title: Evaluation and analysis of dengue virus enhancing and neutralizing activities using simple high-throughput assays.
abstract: The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose-response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.
authors.affiliation: Laboratory of Emerging Infectious Diseases and Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
authors.forename: Xiao-Quan
authors.initials: XQ
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Yan-Fen
authors.initials: YF
authors.lastname: Huang
authors.forename: Xi-Xia
authors.initials: XX
authors.lastname: Ding
authors.forename: Li-Dong
authors.initials: LD
authors.lastname: Liu
authors.forename: Li-Wen
authors.initials: LW
authors.lastname: Qiu
authors.forename: Yu-Xian
authors.initials: YX
authors.lastname: Pan
authors.forename: Yong-Qiang
authors.initials: YQ
authors.lastname: Deng
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Hu
authors.forename: Biao
authors.initials: B
authors.lastname: Di
authors.forename: Cheng-Feng
authors.initials: CF
authors.lastname: Qin
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Che
mesh_terms: D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D019067:Antibody-Dependent Enhancement; D003715:Dengue / Q000175:diagnosis / Q000276:immunology* / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000276:immunology / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D017361:Viral Nonstructural Proteins / Q000276:immunology
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D017361:Viral Nonstructural Proteins