Journal: Journal of Innate Immunity

Article Title: Type I Interferon, Induced by Adenovirus or Adenoviral Vector Infection, Regulates the Cytokine Response to Lipopolysaccharide in a Macrophage Type-Specific Manner

doi: 10.1159/000538282

Figure Lengend Snippet: The ability to produce IL-10 and the height of IL-10R expression regulate the IL-6 response to LPS in mock- and IFN-β-pretreated macrophages. a IL-10 response to LPS: MPI’s, BMM, PC, and AM were pretreated with rmIFN-β, or remained untreated and 6 h later stimulated with LPS. The IL-10 response in culture supernatants was determined 16 h later by ELISA. IFN-β-stimulated BMM produced, on average, 110 pg IL-10/mL, while all other cell types and unstimulated control cells produced none (not shown in the figure). b IL-6 response in the presence of anti-IL-10R. BMM and MPI cells were pretreated with rmIFN-β or remained untreated and, 6 h later, stimulated with LPS in the presence or absence of 1 μg/mL anti-IL-10R or control antibody (anti-HRPO). IL-6 in culture supernatants was determined by a specific ELISA. c LPS-induced IL-6 production in the presence of exogenous IL-10. BMM and MPI cells were stimulated with LPS in the presence or absence of rmIL-10 (0.2 μg/mL). The IL-6 in culture supernatants collected 16 h later was determined by ELISA. d Cell surface IL-10R expression. The detection of IL-10R on cells was analyzed by FACS using anti-IL-10R.PE (filled blue) or isotype control (open black) antibodies. n.d., not detectable.

Article Snippet: To inhibit IL-10 effects in LPS-stimulated cultures, the anti-IL-10R (1B1.3A) and the recommended control antibody (HRPN) from BioXCell were used.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Produced

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: B cells were gated for using 1:20 dilution commercial mouse anti-human CD19-Alexa488 (BioLegend, San Diego, CA, USA) and 1:20 dilution mouse anti-human CD27-PE (stock 50 ug/mL, BioLegend, San Diego, CA, USA), which has been reported as a marker for juvenile B cells. pDCs were gated for flow cytometry with 1:20 dilution commercial mouse anti-human BDCA-2-Alexa488 (stock 200 ug/mL, BioLegend, San Diego, CA, USA) and 1:20 dilution mouse anti-human CD123 (IL-3)-PE (stock 200 ug/mL, BD Biosciences, Franklin Lakes, NJ, USA).

Techniques: Membrane