title: High-throughput assay to identify inhibitors of Vpu-mediated down-regulation of cell surface BST-2.
abstract: Bone marrow stromal cell antigen 2 (BST-2, also known as Tetherin) inhibits HIV-1 release and thereby severely impairs viral replication. HIV-1 accessory protein Vpu induces the down-regulation of cell surface BST-2, and counteracts the antiviral function of BST-2. Blocking Vpu-mediated down-regulation of cell surface BST-2 is viewed as a new opportunity for developing anti-HIV drugs. In this study, we have developed a high-throughput cell-based ELISA to identify small molecules that antagonize HIV-1 Vpu function and consequently inhibit HIV-1 replication through rescuing the antiviral activity of host BST-2. This cell-ELISA shows an excellent correlation with results obtained by flow cytometry (FACS). Under optimal conditions, a Z' factor of 0.605 was achieved in a 96-well format. Together, these results demonstrate that this assay can be used to quantify the cell surface level of BST-2 and be adapted to a high-throughput screening for novel anti-HIV compounds.
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Science, Beijing, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhang
authors.forename: Zhenlong
authors.initials: Z
authors.lastname: Liu
authors.forename: Zeyun
authors.initials: Z
authors.lastname: Mi
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Li
authors.forename: Pingping
authors.initials: P
authors.lastname: Jia
authors.forename: Jinming
authors.initials: J
authors.lastname: Zhou
authors.forename: Xiao
authors.initials: X
authors.lastname: Yin
authors.forename: Xuefu
authors.initials: X
authors.lastname: You
authors.forename: Liyan
authors.initials: L
authors.lastname: Yu
authors.forename: Fei
authors.initials: F
authors.lastname: Guo
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.forename: Chen
authors.initials: C
authors.lastname: Liang
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D015703:Antigens, CD / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D058851:GPI-Linked Proteins / Q000819:agonists / Q000378:metabolism; D020868:Gene Silencing / Q000187:drug effects; D015658:HIV Infections / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology / Q000821:virology; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D057166:High-Throughput Screening Assays; D054298:Human Immunodeficiency Virus Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010957:Plasmids; D011485:Protein Binding / Q000187:drug effects*; D034741:RNA, Small Interfering / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D053585:Virus Attachment / Q000187:drug effects; D014779:Virus Replication / Q000187:drug effects; D000074010:Bone Marrow Stromal Antigen 2; D000086082:Viroporin Proteins
chemical_list: D019380:Anti-HIV Agents; D015703:Antigens, CD; D058851:GPI-Linked Proteins; D054298:Human Immunodeficiency Virus Proteins; D034741:RNA, Small Interfering; D054334:Viral Regulatory and Accessory Proteins; C094572:BST2 protein, human; D000074010:Bone Marrow Stromal Antigen 2; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins