{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T17:51:13Z","timestamp":1790272273485,"version":"4.1.0"},"reference-count":61,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2010,12,1]],"date-time":"2010-12-01T00:00:00Z","timestamp":1291161600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2010,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    HIV replication assays are important tools for HIV drug discovery efforts. Here, we present a full HIV replication system (EASY-HIT) for the identification and analysis of HIV inhibitors. This technology is based on adherently growing HIV-susceptible cells, with a stable fluorescent reporter gene activated by HIV Tat and Rev. A fluorescence-based assay was designed that measures HIV infection by two parameters relating to the early and the late phases of HIV replication, respectively. Validation of the assay with a panel of nine reference inhibitors yielded effective inhibitory concentrations consistent with published data and allowed discrimination between inhibitors of early and late phases of HIV replication. Finer resolution of the effects of reference drugs on different steps of HIV replication was achieved in secondary time-of-addition assays. The EASY-HIT assay yielded high\n                    <jats:italic>Z<\/jats:italic>\n                    \u2032 scores (&gt;0.9) and signal stabilities, confirming its robustness. Screening of the LOPAC\n                    <jats:sup>1280<\/jats:sup>\n                    library identified 10 compounds (0.8%), of which eight were known to inhibit HIV, validating the suitability of this assay for screening applications. Studies evaluating anti-HIV activities of natural products with the EASY-HIT technology led to the identification of three novel inhibitory compounds that apparently act at different steps of HIV-1 replication. Furthermore, we demonstrate successful evaluation of plant extracts for HIV-inhibitory activities, suggesting application of this technology for the surveillance of biological extracts with anti-HIV activities. We conclude that the EASY-HIT technology is a versatile tool for the discovery and characterization of HIV inhibitors.\n                  <\/jats:p>","DOI":"10.1128\/aac.00515-10","type":"journal-article","created":{"date-parts":[[2010,9,28]],"date-time":"2010-09-28T00:44:26Z","timestamp":1285634666000},"page":"5257-5268","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":36,"title":["EASY-HIT: HIV Full-Replication Technology for Broad Discovery of Multiple Classes of HIV Inhibitors"],"prefix":"10.1128","volume":"54","author":[{"given":"Stephan","family":"Kremb","sequence":"first","affiliation":[{"name":"Institute of Virology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 1, D-85764 Neuherberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus","family":"Helfer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 1, D-85764 Neuherberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Werner","family":"Heller","sequence":"additional","affiliation":[{"name":"Institute of Biochemical Plant Pathology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 1, D-85764 Neuherberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dieter","family":"Hoffmann","sequence":"additional","affiliation":[{"name":"Institute of Virology, Technische Universita\u0308t Mu\u0308nchen, Trogerstrasse 30, 81675 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Horst","family":"Wolff","sequence":"additional","affiliation":[{"name":"Institute of Virology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 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Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jo\u0308rg","family":"Durner","sequence":"additional","affiliation":[{"name":"Institute of Biochemical Plant Pathology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 1, D-85764 Neuherberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruth","family":"Brack-Werner","sequence":"additional","affiliation":[{"name":"Institute of Virology, Helmholtz Zentrum Mu\u0308nchen, German Research Center for Environmental Health, Ingolsta\u0308dter Landstrasse 1, D-85764 Neuherberg, 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