Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC
Abstract Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-base...
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| Veröffentlicht in: | Virology (New York, N.Y.) N.Y.), 2010-12, Vol.408 (1), p.1-13 |
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| creator | Edmonds, Tara G Ding, Haitao Yuan, Xing Wei, Qing Smith, Kendra S Conway, Joan A Wieczorek, Lindsay Brown, Bruce Polonis, Victoria West, John T Montefiori, David C Kappes, John C Ochsenbauer, Christina |
| description | Abstract Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env–IMC–LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy. |
| doi_str_mv | 10.1016/j.virol.2010.08.028 |
| format | Article |
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To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env–IMC–LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy.</description><identifier>ISSN: 0042-6822</identifier><identifier>EISSN: 1096-0341</identifier><identifier>DOI: 10.1016/j.virol.2010.08.028</identifier><identifier>PMID: 20863545</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>AIDS Vaccines - immunology ; Antibodies, Neutralizing - immunology ; Assay standardization ; Envelope glycoprotein ; HIV Antibodies - immunology ; HIV neutralization ; HIV reporter virus ; HIV-1 ; HIV-1 - genetics ; HIV-1 - growth & development ; HIV-1 - immunology ; Humans ; Infectious Disease ; Leukocytes, Mononuclear - virology ; Luciferase ; Luciferases, Renilla - biosynthesis ; Luciferases, Renilla - genetics ; Neutralization Tests - methods ; Neutralizing antibody ; PBMC assay ; Recombinant Proteins - biosynthesis ; Recombinant Proteins - genetics ; Staining and Labeling - methods ; Vaccine assessment ; Virology - methods ; Virus Replication</subject><ispartof>Virology (New York, N.Y.), 2010-12, Vol.408 (1), p.1-13</ispartof><rights>Elsevier Inc.</rights><rights>2010 Elsevier Inc.</rights><rights>Copyright © 2010 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c579t-a1a9e0489522f173c948bb4a4c5712f358e966c28608adffe3868dfc27f8ac623</citedby><cites>FETCH-LOGICAL-c579t-a1a9e0489522f173c948bb4a4c5712f358e966c28608adffe3868dfc27f8ac623</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0042682210005581$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20863545$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Edmonds, Tara G</creatorcontrib><creatorcontrib>Ding, Haitao</creatorcontrib><creatorcontrib>Yuan, Xing</creatorcontrib><creatorcontrib>Wei, Qing</creatorcontrib><creatorcontrib>Smith, Kendra S</creatorcontrib><creatorcontrib>Conway, Joan A</creatorcontrib><creatorcontrib>Wieczorek, Lindsay</creatorcontrib><creatorcontrib>Brown, Bruce</creatorcontrib><creatorcontrib>Polonis, Victoria</creatorcontrib><creatorcontrib>West, John T</creatorcontrib><creatorcontrib>Montefiori, David C</creatorcontrib><creatorcontrib>Kappes, John C</creatorcontrib><creatorcontrib>Ochsenbauer, Christina</creatorcontrib><title>Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC</title><title>Virology (New York, N.Y.)</title><addtitle>Virology</addtitle><description>Abstract Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env–IMC–LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy.</description><subject>AIDS Vaccines - immunology</subject><subject>Antibodies, Neutralizing - immunology</subject><subject>Assay standardization</subject><subject>Envelope glycoprotein</subject><subject>HIV Antibodies - immunology</subject><subject>HIV neutralization</subject><subject>HIV reporter virus</subject><subject>HIV-1</subject><subject>HIV-1 - genetics</subject><subject>HIV-1 - growth & development</subject><subject>HIV-1 - immunology</subject><subject>Humans</subject><subject>Infectious Disease</subject><subject>Leukocytes, Mononuclear - virology</subject><subject>Luciferase</subject><subject>Luciferases, Renilla - biosynthesis</subject><subject>Luciferases, Renilla - genetics</subject><subject>Neutralization Tests - methods</subject><subject>Neutralizing antibody</subject><subject>PBMC assay</subject><subject>Recombinant Proteins - biosynthesis</subject><subject>Recombinant Proteins - genetics</subject><subject>Staining and Labeling - methods</subject><subject>Vaccine assessment</subject><subject>Virology - methods</subject><subject>Virus Replication</subject><issn>0042-6822</issn><issn>1096-0341</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkk1v1DAQhiMEotvCL0BCvnHKYjtfzoFKsKK0UhGofFwtxxl3Z3HsYCcr9s4Px7tbKuDCybLnfWfG80yWPWN0ySirX26WWwzeLjlNL1QsKRcPsgWjbZ3TomQPswWlJc9rwflJdhrjhqZ709DH2Qmnoi6qslpkP29gtKjVhN4R7YcRJnATGbwFPVsViLbeQSTekMurrzkj8GMMECO6W3IDDq1VxM4aDQQVgRil0eKkJiDTGohyyu4iHuzKTdj5fkfQrbHDQ0F05OOb96sn2SOjbISnd-dZ9uXi7efVZX794d3V6vV1rqumnXLFVAu0FG3FuWFNodtSdF2pyhRm3BSVgLauNRc1Fao3BgpRi95o3hihdM2Ls-z8mHecuwF6nX4alJVjwEGFnfQK5d8Rh2t567eSt21BBUsJXtwlCP77DHGSA0YNaQgO_BxlU_OyYU0lkrI4KnXwMQYw91UYlXt8ciMP-OQen6RCJnzJ9fzPBu89v3klwaujANKYtghBRo3gNPQYQE-y9_ifAuf_-LVFl_jbb7CDuPFzSMiiZDJySeWn_QbtF4il3amqNIFfB3vFCg</recordid><startdate>20101205</startdate><enddate>20101205</enddate><creator>Edmonds, Tara G</creator><creator>Ding, Haitao</creator><creator>Yuan, Xing</creator><creator>Wei, Qing</creator><creator>Smith, Kendra S</creator><creator>Conway, Joan