Detailed Characterization of Early HIV-1 Replication Dynamics in Primary Human Macrophages.

David Bejarano,Maria Puertas,Barbara Müller,Javier Martinez-Picado,Kathleen Börner,Hans-Georg Kräusslich

doi:10.3390/v10110620

David Bejarano, Maria Puertas + Show 4 more

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https://doi.org/10.3390/v10110620

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Abstract

Macrophages are natural target cells of human immunodeficiency virus type 1 (HIV-1). Viral replication appears to be delayed in these cells compared to lymphocytes; however, little is known about the kinetics of early post-entry events. Time-of-addition experiments using several HIV-1 inhibitors and the detection of reverse transcriptase (RT) products with droplet digital PCR (ddPCR) revealed that early replication was delayed in primary human monocyte-derived macrophages of several donors and peaked late after infection. Direct imaging of reverse-transcription and pre-integration complexes (RTC/PIC) by click-labeling of newly synthesized DNA further confirmed our findings and showed a concomitant shift to the nuclear stage over time. Altering the entry pathway enhanced infectivity but did not affect kinetics of viral replication. The addition of viral protein X (Vpx) enhanced productive infection and accelerated completion of reverse transcription and nuclear entry. We propose that sterile alpha motif (SAM) and histidine/aspartate (HD) domain-containing protein 1 (SAMHD1) activity lowering deoxyribonucleotide triphosphate (dNTP) pools is the principal factor delaying early HIV-1 replication in macrophages.

Highlights

Human immunodeficiency virus type 1 (HIV-1) primarily infects and replicates in cluster of differentiation 4 positive (CD4+ ) T cells
human immunodeficiency virus type 1 (HIV-1) Replication Is Slow in Primary Macrophages
In order to study HIV-1 infection in primary human monocyte-derived macrophages (MDM), terminally differentiated macrophages were prepared from primary monocytes from healthy blood donors

Summary

Introduction

Human immunodeficiency virus type 1 (HIV-1) primarily infects and replicates in cluster of differentiation 4 positive (CD4+ ) T cells. Macrophages constitute a relevant natural target cell of the virus and are a crucial vehicle of viral spread within the infected organism [1]. Kinetics of individual HIV-1 replication steps in T-cell lines and primary CD4+ lymphocytes were previously analyzed in detail (e.g., [6,7,8,9]). A particular focus of interest was on early post-entry events—from cytosolic entry of the viral capsid over reverse transcription of the viral genome to its transport to and integration into the host cell DNA—which establish persistent infection and are key Viruses 2018, 10, 620; doi:10.3390/v10110620 www.mdpi.com/journal/viruses

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