Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 2 Calycosin inhibited group 2 innate lymphoid cell (ILC2) activation in OVA-induced asthmatic mice. (A) The levels of interleukin (IL)-4, (B) IL-5 and (C) IL-13 in the BALF and lung tissues were measured using ELISA and RT-qPCR. (D) ELISA was used to determin the contents of IL-33 in the BALF. (E) Suppression tumorigenicity 2 (ST2) levels in the lung tissues was examined by western blot. The blots were cut prior to hybridization with antibodies during blotting. (F) Immunohistochemistry (IHC) staining was used to detect the expression ST2 and IL-25R. (G) The co-localization of ST2 and CD45 was detected by immunofluorescence (IF). Scale bar = 50 μm. n = 6 for each group. * P< 0.05, **P < 0.01, ***P < 0.001, compared with the sham group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the OVA group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Hybridization, Immunohistochemistry, Expressing, Immunofluorescence

Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 4 ILC2 activation induced by IL-33 was restrained by calycosin in ILC2s. (A) The ILC2 cell viability was examined by CCK-8. (B) Western blot showed the expression of ST2 in ILC2s. The blots were cut prior to hybridization with antibodies during blotting. (C) The secretion of IL-4, (D) IL-5 and (E) IL-13 in ILC2s were detected by ELISA assay. n = 3 for each group. **P < 0.01,***P < 0.001, compared with the control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the IL-33 group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, CCK-8 Assay, Western Blot, Expressing, Hybridization, Enzyme-linked Immunosorbent Assay, Control

Journal: Bone

Article Title: Type 2 innate lymphoid cells inhibit the differentiation of osteoclasts and protect from ovariectomy-induced bone loss.

doi: 10.1016/j.bone.2020.115335

Figure Lengend Snippet: Fig. 1. ILC2 directly suppress osteoclastogenesis in vitro. (a) Bone marrow-derived monocytes (BMM) were co-cultured with innate lymphoid cells type 2 (ILC2; ILC2:BMM ratio = 1:10). Osteoclasts were stained by tartrate acid resistant phosphatase (TRAP) after stimulation with 50 ng/mL RANKL for 5 days. Scale bar = 200 μm, Representative data are from one out of three in- dependent experiments. (b) TRAP-positive cells were counted (Nuclei ≥3). (c) Pit formation assay. BMM were cultured with ILC2 (ILC2:BMM ratio = 1:20 or 1:10). Representative data are from one out of three independent experiments. Scale bar = 200 μm (d) Measurement of resorption pit area. (e) Immunohistochemistry using mouse femur bone sample. Osteoclasts (white arrows with asterisk) were stained by ELF97 substrate and ILC2 (white arrows) were labeled as ST2+IL17RB+lineage (CD3e, CD11b, CD11c, B220, Ly6G) negative cells (n = 6). Scale bar = 50 μm. Data are expressed as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: ILC2 were stained using antibodies against IL17RB (Santa Cruz, sc11754), ST2 (ProSci, 3363), CD3ε (eBioscience, 17-0031), CD11b (R&D systems, 1124), CD11c (Abcam, ab33483), B220 (eBioscience, 14-0452), Ly6G (R&D systems, MAB1037), and DAPI.

Techniques: In Vitro, Derivative Assay, Cell Culture, Staining, Tube Formation Assay, Immunohistochemistry, Labeling

Journal: Bone

Article Title: Type 2 innate lymphoid cells inhibit the differentiation of osteoclasts and protect from ovariectomy-induced bone loss.

doi: 10.1016/j.bone.2020.115335

Figure Lengend Snippet: Fig. 3. Adoptive transfer of ILC2 inhibits osteoclasts in vivo. (a) Light sheet microscopy image of tibia. 1 × 106 sort-purified ILC2 (red cells with white arrows) were antibody labeled and intravenously injected into recipient mice. Asterisks indicate bone tissue. Scale bar indicates 1000 μm. (b) Frequencies of innate lymphoid cells type 2 (ILC2) in the spleen after adoptive transfer. ILC2 were identified as CD5−B220−CD45R−NKp46−CD11b−CD11c−ICOS+ KLRG1+ ST2+ cells. (c) Frequencies of T-bet+ ILC1 and NCR+/−Rorgt+ ILC3 in the spleen of mice with and without adoptive transfer of ILC2s. (d) Histomorphometry of tibial bones showing the number of osteoclasts per bone area (N.Oc/T.Ar) and osteoclast number per bone perimeter (N.Oc/B.Pm) with and without adoptive transfer of ILC2s. (e) μCT analysis of tibial bones showing bone volume per tissue volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and trabecular separation (Tb.Sp) with and without adoptive transfer of ILC2s. (f) Frequencies of T-bet+Th1 cells, Gata3+Th2 cells, Rorgt+Th17 cells and FoxP3+T cells in the spleen of mice with and without adoptive transfer of ILC2s. Data are from two independent experiments. Data are expressed as mean ± SEM. *p < 0.05; **p < 0.01. n = 4–5/group.

Article Snippet: ILC2 were stained using antibodies against IL17RB (Santa Cruz, sc11754), ST2 (ProSci, 3363), CD3ε (eBioscience, 17-0031), CD11b (R&D systems, 1124), CD11c (Abcam, ab33483), B220 (eBioscience, 14-0452), Ly6G (R&D systems, MAB1037), and DAPI.

