Mechanisms of HIV-1 evasion to the antiviral activity of chemokine CXCL12 indicate potential links with pathogenesis

Armani-Tourret, Marie; Zhou, Zhicheng; Gasser, Romain; Staropoli, Isabelle; Cantaloube-Ferrieu, Vincent; Benureau, Yann; García-Pérez, Javier; Pérez‐Olmeda, Mayte; Lorin, Valérie; Puissant‐Lubrano, Bénédicte; Assoumou, Lambert; Delaugerre, Constance; Lelièvre, Jean‐Daniel; Lévy, Yves; Mouquet, Hugo; Martin‐Blondel, Guillaume; Alcamí, José; Arenzana-Seisdedos, Fernando; Izopet, Jacques; Colin, Philippe; Lagane, Bernard

doi:10.1371/journal.ppat.1009526

“…However, our results indicate that these assays, commonly used to discriminate M-tropic and non-M-tropic viruses, fail to recapitulate all the mechanisms by which HIV-1 isolates can infect MΦ. We show that a large panel of CCR5- and/or CXCR4-using viruses acting as non-M-tropic viruses in cell-free virus infection assays ( Fig 1 and refs [31, 56]) can productively infect MΦ when transferred from infected CD4+ T cells, similarly as bona fide M-tropic viruses.…”

Section: Discussionmentioning

confidence: 69%

“…Virus production was quantified by determining the amount of the Gag p24 protein using an enzyme-linked immunosorbent (ELISA) assay (Innogenetics or TaKaRa). For production of HIV-1 Env-pseudotyped viruses in Jurkat cells, we initially produced VSVg-pseudotyped viruses in HEK 293T cells by cotransfection of the pNL-SacII-lacZ/env-Ren proviral vector in combination with pVSVg using the calcium phosphate precipitation technique [97] or PEI [56]. Virus production in the cell-culture supernatant was then quantified as above and, in some cases, titrated on Jurkat cells using flow cytometry, as described [45].…”

Section: Methodsmentioning

confidence: 99%

“…As a first step, we characterized the capacity of a panel of viruses to infect MDMs when used as cell-free viral particles (Fig 1). We used replication competent, NL4-3-derived luciferase reporter viruses pseudotyped with R5 or X4 Envs of biological virus clones isolated from peripheral blood mononuclear cells (PBMCs) of patients at the chronic or AIDS stage of infection [31,56]. We compared the capacity of different amounts of the R5 viruses, generated from HEK 293T cells, to infect PHA/IL-2-activated primary CD4TL from healthy blood donors [23,31], in a virus type-dependent manner (Fig 1B).…”

Section: Cell-free Hiv-1 Particles Expressing Primary Env Inefficient...mentioning

confidence: 99%

“…Cell culture HEK 293T and Jurkat cell lines were obtained and cultured as described [45,56]. PBMCs were isolated from blood of healthy donors by density gradient sedimentation on Histopaque (Sigma) or Human Pancoll (Pan Biotech).…”

mentioning

confidence: 99%

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HIV-1 cell-to-cell spread overcomes the virus entry block of non-macrophage-tropic strains in macrophages

