Mapping of a dengue virus neutralizing epitope critical for the infectivity of all serotypes: insight into the neutralization mechanism

Laboratoire d’Ingénierie des Anticorps 1 , Unité de Biochimie Cellulaire 2 and Unité des Arbovirus et Virus des Fièvres Hémorragiques 3 , Institut Pasteur, Paris, France Département de Biologie des Agents Transmissibles, Centre de Recherche du Service de Santé des Armées, BP 87, 38702 La Tronche Cedex, France 4 Author for correspondence: Philippe Thullier (at Centre de Recherche du Service de Santé des Armées). Fax +33 4 76 63 69 17. e-mail pthullier{at}yahoo.com Dengue virus infections are a growing public health concern and strategies to control the spread of the virus are urgently needed. The murine monoclonal antibody 4E11 might be of interest, since it neutralizes dengue viruses of all serotypes by binding to the 296–400 segment of the major dengue virus envelope glycoprotein (DE). When phage-displayed peptide libraries were screened by affinity for 4E11, phage clone C1 was selected with a 50% frequency. C1 shared three of nine residues with DE 306–314 and showed significant reactivity to 4E11 in ELISA. C1-induced antibodies cross-reacted with DE 296–400 in mice, suggesting that it was a structural equivalent of the native epitope of 4E11 on DE. Accordingly, 4E11 bound to the DE 306–314 synthetic peptide and this reaction was inhibited by DE 296–400 . Moreover, DE 306–314 could block dengue virus infection of target cells in an in vitro assay. A three-dimensional model of DE revealed that the three amino acids shared by DE 296–400 and C1 were exposed to the solvent and suggested that most of the amino acids comprising the 4E11 epitope were located in the DE 306–314 region. Since 4E11 blocked the binding of DE 296–400 to heparin, which is a highly sulfated heparan sulfate (HSHS) molecule, 4E11 may act by neutralizing the interaction of DE 306–314 with target cell-displayed HSHS. Our data suggest that the DE 306–314 segment is critical for the infectivity of all dengue virus serotypes and that molecules that block the binding of DE 306–314 to HSHS may be antiviral reagents of therapeutic interest.