Expression and immunological evaluation of the Escherichia coli-derived hepatitis C virus envelope EI protein

被引:18
作者
Lorenzo, LJ [1 ]
García, O [1 ]
Acosta-Rivero, N [1 ]
Dueñas-Carrera, S [1 ]
Martínez, G [1 ]
Alvarez-Obregón, J [1 ]
Pichardo, D [1 ]
Ramos, A [1 ]
Guerra, I [1 ]
Morales, J [1 ]
机构
[1] Ctr Ingn Genet & Biotecnol, Div Vacunas, Dept Hepatitis C, Havana, Cuba
关键词
EI protein; expression in E. coli; HCV; immunogenicity; purification;
D O I
10.1042/BA20000040
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Immunological response against envelope protein El is very important in natural hepatitis C virus (HCV) infection, although it is insufficient to clear the viraemia, The HCV genomic region encoding the first 149 amino acids of the envelope EI protein (EI340, amino acids 192-340) was expressed in Escherichia coli (to a level of 30% of the whole cellular proteins) and purified to 85%, We measured the immune response in rabbits and mice as well as the reactivity against 37 human sera raised against the whole recombinant protein and EI-encoding peptides, From this, 51.1 % of human sera were found to react with EI340. High-level antibodies against EI340 were obtained in rabbits and mice when immunized. These antibodies had a similar peptide-recognition pattern to that described previously for human sera. The most reactive region was located at the N-terminus of the EI protein. Cellular immunity in mice was evaluated by delayed-type hypersensitivity assay. It revealed the induction of a CD4+ T-cell-mediated response by this protein. This EI340 protein and the animal-derived anti-EI sera are immunological tools that could aid in the monitoring and development of anti-HCV therapies.
引用
收藏
页码:137 / 143
页数:7
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