Ou-Yang P, Chiang B L, Hwang L H, Chen Y G, Yang P M, Chi W K, Chen P J, Chen D S
Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei.
J Med Virol. 1999 Apr;57(4):345-50. doi: 10.1002/(sici)1096-9071(199904)57:4<345::aid-jmv3>3.0.co;2-n.
The nonstructural (NS3) region protein of hepatitis C virus (HCV) possesses major B-cell epitopes that induce antibodies after infection. To elucidate further the characteristics of these B cells and their role in the immune regulation of HCV infection, T9 (portion of NS3 region, amino acids [a.a.] 1188-1493)-specific monoclonal antibodies were derived and mapped for B-cell antigenic determinants with recombinant proteins. A total of 10 T9-specific hybridomas were generated and tested for B-cell antigenic determinants. To analyze the B-cell antigenic determinants, eight recombinant proteins including NS3-e (a.a. 1175-1334), NS3-a' (a.a. 1175-1250), NS3-a (a.a. 1251-1334), NS3-b (a.a. 1323-1412), NS3-c (a.a. 1407-1499), NS3-a/b (a.a. 1251-1412), NS3-bc (a.a. 1323-1499), and NS3-abc (a.a. 1251-1499) encoded by NS3-region internal clones were expressed and tested for immunoblotting. The data suggested IgG hybridomas recognized NS3-a, NS3-a', or NS3-b protein by immunoblotting. By contrast, the NS3-e protein bears the major antigenic determinant recognized by human sera. Half of the hybridomas were found to react with protein NS3-a', which is not a major B-cell antigenic determinant in humans. These data suggested that conformational epitopes in vivo may be important for B-cell recognition.
丙型肝炎病毒（HCV）的非结构（NS3）区蛋白具有主要的B细胞表位，感染后可诱导抗体产生。为了进一步阐明这些B细胞的特征及其在HCV感染免疫调节中的作用，我们制备了T9（NS3区部分，氨基酸[a.a.]1188 - 1493）特异性单克隆抗体，并用重组蛋白对B细胞抗原决定簇进行定位。共产生了10个T9特异性杂交瘤，并对其进行B细胞抗原决定簇检测。为了分析B细胞抗原决定簇，表达了包括NS3 - e（a.a. 1175 - 1334）、NS3 - a'（a.a. 1175 - 1250）、NS3 - a（a.a. 1251 - 1334）、NS3 - b（a.a. 1323 - 1412）、NS3 - c（a.a. 1407 - 1499）、NS3 - a/b（a.a. 1251 - 1412）、NS3 - bc（a.a. 1323 - 1499）和NS3 - abc（a.a. 1251 - 1499）在内的8种由NS3区内部克隆编码的重组蛋白，并进行免疫印迹检测。数据表明，IgG杂交瘤通过免疫印迹识别NS3 - a、NS3 - a'或NS3 - b蛋白。相比之下，NS3 - e蛋白带有被人血清识别的主要抗原决定簇。发现一半的杂交瘤与蛋白NS3 - a'反应，而NS3 - a'并非人类主要的B细胞抗原决定簇。这些数据表明，体内的构象表位可能对B细胞识别很重要。
