Journal: PLOS Pathogens

Article Title: Targeting the MR1-MAIT cell axis improves vaccine efficacy and affords protection against viral pathogens

doi: 10.1371/journal.ppat.1011485

Figure Lengend Snippet: ( A ) B6-MAIT CAST mice were injected i . p . with PR8 (n = 12), 5-OP-RU (n = 12), or both (n = 12). Animals (n = 4/timepoint) were sacrificed 1.5, 6 or 24 hours later, and indicated cytokines were quantified in serum samples. A heatmap was generated to visualize average values at specified timepoints. ( B ) B6-MAIT CAST mice (n = 7/group in four independent experiments) were injected i . p . with 5-OP-RU, recombinant mouse IFN-β (rmIFNβ), 5-OP-RU plus rmIFNβ, or 5-OP-RU plus PR8. ( C ) In two additional experiments, mice (n = 2/group) received 5-OP-RU, rmIFNα1, rmIFNβ and/or PR8 in indicated combinations. Pulmonary MR1 tetramer + cell percentages among TCRβ + events ( B ) and fold changes in MAIT cell frequencies relative to 5-OP-RU treatment alone ( C ) are shown. Black circles and red triangles each represent an individual mouse used in experiment 1 and experiment 2, respectively. ( D ) Additional cohorts (n = 6/group in two independent experiments) received an IFNAR-blocking monoclonal antibody or isotype control twice before and twice after i . p . immunization with PR8 and 5-OP-RU as detailed in Materials and Methods. Peritoneal, pulmonary, hepatic and splenic MAIT cells were enumerated by flow cytometry on day 3 post-immunization. Unpaired t -tests were employed for statistical comparisons, and *, ** and *** denote differences with p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively.

Article Snippet: In a few experiments, mice were injected i . p . with 50 μg of the Toll-like receptor (TLR)3 agonist polyinosinic-polycytidylic acid [poly (I:C)] (InvivoGen), 50 μg of the TLR7 agonist imiquimod (InvivoGen), 10 4 units of recombinant mouse IFN-α1 (rmIFNα1) (R&D Systems), 1 μg of recombinant mouse IFN-β (rmIFNβ) (R&D Systems), or a combination of rmIFNα1 and rmIFNβ, all prepared in PBS.

Techniques: Injection, Generated, Recombinant, Blocking Assay, Control, Flow Cytometry

Journal: Nature communications

Article Title: Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.

doi: 10.1038/s41467-019-13940-6

Figure Lengend Snippet: Fig. 5 Therapeutic RDV reduces replication and pathology. Percent starting weight of 10–12-week-old female Ces1c−/−hDPP4 mice infected with 5E + 04 pfu MERS M35C4 and treated with a subcutaneous vehicle for RDV (N = 13) or remdesivir (RDV, 25 mg/kg, N = 14) BID beginning 1 dpi or b vehicle for LPV/RTV-IFNb (N = 15), LPV/RTV-IFNb low (N = 16) or LPV/RTV-IFNb high (N = 16) beginning 1 dpi. Oral vehicle or lopinavir/ritonavir (160/40 mg/kg) was administered orally once daily. IFNb low (1x human equivalent dose of 1.6 MIU/kg) and high (25x human equivalent dose of 40 MIU/kg) or PBS vehicle were administered via subcutaneous injection every other day. Asterisks indicate statistical differences by two-way ANOVA with Tukey’s multiple comparison test. c Lung hemorrhage 6 dpi for all animals in a, b scored on a scale of 0–4, where 0 is a normal pink healthy lung and 4 is a diffusely discolored dark red lung. d MERS-CoV lung titer 6 dpi in mice as described in a, b. Asterisks indicate statistical significance (N group described in a and b, P < 0.05) by one-way ANOVA with Kruskal–Wallis test for (c, d). Data for a–d are compiled from two independent experiments. For the box and whisker plots, the boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range. e Representative photomicrographs of MERS-CoV antigen (brown) and hematoxylin stained nuclei (blue) in mouse lung tissue sections from 6 dpi. The black bar is 100 µM.

Article Snippet: Recombinant mouse IFNb protein was purchased from R&D Systems (8234-MB/CF, 1.2 × 109 IU/mg calibrated against Murine IFN-beta WHO International Standard) for the in vivo studies and reconstituted in PBS.

Techniques: Infection, Injection, Comparison, Whisker Assay, Staining