Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC

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• 作者：Tara G. Edmonds ; Haitao Ding ; Xing Yuan ; Qing Wei ; Kendra S. Smith ; Joan A. Conway ; Lindsay Wieczorek ; Bruce Brown ; Victoria Polonis ; John T. West ; David C. Montefiori ; John C. Kappes ; Christina Ochse
• 关键词：HIV-1 ; Neutralizing antibody ; PBMC assay ; HIV reporter virus ; Vaccine assessment ; Assay standardization ; HIV neutralization ; Luciferase ; Envelope glycoprotein
• 刊名：Virology
• 出版年：2010
• 出版时间：5 December 2010
• 年：2010
• 卷：408
• 期：1
• 页码：1-13
• 全文大小：1214 K

文摘

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.