The forces driving clonal expansion of the HIV-1 latent reservoir

Liu, Runxia; Simonetti, Francesco R.; Ho, Ya-Chi

doi:10.1186/s12985-019-1276-8

Cited by 63 publications

(70 citation statements)

References 118 publications

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“…Viral sequencing has also revealed many intact proviruses within clonally expanded populations, particularly within the Tem and Th1 subsets, and those expressing Ox40 or CD161 ( De Scheerder et al, 2019 ; Hiener et al, 2017 ; Kuo et al, 2018 ; Lee et al, 2017 ; Li et al, 2019 ). These and many other studies ( Cohn et al, 2020 ; Liu et al, 2020 ) have solidified the view that clonal expansion plays a major role in maintaining the long-lived HIV reservoir.…”

Section: Introductionmentioning

confidence: 54%

Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

Neidleman

Luo

Frouard

et al. 2020

eLife

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The latent reservoir is a major barrier to HIV cure. As latently infected cells cannot be phenotyped directly, the features of the in vivo reservoir have remained elusive. Here, we describe a method that leverages high-dimensional phenotyping using CyTOF to trace latently infected cells reactivated ex vivo to their original pre-activation states. Our results suggest that, contrary to common assumptions, the reservoir is not randomly distributed among cell subsets, and is remarkably conserved between individuals. However, reservoir composition differs between tissues and blood, as do cells successfully reactivated by different latency reversing agents. By selecting 8–10 of our 39 original CyTOF markers, we were able to isolate highly purified populations of unstimulated in vivo latent cells. These purified populations were highly enriched for replication-competent and intact provirus, transcribed HIV, and displayed clonal expansion. The ability to isolate unstimulated latent cells from infected individuals enables previously impossible studies on HIV persistence.

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Section: Introductionmentioning

confidence: 54%

Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

Neidleman

Luo

Frouard

et al. 2020

eLife

View full text Add to dashboard Cite

show abstract

“…Viral sequencing has also revealed many intact proviruses within clonally expanded populations, particularly within the Tem and Th1 subsets, and those expressing Ox40 or CD161 (De Scheerder et al, 2019;Hiener et al, 2017;Kuo et al, 2018;Lee et al, 2017;Li et al, 2019). These and many other studies (Cohn et al, 2020;Liu et al, 2020) have solidified the view that clonal expansion plays a major role in maintaining the longlived HIV reservoir.…”

Section: Introductionmentioning

confidence: 65%

“…5C). To assess the extent of clonal expansion, a major driver of HIV persistence (Cohn et al, 2020;Liu et al, 2020), we determined the proportions of expanded identical sequences (EIS). EIS were prominent in both populations, but the proportions of EIS with intact p24 sequences were consistently higher in the enriched ones ( Fig.…”

Section: Markers Of Knn Latent Cells Enrich For the Replication-compementioning

confidence: 99%

The Atlas of the In Vivo HIV CD4 T Cell Reservoir

Neidleman

Luo

Frouard

et al. 2020

Preprint

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The latent reservoir is a main barrier for curing HIV. But because latently-infected cells cannot be phenotyped directly, the features of the in vivo reservoir have remained elusive. Here, we describe a method that leverages high-dimensional phenotyping using CyTOF to trace latently-infected cells reactivated ex vivo to their original pre-activation states. Our results suggest that contrary to common assumptions, the reservoir is not randomly distributed among cell subsets, and is remarkably conserved between individuals. However, reservoir composition differs between tissues and blood, as do cells successfully reactivated by different latency reversing agents. Most importantly, by selecting 8-10 of our 39 original CyTOF markers, we were able to isolate highly purified populations of unstimulated in vivo latent cells, thereby validating the PP-SLIDE approach for reservoir characterization. These purified populations were highly enriched for replication-competent and intact provirus, transcribed HIV, and displayed clonal expansion. The ability to isolate unstimulated latent cells from infected individuals enables previously impossible studies of HIV persistence.

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“…It is known that mitotic cells with integrated viruses may persist for many years, undergo clonal expansion (61). Clonal expansion seems to be the major mechanism to generate HIV-1 reservoir (62), considered to be established early during primary HIV-1 infection (63).…”

Section: Discussionmentioning

confidence: 99%

Remodeling of the core leads HIV-1 pre-integration complex in the nucleus of human lymphocytes

Blanco-Rodriguez

Gazi²,

Monel

et al. 2020

Preprint

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AbstractRetroviral replication proceeds through obligate integration of the viral DNA into the host genome. To enter the nucleus, the viral DNA must be led through the nuclear pore complex (NPC). During HIV-1 cytoplasmic journey, the viral core acts like a shell to protect the viral genetic material from antiviral sensors and ensure an adequate environment for the reverse transcription. However, the relatively narrow size of the nuclear pore channel requires that the HIV-1 core reshapes into a structure that fits the pore. On the other hand, the organization of the viral CA proteins that remain associated to the pre-integration complex (PIC) during and after nuclear translocation, in particular, in human lymphocytes, the main target cells of HIV-1, is still enigmatic. In this study, we analysed the progressive organizational changes of viral CA proteins within the cytoplasm and the nucleus by immuno-gold labelling. Furthermore, we set up a novel technology, HIV-1 ANCHOR, which enables specific detection of the retrotranscribed DNA by fluorescence microscopy, thereby uncovering the architecture of the potential HIV-1 PIC. Thus, we revealed DNA- and CA-positive complexes by correlated light- and electron microscopy (CLEM). During and after nuclear translocation, HIV-1 appears as a complex of viral DNA decorated by multiple viral CA proteins remodelled in a “pearl necklace” shape. Thus, we observed how CA proteins reshape around the viral DNA to permit the entrance of the HIV-1 in the nucleus. This particular CA protein complex composed by the integrase and the retrotranscribed DNA leads HIV-1 genome inside the host nucleus to potentially replicate.Our findings contribute to the understanding of the early steps of HIV-1 infection and provide new insights into the organization of HIV-1 CA proteins during and after viral nuclear entry.ImportanceHow the reverse transcribed genome reaches the host nucleus remains a main open question related to the infectious cycle of HIV-1. HIV-1 core has a size of ∼100 nm, largely exceeding that of the NPC channel (∼39 nm). Thus, a rearrangement of the viral CA proteins organization is required to achieve effective nuclear translocation. The mechanistic of this process remains undefined due to the lack of a technology capable to visualize potential CA sub-complexes in association with the viral DNA in the nucleus of HIV-1-infected cells.By the means of state-of-the-art technologies (HIV-1 ANCHOR system combined with CLEM), our study shows that remodeled viral complexes retain multiple CA proteins but not intact core or only a single CA monomer. These viral CA complexes associated with the retrotranscribed DNA can be observed in the outer and inner side of the NE, and they represent potential PIC.Thus, our study shed light on critical early steps characterizing HIV-1 infection, thereby revealing novel, therapeutically exploitable points of intervention. Furthermore, we developed and provided a powerful tool enabling direct, specific and high-resolution visualization of intracellular and intranuclear HIV-1 subviral structures.

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The forces driving clonal expansion of the HIV-1 latent reservoir

Cited by 63 publications

References 118 publications

Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

Phenotypic analysis of the unstimulated in vivo HIV CD4 T cell reservoir

The Atlas of the In Vivo HIV CD4 T Cell Reservoir

Remodeling of the core leads HIV-1 pre-integration complex in the nucleus of human lymphocytes

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