A</creator><creator>Wieczorek, Lindsay</creator><creator>Brown, Bruce</creator><creator>Polonis, Victoria</creator><creator>West, John T</creator><creator>Montefiori, David C</creator><creator>Kappes, John C</creator><creator>Ochsenbauer, Christina</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20101205</creationdate><title>Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC</title><author>Edmonds, Tara G ; Ding, Haitao ; Yuan, Xing ; Wei, Qing ; Smith, Kendra S ; Conway, Joan A ; Wieczorek, Lindsay ; Brown, Bruce ; Polonis, Victoria ; West, John T ; Montefiori, David C ; Kappes, John C ; Ochsenbauer, Christina</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c579t-a1a9e0489522f173c948bb4a4c5712f358e966c28608adffe3868dfc27f8ac623</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>AIDS Vaccines - immunology</topic><topic>Antibodies, Neutralizing - immunology</topic><topic>Assay standardization</topic><topic>Envelope glycoprotein</topic><topic>HIV Antibodies - immunology</topic><topic>HIV neutralization</topic><topic>HIV reporter virus</topic><topic>HIV-1</topic><topic>HIV-1 - genetics</topic><topic>HIV-1 - growth & development</topic><topic>HIV-1 - immunology</topic><topic>Humans</topic><topic>Infectious Disease</topic><topic>Leukocytes, Mononuclear - virology</topic><topic>Luciferase</topic><topic>Luciferases, Renilla - biosynthesis</topic><topic>Luciferases, Renilla - genetics</topic><topic>Neutralization Tests - methods</topic><topic>Neutralizing antibody</topic><topic>PBMC assay</topic><topic>Recombinant Proteins - biosynthesis</topic><topic>Recombinant Proteins - genetics</topic><topic>Staining and Labeling - methods</topic><topic>Vaccine assessment</topic><topic>Virology - methods</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Edmonds, Tara G</creatorcontrib><creatorcontrib>Ding, Haitao</creatorcontrib><creatorcontrib>Yuan, Xing</creatorcontrib><creatorcontrib>Wei, Qing</creatorcontrib><creatorcontrib>Smith, Kendra S</creatorcontrib><creatorcontrib>Conway, Joan A</creatorcontrib><creatorcontrib>Wieczorek, Lindsay</creatorcontrib><creatorcontrib>Brown, Bruce</creatorcontrib><creatorcontrib>Polonis, Victoria</creatorcontrib><creatorcontrib>West, John T</creatorcontrib><creatorcontrib>Montefiori, David C</creatorcontrib><creatorcontrib>Kappes, John C</creatorcontrib><creatorcontrib>Ochsenbauer, Christina</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Virology (New York, N.Y.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Edmonds, Tara G</au><au>Ding, Haitao</au><au>Yuan, Xing</au><au>Wei, Qing</au><au>Smith, Kendra S</au><au>Conway, Joan A</au><au>Wieczorek, Lindsay</au><au>Brown, Bruce</au><au>Polonis, Victoria</au><au>West, John T</au><au>Montefiori, David C</au><au>Kappes, John C</au><au>Ochsenbauer, Christina</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC</atitle><jtitle>Virology (New York, N.Y.)</jtitle><addtitle>Virology</addtitle><date>2010-12-05</date><risdate>2010</risdate><volume>408</volume><issue>1</issue><spage>1</spage><epage>13</epage><pages>1-13</pages><issn>0042-6822</issn><eissn>1096-0341</eissn><abstract>Abstract Effective vaccine development for human immunodeficiency virus type 1 (HIV-1) will require assays that ascertain the capacity of vaccine immunogens to elicit neutralizing antibodies (NAb) to diverse HIV-1 strains. To facilitate NAb assessment in peripheral blood mononuclear cell (PBMC)-based assays, we developed an assay-adaptable platform based on a Renilla luciferase (LucR) expressing HIV-1 proviral backbone. LucR was inserted into pNL4-3 DNA, preserving all viral open reading frames. The proviral genome was engineered to facilitate expression of diverse HIV-1 env sequences, allowing analysis in an isogenic background. The resulting Env–IMC–LucR viruses are infectious, and LucR is stably expressed over multiple replications in PBMC. HIV-1 neutralization, targeting TZM-bl cells, was highly correlative comparing virus (LucR) and cell (firefly luciferase) readouts. In PBMC, NAb activity can be analyzed either within a single or multiple cycles of replication. These results represent advancement toward a standardizable PBMC-based neutralization assay for assessing HIV-1 vaccine immunogen efficacy.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>20863545</pmid><doi>10.1016/j.virol.2010.08.028</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Virology (New York, N.Y.), 2010-12, Vol.408 (1), p.1-13 |
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| source | MEDLINE; Elsevier ScienceDirect Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | AIDS Vaccines - immunology Antibodies, Neutralizing - immunology Assay standardization Envelope glycoprotein HIV Antibodies - immunology HIV neutralization HIV reporter virus HIV-1 HIV-1 - genetics HIV-1 - growth & development HIV-1 - immunology Humans Infectious Disease Leukocytes, Mononuclear - virology Luciferase Luciferases, Renilla - biosynthesis Luciferases, Renilla - genetics Neutralization Tests - methods Neutralizing antibody PBMC assay Recombinant Proteins - biosynthesis Recombinant Proteins - genetics Staining and Labeling - methods Vaccine assessment Virology - methods Virus Replication |
| title | Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC |
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