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

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 Ochsenbauer

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)1-13
Number of pages13
JournalVirology
Volume408
Issue number1
DOIs
StatePublished - Dec 5 2010

Fingerprint

Renilla Luciferases
HIV-1
Blood Cells
Clone Cells
Luciferases
Antibodies
Neutralizing Antibodies
Vaccines
Viruses
Firefly Luciferases
Open Reading Frames
Sequence Analysis
Genome
DNA

Keywords

  • Assay standardization
  • Envelope glycoprotein
  • HIV neutralization
  • HIV reporter virus
  • HIV-1
  • Luciferase
  • Neutralizing antibody
  • PBMC assay
  • Vaccine assessment

ASJC Scopus subject areas

  • Virology

Cite this

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

In: Virology, Vol. 408, No. 1, 05.12.2010, p. 1-13.

Research output: Contribution to journalArticle

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 2010, 'Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC', Virology, vol. 408, no. 1, pp. 1-13. https://doi.org/10.1016/j.virol.2010.08.028
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. / Replication competent molecular clones of HIV-1 expressing Renilla luciferase facilitate the analysis of antibody inhibition in PBMC. In: Virology. 2010 ; Vol. 408, No. 1. pp. 1-13.
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