Journal: medRxiv

Article Title: Polymorphism in IFNAR contributes to glucocorticoid response and outcome in ARDS and COVID-19

doi: 10.1101/2022.03.10.22272123

Figure Lengend Snippet: (A ) STAT1 expression in the lung after 4-day culture in the presence of IFN beta with or without hydrocortisone (HC). ( B) pSTAT1 expression in the same specimens as in A. ( C) Example photomicrographs showing higher STAT2 expression in a TT patient than in a CT patient and the effect of HC on its nuclear translocation. Most STAT2 remains in the cytoplasm of the CT patients, whereas nuclear expression is prominent in the TT patient. Indicated insets are shown in the bottom row. Arrows. ( D ) Combined results of all patients noting that two CT samples are excluded in the data as the patients were already under glucocorticoid treatment at the time of sample acquisition. Ns, not significant; *P<0.05; **P<0.01; and ***P<0.001

Article Snippet: The first stage antibodies were anti-alpha chain of the IFN alpha/beta receptor (St John’s Laboratory STJ112765) that was used 1:2000 and 1:5000 and anti-Stat1 (1:400, 9175S), anti-pStat1(1:100, 9167S) and anti-Stat2 (1:200, 72604S) all from Cell Signalling.

Techniques: Expressing, Translocation Assay

Journal: Frontiers in Immunology

Article Title: Adenovirus Encoding Tumor Necrosis Factor Alpha and Interleukin 2 Induces a Tertiary Lymphoid Structure Signature in Immune Checkpoint Inhibitor Refractory Head and Neck Cancer

doi: 10.3389/fimmu.2022.794251

Figure Lengend Snippet: Combining ICI with non-replicative Ad5-CMV-mTNFα/mIL-2 improves trafficking and activity of ICI refractory tumor-infiltrating-lymphocytes and is reflected by changes in the secondary lymphoid organs. (A, C) Flow cytometric quantification of percentages of CD4 + , CD8 + , Foxp3 + , T-bet + expressing tumor-infiltrating lymphocytes is shown for (A). anti-PD-1 (upper panel) and (C) anti-PD-L1 (lower panel) refractory MOC1 tumor-bearing C57BL/6J mice. (B, D) . Bulk tumor proinflammatory cytokine analysis as measured by bead based flex set. Data are presented as logarithmic values. (E, G) Flow cytometric analysis of CD4 + , CD8 + splenic memory compartment (naïve, effector memory [T EM ], central memory [T CM ]) from MOC1 anti-PD1 (upper) and anti-PD-L1 (lower) refractory mice as measured by ± expression of CD44 and CD62L. (F, H) Corresponding treatment group freshly resected whole splenocyte effector function analysis measured by overnight culture with mGranzyme B and mIFN-y dual-ELISpot. Flow cytometric and bulk cytokine analysis data represent two independent experiments. Statistical significance for each data set was evaluated by non-parametric unpaired t test. All data are shown as means ± SEM and significance is represented as *p < 0.05, **p < 0.01, ***p < 0.001, ****p <0.0001, ns, not significant.

Article Snippet: Samples were stained with Cytometric Bead Array Mouse Th1/Th2/Th17 Cytokine kit (560485, BD) and analyzed on BD Accuri C6 Cytometer (BD, Franklin Lakes, NJ, USA) with FCAP Array Software (BD, Franklin Lakes, NJ, USA) according to the manufacturer’s instructions.

Techniques: Activity Assay, Expressing, Enzyme-linked Immunospot