Journal: Pharmacological research

Article Title: Targeting IL-11 system as a treatment of pulmonary arterial hypertension.

doi: 10.1016/j.phrs.2023.106985

Figure Lengend Snippet: Fig. 1. IL-11 and IL-11Rα are increased in whole lung homogenates, isolated pulmonary arteries, and serum of patients with idiopathic pulmonary arterial hypertension (iPAH). Protein expression of (A) IL-11 and (B) IL-11Rα in isolated pulmonary arteries (70–500 µm of internal diameter), (C, D) serum and (E, F) lung tissue homogenates. Protein expression was measured using ELISA kits. (G) CD31 protein expression was measured in isolated pulmonary arteries as endothelial cells marker by ELISA. Data are presented as scatter dot plot with median and interquartile range values. P-values are based on the Mann Whitney test. N indicates number of patients. (H, I) Expression of (H) IL11Rα and (I) TWIST1 in the lungs of PAH patients (All-PAH) compared to lungs of donors (controls) and in the specific subtypes of PAH [idiopathic PAH (IPAH), associated to other systemic diseases (APAH), familial PAH (APAH) and undefined cause PAH (Other)]. (J) Correlation of the IL11Rα and TWIST1 mRNA expression. (H,I)* ** indicates adjusted P < 0.001 vs controls (using Benjamini & Hochberg approach). ## and ### indicate P < 0.01 and P < 0.001 vs controls, respectively. (J) Pearson correlation.

Article Snippet: Protein levels of CD31, IL-11, IL-11RА were measured in isolated human pulmonary arteries, human lung tissue homogenates and serum, using commercially available ELISA kits for CD31 (R&D systems; cat. n. DCD310), IL-11 (Sigma-Aldrich; cat. n. RAB0250) and IL-11RА (Mybiosource; cat. n. MBS047882) according with the manufacturer instructions.

Techniques: Isolation, Expressing, Enzyme-linked Immunosorbent Assay, Marker, MANN-WHITNEY

Journal: Pharmacological research

Article Title: Targeting IL-11 system as a treatment of pulmonary arterial hypertension.

doi: 10.1016/j.phrs.2023.106985

Figure Lengend Snippet: Fig. 2. IL-11 and IL-11Rα expression localizes in pulmonary artery endothelial and smooth muscle cells of patients with idiopathic pulmonary artery hypertension (iPAH). (A, B, C, D) Human lung tissue from control subjects and iPAH patients was immune-stained with IL-11, IL-11Rα and alpha smooth muscle actin (αSMA) antibodies. (A) Representative images are showed from lung areas with pulmonary artery remodelling. (B) Vascular wall thickening was quantified in a total of 20–30 pulmonary arteries per patient. (C, D) Immunohistochemical score quantification of IL-11 and IL-11Rα in a total of 20–30 pulmonary arteries per patient. Scale bar: 100 µm. Data are presented as scatter dot plot with median and interquartile range values. P-values are based on the Mann Whitney test analysis. Bar dots indicates the number of patients in each graph. (E) Human lung tissue from iPAH patients was immune-stained with IL-11, IL-11Rα, αSMA and CD31 antibodies and with secondary fluorescence antibodies. Representative images are shown. White colour represents co-localization of both antibodies. (F) human pulmonary artery endothelial cells (HPAECs) and (G) human pulmonary artery smooth muscle cells (HPASMCs) were isolated from pulmonary arteries of control subjects and iPAH patients and cultured until passage 1. Cell culture supernatants were collected to measure IL-11 by ELISA. Data are presented as scatter dot plot with median and interquartile range values of n = 6 patients in each group run in triplicate. P-values are based on the Mann Whitney test analysis.

Article Snippet: Protein levels of CD31, IL-11, IL-11RА were measured in isolated human pulmonary arteries, human lung tissue homogenates and serum, using commercially available ELISA kits for CD31 (R&D systems; cat. n. DCD310), IL-11 (Sigma-Aldrich; cat. n. RAB0250) and IL-11RА (Mybiosource; cat. n. MBS047882) according with the manufacturer instructions.

Techniques: Expressing, Control, Staining, Immunohistochemical staining, MANN-WHITNEY, Fluorescence, Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Pharmacological research

Article Title: Targeting IL-11 system as a treatment of pulmonary arterial hypertension.

doi: 10.1016/j.phrs.2023.106985

Figure Lengend Snippet: Fig. 6. rhIL-11 and soluble rhIL-11Rα are downstream effectors of different fibrotic mediators promoting endothelial to mesenchymal transition. (A,B) Human pulmonary arterial endothelial cells (HPAEC) were isolated from healthy subjects and idiopathic pulmonary arterial hypertension (iPAH). (A) Supernatant levels of IL-11 and soluble IL-11Rα were measured by ELISA in presence or absence of stimulation with TGFβ1, EDN-1 or PDGF during 48 h (n = 4 patients in triplicate). (B-D) Neutralizing mAb-IL-11 or mAb-IL-1Rα were added 1 h before (A) TGFβ1, (B) EDN-1 or (C) PDGF stimulation during 48 h in HPAEC from iPAH patients (n = 4 patients in triplicate). Mesenchymal markers αSMA, collagen type I and fibronectin, as well as endothelial eNOS were measured by qRT-PCR. Data are presented as scatter dot plot with mean and standard error of experiments from cells of 4 different iPAH patients run in triplicate. P-values are based on two-way ANOVA followed by post hoc Bonferroni tests for multiple comparison. * P < 0.05, * * P < 0.01, * ** P < 0.001, * ** * P < 0.0001 vs control group; #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001 vs stimulus.

Article Snippet: Protein levels of CD31, IL-11, IL-11RА were measured in isolated human pulmonary arteries, human lung tissue homogenates and serum, using commercially available ELISA kits for CD31 (R&D systems; cat. n. DCD310), IL-11 (Sigma-Aldrich; cat. n. RAB0250) and IL-11RА (Mybiosource; cat. n. MBS047882) according with the manufacturer instructions.

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Comparison, Control