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PMID: 15542643 Published · ppublish English Journal Article

Chimpanzee Fab fragments and a derived humanized immunoglobulin G1 antibody that efficiently cross-neutralize dengue type 1 and type 2 viruses.

Journal of virology ·Vol. 78 ·No. 23 ·2004-12-00 ·Pages 12910-8

Goncalvez AP, Men R, Wernly C, Purcell RH, Lai CJ

Abstract

Passive immunization with monoclonal antibodies from humans or nonhuman primates represents an attractive alternative to vaccines for prevention of illness caused by dengue viruses (DENV) and other flaviviruses, including the West Nile virus. In a previous study, repertoire cloning to recover Fab fragments from bone marrow mRNA of chimpanzees infected with all four DENV serotypes (dengue virus serotype 1 [DENV-1] to DENV-4) was described. In that study, a humanized immunoglobulin G1 (IgG1) antibody that efficiently neutralized DENV-4 was recovered and characterized. In this study, the phage library constructed from the chimpanzees was used to recover Fab antibodies against the other three DENV serotypes. Serotype-specific neutralizing Fabs were not identified. Instead, we recovered DENV-neutralizing Fabs that specifically precipitated the envelope protein and were cross-reactive with all four DENV serotypes. Three of the Fabs competed with each other for binding to DENV-1 and DENV-2, although each of these Fabs contained a distinct complementarity determining region 3 (CDR3)-H sequence. Fabs that shared an identical or nearly identical CDR3-H sequences cross-neutralized DENV-1 and DENV-2 at a similar high 50% plaque reduction neutralization test (PRNT(50)) titer, ranging from 0.26 to 1.33 microg/ml, and neutralized DENV-3 and DENV-4 but at a titer 10- to 20-fold lower. One of these Fabs, 1A5, also neutralized the West Nile virus most efficiently among other flaviviruses tested. Fab 1A5 was converted to a full-length antibody in combination with human sequences for production in mammalian CHO cells. Humanized IgG1 1A5 proved to be as efficient as Fab 1A5 for cross-neutralization of DENV-1 and DENV-2 at a titer of 0.48 and 0.95 microg/ml, respectively. IgG1 1A5 also neutralized DENV-3, DENV-4, and the West Nile virus at a PRNT(50) titer of approximately 3.2 to 4.2 microg/ml. This humanized antibody represents an attractive candidate for further development of immunoprophylaxis against DENV and perhaps other flavivirus-associated diseases.

MeSH Terms
Amino Acid Sequence Animals Antibodies, Viral/immunology Cross Reactions Dengue Virus/classification,immunology Enzyme-Linked Immunosorbent Assay Epitopes Humans Immunoglobulin Fab Fragments/immunology Immunoglobulin G/immunology Immunoglobulin Heavy Chains/chemistry Immunoglobulin Light Chains/chemistry Immunoglobulin Variable Region/chemistry Molecular Sequence Data Pan troglodytes West Nile virus/immunology
Chemicals
Antibodies, Viral Epitopes Immunoglobulin Fab Fragments Immunoglobulin G Immunoglobulin Heavy Chains Immunoglobulin Light Chains Immunoglobulin Variable Region
Authors & Affiliations
5 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.
Men Ruhe
Wernly Claire
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.78.23.12910-12918.2004
Published
2004-12-00
Pages
12910-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC525007
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.78.23.12910-12918.2004 PMC Full Text View journal details
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