Faculty Opinions recommendation of Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing.

Abstract We employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5′ and 3′ LTRs of integrated HIV-1 proviral DNA copies from latently infected human CD4+ T-cells. Comprehensive assessment of whole-genome sequencing of HIV-1 eradicated cells ruled out any off-target effects by our CRISPR/Cas9 technology that might compromise the integrity of the host genome and further showed no effect on several cell health indices including viability, cell cycle and apoptosis. Persistent co-expression of Cas9 and the specific targeting guide RNAs in HIV-1-eradicated T-cells protected them against new infection by HIV-1. Lentivirus-delivered CRISPR/Cas9 significantly diminished HIV-1 replication in infected primary CD4+ T-cell cultures and drastically reduced viral load in ex vivo culture of CD4+ T-cells obtained from HIV-1 infected patients. Thus, gene editing using CRISPR/Cas9 may provide a new therapeutic path for eliminating HIV-1 DNA from CD4+ T-cells and potentially serve as a novel and effective platform toward curing AIDS.

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Characterization of interactions between transcription factors and a regulatory region spanning nt −320 to −281 of the HIV-1 LTR in T-lymphoid and non-T-lymphoid cells

Journal of Biomedical Science ◽

10.1007/bf02256580 ◽

2002 ◽

Vol 9 (1) ◽

pp. 68-81

Author(s):

Lloyd A. Pereira ◽

Melissa J. Churchill ◽

Andrew G. Elefanty ◽

Theo Gouskos ◽

Paul F. Lambert ◽

...

Keyword(s):

Transcription Factors ◽

Regulatory Region ◽

Lymphoid Cells ◽

Hiv 1

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HIV-1-induced cytokines deplete homeostatic ILCs and expand TCF7-dependent memory NK cells: Supplemental Figures and Tables

10.1101/221010 ◽

2017 ◽

Cited By ~ 1

Author(s):

Yetao Wang ◽

Kyle Gellatly ◽

Sean McCauley ◽

Pranitha Vangala ◽

Kyusik Kim ◽

...

Keyword(s):

Nk Cells ◽

Inflammatory Cytokines ◽

Nk Cell ◽

Ex Vivo ◽

Developmental Trajectory ◽

Lymphoid Cells ◽

Infected People ◽

Cell Induction ◽

Hiv 1 ◽

Memory Nk Cells

HIV-1-infected people who take medications that suppress viremia, preserve CD4+ T cells, and prevent AIDS, have chronic inflammation with increased cardiovascular mortality. To investigate the etiology of this inflammation, the effect of HIV-1 on innate lymphoid cells (ILCs) and NK cells was examined. Homeostatic ILCs in blood and intestine were depleted permanently. NK cells were skewed towards a memory subset. Cytokines that are elevated during HIV-1 infection reproduced both abnormalities ex vivo. Pseudotime analysis of single NK cell transcriptomes revealed a developmental trajectory towards a subset with expression profile, chromatin state, and biological function like memory T lymphocytes. Expression of TCF7, a WNT transcription factor, increased over the course of the trajectory. TCF7 disruption, or WNT inhibition, prevented memory NK cell induction by inflammatory cytokines. These results demonstrate that inflammatory cytokines associated with HIV-1 infection irreversibly disrupt homeostatic ILCs and cause developmental shift towards TCF7+ memory NK cells.

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Inhibition of HIV-1 infection of primary CD4+ T-cells by gene editing of CCR5 using adenovirus-delivered CRISPR/Cas9

Journal of General Virology ◽

10.1099/vir.0.000139 ◽

2015 ◽

Vol 96 (8) ◽

pp. 2381-2393 ◽

Cited By ~ 105

Author(s):

Chang Li ◽

Xinmeng Guan ◽

Tao Du ◽

Wei Jin ◽

Biao Wu ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Gene Editing ◽

Hiv 1

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Characterization of Simian Immunodeficiency Virus SIVSM/Human Immunodeficiency Virus Type 2 Vpx Function in Human Myeloid Cells

Journal of Virology ◽

10.1128/jvi.01181-08 ◽

2008 ◽

Vol 82 (24) ◽

pp. 12335-12345 ◽

Cited By ~ 95

Author(s):

Caroline Goujon ◽

Vanessa Arfi ◽

Thomas Pertel ◽

Jeremy Luban ◽

Julia Lienard ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Simian Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Intracellular Localization ◽

Lymphoid Cells ◽

Viral Dna ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Human immunodeficiency virus type 2 (HIV-2)/simian immunodeficiency virus SIVSM Vpx is incorporated into virion particles and is thus present during the early steps of infection, when it has been reported to influence the nuclear import of viral DNA. We recently reported that Vpx promoted the accumulation of full-length viral DNA following the infection of human monocyte-derived dendritic cells (DCs). This positive effect was exerted following the infection of DCs with cognate viruses and with retroviruses as divergent as HIV-1, feline immunodeficiency virus, and even murine leukemia virus, leading us to suggest that Vpx counteracted an antiviral restriction present in DCs. Here, we show that Vpx is required, albeit to a different extent, for the infection of all myeloid but not of lymphoid cells, including monocytes, macrophages, and monocytoid THP-1 cells that had been induced to differentiate with phorbol esters. The intracellular localization of Vpx was highly heterogeneous and cell type dependent, since Vpx localized differently in HeLa cells and DCs. Despite these differences, no clear correlation between the functionality of Vpx and its intracellular localization could be drawn. As a first insight into its function, we determined that SIVSM/HIV-2 and SIVRCM Vpx proteins interact with the DCAF1 adaptor of the Cul4-based E3 ubiquitin ligase complex recently described to associate with HIV-1 Vpr and HIV-2 Vpx. However, the functionality of Vpx proteins in the infection of DCs did not strictly correlate with DCAF1 binding, and knockdown experiments failed to reveal a functional role for this association in differentiated THP-1 cells. Lastly, when transferred in the context of a replication-competent viral clone, Vpx was required for replication in DCs.

