不同基因型戊型肝炎病毒存在多种类型抗原表位  

Hepatitis E virus of different genotypes contains multiple-type antigenic epitopes

作　　者：邓蕾[1] 孟继鸿[1] 赵宇[1] 张红梅[1] 戴星[2] 

机构地区：[1]东南大学基础医学院病原生物学与免疫学系,南京210009 [2]东南大学医学院附属中大医院皮肤科,南京210009

出　　处：《微生物学报》2006年第1期120-126,共7页Acta Microbiologica Sinica

基　　金：国家自然科学基金(30271231;30271212);江苏省自然科学基金(BK2002053);江苏省卫生厅医学科技发展基金(H200115);教育部留学回国人员科研启动基金(2004176)~~

摘　　要：以戊型肝炎病毒(HEV)ORF2重组蛋白p166Us为免疫原制备单克隆抗体(McAbs),采用间接ELISA和免疫印迹法,检测McAbs与不同基因型和亚型HEV重组蛋白p166Bur(Ⅰa型)、p166Pak(Ⅰb型)、p166Mor(Ⅰc型)、p166Mex(Ⅱ型)、p166Us(Ⅲ型)、p166Nz(猪HEV,Ⅲ型)和p166Chn(Ⅳ型)的反应性,采用抗原或抗体竞争ELISA分析p166蛋白与天然HEV颗粒之间抗原表位的关系。结果获得4D3、2E3、11E11、12H5、3A3和1F16株稳定分泌McAbs的杂交瘤细胞株。4D3分泌的McAb与7种p166均发生反应,其与免疫原p166Us的结合可被Ⅰ、Ⅱ、Ⅲ或Ⅳ型天然HEV颗粒或病人血清竞争抑制。2E3、11E11和12H5分泌的McAbs只与p166Us、p166Nz和p166Chn发生反应,它们与p166Us的结合仅能被Ⅲ和IV型病毒或血清所抑制。3A3分泌的McAb只与p166Us及p166Nz结合,1F1分泌的McAb只与p166Us结合,两者均能被Ⅲ型美国株竞争抑制,而Ⅰ、Ⅱ、Ⅳ型不能抑制它们与p166Us的结合。由此可见,不同基因型和亚型HEV ORF2编码蛋白p166上存在多种类型抗原表位,其中包括Ⅰ、Ⅱ、Ⅲ、Ⅳ基因型共同的,Ⅲ、Ⅳ基因型共有的和第Ⅲ基因型特异的等,这些表位与天然HEV颗粒上的抗原表位具有相同的免疫学特征。Monoclonal antibodies （McAbs） were prepared against a recombinant protein p166Us derived from US-1 strain of hepatitis E virus （HEV）. The immune reactivity of the McAbs to seven p166s derived from different genotypes and subtypes of HEV, which included p166Bur （genotype Ⅰ a）, pl66Pak （genotype Ⅰ b）, p166Mor （genotype Ⅰ c）, p166Mex （genotype Ⅱ ）, p166Us （genotype Ⅲ）, p166Nz （swine HEV, genotype Ⅲ ） and p166Chn （genotype Ⅳ ）, was tested by an indirect enzyme-linked immunosorbent assay （ELISA） and a Western blotting assay. The immunological relationship between the McAbs and native HEV particles or anti-HEV positive serum samples was analyzed by an antigen-competitive or antibody-competitive ELISA. Totally, six McAb-producible hybridoma cell lines, designated by the name of 4D3,2E3, 11E11, 12H5,3A3 and 1F1 respectively, were cloned and obtained in this study. The McAb of 4D3 could react to all of the seven p166 recombinant proteins. This kind of reaction could be inhibited by each of 4 genotypes of native HEV particles or anti-HEV positive serum samples. The McAbs produced by 2E3,11E11 and 12H5 reacted to p166Us, p166Nz and p166Chn, but did not react to p166Bur, p166Pak, p166Mor and p166Mex. The reaction of McAb of 2E3, as an example of the McAbs of 2E3, liEn and 12H5, could be only inhibited by genotype Ⅲ and Ⅳ HEV or anti-HEV positive serum. The McAb of 3A3 could bind to p166Us as well as p166Nz. The McAb produced by 1F1 was reactive to the p166Us only. However, neither of Ⅰ , Ⅱ , Ⅳ genotype HEV particles or antisera could inhibit both of their reactions to p166Us. The data as mentioned above suggested that there are multiple-type antigenic epitopes such as genotype Ⅰ , Ⅱ, Ⅲ and iV common, Ⅲ and Ⅳ common, and Ⅲ specific proteins of different genotypes and subtypes of HEV. Moreover, the antigenic these on native HEV particles possess identical immunological characteristics epitopes within HEV ORF2 encoded p166 epitopes on recombinant protei

关 键 词：戊型肝炎病毒 抗原表位 单克隆抗体 重组蛋白 基因型 

分 类 号：R392.1[医药卫生—免疫学]