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.
journal_name
Virologyjournal_title
Virologyauthors
Wieczorek L,Brown BK,Delsarto Macedo C,Wesberry-Schmierer M,Ngauy V,Rosa Borges A,Michael NL,Marovich MA,Montefiori DC,Polonis VRdoi
10.1016/j.virol.2013.09.014subject
Has Abstractpub_date
2013-12-01 00:00:00pages
240-8issue
1-2eissn
0042-6822issn
1096-0341pii
S0042-6822(13)00546-1journal_volume
447pub_type
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