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李辉文,何韦韦,谭文婷,王莉,陈晓玲,邓国宏.HLA-DRB1*0803与HLA-DRB1*0901等位特异性HBcAg表位肽的筛查与鉴定[J].陆军军医大学学报(原第三军医大学学报),2015,37(09):833-838.
Li Huiwen,He Weiwei,Tan Wenting,Wang Li,Chen Xiaoling,Deng Guohong.Screening and identification of HLA-DRB1*0803 and HLA-DRB1*0901 restricted epitopes from hepatitis B core antigen[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(09):833-838.
李辉文,何韦韦,谭文婷,王莉,陈晓玲,邓国宏.HLA-DRB1*0803与HLA-DRB1*0901等位特异性HBcAg表位肽的筛查与鉴定[J].陆军军医大学学报(原第三军医大学学报),2015,37(09):833-838. DOI:
Li Huiwen,He Weiwei,Tan Wenting,Wang Li,Chen Xiaoling,Deng Guohong.Screening and identification of HLA-DRB1*0803 and HLA-DRB1*0901 restricted epitopes from hepatitis B core antigen[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(09):833-838. DOI:
目的 鉴定HLA-DRB1
*
0803与HLA-DRB1
*
0901等位特异性HBV表位,进一步认识HLA-DR不同等位限制性表位的差异。 方法 BLCL细胞表面高表达HLA-DR分子,从HBcAg肽池负载的特征HLA-DR等位的BLCL细胞表面上收集相关多肽,多肽经过过滤、脱盐、浓缩等处理步骤后对其进行液质联用质谱分析(LC-MS),再结合生物信息学预测工具NetMHCⅡpan 3.0 Server初步确定其潜在候选表位。 结果 筛选出HLA-DRB1
*
0803限制性表位HBc17-31、HBc22-36、HBc27-41、HBc82-96、HBc87-101、HBc92-106、HBc117-131、HBc122-136共8条,HLA-DRB1
*
0901限制性表位HBc2-16、HBc7-21、HBc12-26、HBc27-41、HBc82-96、HBc87-101、HBc122-136共7条,其中相同的有4条。 结论 发现了HBc7-21、HBc22-36、HBc27-41、HBc82-96、HBc87-101、HBc92-106新的HLA-Ⅱ类分子表位,不同的HLA-DR等位限制的HBcAg表位肽存在差异。
Objective To identify the specific epitopes of HLA-DRB1
*
0803 and HLA-DRB1
*
0901 alleles from hepatitis B virus for further understanding the difference of the epitopes restricted to the different HLA-DR alleles. Methods After incubating the peptide pool of pre-core and core sequence of hepatitis B virus with a B lymphoblastoid cell line (BLCL
homozygous human B cell lines of allele
HLA-DRB1
*
0803 or HLA-DRB1
*
0901 with cell surface highly expressing HLA-DR molecules)
the peptides were eluted and collected from the BLCL surface without affecting cell viability. Then the peptides were filtered
desalted
concentrated and analyzed by liquid chromatography-mass spectrometry (LC-MS). Bioinformatics predictive tool NetMHCⅡpan 3.0 Server was employed to confirm the candidate epitopes. Results There were 8 potential epitopes restricted to HLA-DRB1
*
0803 finally identify from HBV
that is
HBc17-31
HBc22-36
HBc27-41
HBc82-96
HBc87-101
HBc92-106
HBc117-131
and HBc122-136
and 7 potential epitopes restricted to HLA-DRB1
*
0901
that is
HBc2-16
HBc7-21
HBc12-26
HBc27-41
HBc82-96
HBc87-101
and HBc122-136. Four of them were same epitopes. Conclusion We have found 6 new epitopes to HLA class Ⅱ
ie
HBc7-21
HBc22-36
HBc27-41
HBc82-96
HBc87-101 and HBc92-106
and there are some differences among the HBcAg epitopes restricted to different HLA-DR alleles.
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