Journal: Structure (London, England : 1993)

Article Title: Structural insights into the binding of SARS-CoV-2, SARS-CoV, and hCoV-NL63 spike receptor-binding domain to horse ACE2

doi: 10.1016/j.str.2022.07.005

Figure Lengend Snippet:

Article Snippet: Trichoplusia ni cells , ATCC , PTA-9384.

Techniques: Recombinant, Luciferase, Expressing, Plasmid Preparation, Software, Spectrophotometry

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: ( A ) Differentiated THP-1 (dTHP1) cells were infected with BCG at the MOI of 1 and then stimulated or not with 0.5, 5, 50 μg/ml of MSU for 3 and 5 days. The results are expressed as means ± Standard Deviation (SD) of CFU values performed in triplicate and are representative of three independent experiments. * p ≤ 0.001 in comparison with non-stimulated control cells. ( B ) Stimulation of human macrophages with MSU enhances phagocytosis of BCG. Differentiated THP-1 cells were exposed to BCG at the MOI of 1 for 3 hour in the presence or not of 0.05, 0.5, 5 μg/ml MSU. Results are expressed as mean ± SD of CFU values performed in triplicate and are representative of two independent experiments. * p < 0.05 in comparison with non-stimulated control cells.

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: Infection, Standard Deviation

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: ( A ) dTHP-1 cells were incubated with 3 μM Fluo-3/AM at 37°C for 1 hour in the dark and were stimulated with 5 μg/ml MSU. After stimulation, fluorescence emission was continuously monitored for 30 minutes and expressed as to determine relative alteration in intensity. ( B ) dTHP-1 cells were incubated for 1 hour at the dark with 10 μM DCF, or with 3 μM Fluo-3/AM (inset of the figure), and were stimulated with 5 μg/ml MSU. Ca 2+ dependence ROS generation was assessed by adding 20 μM BAPTA-AM or 3 mM EGTA 30 minutes and 15 minutes before MSU addition, respectively. Fluorescence emission was monitored at 20 minutes after stimulation. Results are expressed as mean ± SD of arbitrary fluorescence units performed in triplicate and are representative of three separate experiments. * p < 0.01 in comparison with non-stimulated control cells

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: Incubation, Fluorescence

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: ( A ) dTHP-1 cells were infected with NHS labelled BCG at the MOI of 1 and then stimulated overnight with 5 μg/ml of MSU, in the presence or absence 10 μM Chloroquine. Results are expressed in terms of mean ± SD of arbitrary fluorescence units of triplicate values and are representative of two independent experiments. * p < 0.001 in comparison with non-stimulated control cells, ° p < 0.001 in comparison with MSU stimulated cells. ( B ) dTHP-1 cells were infected with BCG, stimulated overnight with 5 μg/ml of MSU, in the presence or absence of 10 μM Chloroquine (Cq), and then labelled with 1 μM Lysosensor green DND 189. Results are expressed as mean ± SD of pH values from cultures performed in triplicate and are representative of two independent experiments. * p < 0.001 in comparison with non-stimulated control cells, ° p < 0.001 in comparison with BCG infected MSU stimulated cells ( C ) Protease activity of BCG infected dTHP-1 cells stimulated overnight with 5 μg/ml of MSU was analysed by loading of 10 μg/ml DQ red BSA for 2h at 37°C. Results are expressed as mean ± SD of triplicate values and are representative of three independent experiments. * p ≤ 0.01 in comparison with non-stimulated control cells. ( D ) Confocal microscopy representative images out of 10 per condition showing the increase of Auramine-stained BCG (green) residing in LAMP-3 positive vacuoles (red) after stimulation with 5 μg/ml of MSU. One representative experiment out of three is shown. ( E ) Summary of the mean percentage ± standard deviation (SD) of BCG co-localizing in LAMP-3-positive vacuoles determined by acquiring at least 10 images per condition and by counting ≥ 50 dTHP-1 cells per sample, after stimulation or not with MSU. Three different experiments were assessed. * p < 0.05 in comparison with BCG-infected cells (data were analyzed using the unpaired Student’s t -test).

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: Infection, Fluorescence, Activity Assay, Confocal Microscopy, Staining, Standard Deviation

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: dTHP-1 cells were infected or not with BCG at the MOI of 5 and then labelled with 10 μM DCF for 60 min. Thereafter, cells were washed twice, stimulated or not overnight with 5 μg/ml MSU in the presence or absence of 10 μM Chloroquine (Cq). Results are expressed as means ± SD of arbitrary fluorescence units of triplicate values and are representative of two independent experiments. * p < 0.001 in comparison with non-stimulated control cells. ° p < 0.001 in comparison with MSU stimulated cells or with MSU-stimulated BCG-infected cells.

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: Infection, Fluorescence

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: BCG infected dTHP-1 cells were stimulated or not with 5 μg/ml MSU and CFU assays were performed at day 3 post-infection. ( A ) In order to ascertain whether phagolysosome maturation was responsible for intracellular mycobacterial killing, 10 μM chloroquine (Cq) or 20 mM NH 4 Cl was added to BCG-infected cells together with MSU. Results are expressed as mean ± SD of CFU values performed in triplicate and are representative of two independent experiments. * p < 0.001 in comparison with non-stimulated control cells. ( B ) In order to ascertain the role of ROS in intracellular mycobacterial killing, 100 U/ml PEG-catalase was added to BCG infected cells together with MSU. Results are expressed as mean ± SD of CFU values performed in triplicate and are representative of two independent experiments. * p < 0.001 in comparison with non-stimulated control cells.

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: Infection

Journal: PLoS ONE

Article Title: Monosodium Urate Crystals Promote Innate Anti-Mycobacterial Immunity and Improve BCG Efficacy as a Vaccine against Tuberculosis

doi: 10.1371/journal.pone.0127279

Figure Lengend Snippet: ( A ) Diagram showing the course of in vitro preincubation experiment. THP-1 cells were cultured with 5 or 50 μg/ml MSU for 3 days. Thereafter, the medium was changed to remove MSU stimulus and cells cultured for further 4 days. Finally, cells were exposed to BCG at the MOI of 10 for 3 hours (T0), washed and cultured for further 3 days (T3). ( B ) Intracellular mycobacterial growth was monitored in BCG infected THP-1 cells prestimulated or not with 5 or 50 μg/ml MSU. Results are shown as mean ± SD of CFU values performed in triplicate and are representative of three independent experiments. ° p < 0.05 and * p < 0.001 in comparison with same time non pre-stimulated control cells. ( C ) Intracellular mycobacterial growth was monitored in THP-1 cells, pre-stimulated or not with 50 μg/ml MSU, and exposed or not to 10 μM chloroquine after BCG infection for 3 days. Results are shown as mean ° SD of CFU values performed in triplicate. * p < 0.05 and ° p < 0.01 in comparison with same time non pre-stimulated control cells. ( D ) IL-1β production in the supernatant of BCG infected THP-1 cells pre-stimulated or not with either 5 or 50 μg/ml MSU. Results are shown as mean ± SD of values performed in triplicate and are representative of three independent experiments. * p < 0.05 in comparison with same time non pre-stimulated control cells.

Article Snippet: After removal of extra-cellular bacilli, cells were stimulated or not with MSU (Enzo Life Sciences Inc.) at the concentration of 0.5, 5, 50 μg/ml and colony forming unit (CFU) assays were performed at day 3 and 5 post-infection, as previously described [ ].

Techniques: In Vitro, Cell Culture, Infection