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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Edmonds, TG; Ding, H; Yuan, X; Wei, Q; Smith, KS; Conway, JA; Wieczorek, L; Brown, B; Polonis, V; West, JT; Montefiori, DC; Kappes, JC; Ochsenbauer, C

Published Date

  • December 5, 2010

Published In

Volume / Issue

  • 408 / 1

Start / End Page

  • 1 - 13

PubMed ID

  • 20863545

Pubmed Central ID

  • 20863545

Electronic International Standard Serial Number (EISSN)

  • 1096-0341

Digital Object Identifier (DOI)

  • 10.1016/j.virol.2010.08.028

Language

  • eng

Conference Location

  • United States