Mitigation of variation observed in a peripheral blood mononuclear cell (PBMC) based HIV-1 neutralization assay by donor cell pooling

Lindsay Wieczorek, Bruce K. Brown, Camila DelSarto Macedo, Maggie Wesberry-Schmierer, Viseth Ngauy, Andrew Rosa Borges, Nelson L. Michael, Mary A. Marovich, David C. Montefiori, Victoria R. Polonis*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Cultured primary peripheral blood mononuclear cells (PBMC) represent a potentially physiologic in vitro model of HIV-1 infection, but assessment of antibody-mediated HIV-1 neutralization using PBMC has been hindered by donor variability and lack of a sustainable individual PBMC source. To advance this model for HIV vaccine evaluation, intra- and inter-assay variability were assessed using monoclonal and polyclonal antibodies and PBMC targets from multiple HIV-seronegative donors. Inter-assay variability was introduced by using different PBMC for virus propagation, and more substantially, for assay targets. Neutralization titers varied by as much as 4 logs when using different individual donor PBMC as targets; variability was antibody-specific, with the greatest variation observed using an individual polyclonal plasma. Pooling of multiple PBMC donors significantly reduced median inter-assay variation to the level of intra-assay variation, suggesting a pathway forward for establishing a uniform, sustainable and standardized approach to the assessment of antibody function using a PBMC model.

Original languageEnglish
Pages (from-to)240-248
Number of pages9
JournalVirology
Volume447
Issue number1-2
DOIs
StatePublished - Dec 2013
Externally publishedYes

Keywords

  • HIV-1 neutralization
  • Infectious molecular clones
  • Monoclonal antibodies
  • PBMC targets
  • Peripheral blood mononuclear cells
  • Primary isolates
  • Standardization

Fingerprint

Dive into the research topics of 'Mitigation of variation observed in a peripheral blood mononuclear cell (PBMC) based HIV-1 neutralization assay by donor cell pooling'. Together they form a unique fingerprint.

Cite this