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

Virology. 2010 Dec 5;408(1):1-13. doi: 10.1016/j.virol.2010.08.028. Epub 2010 Sep 21.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / immunology
  • Antibodies, Neutralizing / immunology
  • HIV Antibodies / immunology*
  • HIV-1 / genetics
  • HIV-1 / growth & development*
  • HIV-1 / immunology
  • Humans
  • Leukocytes, Mononuclear / virology*
  • Luciferases, Renilla / biosynthesis*
  • Luciferases, Renilla / genetics
  • Neutralization Tests / methods
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Staining and Labeling / methods*
  • Virology / methods*
  • Virus Replication*

Substances

  • AIDS Vaccines
  • Antibodies, Neutralizing
  • HIV Antibodies
  • Recombinant Proteins
  • Luciferases, Renilla