vvha1, Vvha2, Vvha3 and Vvha4 with partial Vibrio vulnificushemolysin (VvhA) gene fragment were successfully screened by bioinformatics prediction technique. The epitope peptides M13KE-Vvha1, M13KE-Vvha2, M13KE-Vvah3 and M13KE-Vvha4, and the M13KE-Vvah2 and M13KE-Vvha4 antigens are more reactive than M13KE-Vvha1 and M13KE-Vvha3. Four kinds of antigenic peptides were injected intraperitoneally into ICR mice. The CD4 ~ + T lymphocyte subsets and IL-2, IL-4, IL-6 and IFN-γ cells were detected by flow cytometry and ELISA The results showed that the changes of CD4 ~ + T lymphocyte subsets in M13KE-Vvha1, M13KE-Vvha2 and M13KE-Vvha4 mice were more obvious than M13KE-Vvha3. Compared with the normal group, the secretion of IL-2, IL-4, IL-6 and IFN-γ in the three groups of mice had statistical significance; while the mice treated with M13KE-Vvha3 had only IL -4 changes with significant statistical significance. Therefore, we successfully screened T cells and B cell epitopes Vvha2 and Vvha4 containing partial VvhA fragments, which laid the foundation for the further development of multipleantigenic peptide (MAP) vaccine.