Journal: NPJ Vaccines

Article Title: Adenovectors encoding RSV-F protein induce durable and mucosal immunity in macaques after two intramuscular administrations

doi: 10.1038/s41541-019-0150-4

Figure Lengend Snippet: RSV-F-specific CD8+ T cells a – d and CD4+ T cells e–h expressing IFNγ a , e , TNFα b , f , or IL-2 c , g were determined in cells extracted from BAL, using polychromatic flow cytometry, after stimulation with a pool of 15-mer peptides overlapping by 11 amino acids, covering the RSV-F protein sequence. Shown are arithmetic group means with upper 95% confidence limit (upper whisker) over time, of individually background subtracted values. Animals received a homologous prime-boost with the Ad26/Ad35 mix (light gray), a homologous prime-boost with Ad26 (dark gray), or a heterologous prime-boost with Ad26 and Ad35 (black). Arrows indicate immunization time points. Polyfunctional composition of the cytokine response in CD8+ T cells e and CD4+ T cells h of individual animals 4 weeks post boost (study week 16) was determined as described before and is summarized in the pie charts. Responders as defined by Fisher’s exact test are indicated by an asterisk. Data from individual animals in Figures d , h are shown in corresponding order.

Article Snippet: Cells were washed in FACS staining buffer, then surface stained for 30 min at RT in the dark with 100 µl of a cocktail containing antibodies targeting CD3 (BD Biosciences, clone SP34-2, catalog number 557917, 1 µl), CD4 (BD Biosciences, catalog number 562843, clone L200, 0.5 µl), CD8 (BD Biosciences, catalog number 557834, clone SK1, 5 µl), CD28 (BD Biosciences, clone L293, catalog number 337181, 20 µl), and CD95 (clone DX2, catalog number 561978, 20 µl).

Techniques: Expressing, Flow Cytometry, Sequencing, Whisker Assay

Journal: bioRxiv

Article Title: Plasmacytoid dendritic cells (pDCs) display surface but not membrane-bound IgE across a broad range of total serum IgE levels

doi: 10.1101/2023.11.30.569292

Figure Lengend Snippet: B-cell (A-B) and pDC (C-D) gating strategies. All cells of interest were initially filtered using forward scatter (FSC) and side scatter (SSC) gating based on previously published location of B-cells and pDCs among PBMCs. (A) B cells were then identified with CD19 (Alexa488) and CD27 (PE) indicated by a red rectangular gate. (B) CD19 + B cells were then sub-selected for CD27 (PE) and Quilizumab in an effort to identify possible membrane bound IgE bearing B cells (upper right quadrant). In a similar manner (C) pDC were first identified with BDCA2 (Alexa488) and CD123 (PE) indicated by a red circle. (D) Finally, BDCA + pDC were then sub-selected for CD123 + (PE) and Quilizumab to identify possible membrane bound IgE bearing pDC (upper right quadrant).

Article Snippet: Target antibodies included quilizumab (mouse anti-human IgE, used at 10 µg/ml and obtained courtesy of Genentech, Inc. (San Francisco, CA, USA), anti-human FcεRI (0.5 µg/mL, AER-37, eBioscience/Thermo Fisher, Waltham, MA, USA), and appropriate isotype controls.

Techniques: Membrane

Journal: European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V

Article Title: Chip-based human liver–intestine and liver–skin co-cultures – A first step toward systemic repeated dose substance testing in vitro

doi: 10.1016/j.ejpb.2015.03.002

Figure Lengend Snippet: Performance of liver–skin co-cultures in endothelialized MOCs over 14 days. (A) Calcein AM staining (red) showed viable and evenly distributed HDMECs in all areas of the microfluidic circuit. (B) Higher magnification view of HDMEC monolayer. (C) HDMEC-liver tissue interaction shown by cytokeratin 8/18 (red) staining of hepatocytes and vWF (green) staining of HDMECs. (D) Skin biopsy stained for cytokeratin 15 (red) and cytokeratin 10 (green) showing a preserved continuous layer of cells. Nuclei were stained with Hoechst (blue). Scale bars 100 μm. (E) Glucose and lactate metabolic activity of co-cultures of skin, liver and endothelial cells over 14 days.

Article Snippet: Skin tissues were stained for mouse anti-human cytokeratin 10 (Merck-Millipore, Darmstadt, Germany) and rabbit anti-human cytokeratin 15 (Abcam).

Techniques: Staining, Activity Assay