Journal: bioRxiv

Article Title: Interleukin-27-induced HIV-resistant dendritic cells suppress reveres transcription following virus entry in an SPTBN1, Autophagy, and YB-1 independent manner

doi: 10.1101/2023.06.12.544550

Figure Lengend Snippet: (A) iDC or 27DC were either mock-infected or infected with HIV-1 AD8 virion containing Vpr-Blam for 2 hours. The cells were loaded with the fluorescent dye CCF2. The cleaved substrate of CCF2-AM by Blam was measured by excitation of the substrate at 410 nm and emission shift from green (520nm) to blue (450 nm) dye. As a positive control for suppression of infection/fusion, cells were treated with 10 nM TAK779 before adding HIV and then infected with the HIV. The dot plot is representative of 4 independent experiments. (B) Percentages of fusion were quantified by detecting CCF2, and the results shown are mean ± SE from four independent studies. (C) iDC and 27DC were infected with pseudotyped HIV-Luc-VSVG for 2 hours and then incubated for 48 hours. Luciferase activity in HIV-Luc-infected iDC and 27DC was measured. The data shows mean ± SD. (D and E) HIV-1 AD8 -infected iDC and 27DC were cultured for 24 hours at 37°C. Genomic DNA was extracted and subjected to quantify proviral DNA copy numbers. Primer/probes against the early RT products (D) and the late RT products (E) were utilized . The result is representative of three different donors; data indicates means ± SD (n=3). *: p <0.05, **: p <0.01, and ***: p <0.001.

Article Snippet: To suppress the virus binding TAK779 (Bio-Techne Corp., Minneapolis, MN, USA) was supplemented during the infection.

Techniques: Infection, Positive Control, Incubation, Luciferase, Activity Assay, Cell Culture

Journal:

Article Title: Inhibition of Human Immunodeficiency Virus Replication by a Dual CCR5/CXCR4 Antagonist

doi: 10.1128/JVI.78.23.12996-13006.2004

Figure Lengend Snippet: PCR-based detection of virus (HIV-1 NL4.3 and HIV-1 ADA) entry into U87.CD4.CXCR4 and U87.CD4.CCR5 cells. Two hours after infection with HIV-1, total DNA was extracted from the cells and analyzed by PCR with HIV-1-specific primers from the LTR R/U5 region. Lane 1, positive control; lane 2, 200 μM AMD3451; lane 3, 1 μM AMD3100; lane 4, 2 μM TAK-779. DNA recovery was controlled by PCR with β-actin-specific primers. The data shown are from one representative experiment out of two.

Article Snippet: As a control for the viral entry assays, AMD3100 (1 μM) (but not TAK-779 [Takeda, Osaka, Japan]) inhibited the entry of the X4 virus, whereas TAK-779 (2 μM) (but not AMD3100) inhibited the entry of R5 virus (Fig. ).

Techniques: Infection, Positive Control

Journal:

Article Title: Inhibition of Human Immunodeficiency Virus Replication by a Dual CCR5/CXCR4 Antagonist

doi: 10.1128/JVI.78.23.12996-13006.2004

Figure Lengend Snippet: (A) Opposite effects of AMD3451 and AMD3100 on the binding of anti-CXCR4 (12G5) MAb to Molt-4 cells. After preincubation for 30 min on ice with or without the compounds at the indicated concentrations, the cells were stained with phycoerythrin-conjugated 12G5 MAb and analyzed by flow cytometry. The upper panel shows the aspecific background fluorescence of Molt-4 cells, as determined by an isotype control MAb. The second panel shows Molt-4 cells stained with the antibody in the absence of any compound. The MFI of the cell population is indicated in each histogram. The data shown are from one representative experiment out of four. (B) Effect of AMD3451 on competition binding of anti-CXCR4 antibody 12G5. The binding assay was performed with COS-7 cells transfected with wild-type (wt) CXCR4 with 125I-12G5 as a radioligand. The data are shown as means ± standard errors of the means (n = 3).

Article Snippet: As a control for the viral entry assays, AMD3100 (1 μM) (but not TAK-779 [Takeda, Osaka, Japan]) inhibited the entry of the X4 virus, whereas TAK-779 (2 μM) (but not AMD3100) inhibited the entry of R5 virus (Fig. ).

