Journal: bioRxiv

Article Title: Distinct roles for thymic stromal lymphopoietin (TSLP) and IL-33 in experimental eosinophilic esophagitis

doi: 10.1101/2025.02.25.640192

Figure Lengend Snippet: Experimental EoE was induced in wild type (WT) mice using oxazolone (OXA). Starting on day 19, the mice received two intraperitoneal injections per week of isotype control antibodies or anti-TSLP (clones M702 or clone 28F12, A). On day 36, the mice were euthanized, and esophageal tissues were fixed, paraffin embedded, and slides were generated. The slides were stained with anti-major basic protein (MBP, B) and esophageal eosinophils were quantified (C). Epithelial cell proliferation was determined using anti-Ki67 staining (D, E). H&E-stained slides (G) were analysed for lamina propria (F) as well as epithelial (H) thickness. Esophageal vascularization was determined using anti-CD31 staining (I, J). Single cell suspensions of esophageal tissue were obtained from oxazolone-challenged isotype control or anti-TSLP (M702)-treated mice using enzymatic digestion and the levels of eosinophils and CD4 + T cells determined by flow cytometry (K, L). Representative photomicrographs of anti-MBP (B), anti-Ki-67 (D), H&E (G) and anti-CD31 (I) are shown. Data are presented as mean ± SEM and are representative of n=3 experiments conducted with 10-12 mice per group, ns-nonsignificant, *-p<0.05, **-p<0.01, ***-p<0.001, ****-p<0.0001 .

Article Snippet: To substantiate these findings, TSLP was neutralized using an additional clone of anti-TSLP (BioXcell, clone 28F12).

Techniques: Control, Clone Assay, Generated, Staining, Flow Cytometry

Journal: bioRxiv

Article Title: Distinct roles for thymic stromal lymphopoietin (TSLP) and IL-33 in experimental eosinophilic esophagitis

doi: 10.1101/2025.02.25.640192

Figure Lengend Snippet: Experimental EoE was induced in wild type (WT) mice. Starting on day 19, the mice received two intraperitoneal injections per week of isotype control antibodies or anti-TSLP (clones M702 or clone 28F12). On day 36, the mice were euthanized, and esophageal tissue was obtained and subjected to bulk RNA sequencing. PCA plot of vehicle- and oxazolone (OXA)-challenged mice treated with either isotype control or anti-TSLP antibodies (clones M702 and/or 28F12) is shown (A). Heat plot analysis of all differentially expressed genes (absolute fold change > 2, p<0.05) (B) is presented. Analysis of the top differentially expressed secreted factors, cytokine receptors, adhesion molecules and extracellular matrix components is shown (C-D). Gene Set Enrichment Analysis (GSEA) and STRING analysis were performed on the DEGs, and comparison of OXA-challenged vehicle-treated mice vs. OXA-challenged anti-TSLP treated mice was performed (E-H). The top up and down regulated pathways, which were regulated by TSLP are presented (E-F). In C, D and E, each lane represents a different mouse.

Article Snippet: To substantiate these findings, TSLP was neutralized using an additional clone of anti-TSLP (BioXcell, clone 28F12).

Techniques: Control, Clone Assay, RNA Sequencing, Comparison

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Interleukin-17 receptor A maintains and protects the skin barrier to prevent allergic skin inflammation 1

doi: 10.4049/jimmunol.1602185

Figure Lengend Snippet: A. Representative pictures of 12-week old Il17ra-/-/Flgft/ft and Il17ra-/- mice compared to sex- and age-matched Flgft/ft and WT control mice. B. Clinical skin severity score. C. Total serum levels of IgE at 12-weeks of age. D. Measurement of trans epidermal water loss (TEWL) of mice as indicated (5-6 mice per group) at the specified time points. E. Skin cytokine protein levels for IL-4, IL-13 and IL-5 prior to clinical skin inflammation, and F levels for TSLP, IL-33 and IL-25, determined by ELISA and expressed as pg of cytokine per mg of total skin protein. G. Frequency of T cells and eosinophils in the skin expressed as a percentage of CD45+ cells. Data are presented as mean and SEM, from 4-9 mice per group, and statistical differences between groups were determined using Student's t test.

Article Snippet: Real-time quantitative PCR was performed on an AB StepOnePLus Real-time PCR system (Life Technologies, UK), using predesigned TaqMan gene expression assays specific for murine Tslp (Mm00498739_m1), Il25 (Mm00499822_m1), Il33 (Mm01195784_m1), S100a7 (Mm01218201_m1), S100a8 (Mm00496696_m1), S100a9 (Mm00656925_m1) Defb2 (Mm00657074_m1) and Cramp (Mm00438285_m1).

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology

Article Title: Inflammatory cytokine TSLP stimulates platelet secretion and potentiates platelet aggregation via a TSLPR-dependent PI3K/Akt signaling pathway.

doi: 10.1159/000369684

Figure Lengend Snippet: Fig. 2. Expression of P-selectin and activated integrin αIIbβ3 in human platelets after TSLP treatment. A. Flow cytometry of P-selectin expression in human platelets after TSLP stimulation (ng/ml). ADP (5 μM) and thrombin (0.01U/ml) served as positive controls. Arithmetic means ± SEM (n=8) are shown. *(p<0.05) and **(p<0.01) indicate significant differences compared with resting platelets. B. Flow cytometry of activated integrin αIIbβ3 (PAC-1) expression in human platelets after TSLP stimulation (ng/ml). ADP (5 μM) and thrombin (0.01U/ml) served as positive controls. Arithmetic means ± SEM (n=8) are shown. **(p<0.01) indicates a significant difference.

Article Snippet: The platelets were activated using recombinant human or murine TSLP (R&D Systems).

Techniques: Expressing, Flow Cytometry