人巨细胞病毒临床株US3基因HLA-A*0201限制性CTL表位预测

目的分析晚期黄疸新生儿人巨细胞病毒(HCMV)感染患儿临床株US3基因序列,预测HCMV US3基因HLA-A*0201限制性CTL表位。方法应用巢式PCR法检测20例HCMV感染新生儿标本US3基因,结果进行双向DNA测序。SYFPEITHI和NetCTL方案预测HCMV临床株US3基因HLA-A*0201限制性CTL表位。结果①以Towne作为参考株,序列比对分析显示,20株临床分离株US3的ORF长度均与参考株相同,为561 bp,编码186个氨基酸蛋白。US3氨基酸序列高度保守,仅几个位点在少数分离株中存在变异,变异主要集中在序列的N端,大部分是同义突变。②通过预测获得5个HCMV US3基因HLA-A*0201限制性CTL表位:FTEKHFVSV、RMDYSSQTI、RMDYSSHTI、SIRDDNWGL和SMRDDNWGL。不同临床株US3基因HLA-A*0201限制性CTL表位存在差异。结论初步筛选出人巨细胞病毒临床株US3基因HLA-A*0201限制性CTL表位,为进一步研究HCMV感染的免疫机制提供实验依据。

Prediction of HLA-A*0201 Restricted CTL Epitopes of Protein Expressed by Human Cytomegalovirus Gene US3 Open Reading Frame

Objective To analyze the genetic variations of cytomegalovirus US3 gene in clinical strains,and to predict the HLA-A*0201 restricted CTL epitopes of the protein in late jaundice infants. Methods Nest PCR was performed to amplify the entire HCMV-US3 gene region of clinical strains collected from 20 HCMV infected infants.The PCR products were sequenced and the SYFPEITHI and NetCTL were used to predict the potential T cell CTL epitopes of US3 protein. Results By comparison with Towne sequence,the length of US3...