Journal: The Journal of Clinical Investigation

Article Title: Tumor-intrinsic PIK3CA represses tumor immunogenicity in a model of pancreatic cancer

doi: 10.1172/JCI123540

Figure Lengend Snippet: Flow cytometric analysis of cell surface levels of ( A ) H-2K b and ( B ) CD80 in WT and αKO cells. The left panels show representative histograms and the right graphs show the mean ± SEM of the geometric means (geo. mean) of each flow cytometry distribution. H-2K b , n = 4; CD80, n = 3. *** P = 0.0007 (unpaired t test). ( C ) qRT-PCR analysis of mRNA expression of H-2K b , B2m, and CD80 in WT and αKO cells. Gene expression changes were normalized to Hprt . Graph shows mean ± SEM ( n = 6). **** P < 0.0001 (unpaired t test). ( D – F ) B6 mice were implanted with 0.5 million αKO/shB2m, αKO/CD80KO, or αKO/CD80KO+shB2m cells in the head of the pancreas and tumor growth was monitored by IVIS imaging. ( D ) Kaplan-Meier survival curves for αKO/shB2m, αKO/CD80KO, and αKO/CD80KO+shB2m mice. *A single mouse was euthanized for histology and removed from the study. Median survival: αKO/shB2m, 110.5 days; αKO/CD80KO+shB2m, 87.5 days. All αKO/CD80KO mice were alive at 115 days. P = 0.0026 (log-rank test). ( E ) Representative H&E and IHC images of pancreatic sections from αKO/shB2m or αKO/CD80KO+shB2m mice that died of tumor progression. Scale bar: 100 μm. ( F ) Representative H&E-stained pancreatic sections from a single αKO/shB2m mouse or αKO/CD80KO mice euthanized at 101 or 160 days, respectively. Scale bar: 100 μm.

Article Snippet: Single cell clones were generated by serial dilution and confirmed by flow cytometry. αKO cells with reduced levels of B2m (referred to as αKO/shB2m) were generated by infecting αKO cells with B2m shRNA lentiviral particles (Santa Cruz Biotechnology, sc-29593-V).

Techniques: Flow Cytometry, Quantitative RT-PCR, Expressing, Gene Expression, Imaging, Staining

Journal: The Lancet Regional Health - Europe

Article Title: Efficacy of repeated immunoadsorption in patients with post-COVID myalgic encephalomyelitis/chronic fatigue syndrome and elevated β2-adrenergic receptor autoantibodies: a prospective cohort study

doi: 10.1016/j.lanepe.2024.101161

Figure Lengend Snippet: Course of immunoglobulin and autoantibody levels over the study period ( n = 20), the duration of IA therapy is indicated by the green bar. Error bars represent 95% confidence intervals. Data were analyzed using a linear mixed-effects model fitted by restricted maximum likelihood (REML), with t-tests computed using Satterthwaite's method for degrees of freedom with significance levels indicated as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. From left to right, the panels display the trajectories of: a) immunoglobulin G (IgG), b) immunoglobulin A (IgA), c) immunoglobulin M (IgM), d) β1 adrenergic receptor autoantibodies (β1 AR-AB), e) β2 adrenergic receptor autoantibodies (β2 AR-AB), f) M3 acetylcholine receptor autoantibodies (M3 AchR-AB), and g) M4 acetylcholine receptor autoantibodies (M4 AchR-AB).

Article Snippet: Charité holds, together with CellTrend, a patent for the diagnostic use of autoantibodies against β2 AR-AB.

Techniques:

Journal: Frontiers in Oncology

Article Title: Propranolol Sensitizes Vascular Sarcoma Cells to Doxorubicin by Altering Lysosomal Drug Sequestration and Drug Efflux

doi: 10.3389/fonc.2020.614288

Figure Lengend Snippet: Expression of β-ARs in primary hemangiosarcomas and hemangiosarcoma cell lines. (A) Representative images of β1-AR, β2-AR, and β3-AR expression in visceral hemangiosarcomas (n = 10) from dogs. IHC; horseradish peroxidase; counter stain = hematoxylin. (B) Box and whiskers plot of β1-AR, β2-AR, and β3-AR IHC scores from the analyzed tissues and the expression in canine splenic hematomas as a control (n = 5). (C) Expression of β-ARs by canine hemangiosarcoma (COSB, DD-1), human angiosarcoma (ISO-HAS), and mouse angiosarcoma (SVR) cell lines. Proteins were detected in cell lysates by immunoblotting. β-actin was used as a gel-loading control.

Article Snippet: After blocking, the membranes were incubated overnight at 4°C with one of the following antibodies diluted in Odyssey ® Blocking Buffer or TBST +3% BSA: anti-β1-AR (1:1000, #250919, Abbiotec), anti-β2-AR (1:1000, #251604, Abbiotec), anti-β3-AR (1:1000, #251434, Abbiotec), anti-phospho-checkpoint kinase 1 (CHK1) (Ser345) (1:1000, #2348, Cell Signaling, Danvers, MA, USA), anti-phospho-checkpoint kinase 2 (CHK2) (Thr68) (1:1000, #2197, Cell Signaling), anti-phospho-ataxia telangiectasia- and Rad3-related protein (ATM) (Ser428) (1:1000, #2853, Cell Signaling), anti-phospho-protein kinase ataxia-telangiectasia mutated protein (ATR) (Ser1981) (1:1000, #5883, Cell Signaling), and anti-β-actin (1:5000, #sc8432, Santa Cruz Biotechnology, Dallas, TX, USA; 1:5000, #A5441, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Expressing, Staining, Western Blot