Techniques: Adoptive Transfer Assay, In Vivo, Microscopy, Purification, Labeling, Injection

Journal: Mediators of Inflammation

Article Title: Serum Soluble ST2 Correlated with Symptom Severity and Clinical Response of Sublingual Immunotherapy for House Dust Mite-Induced Allergic Rhinitis Patients

doi: 10.1155/2021/5576596

Figure Lengend Snippet: Serum sST2 concentrations were increased in AR patients than in healthy controls. (a) sST2 levels were elevated in the AR group in comparison with the HC group; (b) the serum sST2 levels were higher in the MSAR group than in the MAR and the HC group. sST2: soluble suppressor of tumorigenicity 2; AR: allergic rhinitis; HC: healthy control; MAR: mild allergic rhinitis; MSAR: moderate-severe allergic rhinitis.

Article Snippet: Serum sST2 measurement was performed by an Enzyme-Linked Immunosorbent Assay- (ELISA-) utilizing human sST2 ELISA kit (Proteintech, Rosemont, USA) referring to the manufacturer's instructions.

Techniques: Comparison, Control

Journal: Mediators of Inflammation

Article Title: Serum Soluble ST2 Correlated with Symptom Severity and Clinical Response of Sublingual Immunotherapy for House Dust Mite-Induced Allergic Rhinitis Patients

doi: 10.1155/2021/5576596

Figure Lengend Snippet: Correlation between serum sST2 levels and TNSS (a), VAS (b), HDM-specific IgE (c), and total IgE (d). sST2: soluble suppressor of tumorigenicity 2; HDM: house dust mite; TNSS: total nasal symptom score; VAS: visual analogue scale.

Article Snippet: Serum sST2 measurement was performed by an Enzyme-Linked Immunosorbent Assay- (ELISA-) utilizing human sST2 ELISA kit (Proteintech, Rosemont, USA) referring to the manufacturer's instructions.

Techniques:

Journal: Mediators of Inflammation

Article Title: Serum Soluble ST2 Correlated with Symptom Severity and Clinical Response of Sublingual Immunotherapy for House Dust Mite-Induced Allergic Rhinitis Patients

doi: 10.1155/2021/5576596

Figure Lengend Snippet: The serum levels sST2 in the effective group and ineffective group. (a) The sST2 levels were lower in the effective group than in the ineffective group; (b) patients in the effective group exhibited significantly lower sST2 levels post-SLIT than pre-SLIT; (c) no statistical difference was observed in the ineffective group between post-SLIT and pre-SLIT. (d) the change of sST2 levels was higher in the effective group than in the ineffective group. sST2: soluble suppressor of tumorigenicity 2; SLIT: sublingual immunotherapy.

Article Snippet: Serum sST2 measurement was performed by an Enzyme-Linked Immunosorbent Assay- (ELISA-) utilizing human sST2 ELISA kit (Proteintech, Rosemont, USA) referring to the manufacturer's instructions.

Techniques:

Journal: Mediators of Inflammation

Article Title: Serum Soluble ST2 Correlated with Symptom Severity and Clinical Response of Sublingual Immunotherapy for House Dust Mite-Induced Allergic Rhinitis Patients

doi: 10.1155/2021/5576596

Figure Lengend Snippet: ROC curve analysis of serum sST2 in predicting the efficacy of SLIT. ROC: receiver operating characteristic; sST2: soluble suppressor of tumorigenicity 2; SLIT: sublingual immunotherapy.

Article Snippet: Serum sST2 measurement was performed by an Enzyme-Linked Immunosorbent Assay- (ELISA-) utilizing human sST2 ELISA kit (Proteintech, Rosemont, USA) referring to the manufacturer's instructions.

Techniques:

Journal: Mediators of Inflammation

Article Title: Critical Roles of IL-33/ST2 Pathway in Neurological Disorders

doi: 10.1155/2018/5346413

Figure Lengend Snippet: Releases and signalling of IL-33. Full length of IL-33 is predominantly expressed by stromal cells, endothelial and epithelial cells, macrophages, and fibroblasts. In necrosis, IL-33 is released and binds to ST2L/IL1-RAcP heterodimer receptor, which induces signalling via TIR domain of ST2L/IL1-RAcP and recruits MyD88 and followed by IRAK1/4 and TRAF6. TRAF6 further induces either MAPK which in turns activating P38, ERK and JNK, and/or IKK pathway which activate NF- κ B. Both pathways lead to the recruitment of transcription factors and proinflammatory cytokines, which consequently drives the regulatory transcription factors of Th2 cytokines and chemokines. Alternatively, binding of IL-33 to sST2 neutralises the proinflammatory effect of IL-33 as well as with SIGIRR which negatively regulates ST2L signalling pathway. In apoptosis, IL-33 is cleaved and deactivated by caspase-3/7.

Article Snippet: There are variants of ST2 receptors, namely, ST2L, a membrane-anchored receptor that responds to IL-33 by heterodimerising with IL-1RAcP [ , ]; sST2, a decoy and soluble isoform receptor identical to ST2L [ , ]; ST2V, an isoform receptor similar to sST2 with exception of the third extracellular immunoglobulin domain and can be found in human [ , ]; and ST2LV, similar with ST2L without transmembrane domain and currently described in chicken, Gallus gallus , solely [ ].

Techniques: Binding Assay