enzyme linked immunosorbent assay elisa  (R&D Systems)

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: Effect of obesity and sex on the circulating levels of interleukin (IL)-33 and soluble suppression of tumorigenicity 2 (sST2). Fasting serum concentrations of ( A ) IL-33, ( C ) sST2 and ( E ) IL-33/sST2 in normal-weight (NW-NG) volunteers ( n = 22), patients with obesity with normoglycemia (OB-NG) ( n = 24) and with obesity-associated type 2 diabetes (OB-T2D) ( n = 27). Circulating concentrations of ( B ) IL-33, ( D ) sST2 and ( F ) IL-33/sST2 in males ( n = 26) and females ( n = 47) volunteers. Data are mean ± SEM. Differences between groups were analyzed by one-way ANOVA followed by Tukey’s tests or by unpaired two-tailed Student’s t test as appropriate. * P < 0.05, ** P < 0.01 and *** P < 0.001

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Two Tailed Test

Journal: Molecular Medicine

Article Title: Dual and context-dependent role of the interleukin-33/soluble suppression of tumorigenicity 2 axis in obesity and adipose tissue inflammation

doi: 10.1186/s10020-026-01454-z

Figure Lengend Snippet: The increased circulating levels of IL-33 in obesity together the decrease in its soluble decoy receptor sST2 may be potentially enhancing IL-33 bioactivity. In addition, gene and protein expression levels of IL-33 in visceral adipose tissue (VAT) are upregulated in obesity in both adipocytes and stromal vascular fraction cells. IL-33 modulates adipocyte function by upregulating both anti-inflammatory mediators ( ADIPOQ , ITLN1 and IL13 ) and pro-inflammatory cytokines ( TNF , IL8 ). Under inflammatory conditions, such as LPS stimulation, IL-33 attenuates excessive pro-inflammatory responses and promotes protective adipokine expression in visceral adipocytes. In THP-1-derived macrophages, adipocyte-derived factors suppress IL33 expression, whereas IL-33 attenuates macrophage pro-inflammatory activation and enhances IL4 expression only in the context of adipocyte-conditioned medium. Together, these findings suggest the dual and context-dependent nature of IL-33 in obesity, acting as a compensatory mediator to maintain tissue homeostasis while potentially contributing to chronic low-grade inflammation when elevated chronically. ADIPOQ, adiponectin; IL , interleukin; ITLN1 , omentin; TNF , tumor necrosis factor α

Article Snippet: Circulating concentrations of IL-33 (D3300B) and sST2 (DST200) were quantified using commercial ELISA kits (R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Expressing, Derivative Assay, Activation Assay

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 protects from IL (interleukin)-33–induced endothelial-to-mesenchymal transition (EndMT) and induces sST2 (soluble supression of tumorigenicity 2) expression in control pulmonary arterial endothelial cells (PAECs) in vitro. A , Representative immunofluorescent staining of PAEC for CD31 (endothelial marker), SM22α (smooth muscle protein 22-alpha; mesenchymal marker), and 4′,6-diamidino-2-phenylindole (DAPI; nuclei). Cells were treated with BMP9 (1 ng/mL), IL-33 (100 ng/mL), both, or left untreated (control [CTR]) for 3 days. Bar graphs show CD31 and SM22α intensity quantification (n=3). B , sST2 , ST2L , CTGF , and PAI-1 gene expressions in PAECs after 16-hour stimulation with TGF (transforming growth factor)-β (1 ng/mL), activin A (50 ng/mL), or untreated (CTR; technical replicates [t.n.]=3). C , sST2 , ST2L , ID1 , and ID3 gene expressions in PAECs after 3-hour stimulation with BMP4 (50 ng/mL), BMP6 (50 ng/mL), BMP9 (1 ng/mL), BMP10 (1 ng/mL), or untreated (CTR; t.n.=3). Statistical analysis: 1-way ANOVA with the Tukey post hoc test; * P <0.05, ** P <0.01, and **** P <0.0001. Data are shown as mean±SD.

Article Snippet: Cytokine and sST2 levels were measured using human IL-33 DuoSET ELISA (R&D Systems, DY3625B) and human sST2 ELISA (Elabscience, E-EL-H6082) per manufacturer’s instructions. sST2 and BMP9 levels in the plasma of patients with PAH were quantified using ELISA kits from Elabscience (E-EL-H6082) and R&D Systems (DY3209), respectively.

