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目的预测和初步鉴定鼠Ⅲ型肝炎病毒(MHV-3)的CTL表位,为基于慢性肝炎病毒抗原表位的免疫治疗奠定理论基础。方法利用超基序、量化基序及人工神经网络方案相结合预测MHV-3 S蛋白的H-2K~K限制性CTL表位;用分子模拟对候选表位肽做进一步筛选;人工合成相关待测表位肽,利用T2-K~K细胞株测定各肽与H-2K~K分子的结合力。结果结合超基序、量化基序、人工神经网络预测结果及分子模拟结果,预测出了4条候选表位肽。MHC亲和力实验表明,在候选的4条表位肽中,IEPYNGVI(141-148)、YELSGYTV(306-313)与H-2K~K呈中等亲和力结合,荧光系数分别为1.25及1.04。结论表位预测的结果与亲和力分析结果一致性较好,两者联合应用初步认为IEPYNGVI(141-148)及YELSGYTV(306-313)为MHV-3 S蛋白H-2K~K限制性CTL表位的可能性最大,为下一步体内鉴定及基于小鼠肝炎病毒抗原表位的免疫治疗奠定基础。
Objective To predict and preliminarily identify the CTL epitope of murine hepatitis virus type 3 (MHV-3), and lay the theoretical foundation for the immunotherapy based on the antigenic epitopes of chronic hepatitis B virus. Methods The H-2K-K restricted CTL epitopes of MHV-3 S protein were predicted by combining the super-motifs, the quantitative motifs and the artificial neural network. The candidate epitope peptides were further screened by molecular simulation. Epitope peptides were measured and the binding affinity of each peptide to H-2K ~ K molecules was measured using T2-K ~ K cell lines. Results Four candidate epitope peptides were predicted based on the prediction of super-motif, quantified motif, artificial neural network and molecular simulation. MHC affinity experiments showed that IEPYNGVI (141-148) and YELSGYTV (306-313) had moderate affinities to H-2K ~ K among the four epitope peptides with fluorescence coefficients of 1.25 and 1.04, respectively. Conclusions The results of epitope prediction are in good agreement with the results of affinity analysis. The combination of the two suggested that IEPYNGVI (141-148) and YELSGYTV (306-313) were the H-2K ~ K restricted CTL epitopes of MHV-3 S protein The most likely to lay the foundation for the next in vivo identification and immunotherapy based on mouse hepatitis antigen epitopes.