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Biotechnology and Applied Biochemistry (2000) 32, (137–143) (Printed in Great Britain)

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Expression and immunological evaluation of the Escherichia coli-derived hepatitis C virus envelope E1 protein

Lázaro J. Lorenzo¹, Odalys García, Nelson Acosta-Rivero, Santiago Dueñas-Carrera, Gillian Martínez, Julio Álvarez-Obregón, Dagmara Pichardo, Astrid Ramos, Ivis Guerra and Juan Morales

Departamento de Hepatitis C, División de Vacunas, Centro de Ingeniería Genética y Biotecnología, P.O. Box 6162, La Habana, Cuba

Abbreviations used: aa, amino acids; HCV, Hepatitis C virus; DTH, delayed-type hypersensitivity; rHBsAg, recombinant hepatitis B surface antigen.

¹ To whom correspondence should be addressed (e-mail juan.morales@cigb.edu.cu).

Immunological response against envelope protein E1 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 E1 protein (E1₃₄₀, 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 E1-encoding peptides. From this, 51.1% of human sera were found to react with E1₃₄₀. High-level antibodies against E1₃₄₀ 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 E1 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 E1₃₄₀ protein and the animal-derived anti-E1 sera are immunological tools that could aid in the monitoring and development of anti-HCV therapies.

Received 9 May 2000/12 June 2000; accepted 20 June 2000

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