Journal: Nature

Article Title: Spatial proteomics identifies JAKi as treatment for a lethal skin disease.

doi: 10.1038/s41586-024-08061-0

Figure Lengend Snippet: Fig. 5 | JAK/STAT inhibition reduces severity of TEN in vitro and in vivo. a,b, Schematic of live-cell imaging assay (a) with representative images of labelled keratinocytes (arrows) and unlabelled PBMCs (arrowhead) in co-culture for the indicated duration, with or without 50 nM tofacitinib to measure cytotoxicity over time (b). r, resting; a, activated. n = 6 biological replicates per condition, 4 field of views per well. Data are mean ± s.e.m. c, Schematic of oral JAKi treatment in the smac-mimetic-induced model of TEN. Oral JAKi (tofacitinib (tofa), 30 mg kg−1; baricitinib (bari), 10 mg kg−1; abrocitinib (abr), 20 mg kg−1) was started one day before (‘pre-treat model’) or 3 h after (‘treatment model’) subcutaneous injection of smac mimetic. Outcome assessment was performed on day 1 (histology) and day 3 (clinical score). d–j, Clinical assessment (d,g,h), histology (e), dermal thickness (f; 15 measurements per mouse),

Article Snippet: Tofacitinib (CST, 14703) or control (DMSO) was added in the indicated concentration as well as Celltox Green (Promega, G8731, 1:8,000 final dilution) to reach a final volume of 200 μl per well.

Techniques: Inhibition, In Vitro, In Vivo, Live Cell Imaging, Co-Culture Assay, Injection

Journal: Science translational medicine

Article Title: A gut-restricted glutamate carboxypeptidase II inhibitor reduces monocytic inflammation and improves preclinical colitis

doi: 10.1126/scitranslmed.abn7491

Figure Lengend Snippet: (A) Synthetic scheme for generation of GCPII inhibitor, 2-PMPA, and LCA (8a), DCA (8b), or UDCA (8c) conjugates. (B to D) Efficacy evaluation in the 2.5% DSS colitis mouse model at a 10 mg/kg of 2-PMPA molar equivalent (eq.) dose. Body weight, stool consistency, and rectal bleeding were scored daily and summated to calculate the disease activity index (DAI). DCA–2-PMPA (IBD3540) was identified as an active compound and was evaluated head to head versus its constituents, (E) 2-PMPA and DCA, and clinical agents (F) sulfasalazine and (G) tofacitinib, at doses indicated. All test articles were administered once daily by oral gavage beginning on study day 0, except for tofacitinib, which was administered twice daily. (B to G) n = 5 to 15 per group; (B to D and G) repeated measures (RM) two-way ANOVA with post hoc Bonferroni’s multiple comparisons test; (E and F) mixed effects model (REML, restricted maximum likelihood) with post hoc Bonferroni’s multiple comparisons test; *P < 0.05, **P < 0.01, and ***P < 0.001. Studies were performed in duplicate with representative data shown. RT, room temperature; DCM, dichloromethane; DMF, N,N-dimethylformamide; THF, tetrahydrofuran.

Article Snippet: Treatments included vehicle (5% ethanol, 5% Tween 80, and 90% 50 mM HEPES SID), test articles (as indicated), sulfasalazine (50 mg/kg SID; Sigma-Aldrich, S0883), or tofacitinib (50 mg/kg BID; BOC Sciences, B0084-453466).

Techniques: Activity Assay

Journal: Cardiovascular Research

Article Title: Interleukin-7 and interleukin-15 drive CD4 + CD28 null T lymphocyte expansion and function in patients with acute coronary syndrome

doi: 10.1093/cvr/cvaa202

Figure Lengend Snippet: Blockade of IL-7/IL-15 signalling with the JAK1/JAK3 selective inhibitor Tofacitinib prevents CD28 null T-cell expansion. Fresh peripheral blood samples from ACS patients ( n = 10) were stained with CD4, CD28, CD122, CD127, CD132, and CD215 monoclonal antibodies. Illustrative dot plots ( A ) and graphs ( B ) show the percentage of CD28 null and CD28 + T cells expressing CD122, CD132, CD127, and CD215; dashed gates, isotype control antibody (Ctrl). CD4 + T cells from ACS patients ( n = 10) were cultured alone (w/o) or treated with 50 ng/mL IL-7 or IL-15 for 3–4 days and expression of CD127 and CD215 was analysed on CD28 null and CD28 pos T cells. Illustrative dot plots and graphs display the percentage of CD127 + cells ( C and D ), and CD215 + cells ( E and F ) in CD28 null and CD28 pos T cells; dashed gates, isotype control antibody (Ctrl). ( G ) Baseline levels of phosphorylated STAT5 (pSTAT5) were quantified using the PhosFlow Method (described in Methods); black and dashed histograms, isotype control antibody; the numbers indicate the mean fluorescence intensity (MFI). Illustrative histograms and bar graph display the expression levels (MFI) of pSTAT5 in CD28 null and CD28 pos T cells ( n = 6; mean±SEM). ( H ) CD4 + T cells from ACS patients ( n = 10) were cultured alone (w/o) or treated with 50 ng/mL IL-7 or IL-15 in the presence or absence of 100 nM Tofacitinib (Tofa) for 7 days. Graphs show the fold change in the percentage of CD28 null or CD28 pos T cells (mean±SEM). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. ( B ) Two-tailed Mann–Whitney test; ( D and F ) two-tailed Wilcoxon matched-pairs signed-rank test; ( G ) paired two-tailed Student’s t -test; ( H ) two-way ANOVA with post-test Bonferroni for multiple comparisons.

Article Snippet: Where indicated, cells were stimulated with IL-7/IL-15 in the presence or absence of 100 nM Tofacitinib (CP 690550 citrate, Tocris Bioscience).

Techniques: Staining, Bioprocessing, Expressing, Control, Cell Culture, Fluorescence, Two Tailed Test, MANN-WHITNEY