{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,26]],"date-time":"2025-09-26T13:07:53Z","timestamp":1758892073040},"reference-count":23,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2015,11,9]],"date-time":"2015-11-09T00:00:00Z","timestamp":1447027200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Italian Ministry for Education, University and Research, Research Projects of National Interest"},{"name":"Italian Ministry of Health, ISS, AIDS Project"},{"name":"University of Rome \u201cTor Vergata\u201d."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2016,6]]},"abstract":"<jats:sec><jats:label \/><jats:p>An assay, specifically optimized to evaluate the anti\u2010HIV activity of antiretrovirals by flow cytometry analysis, is described. As widely used anti\u2010HIV agents, zidovudine (AZT), abacavir (ABC), 2\u2032,3\u2032\u2010dideoxyinosine (DDI), lamivudine (3TC), nevirapine (NVP), and efavirenz (EFV), and as drugs of recent approval raltegravir (RAL), etravirine (ETR), and rilpivirine (RPV), were utilized as reference drugs. HIV\u20101 NL4\u20103 virus was prepared by transfection of HEK293T cells with purified plasmid DNA and quantified by p24 antigen\u2010capture assay. For infection, CEM\u2010GFP cells were exposed to vehicle or to several concentrations of the drugs for 2\u2009hr at 37\u00b0C before HIV\u20101 NL4\u20103 was added to each sample. The adsorption was prolonged for 3\u2009hr at 37\u00b0C. After 72\u2009hr of incubation, HIV\u2010induced GFP expression in infected CEM\u2010GFP cells was assessed by flow cytometry analysis and expressed as % positive cells. For comparison, p24 production in supernatants was assessed by a commercial ELISA kit. On the basis of IC<jats:sub>50<\/jats:sub> values, the anti\u2010HIV activity, as assayed by this method, was EFV\u2009&gt;\u20093TC\u2009&gt;\u2009AZT\u2009&gt;\u2009NVP\u2009&gt;\u2009DDI\u2009&gt;\u2009ABC and ETR\u2009&gt;\u2009RPV\u2009&gt;\u2009RAL. The comparison between the IC<jats:sub>50<\/jats:sub> values calculated through flow cytometry and p24 production revealed overlapping results, showing that the optimized protocol of CEM\u2010GFP infection with HIV NL4\u20103 is a suitable method to perform quantitative, rapid and low\u2010expensive screening tests to evaluate the in vitro effect of new candidate anti\u2010HIV drugs. <jats:bold><jats:italic>J. Med. Virol. 88:979\u2013986, 2016<\/jats:italic><\/jats:bold>. \u00a9 2015 Wiley Periodicals, Inc.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jmv.24418","type":"journal-article","created":{"date-parts":[[2015,10,31]],"date-time":"2015-10-31T23:08:48Z","timestamp":1446332928000},"page":"979-986","source":"Crossref","is-referenced-by-count":7,"title":["Testing anti\u2010HIV activity of antiretroviral agents in vitro using flow cytometry analysis of CEM\u2010GFP cells infected with transfection\u2010derived HIV\u20101 NL4\u20103"],"prefix":"10.1002","volume":"88","author":[{"given":"Caterina","family":"Frezza","sequence":"first","affiliation":[{"name":"Department of Systems Medicine, University of Rome \u201cTor Vergata\u201d, Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandro","family":"Grelli","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine and Surgery, University of Rome \u201cTor Vergata\u201d, Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maurizio","family":"Federico","sequence":"additional","affiliation":[{"name":"Division of Pathogenesis of Retroviruses, National AIDS Center, Istituto Superiore di Sanit\u00e0, Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Marino\u2010Merlo","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences, University of Messina, Messina Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Antonio","family":"Mastino","sequence":"additional","affiliation":[{"name":"Department of Biological and Environmental Sciences, University of Messina, Messina Italy"},{"name":"Institute of Translational Pharmacology, CNR, Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beatrice","family":"Macchi","sequence":"additional","affiliation":[{"name":"Department of Systems Medicine, University of Rome \u201cTor Vergata\u201d, Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,11,9]]},"reference":[{"key":"e_1_2_5_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.59.2.284-291.1986"},{"key":"e_1_2_5_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2010.03.015"},{"key":"e_1_2_5_4_1","doi-asserted-by":"publisher","DOI":"10.1186\/1742-4690-11-46"},{"key":"e_1_2_5_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2011.01.024"},{"key":"e_1_2_5_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2010.12.031"},{"key":"e_1_2_5_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2011.01.016"},{"key":"e_1_2_5_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2010.06.052"},{"key":"e_1_2_5_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2010.01.023"},{"key":"e_1_2_5_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.23748"},{"key":"e_1_2_5_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.9.4653"},{"key":"e_1_2_5_12_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M105582200"},{"key":"e_1_2_5_13_1","doi-asserted-by":"publisher","DOI":"10.3390\/v6104095"},{"key":"e_1_2_5_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.37.5.1095"},{"key":"e_1_2_5_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejmech.2011.01.037"},{"key":"e_1_2_5_16_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.cdd.4401668"},{"key":"e_1_2_5_17_1","doi-asserted-by":"publisher","DOI":"10.1097\/QCO.0b013e328332c54d"},{"key":"e_1_2_5_18_1","first-page":"540","article-title":"Ellagic acid & gallic acid from Lagerstroemia speciosa L. inhibit HIV\u20101 infection through inhibition of HIV\u20101 protease & reverse transcriptase activity","volume":"137","author":"Modi M","year":"2013","journal-title":"Indian J Med Res"},{"key":"e_1_2_5_19_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6882-13-158"},{"key":"e_1_2_5_20_1","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/165871"},{"key":"e_1_2_5_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/0166-0934(88)90134-6"},{"key":"e_1_2_5_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01044-12"},{"key":"e_1_2_5_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02038-13"},{"key":"e_1_2_5_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neures.2012.05.014"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjmv.24418","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.24418","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,4]],"date-time":"2023-10-04T05:10:41Z","timestamp":1696396241000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.24418"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,11,9]]},"references-count":23,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2016,6]]}},"alternative-id":["10.1002\/jmv.24418"],"URL":"https:\/\/doi.org\/10.1002\/jmv.24418","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,11,9]]}}}