Han

¹

,

Cantaloube-Ferrieu

²

,

Xie

³

et al. 2022

Preprint

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Macrophages (MΦ) are increasingly recognized as HIV-1 target cells involved in the pathogenesis and persistence of infection. Paradoxically, in vitro infection assays suggest that virus isolates are mostly T-cell-tropic and rarely MΦ-tropic. The latter are assumed to emerge under CD4+ T-cell paucity in tissues such as the brain or at late stage when the CD4 T-cell count declines. However, assays to qualify HIV-1 tropism use cell-free viral particles and may not fully reflect the conditions of in vivo MΦ infection through cell-to-cell viral transfer. Here, we investigated the capacity of viruses expressing primary envelope glycoproteins (Envs) with CCR5 and/or CXCR4 usage from different stages of infection, including transmitted/founder Envs, to infect MΦ by a cell-free mode and through cell-to-cell transfer from infected CD4+ T cells. The results show that most viruses were unable to enter MΦ as cell-free particles, in agreement with the current view that non-M-tropic viruses inefficiently use CD4 and/or CCR5 or CXCR4 entry receptors on MΦ. In contrast, all viruses could be effectively cell-to-cell transferred to MΦ from infected CD4+ T cells. We further showed that viral transfer proceeded through Env-dependent cell-cell fusion of infected T cells with MΦ targets, leading to the formation of productively infected multinucleated giant cells. Compared to cell-free infection, infected T-cell/MΦ contacts showed enhanced interactions of R5 M- and non-M-tropic Envs with CD4 and CCR5, resulting in a reduced dependence on receptor expression levels on MΦ for viral entry. Altogether, our results show that virus cell-to-cell transfer overcomes the entry block of isolates initially defined as non-macrophage-tropic, indicating that HIV-1 has a more prevalent tropism for MΦ than initially suggested. This sheds light into the role of this route of virus cell-to-cell transfer to MΦ in CD4+ T cell rich tissues for HIV-1 transmission, dissemination and formation of tissue viral reservoirs.

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“…As a first step, we characterized the capacity of a panel of viruses to infect MDMs when used as cell-free viral particles ( Fig 1 ). We used replication competent, NL4-3-derived luciferase reporter viruses pseudotyped with R5 or X4 Envs of biological virus clones isolated from peripheral blood mononuclear cells (PBMCs) of patients at the chronic or AIDS stage of infection [ 31 , 56 ]. We compared the capacity of different amounts of the R5 viruses, generated from HEK 293T cells, to infect PHA/IL-2-activated primary CD4TL from healthy blood donors ( Fig 1A ).…”

Section: Resultsmentioning

confidence: 99%

HIV-1 cell-to-cell spread overcomes the virus entry block of non-macrophage-tropic strains in macrophages

Han

¹

,

Cantaloube-Ferrieu

²

,

Xie

³

et al. 2022

Self Cite

View full text Add to dashboard Cite

Macrophages (MΦ) are increasingly recognized as HIV-1 target cells involved in the pathogenesis and persistence of infection. Paradoxically, in vitro infection assays suggest that virus isolates are mostly T-cell-tropic and rarely MΦ-tropic. The latter are assumed to emerge under CD4+ T-cell paucity in tissues such as the brain or at late stage when the CD4 T-cell count declines. However, assays to qualify HIV-1 tropism use cell-free viral particles and may not fully reflect the conditions of in vivo MΦ infection through cell-to-cell viral transfer. Here, we investigated the capacity of viruses expressing primary envelope glycoproteins (Envs) with CCR5 and/or CXCR4 usage from different stages of infection, including transmitted/founder Envs, to infect MΦ by a cell-free mode and through cell-to-cell transfer from infected CD4+ T cells. The results show that most viruses were unable to enter MΦ as cell-free particles, in agreement with the current view that non-M-tropic viruses inefficiently use CD4 and/or CCR5 or CXCR4 entry receptors on MΦ. In contrast, all viruses could be effectively cell-to-cell transferred to MΦ from infected CD4+ T cells. We further showed that viral transfer proceeded through Env-dependent cell-cell fusion of infected T cells with MΦ targets, leading to the formation of productively infected multinucleated giant cells. Compared to cell-free infection, infected T-cell/MΦ contacts showed enhanced interactions of R5 M- and non-M-tropic Envs with CD4 and CCR5, resulting in a reduced dependence on receptor expression levels on MΦ for viral entry. Altogether, our results show that virus cell-to-cell transfer overcomes the entry block of isolates initially defined as non-macrophage-tropic, indicating that HIV-1 has a more prevalent tropism for MΦ than initially suggested. This sheds light into the role of this route of virus cell-to-cell transfer to MΦ in CD4+ T cell rich tissues for HIV-1 transmission, dissemination and formation of tissue viral reservoirs.

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