Journal: bioRxiv

Article Title: A Two-Step Activation Mechanism Enables Mast Cells to Differentiate their Response between Extracellular and Invasive Enterobacterial Infection

doi: 10.1101/2023.08.31.555657

Figure Lengend Snippet: A : RNA sequencing of BMMCs left uninfected, or infected with MOI 50 of S. Tm wt or S .Tm Δ invG SL1344 for 4h, presented as the top 30 significantly upregulated genes between S .Tm wt -infected and uninfected control, displayed as a z-score-transformed heatmap. Genes are sorted by the formula “ S .Tm wt - S .Tm Δ invG ” to highlight differences between those two groups. B : Heatmap of relative log2 fold changes between the indicated groups, derived from a cytokine array of 24h supernatants from BMMCs infected with MOI 50 of S. Tm wt or S .Tm Δ invG SL1344. C : Representative IF images of MCs in close contact to CD45+ and CD18+ immune cells in the S. Tm wt SL1344-infected intestinal mucosa and lumen at 48h p.i.. Arrows indicate MCs, scale bars: 10 µm. D : Trypan blue-based live cell counts of bone marrow nucleated cells, cultured for 7 days in base medium supplemented with either medium alone, 24h uninfected BMMC supernatant, S. Tm inoculum-conditioned supernatant, or supernatant of BMMCs infected with S. Tm wt for 24h. E-F : Similar setup as in C, but quantification of the total number of CD45 + CD11b + CD11c − cells (containing monocytes) ( E ) and CD45 + F4/80 + cells (containing macrophages) ( F ) in bone marrow cultures treated with the indicated supernatants, or base medium alone. For C-E, data was statistically analyzed with ANOVA and Dunnet’s posthoc test, using “BMMC+ S .Tm” for comparisons to all other groups.

Article Snippet: Proteome Profiler Mouse XL Cytokine Array (Bio-Techne, #ARY028) was used, following the manufacturer’s instruction for fluorescent detection with IRDye® 800CW Streptavidin (LI-COR, #926-32230) on an Odyssey CLx Infrared Imager.

Techniques: RNA Sequencing Assay, Infection, Transformation Assay, Derivative Assay, Cell Culture

Journal:

Article Title: Association of Gamma Interferon and Interleukin-17 Production in Intestinal CD4 + T Cells with Protection against Rotavirus Shedding in Mice Intranasally Immunized with VP6 and the Adjuvant LT(R192G) ▿

doi: 10.1128/JVI.01877-06

Figure Lengend Snippet: Cytokine secretion after in vitro stimulation of lymphocytes isolated prior to challenge (time zero) from the MLN, PP, IEL, and LP of naive, VP6-immunized, LT(R192G)-immunized, and VP6/LT(R192G)-immunized mice. Lymphocytes isolated from 12 animals were pooled and placed in culture and stimulated with VP6 or left unstimulated for 18 h. Supernatants were harvested, and the cytokine concentrations were determined by ELISA. Values are expressed as the amount of cytokine secreted in the supernatants of the stimulated cultures minus the amount secreted in the unstimulated cultures (in pg/ml). The data are representative of one of three independent experiments.

Article Snippet: The GEArray Q series mouse common cytokine gene array (SuperArray Bioscience Corp., Frederick, MD) was used to determine mRNA expression of cytokines in all samples.

Techniques: In Vitro, Isolation, Enzyme-linked Immunosorbent Assay