dengue 317突变无影响.pdf

JOURNAL OF VIROLOGY, Dec. 2004, p. 12919–12928 Vol. 78, No. 23 0022-538X/04/$08.00 0 DOI: 10.1128/JVI.78.23.12919–12928.2004 Epitope Determinants of a Chimpanzee Fab Antibody That Efficiently Cross-Neutralizes Dengue Type 1 and Type 2 Viruses Map to Inside and in Close Proximity to Fusion Loop of the Dengue Type 2 Virus Envelope Glycoprotein 1 2 1 Ana P. Goncalvez, Robert H. Purcell, and Ching-Juh Lai * Molecular Viral Biology Section 1 and Hepatitis Viruses Section,2 Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland Received 18 May 2004/Accepted 19 July 2004 The epitope determinants of chimpanzee Fab antibody 1A5, which have been shown to be broadly reactive to flaviviruses and efficient for cross-neutralization of dengue virus type 1 and type 2 (DENV-1 and DENV-2), were studied by analysis of DENV-2 antigenic variants. Sequence analysis showed that one antigenic variant contained a Gly-to-Val substitution at position 106 within the flavivirus-conserved fusion peptide loop of the envelope protein (E), and another variant contained a His-to-Gln substitution at position 317 in E. Substitu- tion of Gly106Val in DENV-2 E reduced the binding affinity of Fab 1A5 by approximately 80-fold, whereas sub- stitution of His317Gln had little or no effect on antibody binding compared to the parental virus. Treatment of DENV-2 with -mercaptoethanol abolished binding of Fab 1A5, indicating that disulfide bridges were