Measuring antibody neutralization of dengue virus (DENV) using a flow cytometry-based technique.
Dengue virus (DENV) is an emerging virus that threatens over two-third of the world's population. The specific diagnosis of dengue infection by serology is based on assays that detect DENV-specific antibodies including neutralizing antibodies (Abs). Neutralizing Abs are an important, if not the main, mechanism of protection from natural dengue virus (DENV) infection as well. The current gold-standard assay for measuring neutralizing Ab responses against DENV is the plaque reduction neutralization assay (PRNT). However, this assay is slow and laborious and utilizes physiologically irrelevant cell lines. Here, we describe a relatively high-throughput, flow cytometry-based neutralization assay for DENV that has been optimized for use with a human monocytic suspension cell line, U937 + DC-SIGN, or the more commonly used adherent monkey kidney cells, Vero-81.
Department of Microbiology and Immunology, and the Southeast Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
Ruklanthi
R
de Alwis
Aravinda M
AM
de Silva
D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism; D009500:Neutralization Tests / Q000379:methods*; D011956:Receptors, Cell Surface / Q000378:metabolism; D012692:Serial Passage; D012703:Serotyping; D013194:Staining and Labeling; D013223:Statistics as Topic; D020298:U937 Cells; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin