HPV16 E5多表位肽免疫原性研究

Immunogenicity of HPV16 E5 polyepitope peptides

目的筛选人乳头瘤病毒16型(HPV16)E5蛋白多表位肽段,并评估其诱导小鼠产生特异性体液免疫和细胞免疫的免疫原性反应。方法通过Uniprot获得HPV16 E5蛋白的氨基酸序列,采用EXPASY和SYFPEITHI等软件预测HPV16 E5蛋白的表位,筛选出富含B细胞和CTL表位的氨基酸肽段——HPV16 E5蛋白N端处aa28-46(LIRPLLLSVSTYTSLIILV),进一步用该多表位肽免疫C57BL/6小鼠,评估其诱导产生特异性体液免疫和细胞免疫的反应。结果该肽段可诱导小鼠产生较高水平的特异性IgG抗体,且抗体滴度随着免疫次数增加而升高,同时可诱导小鼠产生特异性的CTL反应,在效靶比为40∶1时,对靶细胞产生显著的特异性杀伤作用。结论HPV16 E5蛋白多表位肽aa28-46(LIRPLLLSVSTYTSLIILV)具有良好的免疫原性,可为基于HPV16 E5蛋白的疫苗研究提供基础。

This study was performed to predict and screen the multi epitope peptide of human papilloma virus type 16(HPV16)E5 protein,and analyze its specific humoral and cellular immunity in mice.The amino acid sequence of HPV16 E5 protein was obtained by Uniprot,then EXPASY and SYFPEITHI softwares were used to predict the epitopes of HPV16 E5 protein.The amino acid peptides rich in B cell and CTL epitopes were screened out:HPV16 E5 N-terminal aa28-46(LIRPLLLSVSTYTSLIILV).C57BL/6 mice were further immunized with this multi-epitope peptide to evaluate the specific humoral and cellular immune responses.Data showed that the peptide could induce high levels of specific IgG antibody against HPV16 E5,and the antibody titer increased with the increase of immunization times.In addition,the specific cytotoxic T lymphocyte(CTL)could be effectively induced from the splenic lymphocytes of the mice immunized with E5 multiple epitopes and show the highest cytotoxicity when the effector∶target ratio was 40∶1.In conclusion,HPV16 E5 protein multi-epitope peptide aa28-46(LIRPLLLSVSTYTSLIILV)has good immunogenicity,which provide a practical strategy for developing HPV vaccine based on HPV16 E5 protein.