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Analysis of C-terminally truncated dengue 2 and dengue 3 virus envelope glycoproteins: processing in insect cells and immunogenic properties in mice

1 Department of Virology and 2 Department of Biotechnology, Institute Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France We constructed two recombinant Autographa californica nuclear polyhedrosis baculoviruses. Spodoptera frugiperda (Sf9) cells containing these constructs produce carboxy-termi...

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Bibliographic Details
Published in:Journal of general virology 1994-07, Vol.75 (7), p.1569-1578
Main Authors: Delenda, Christophe, Staropoli, Isabelle, Frenkiel, Marie-Pascale, Cabanie, Lucien, Deubel, Vincent
Format: Article
Language:English
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Summary:1 Department of Virology and 2 Department of Biotechnology, Institute Pasteur, 25 rue du Dr Roux, 75724 Paris Cedex 15, France We constructed two recombinant Autographa californica nuclear polyhedrosis baculoviruses. Spodoptera frugiperda (Sf9) cells containing these constructs produce carboxy-terminally truncated envelope E proteins representing dengue (DEN) virus serotypes 2 and 3. The two recombinant proteins contained their homologous signal sequences at the N terminus and were truncated by 71 and 74 amino acids at the C terminus, respectively. This allowed the translocation of the recombinant proteins to the endoplasmic reticulum followed by glycosylation processing and secretion into the extracellular medium. An additional unglycosylated form which was not secreted was detected inside the infected Sf9 cells. Sera from Swiss mice immunized with the infected Sf9 cell lysates gave a DEN cross-reactive response in ELISA and substantial amounts of neutralizing antibodies to the homologous virus. Similar antibody titres were obtained when the two recombinant proteins were inoculated concomitantly. BALB/c mice were vaccinated with three doses of the recombinant E proteins, taken as monovalent or bivalent immunogens, and challenged with mouse-adapted DEN-2 virus. DEN-2 E protein induced a good protection (90%) against lethal encephalitis and recombinant DEN-3 E protein gave a substantial cross-protection (54%). Eighty-two percent of the mice immunized with a mixture of both recombinant E proteins survived the DEN-2 virus challenge. Received 20 October 1993; accepted 2 February 1994.
ISSN:0022-1317
1465-2099
DOI:10.1099/0022-1317-75-7-1569