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Differential reactivity of mouse monoclonal anti-HBs antibodies with recombinant mutant HBs antigens

World Journal of Gastroenterology 2006年第33期12卷 5368-5374页

作者： Azam Roohi Yaghoub Yazdani jalal khoshnoodi Seyed Mohammad Jazayeri William F Carman Mahmood Chamankhah Manley Rashedan Fazel Shokri Department of Immunology School of Public Health Tehran University of Medical Sciences Tehran Iran Department of Virology School of Public Health Tehran University of Medical Sciences Tehran Iran Division of Virology Institute of Biomedical and Life Science University of Glasgow Glasgow United Kingdom Monoclonal Antibody Research Center Avesina Research Center Tehran Iran

AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutions mostly located within the “a” ***: The entire HBs genes previously cloned into a mammalian expression vector were transiently transfected into COS7 cells. Two standard unmutated sequences of the ayw and adw subtypes served as controls. Secreted mutant proteins were collected and measured by three commercial diagnostic immunoassays to assess transfection efficiency. Reactivity of anti-HBs mAbs with mutated HBsAgs was determined by sandwich enzyme-linked immunosorbent assay (ELISA).RESULTS: Reactivity of anti-HBs mAbs with mutated HBsAgs revealed different patterns. While three mutants reacted strongly with all mAbs, two mutants reacted weakly with only two mAbs and the remaining proteins displayed variable degrees of reactivity towards different mAbs. Accordingly, four groups of mAbs with different but overlapping reactivity patterns could be envisaged. One group consisting of two mAbs (37C5-S7 and 35C6-S11) was found to recognize stable linear epitopes conserved in all mutants. Mutations outside the “a” determinant at positions 120 (P→S), 123(T→N) and 161(M→T) were found to affect reactivity of these ***: Our findings could have important implications for biophysical studies, vaccination strategies and immunotherapy of hepatitis B virus (HBV) mutants.

关键词： Hepatitis B surface antigen Hepatitis B virus Mutant Epitope mapping Vaccination Monoclonal antibody

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