TY - JOUR
T1 - Mitigation of variation observed in a peripheral blood mononuclear cell (PBMC) based HIV-1 neutralization assay by donor cell pooling
AU - Wieczorek, Lindsay
AU - Brown, Bruce K.
AU - DelSarto Macedo, Camila
AU - Wesberry-Schmierer, Maggie
AU - Ngauy, Viseth
AU - Rosa Borges, Andrew
AU - Michael, Nelson L.
AU - Marovich, Mary A.
AU - Montefiori, David C.
AU - Polonis, Victoria R.
N1 - Funding Information:
This work was supported by a cooperative agreement ( W81XWH-07-2-0067 ) between the Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., and the U.S. Department of Defense (DOD) (U.S. Army Medical Research and Material Command) , working together with the Division of AIDS, National Institute for Allergy and Infectious Diseases National Institutes of Health. The views expressed here are the private opinions of the authors and are not to be considered as official or reflecting the views of the U.S. Army, the U.S. Department of Defense, or the U.S. Government. Use of trade names is for identification only and does not imply endorsement by the U.S. government.
PY - 2013/12
Y1 - 2013/12
N2 - 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.
AB - 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.
KW - HIV-1 neutralization
KW - Infectious molecular clones
KW - Monoclonal antibodies
KW - PBMC targets
KW - Peripheral blood mononuclear cells
KW - Primary isolates
KW - Standardization
UR - http://www.scopus.com/inward/record.url?scp=84885082385&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2013.09.014
DO - 10.1016/j.virol.2013.09.014
M3 - Article
C2 - 24210120
AN - SCOPUS:84885082385
SN - 0042-6822
VL - 447
SP - 240
EP - 248
JO - Virology
JF - Virology
IS - 1-2
ER -