title: Development and evaluation of a FACS-based medium-throughput assay for HCV entry inhibitors.
abstract: The interaction between the hepatitis C virus (HCV) envelope glycoprotein E2 and the human tetraspanin protein CD81 is one of the key events involved in HCV cell entry. Therefore, compounds that interfere with this interaction may be useful tools for basic research and potential drugs for the treatment of HCV infection. The authors describe a medium-throughput assay for ligands of the E2 binding site on the CD81 receptor. In the assay, human hepatoma cells are incubated with the test compounds and stained with a fluorescently labeled anti-CD81 antibody (JS81). Flow cytometry is used to detect the level of bound antibody, reflecting the inhibitory potencies of the compounds. Eighty percent of compounds active in the assay show efficacy in an infection assay using luciferase reporter genome in cell culture. Thus, the assay can be used as a fast screening system for inhibitors of interaction of viral E2 to host cell CD81-LELs.
authors.affiliation: Pharmaceutical and Medicinal Chemistry, Saarland University, Saarbrücken, Germany.
authors.forename: Sigrid
authors.initials: S
authors.lastname: Ziegler
authors.forename: Bernd
authors.initials: B
authors.lastname: Kronenberger
authors.forename: Beatrice A-M
authors.initials: BA
authors.lastname: Albrecht
authors.forename: Artur
authors.initials: A
authors.lastname: Kaul
authors.forename: Anna-Lena
authors.initials: AL
authors.lastname: Gamer
authors.forename: Christian D
authors.initials: CD
authors.lastname: Klein
authors.forename: Rolf W
authors.initials: RW
authors.lastname: Hartmann
mesh_terms: D000906:Antibodies; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D016174:Hepacivirus / Q000187:drug effects*; D006801:Humans; D010799:Phycoerythrin / Q000378:metabolism; D013194:Staining and Labeling; D013696:Temperature; D013997:Time Factors; D053586:Virus Internalization / Q000187:drug effects*
chemical_list: D000906:Antibodies; D000998:Antiviral Agents; D010799:Phycoerythrin