title: Development of a FACS-based assay for evaluating antiviral potency of compound in dengue infected peripheral blood mononuclear cells.
abstract: Dengue fever is the most important arthropod-transmitted viral disease affecting humans. It is a blood-borne disease characterized by persistent fever and joint pain. In the blood, primary peripheral blood mononuclear cells (PBMCs), in particular monocytes, are the main target of the dengue virus (DENV). These cells are poorly permissive for in vitro dengue virus infection and their infectivity varies from donor to donor. To overcome this barrier, an anti-dengue antibody was used to improve the infectivity of DENV-2 clinical isolates to PBMCs, the monocytic leukemia cell line, THP-1 and the granulocyte cell line, KU812. A higher throughput 96-well-format assay based on a fluorescent-activated cell sorter could potentially be developed to evaluate the antiviral potency of compounds in DENV-infected PBMCs in vitro. The results correlate well with data obtained by a standard plaque assay. Altogether, an assay has been developed that enables evaluation of the antiviral activity of test compounds in a physiologically-relevant cell system (PBMCs). These screening processes are urgently needed for dengue drug discovery.
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore.
authors.forename: Yilong
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore. Electronic address: yen_liang.chen@novartis.com.
authors.forename: Yen-Liang
authors.initials: YL
authors.lastname: Chen
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore.
authors.forename: Maxime
authors.initials: M
authors.lastname: Herve
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore.
authors.forename: Feng
authors.initials: F
authors.lastname: Gu
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore.
authors.forename: Pei-Yong
authors.initials: PY
authors.lastname: Shi
authors.affiliation: Novartis Institute for Tropical Diseases, Singapore, Singapore.
authors.forename: Francesca
authors.initials: F
authors.lastname: Blasco
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification* / Q000494:pharmacology; D002460:Cell Line; D003715:Dengue / Q000821:virology*; D003716:Dengue Virus / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006098:Granulocytes / Q000821:virology*; D006801:Humans; D009000:Monocytes / Q000821:virology*
chemical_list: D000998:Antiviral Agents
keywords: Dengue; FACS; Screening