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Characterization of monoclonal antibodies against hepatitis C virus nonstructural protein 3: Different antigenic determinants from human B cells
Author(s) -
OuYang Pu,
Chiang BorLuen,
Hwang LihHwa,
Chen YiGuang,
Yang PeiMing,
Chi WeiKuang,
Chen PeiJer,
Chen DingShinn
Publication year - 1999
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/(sici)1096-9071(199904)57:4<345::aid-jmv3>3.0.co;2-n
Subject(s) - ns3 , epitope , antigen , virology , monoclonal antibody , biology , antibody , recombinant dna , epitope mapping , hepatitis c virus , b cell , microbiology and biotechnology , virus , immunology , biochemistry , gene
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 pro‐ tein bears the major antigenic determinant recognized by human sera. Half of the hybrid‐ omas 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. J. Med. Virol. 57:345–350, 1999. © 1999 Wiley‐Liss, Inc.