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COMMD1/Murr1 Reinforces HIV-1 Latent Infection through IκB-α Stabilization

Journal of Virology ◽

10.1128/jvi.03105-14 ◽

2014 ◽

Vol 89 (5) ◽

pp. 2643-2658 ◽

Cited By ~ 11

Author(s):

Manabu Taura ◽

Eriko Kudo ◽

Ryusho Kariya ◽

Hiroki Goto ◽

Kouki Matsuda ◽

...

Keyword(s):

Transcription Initiation ◽

Primary Infection ◽

Latent Infection ◽

Myeloid Cells ◽

Innate Immune ◽

Lymphoid Cells ◽

Inhibition Mechanism ◽

Major Barrier ◽

Infected Cells ◽

Hiv 1

ABSTRACTThe transcription factor NF-κB is important for HIV-1 transcription initiation in primary HIV-1 infection and reactivation in latently HIV-1-infected cells. However, comparative analysis of the regulation and function of NF-κB in latently HIV-1-infected cells has not been done. Here we show that the expression of IκB-α, an endogenous inhibitor of NF-κB, is enhanced by latent HIV-1 infection via induction of the host-derived factor COMMD1/Murr1 in myeloid cells but not in lymphoid cells by using four sets of latently HIV-1-infected cells and the respective parental cells. IκB-α protein was stabilized by COMMD1, which attenuated NF-κB signaling during Toll-like receptor ligand and tumor necrosis factor alpha treatment and enhanced HIV-1 latency in latently HIV-1-infected cells. Activation of the phosphoinositol 3-kinase (PI3K)–JAK pathway is involved in COMMD1 induction in latently HIV-1-infected cells. Our findings indicate that COMMD1 induction is the NF-κB inhibition mechanism in latently HIV-1-infected cells that contributes to innate immune deficiency and reinforces HIV-1 latency. Thus, COMMD1 might be a double-edged sword that is beneficial in primary infection but not beneficial in latent infection when HIV-1 eradication is considered.IMPORTANCEHIV-1 latency is a major barrier to viral eradication in the era of combination antiretroviral therapy. In this study, we found that COMMD1/Murr1, previously identified as an HIV-1 restriction factor, inhibits the proteasomal degradation of IκB-α by increasing the interaction with IκB-α in latently HIV-1-infected myeloid cells. IκB-α protein was stabilized by COMMD1, which attenuated NF-κB signaling during the innate immune response and enhanced HIV-1 latency in latently HIV-1-infected cells. Activation of the PI3K-JAK pathway is involved in COMMD1 induction in latently HIV-1-infected cells. Thus, the host-derived factor COMMD1 is beneficial in suppressing primary infection but enhances latent infection, indicating that it may be a double-edged sword in HIV-1 eradication.

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Expression of the Human Immunodeficiency Virus-Tat Gene in Lymphoid Tissues of Transgenic Mice Is Associated With B-Cell Lymphoma

Blood ◽

10.1182/blood.v94.1.275.413a30_275_282 ◽

1999 ◽

Vol 94 (1) ◽

pp. 275-282 ◽

Cited By ~ 43

Author(s):

Ramendra K. Kundu ◽

Frank Sangiorgi ◽

Lan-Ying Wu ◽

Paul K. Pattengale ◽

David R. Hinton ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Transgenic Mice ◽

B Cell ◽

Cultured Cells ◽

B Cell Lymphoma ◽

Lymphoid Cells ◽

Lymphoid Tissues ◽

Genetic Event ◽

Immunodeficiency Virus ◽

Hiv 1

The human immunodeficiency virus type 1 (HIV-1) Tat gene, a potent transactivator of viral and cellular genes, has been proposed as a key agent in the pathogenesis of acquired immune deficiency syndrome related disorders, including nonHodgkin’s lymphoma. In cultured cells, the HIV-1 Tat protein can induce the expression of the cytokines interleukin-6 (IL-6) and IL-10, which are known to induce proliferation and differentiation of lymphoid cells. Such alterations in cytokine expression, together with a secondary genetic event, are thought to ultimately lead to oncogenic transformation. To address the influence of Tat on lymphoid development in the context of the whole organism, we produced several transgenic mouse lines that express the Tat gene under the control of an actin promoter. We show here that this promoter directs expression to a variety of sites, including spleen, bone marrow, and lymph nodes. Approximately 25% to 30% of the Tat-transgenic population developed enlarged spleens within 1 year after birth. On histological examination, a significant number of spleens from Tat-transgenic mice exhibited malignant lymphoma of B-cell origin. IgG heavy chain rearrangement confirmed the clonal B-cell nature of these lymphoproliferations. In contrast, T-cell receptor genes exhibited a germline (unrearranged) structure. Reverse transcription polymerase chain reaction analysis of transgenic spleens revealed that mRNA encoding cytokines IL-6 and IL-10 was upregulated, suggesting a possible mechanism for the B-cell expansion in vivo.

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