Techniques: Expressing, Control, In Vitro, Staining, Marker

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 induces sST2 (soluble supression of tumorigenicity 2) expression in vitro in pulmonary arterial endothelial cells (PAECs) via ALK1 (Activin Receptor-like Kinase) signaling in a dose- and time-dependent manner. A , Secreted protein levels of sST2 and IL (interleukin)-33 in PAEC supernatants after 24-hour BMP9 (1-ng/mL) stimulation compared with unstimulated controls. Each data point represents 3 biological replicates per donor (n=4). B , Dose-dependent increase in sST2 protein secretion following 24-hour stimulation with 0.1-, 1-, or 5-ng/mL BMP9 (technical replicates [t.n.]=3). C , Time-dependent induction of sST2 mRNA expression after 1, 3, or 24 hours of BMP9 stimulation (1 ng/mL; t.n.=3). D , sST2 mRNA expression in PAECs pretreated with LDN-193189 (120 nmol/L, 30 minutes) followed by BMP9 (1-ng/mL) stimulation for 3 or 24 hours (t.n.=3). E , Secreted sST2 protein levels following 24-hour stimulation with increasing concentrations of BMP9 in the presence or absence of LDN-193189 (120 nmol/L; t.n.=3). F , sST2 mRNA expression in PAECs transfected with siRNA targeting ALK1 or ENG (endoglin) and stimulated with BMP9 (1 ng/mL, 3 hours; t.n.=3). Statistical analysis: ( A ) unpaired Student t test, ( B , E , and F ) 1-way ANOVA with the Tukey post hoc test for multiple comparisons, and ( C and D ) 2-way ANOVA with the Tukey post hoc test for multiple comparisons. * P <0.05, ** P <0.01, *** P <0.001, and **** P <0.0001. Data are shown as ( A ) mean±SEM and ( B – F ) mean±SD. ns indicates not significant.

Article Snippet: Cytokine and sST2 levels were measured using human IL-33 DuoSET ELISA (R&D Systems, DY3625B) and human sST2 ELISA (Elabscience, E-EL-H6082) per manufacturer’s instructions. sST2 and BMP9 levels in the plasma of patients with PAH were quantified using ELISA kits from Elabscience (E-EL-H6082) and R&D Systems (DY3209), respectively.

Techniques: Expressing, In Vitro, Transfection

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 prevents IL (interleukin)-33–induced endothelial-to-mesenchymal transition (EndMT) in pulmonary arterial endothelial cells (PAECs) in vitro, similar to rsST2 (recombinant soluble ST2), thereby inhibiting IL-33 target gene expression. A , Representative immunofluorescent staining of for CD31 (endothelial marker), SM22α (smooth muscle protein 22-alpha; mesenchymal marker), and 4′,6-diamidino-2-phenylindole (DAPI; nuclei). Cells were pretreated with BMP9 (1 ng/mL, 3 hours) or rsST2 (1 μg/mL, 30 minutes) before IL-33 (100-ng/mL) stimulation for 3 days. Bar graphs show CD31 and SM22α intensity quantification (n=3). B and C , Representative immunoblots of p-p38 (phosphorylated p38 mitogen-activated protein kinase) and p-IκBα (phosphorylated inhibitor of nuclear factor kappa-B alpha) in PAECs pretreated overnight with BMP9 (1 ng/mL), rST2 (1 μg/mL, 30 minutes), or left untreated before IL-33 (100-ng/mL) stimulation for 5, 15, or 60 minutes. Densitometric analysis was performed using ImageJ, with protein levels normalized to vinculin and expressed as fold change relative to the 0-minute control (n=3). D , MyD88 , IL1RAP , and IL-8 gene expressions in PAEC pretreated with BMP9 (1 ng/mL, 3 hours) before IL-33 (100 ng/mL, 24 hours) stimulation or left untreated (control [CTR]; n=3). Statistical analysis: 1-way ANOVA ( A and D ) or 2-way ANOVA ( B and C ) with the Tukey post hoc test; P <0.05, * P <0.01, and ** P <0.001. Data are shown as mean±SD ( A and D ) or mean±SEM ( B and C ). ns indicates not significant.

