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PMID: 15542644 Published · ppublish English Journal Article

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.

Journal of virology ·Vol. 78 ·No. 23 ·2004-12-00 ·Pages 12919-28

Goncalvez AP, Purcell RH, Lai CJ

Abstract

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. Substitution of Gly(106)Val in DENV-2 E reduced the binding affinity of Fab 1A5 by approximately 80-fold, whereas substitution of His(317)Gln had little or no effect on antibody binding compared to the parental virus. Treatment of DENV-2 with beta-mercaptoethanol abolished binding of Fab 1A5, indicating that disulfide bridges were required for the structural integrity of the Fab 1A5 epitope. Binding of Fab 1A5 to DENV-2 was competed by an oligopeptide containing the fusion peptide sequence as shown by competition enzyme-linked immunosorbent assay. Both DENV-2 antigenic variants were shown to be attenuated, or at least similar to the parental virus, when evaluated for growth in cultured cells or for neurovirulence in mice. Fab 1A5 inhibited low pH-induced membrane fusion of mosquito C6/36 cells infected with DENV-1 or DENV-2, as detected by reduced syncytium formation. Both substitutions in DENV-2 E lowered the pH threshold for membrane fusion, as measured in a fusion-from-within assay. In the three-dimensional structure of E, Gly(106) in domain II and His(317) in domain III of the opposite E monomer were spatially close. From the locations of these amino acids, Fab 1A5 appears to recognize a novel epitope that has not been mapped before with a flavivirus monoclonal antibody.

MeSH Terms
Amino Acid Sequence Animals Antibodies, Viral/immunology Dengue Virus/growth & development,immunology Epitope Mapping Immunoglobulin Fab Fragments/immunology Membrane Fusion Mice Molecular Sequence Data Neutralization Tests Pan troglodytes Viral Envelope Proteins/chemistry,immunology
Chemicals
Antibodies, Viral E-glycoprotein, Dengue virus type 2 Immunoglobulin Fab Fragments Viral Envelope Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Goncalvez Ana P
Molecular Viral Biology Section, Laboratory of Infectious Diseases, NIAID, NIH, Building 50, Room 6349, 50 South Dr., MSC 8009, Bethesda, MD 20892, USA.
Purcell Robert H
Lai Ching-Juh
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.78.23.12919-12928.2004
Published
2004-12-00
Pages
12919-28
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC525008
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.78.23.12919-12928.2004 PMC Full Text View journal details
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