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. 2018 Jul 1;78(3):348-355.
doi: 10.1097/QAI.0000000000001675.

Neutralization Sensitivity of a Novel HIV-1 CRF01_AE Panel of Infectious Molecular Clones

Agnes-Laurence Chenine  1   2   3 , Melanie Merbah  1   2 , Lindsay Wieczorek  1   2 , Sebastian Molnar  1   2 , Brendan Mann  1   2 , Jenica Lee  1   2 , Anne-Marie OʼSullivan  1   2 , Meera Bose  1   2 , Eric Sanders-Buell  1   2 , Gustavo H Kijak  1   2   4 , Carolina Herrera  5 , Robert McLinden  1   2   6 , Robert J OʼConnell  7 , Nelson L Michael  1 , Merlin L Robb  1   2 , Jerome H Kim  1   8 , Victoria R Polonis  1 , Sodsai Tovanabutra  1   2
Affiliations

Neutralization Sensitivity of a Novel HIV-1 CRF01_AE Panel of Infectious Molecular Clones

Agnes-Laurence Chenine et al. J Acquir Immune Defic Syndr. .

Abstract

Background: HIV-1 CRF01_AE is dominant in Thailand where RV144 vaccine trial was conducted. To study immune correlates of protection in ongoing trials, CRF01_AE-derived reagents are essential. Here, we present a panel of 14 HIV-1 infectious molecular clones (IMCs) identified from different stages of infection and characterization of their neutralization sensitivity using 2 standard assays.

Methods: One full-length IMC was constructed using a transmitted-founder virus to express Renilla luciferase (LucR) reporter gene and full-length envelopes (envs) of exogenous HIV-1. A panel of IMCs was generated, expressing envs of viruses from acute (Fiebig stages I/II and I-IV) and chronic (>Fiebig VI) infection. Neutralization assays were performed using TZM-bl or A3R5 cell lines, and sera or monoclonal antibodies (mAbs). Wilcoxon matched-paired test was used to assess neutralization differences between assays and reagents; correlation coefficients were evaluated by linear regression.

Results: Neutralization potency observed was significantly higher in the A3R5 assay when testing mAbs and serum pools (P < 0.0001); the stage of infection from which env was derived did not associate with IMC neutralization sensitivity. Neutralization values from A3R5 and TZM-bl assays were strongly correlated when mAbs were tested (R = 0.7, P < 0.0001), but a weaker association was seen with serum pools (R = 0.17, P = 0.03).

Conclusions: This novel panel of CRF01_AE reporter IMC is useful for assessing vaccine-induced neutralizing antibodies in multiple assays, including those using primary cell targets. The significant differences in TZM-bl and A3R5 neutralization sensitivity, as well as the poor association when using polyclonal sera indicates the need for caution in choosing one specific platform.

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Conflict of interest statement

Conflicts of Interest: The authors have declared that no conflicts of interest exist.

Figures

Figure 1
Figure 1
Neutralization sensitivity of CRF01_AE viruses against mAbs (A) and sera (B) measured in A3R5 and TZM-bl cells by luminescence readouts. Linear regression to compare neutralization sensitivity against mAbs (C) and sera (D) between both cell lines were performed.
Figure 2
Figure 2
Neutralization sensitivity against the four different antigenic regions on CRF01_AE envelopes measured in A3R5 and TZM-bl cells using specific mAbs (A). Neutralization susceptibility against CRF01_AE and subtype B pooled sera was evaluated in A3R5 and TZM-bl cells (B). Neutralization sensitivity of CRF01_AE viruses isolated at different Fiebig stages against mAbs (C) and CRF01_AE and subtype B pooled sera (D) measured in A3R5 and TZM-bl cells by luminescence readouts.
Figure 3
Figure 3
Neutralization sensitivity of individual CRF01_AE virus against the panel of mAbs measured in A3R5 and TZM-bl cells.
See this image and copyright information in PMC

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