Article Snippet: Cytokine and sST2 levels were measured using human IL-33 DuoSET ELISA (R&D Systems, DY3625B) and human sST2 ELISA (Elabscience, E-EL-H6082) per manufacturer’s instructions. sST2 and BMP9 levels in the plasma of patients with PAH were quantified using ELISA kits from Elabscience (E-EL-H6082) and R&D Systems (DY3209), respectively.

Techniques: In Vitro, Recombinant, Targeted Gene Expression, Staining, Marker, Western Blot, Control

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: BMP (bone morphogenetic protein) 9 protects from IL (interleukin)-33–induced endothelial-to-mesenchymal transition (EndMT) and induces sST2 (soluble supression of tumorigenicity 2) expression in pulmonary arterial endothelial cells (PAECs) from patients with pulmonary arterial hypertension (PAH) in vitro. A , Representative immunofluorescent staining of PAH PAEC for CD31 (endothelial marker), SM22α (smooth muscle protein 22-alpha; mesenchymal marker), and 4′,6-diamidino-2-phenylindole (DAPI). Cells were treated with BMP9 (1 ng/mL), IL-33 (100 ng/mL), both, or left untreated (control [CTR]) for 3 days. Bar graphs show quantification of CD31 and SM22α intensity (technical replicates [t.n.]=3). B , sST2 , ST2L , CTGF , and Pai-1 gene expressions in PAH PAECs s after 16-hour stimulation with TGF (transforming growth factor)-β (1 ng/mL), activin A (50 ng/mL), or untreated (CTR; t.n.=3). C , sST2 , ST2L , ID1 , and ID3 gene expressions in PAH PAECs after 3-hour stimulation with BMP4 (50 ng/mL), BMP6 (50 ng/mL), BMP9 (1 ng/mL), BMP10 (1 ng/mL), or untreated (CTR; t.n.=3). Statistical analysis: 1-way ANOVA with the Tukey post hoc test; P <0.05, * P <0.01, ** P <0.001, and *** P <0.0001. Data are shown as mean±SD.

Article Snippet: Cytokine and sST2 levels were measured using human IL-33 DuoSET ELISA (R&D Systems, DY3625B) and human sST2 ELISA (Elabscience, E-EL-H6082) per manufacturer’s instructions. sST2 and BMP9 levels in the plasma of patients with PAH were quantified using ELISA kits from Elabscience (E-EL-H6082) and R&D Systems (DY3209), respectively.

Techniques: Expressing, In Vitro, Staining, Marker, Control

Journal: Hypertension (Dallas, Tex. : 1979)

Article Title: BMP9 Modulates IL-33 Signaling to Mitigate EndMT in Pulmonary Arterial Hypertension

doi: 10.1161/HYPERTENSIONAHA.125.24916

Figure Lengend Snippet: IL (interleukin)-33 expression is upregulated in pulmonary vessels of patients with pulmonary arterial hypertension (PAH), and circulating sST2 (soluble supression of tumorigenicity 2) correlates with BMP (bone morphogenetic protein) 9 levels in stratified PAH groups. A , Circulating levels of sST2 in patients with PAH stratified by sex, age, and New York Heart Association class (n=79). B , Correlation between circulating sST2 and BMP9 in substratified groups with high circulating sST2 levels. C , Representative images of immunohistochemical staining for IL-33, CD31, α-SMA (α-smooth muscle actin), and 4′,6-diamidino-2-phenylindole (DAPI) in lung sections from patients with PAH or healthy controls (n=4). D , Quantification of IL-33⁺ endothelial cells (ECs) per vessel, normalized to total ECs, from ≥30 vessels across 4 individuals per condition. Statistical analysis: ( A ) the Mann-Whitney U test on log-transformed values, ( B ) the Spearman correlation, and ( D ) the unpaired t test; P <0.05, * P <0.01, ** P <0.001, and *** P <0.0001. Data are shown as mean±SD.

Article Snippet: Cytokine and sST2 levels were measured using human IL-33 DuoSET ELISA (R&D Systems, DY3625B) and human sST2 ELISA (Elabscience, E-EL-H6082) per manufacturer’s instructions. sST2 and BMP9 levels in the plasma of patients with PAH were quantified using ELISA kits from Elabscience (E-EL-H6082) and R&D Systems (DY3209), respectively.

Techniques: Expressing, Immunohistochemical staining, Staining, MANN-WHITNEY, Transformation Assay