Techniques: Binding Assay, Staining, Flow Cytometry, Fluorescence, Transfection

Journal:

Article Title: Analysis of Binding Sites for the New Small-Molecule CCR5 Antagonist TAK-220 on Human CCR5

doi: 10.1128/AAC.49.11.4708-4715.2005

Figure Lengend Snippet: Molecular models of TAK-220-huCCR5 and TAK-779-huCCR5 complexes and chemical structures of the antagonists. The binding pocket viewed from the extracellular side of the receptor is shown for each complex. The key binding-site residues are represented by Corey-Pauling-Koltun (CPK) models with residue names. Main chain atoms are colored green, and acidic, basic, hydrophilic, and hydrophobic side chains are colored red, cyan, magenta, and yellow, respectively. TAK-220 and TAK-779 are color-coded for each atom (white for carbon, red for oxygen, and cyan for nitrogen).

Article Snippet: The initial structures of TAK-220 and TAK-779 were constructed with Insight II (version 2000.1, Accelrys Inc., San Diego, CA) and subjected to energy minimization with Discover.

Techniques: Binding Assay

Journal:

Article Title: Analysis of Binding Sites for the New Small-Molecule CCR5 Antagonist TAK-220 on Human CCR5

doi: 10.1128/AAC.49.11.4708-4715.2005

Figure Lengend Snippet: Dose-dependent entry inhibition by TAK-220 and TAK-779. A pseudotyped reporter virus infection assay was performed as described in the legend of Fig. ​Fig.2A2A except for the addition of CCR5 antagonists (TAK-220 or TAK-779) at indicated concentrations. Data are shown as average ± SEM from multiple experiments, and the number of experiments is indicated in parentheses. Asterisks indicate a statistically significant decrease relative to virus entry without CCR5 antagonists (*, P <0.05; **, P <0.01; Student's t test).

Article Snippet: The initial structures of TAK-220 and TAK-779 were constructed with Insight II (version 2000.1, Accelrys Inc., San Diego, CA) and subjected to energy minimization with Discover.

Techniques: Inhibition, Infection

Journal:

Article Title: Analysis of Binding Sites for the New Small-Molecule CCR5 Antagonist TAK-220 on Human CCR5

doi: 10.1128/AAC.49.11.4708-4715.2005

Figure Lengend Snippet: Effects of mouse-type mutations in huCCR5 on entry inhibition of pseudotyped reporter virus by TAK-220 and TAK-779. U87-CD4 cells transiently expressing wild-type huCCR5 or huCCR5 mutants were infected as described in Fig. ​Fig.2A2A except for the presence or absence of CCR5 antagonists (TAK-220 or TAK-779) at the concentrations indicated. Results are expressed as in Fig. ​Fig.33.

Article Snippet: The initial structures of TAK-220 and TAK-779 were constructed with Insight II (version 2000.1, Accelrys Inc., San Diego, CA) and subjected to energy minimization with Discover.

Techniques: Inhibition, Expressing, Infection

Journal:

Article Title: Analysis of Binding Sites for the New Small-Molecule CCR5 Antagonist TAK-220 on Human CCR5

doi: 10.1128/AAC.49.11.4708-4715.2005

Figure Lengend Snippet: Effects of Ala substitutions in huCCR5 on entry inhibition of pseudotyped reporter virus by TAK-220 and TAK-779. U87-CD4 cells transiently expressing wild-type huCCR5 or huCCR5 mutants were infected as described in Fig. ​Fig.2A2A except for the presence or absence of CCR5 antagonists (TAK-220 or TAK-779) at the concentrations indicated. Luciferase activity was measured as described in Fig. ​Fig.2A.2A. Results are expressed as in Fig. ​Fig.33.

Article Snippet: The initial structures of TAK-220 and TAK-779 were constructed with Insight II (version 2000.1, Accelrys Inc., San Diego, CA) and subjected to energy minimization with Discover.

Techniques: Inhibition, Expressing, Infection, Luciferase, Activity Assay