Amplification of Replication Competent HIV-1 by Adoptive Transfer of Human Cells From Infected Humanized Mice.

Hang Su,Sruthi Sravanam,Kamel Khalili,Rafal Kaminski,Santhi Gorantla,Howard E Gendelman,Prasanta K Dash,Larisa Poluektova

doi:10.3389/fcimb.2020.00038

Abstract

Detection of latent human immunodeficiency virus type 1 (HIV-1) in “putative” infectious reservoirs is required for determining treatment efficiency and for viral elimination strategies. Such tests require induction of replication competent provirus and quantitative testing of viral load for validation. Recently, humanized mice were employed in the development of such tests by employing a murine viral outgrowth assay (mVOA). Here blood cells were recovered from virus infected antiretroviral therapy suppressed patients. These cells were adoptively transferred to uninfected humanized mice where replication competent virus was recovered. Prior reports supported the notion that an mVOA assay provides greater sensitivity than cell culture-based quantitative VOA tests for detection of latent virus. In the current study, the mVOA assays was adapted using donor human hematopoietic stem cells-reconstituted mice to affirm research into HIV-1 elimination. We simulated an antiretroviral therapy (ART)-treated virus-infected human by maintaining the infected humanized mice under suppressive treatment. This was operative prior to human cell adoptive transfers. Replication-competent HIV-1 was easily detected in recipient animals from donors with undetectable virus in plasma. Moreover, when the assay was used to investigate viral presence in tissue reservoirs, quantitative endpoints were determined in “putative” viral reservoirs not possible in human sample analyses. We conclude that adoptive transfer of cells between humanized mice is a sensitive and specific assay system for detection of replication competent latent HIV-1.

Highlights

Antiretroviral therapy (ART) has transformed human immunodeficiency virus type one (HIV1) infection from a life-threatening disease into a life-long chronic condition
The results showed that virus can be readily recovered demonstrating that adoptive transfer between humanized mice is an effective means to recover residual human immunodeficiency virus type 1 (HIV-1) that includes suppressive ART treatments
While tissue HIV-1 remained below the limits of detection in M3319 and M3336, viral DNA and RNA was observed in M3324, demonstrating that HIV-1 persisted in tissues even when peripheral viral amplification was not observed (Figures 1E,F)

Summary

Introduction

Antiretroviral therapy (ART) has transformed human immunodeficiency virus type one (HIV1) infection from a life-threatening disease into a life-long chronic condition. The major barrier for “cure” rests in the ability to eliminate HIV-1 latency This will require the clearance of few numbers of infected cells present during the disease course following treatment where cell populations emerge that contain replication-competent latent HIV-1 DNA. While research into viral elimination strategies show promise to eliminate the latent viral reservoir, highly sensitive and specific validations are required for HIV-1 detection Such tests enable measurements of the size, location and distribution of any residual virus in tissues, in cells and in body fluids (Laird et al, 2013; Rouzioux and Richman, 2013; Gupta et al, 2017; Sanyal et al, 2017). This has led to the development of alternative viral detection systems (Metcalf Pate and Blankson, 2017; Schmitt and Akkina, 2018)

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Results

Conclusion