HIV-1 envelope-CXCR4 signaling triggers cofilin activation to promote cortical actin dynamics and HIV nuclear migration.

<p>(A) HIV-1 envelope binding to CD4 and the chemokine coreceptor, CXCR4, triggers membrane fusion and signaling transduction. The initial viral contact with CD4 and then CXCR4 may trigger rapid actin polymerization to facilitate CD4/CXCR4 cocapping for fusion and entry. Following fusion, the viral preintegration complex (PIC) may be directly anchored onto F-actin to facilitate reverse transcription. Subsequent actin activity mediated by cofilin activation through CXCR4 promotes viral nuclear migration. (B) Model of HIV PIC migration along the cortical actin filaments. It is possible that cofilin activation increases actin treadmilling, which promotes the movement of the viral PIC across the cortical actin barrier, allowing PIC to gain access to the perinuclear or nuclear region. The number is arbitrarily assigned to an actin monomer to demonstrate the actin movement during treadmilling. (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000520#ppat-1000520-g003" target="_blank">Figure 3</a> is modified from <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000520#ppat.1000520-Yoder1" target="_blank">[44]</a> with permission).</p

Chemokine receptor signaling pathways.

<p>SDF-1 binding to CXCR4 or RANTES/MIP-1α/MIP-1β binding to CCR5 activates G proteins (Gα particularly Gα_i, Gα_q, and Gβγ) and multiple downstream pathways. (A) Gα_q activates phospholipases such as phospholipase C-γ (PLC-γ), which hydrolyzes phosphatidylinositol-4,5-biphosphate (PIP2) to generate inositol triphosphate (IP3) and diacylglyerol (DAG), triggering calcium influx and the activation of kinases such as protein kinase C (PKC). (B) Gα_i activates phospholipases, phosphodiesterases, and the lipid kinase PI3K via Src-family kinases. Gβγ also activates PI3Kγ. PI3K activation stimulates downstream targets such as protein kinase B (PKB/Akt), NF-κB, mitogen/extracellular signal-regulated kinase (MEK-1), and extracellular signal-regulated kinase (ERK1/2). PI3K also triggers the tyrosine phosphorylation of focal adhesion complex components such as proline-rich tyrosine kinase (Pyk2), paxillin, Crk, and p130Cas. (C) GTP-bound Gβγ stimulates guanine nucleotide exchange factors (GEFs) such as TIAM1 and PREX1 specific for the Rho family GTPases (Rac/CDC42/RhoA). These GTPases activate pathways regulating cytoskeleton: Rac activates p21-activated kinase (PAK), which then activates LIM kinase (LIMK), leading to cofilin phosporylation and actin polymerization. CDC42 promotes actin assembly through the Wiskott-Aldrich Syndrome family protein (WASP) and actin-nucleating protein Arp2/3. RhoA activates Rho kinase (ROCK) , leading to myosin light-chain (MLC) phosporylation and microtubule rearrangement. (D) SDF-1 may also trigger Gα_i-independent activation of the JAK-STAT pathways.</p

Components of the chemokine coreceptor signaling pathways activated by HIV-1 envelope.

<p>HIV-1 gp120 binding to CXCR4 or CCR5 activates a number of signaling molecules common to chemokine-mediated signaling pathways, including (A) PLC-γ-dependent calcium flux and NFAT nuclear translocation; (B) PI3K-dependent activation of FAK, PyK2, AKT, and ERK1/2; (C) the downstream targets of the Rho family GTPases such as LIMK1 and cofilin for actin rearrangement.</p

Additional file 1 of Cumulative triglyceride-glucose index is a risk for CVD: a prospective cohort study

Additional file 1: Fig. S1. Kaplan-Meier incidence rate of stroke and MI by TyG index. a quartiles of cumulative TyG index for stroke. b quartiles of cumulative TyG index for MI. c exposure duration with a higher TyG index for stroke. d exposure duration with a higher TyG index for MI

Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from -3

F AnlO-mediated cytolysis by 6-BOCD. HEK293T cells were cotransfected with pCMVΔR8.2 and either pNL-GFP-RRE-SA (labeled as GFP) or pNL-AnlO-GFP-RRE-SA (labeled as AnlO-GFP) in the absence or presence of various doses of 6-BOCD. Increases in viable GFP cells were measured by flow cytometry. (C) Lack of effect of 6-BOCD on α-hemolysin. Cells were similarly cotransfected with pCMVΔR8.2 and pNL-α-HL-GFP-RRE-SA (labeled as α-HL-GFP) in the absence or presence of various doses of 6-BOCD. (D) No inhibition of 6-BOCD on viral infectivity. Lentiviral particles, vNL-GFP-RRE-SA, were generated by cotransfection of HEK293T cells with pNL-GFP-RRE-SA, pCMVΔR8.2, and pCMV-VSV-G in the absence or presence of different concentrations of 6-BOCD. The resulting viral particles were used to infect an HIV-1-positive cell, J1.1 (using an equal p24 level of viruses, 150 ng p24 per million cells). The percentage of GFP positive J1.1 cells was used as an indicator for viral infectivity. (E) The Rev-dependent AnlO lentiviral vector is suicidal in HIV-1-positive cells. The HIV-1-positive cell, J1.1, was infected with vNL-AnlO-RRE-SA (300 ng p24 per million cells). As a control, HIV-1-negative Jurkat cells were identically infected. Following infection, cells were harvested at different times and total cellular DNA was extracted and PCR amplified with primers for the gene (AnlO). As a control, the DNA was also amplified with primers for the cellular β-actin pseudogene (β-actin) to ensure that the same number of cells was used.<p><b>Copyright information:</b></p><p>Taken from "Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from "</p><p>http://www.retrovirology.com/content/5/1/36</p><p>Retrovirology 2008;5():36-36.</p><p>Published online 25 Apr 2008</p><p>PMCID:PMC2391154.</p><p></p

Neutralization of ROS with MnTBAP abrogated active TGF-β and reduced CD25Foxp3T cells

<p><b>Copyright information:</b></p><p>Taken from "Endogenous TGF-β activation by reactive oxygen species is key to Foxp3 induction in TCR-stimulated and HIV-1-infected human CD4CD25T cells"</p><p>http://www.retrovirology.com/content/4/1/57</p><p>Retrovirology 2007;4():57-57.</p><p>Published online 9 Aug 2007</p><p>PMCID:PMC2096626.</p><p></p> CD4CD25T cells were cultured with anti-CD3 and anti-CD28 antibodies in the presence or absence of MnTBAP (100 μM) for 3 and 5 days. The intracellular ROS production was determined by DHE staining. Active TGF-β in the supernatants was determined with ELISA. The intracellular Foxp3 protein was determined by FACS staining. . A representative overlay of histograms of ROS in the cultured T cells with (+MnTBAP) and without (-MnTBAP) MnTBAP at days 3 and 5. The filled histograms were from unlabeled cells as negative control for DHE staining. The experiment was repeated three times with similar results. . The values are shown as the Mean ± SD of active TGF-β1 in the culture supernatants at days 3 and 5 (n = 2). . MnTBAP reduced CD25Foxp3T cells. Each symbol represents one individual

Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from -4

K293T cells were cotransfected with pNL-AnlO-GFP-RRE-SA, pCMVΔ8.2, and the M-tropic envelope construct, pCAGGSSF162gp160, in the presence of 6-BOCD. Viruses were harvested, concentrated, and used to infect human macrophages. (B) Specific killing of HIV-1-positive macrophages by the Rev-dependent AnlO lentiviral vector. Human macrophages were derived from peripheral monocytes. Cells were not infected (Cell) or infected with NL4-3.HSA.R+E-(VSV-G) (m.o.i. 0.1). Following HIV infection, at 24 hours, HIV-1-infected cells were super-infected with the lentivirus vNL-AnlO-RRE-SA (approximate m.o.i. 0.5 – 1) or with the same lentivirus using a 10-fold higher dosage (*). HIV-1-infected cells were stained with a PE-labeled rat monoclonal antibody against mouse CD24 (HSA) and analyzed by flow cytometry at 10 days post infection with HIV-1. (C) Undetectable cytolytic activity of the Rev-dependent AnlO lentiviral vector in un-infected macrophages. To determine whether vNL-AnlO-RRE-SA can non-specifically kill un-infected macrophages, cells were similarly infected with highly concentrated virus (m.o.i. 5 – 10). Following infection for two weeks, cells were harvested at different times and analyzed by propidium iodide (PI) staining and flow cytometry for cytolysis. As controls, cells were also mock infected with medium, or the same dose of an empty vector virus, vNL-RRE-SA. Cells were also treated with puromycin (500 ng/ml) to induce non-specific cytolysis for the validation of PI staining and flow cytometry analysis.<p><b>Copyright information:</b></p><p>Taken from "Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from "</p><p>http://www.retrovirology.com/content/5/1/36</p><p>Retrovirology 2008;5():36-36.</p><p>Published online 25 Apr 2008</p><p>PMCID:PMC2391154.</p><p></p

Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from -7

293T cells were cotransfected with pNL-GFP-RRE-SA, pCMVΔ8.2, and the VSV-G construct. Viruses were harvested, concentrated, and used to infect a human T cell line, CEM-SS, as well as primary human macrophages. (B) Specificity of the Rev-dependent lentiviral vector in HIV-1-positive T cells. CEM-SS cells were not infected () or infected with NL4-3.HSA.R+E-(VSV-G) (, 500 ng p24 per million cells), a VSV-G pseudotyped HIV-1 strain with the murine heat-stable antigen CD24 (HSA) gene inserted into the region that allows HIV-1-positive cells to be monitored by surface staining of HSA. At 24 hours, cells were superinfected with lentivirus vNL-GFP-RRE-SA (, m.o.i. 10). For comparison, cells were also singly infected with either vNL-GFP- RRE-SA () or NL4-3.HSA.R+E-(VSV-G) (). At 72 hours, cells were harvested, stained with a PE-labeled rat monoclonal antibody against mouse CD24 (HSA), and then analyzed on a flow cytometer for both HSA and GFP expression. Isotype staining was not shown. (C) Specificity of the Rev-dependent lentiviral vector in HIV-1-positive macrophages. Human macrophages were derived from peripheral monocytes by culturing in 10 ng/ml M-CSF for two weeks. Cells were not infected () or infected with HIV- 1(AD8) (, 380 ng p24 per million). At 24 hours, cells were superinfected with lentivirus vNL-GFP-RRE-SA (, m.o.i. 10). For comparison, cells were also singly infected with either vNL-GFP-RRE-SA () or HIV-1(AD8) (). At 72 hours, cells were harvested and analyzed on a flow cytometer for GFP expression. (D) Fluorescent microscopy of GFP expression in macrophages infected with HIV-1 and the Rev-dependent GFP lentiviral vector. Cells in (C, h) were also examined with fluorescent microscope. The left and right panels show the bright and green fluorescent fields of the same cells. Red arrows indicate an HIV-1-infected cell expressing the GFP protein.<p><b>Copyright information:</b></p><p>Taken from "Selective killing of HIV-1-positive macrophages and T cells by the Rev-dependent lentivirus carrying from "</p><p>http://www.retrovirology.com/content/5/1/36</p><p>Retrovirology 2008;5():36-36.</p><p>Published online 25 Apr 2008</p><p>PMCID:PMC2391154.</p><p></p

The cell-free supernatant from TCR stimulated CD4CD25T cell culture contained ROS

<p><b>Copyright information:</b></p><p>Taken from "Endogenous TGF-β activation by reactive oxygen species is key to Foxp3 induction in TCR-stimulated and HIV-1-infected human CD4CD25T cells"</p><p>http://www.retrovirology.com/content/4/1/57</p><p>Retrovirology 2007;4():57-57.</p><p>Published online 9 Aug 2007</p><p>PMCID:PMC2096626.</p><p></p> CD4CD25T cells were cultured with anti-CD3 and anti-CD28 for the indicated time points and ROS in the culture supernatant was detected using DCFH-DA as described in the Method section. Oxidation of DCFH-DA was measured using a spectrofluorometer at wavelength 485/535 nm and is represented as fluorescent units. The experiment was repeated twice with similar results