TY - JOUR
T1 - Comparison of p24 measurement by ELISA versus indicator cells for detecting residual HIV infectivity in vitro
AU - Herbein, Georges
AU - Illei, Peter
AU - Montaner, Luis J.
AU - James, William
AU - Gordon, Siamon
N1 - Funding Information:
We acknowledget he help of Dr C. Entwistle and the staff of the Oxford Regional Transfusion Servicef or providing buffy coats. This work was supportedb y researchg rants from the Medical ResearchC ouncil (AIDS Directed Program). G. Herbein was supported by a MRC-INSERM/ AIDS award and L.J. Montaner by a Marshall Scholarship.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1996/4/26
Y1 - 1996/4/26
N2 - Inactivation of HIV-1 contaminated materials or biological samples is of great importance and requires the use of a reliable assay to detect residual infectivity. In this study we treated cell-free or cell-associated (monocyte- derived macrophages) HIV-1 with two chemicals known for their antiviral activities, β-propiolactone (βPL) and formaldehyde (FO), and tested it for the presence of residual infectivity. HIV-1 infected primary monocyte- derived macrophages (MDM) or cell-free HIV-1 were fixed with increasing concentrations of either βPL or FO for 1 day at 4°C. Then either fresh primary MDM or fresh medium was added, and the supernatant p24 levels were assayed up to 12 days after infection. All the supernatants harvested were added to indicator cells, fresh primary MDM, to assess for residual infectivity. The results show that p24 measurement is not a reliable assay for the detection of residual infectious virions after chemical fixation of HIV-infected primary MDM. In contrast, the use of indicator primary cells (MDM) is a much more sensitive and reliable assay. By performing an indicator cell assay we showed that FO efficiently inactivates cell-associated and cell-free HIV-1 at concentrations as low as 1% v/v. In contrast βPL is more efficient in inactivating cell-free than cell-associated virus and does not inactivate cell-associated HIV-1 at concentrations as high as 1% v/v.
AB - Inactivation of HIV-1 contaminated materials or biological samples is of great importance and requires the use of a reliable assay to detect residual infectivity. In this study we treated cell-free or cell-associated (monocyte- derived macrophages) HIV-1 with two chemicals known for their antiviral activities, β-propiolactone (βPL) and formaldehyde (FO), and tested it for the presence of residual infectivity. HIV-1 infected primary monocyte- derived macrophages (MDM) or cell-free HIV-1 were fixed with increasing concentrations of either βPL or FO for 1 day at 4°C. Then either fresh primary MDM or fresh medium was added, and the supernatant p24 levels were assayed up to 12 days after infection. All the supernatants harvested were added to indicator cells, fresh primary MDM, to assess for residual infectivity. The results show that p24 measurement is not a reliable assay for the detection of residual infectious virions after chemical fixation of HIV-infected primary MDM. In contrast, the use of indicator primary cells (MDM) is a much more sensitive and reliable assay. By performing an indicator cell assay we showed that FO efficiently inactivates cell-associated and cell-free HIV-1 at concentrations as low as 1% v/v. In contrast βPL is more efficient in inactivating cell-free than cell-associated virus and does not inactivate cell-associated HIV-1 at concentrations as high as 1% v/v.
KW - Formaldehyde
KW - HIV, β propiolactone
KW - Indicator cells
KW - Residual infectivity
KW - p24
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U2 - 10.1016/0166-0934(96)02007-1
DO - 10.1016/0166-0934(96)02007-1
M3 - Article
C2 - 8783162
AN - SCOPUS:0030049410
SN - 0166-0934
VL - 58
SP - 167
EP - 173
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1-2
ER -