Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Central role of T helper 17 cells in chronic hypoxia-induced pulmonary hypertension

doi: 10.1152/ajplung.00531.2016

Figure Lengend Snippet: CH leads to an increase in lung T helper 17 (TH17) cells without T regulatory cells being affected. Lungs were removed from vehicle- and SR1001 (25 mg·kg−1·day sc−1)-treated mice after 5 days of normoxic or CH (CH 5d) exposure. Single-cell suspensions from the lungs were labeled with anti-CD3, anti-CD4, and anti-IL-17A or FoxP3. A: representative scatter plots. Cells were gated based on forward and side scatter and CD3 CD4. B: summary of the %CD4+ IL-17A+ or CD4+ FoxP3+ cells relative to total CD3+ cells normalized to the average of the vehicle-treated normoxic group of each set of experiments. C: representative scatter plots of CD3+, CD4+ T helper cells from lungs from mice exposed to normoxia or 5 days of CH. D: summary of the %CD3+ T cells relative to total cells and CD3+, CD4+, and T helper cells relative to total CD3+ cells. Values are means ± SE; n = no. of animals, *P < 0.05 vs. normoxia vehicle; #P < 0.05 vs. CH vehicle, analyzed by 2-way ANOVA, followed by multiple-comparison Student-Newman-Keuls test.

Article Snippet: Cells were washed and stained with PerCP/Cy5.5 anti-mouse CD4 (clone GK1.5, 100432; BioLegend, San Diego, CA), APC/Cy7 anti-mouse CD3ε (clone 145-2C11, 100330; BioLegend), and PE anti-mouse IL-17A (clone TC11-18H10.1, 506910; BioLegend) or PE anti-mouse/rat/human FoxP3 (clone 150D, 32008, BioLegend) for 1 h at 4°C in the dark.

Techniques: Labeling, Comparison

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Central role of T helper 17 cells in chronic hypoxia-induced pulmonary hypertension

doi: 10.1152/ajplung.00531.2016

Figure Lengend Snippet: The adoptive transfer of TH17 cells causes spontaneous PH in RAG1−/− mice. CD4+ T cells were purified from IL-17A enhanced green fluorescent protein (EGFP) mice and cultured under TH17-polarizing conditions. TH17 cells were purified by fluorescence-activated cell sorting and injected into RAG1−/− mice. After 14 days, mice remained in normoxia or were exposed to CH for 21 days and compared with mice that did not receive TH17 cells. A: RVSP. B: %wall thickness. C: Fulton’s index. D: hematocrit. E: representative images of lung sections immunostained for CD3 from mice receiving adoptive transfer or a RAG1−/− mouse that received no adoptive transfer. Scale bars, 100 µm. F: flow cytometric analysis of TH17 cells from the spleens of mice that received an adoptive transfer. Values are means ± SE; n = no. of animals. *P < 0.05 vs. normoxia; #P < 0.05 vs. normoxia No AT; &P < 0.05 vs. CH No AT. Analyzed by 2-way ANOVA, followed by multiple comparisons Student-Newman-Keuls test.

Article Snippet: Cells were washed and stained with PerCP/Cy5.5 anti-mouse CD4 (clone GK1.5, 100432; BioLegend, San Diego, CA), APC/Cy7 anti-mouse CD3ε (clone 145-2C11, 100330; BioLegend), and PE anti-mouse IL-17A (clone TC11-18H10.1, 506910; BioLegend) or PE anti-mouse/rat/human FoxP3 (clone 150D, 32008, BioLegend) for 1 h at 4°C in the dark.

Techniques: Adoptive Transfer Assay, Purification, Cell Culture, Fluorescence, FACS, Injection

Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Central role of T helper 17 cells in chronic hypoxia-induced pulmonary hypertension

doi: 10.1152/ajplung.00531.2016

Figure Lengend Snippet: IL-17A stimulates migration but does not affect mouse pulmonary arterial smooth muscle cell number in culture. A: %wound density in the presence or absence of IL-17A (100 ng/ml). B: confluency density normalized to time 0 in the presence or absence of IL-17A (100 ng/ml) in serum-starved cells. Growth medium was used a positive control. *P < 0.05, cells from n = 4 mice, 2-way repeated-measures ANOVA, followed by multiple-comparison Student-Newman-Keuls test.

Article Snippet: Cells were washed and stained with PerCP/Cy5.5 anti-mouse CD4 (clone GK1.5, 100432; BioLegend, San Diego, CA), APC/Cy7 anti-mouse CD3ε (clone 145-2C11, 100330; BioLegend), and PE anti-mouse IL-17A (clone TC11-18H10.1, 506910; BioLegend) or PE anti-mouse/rat/human FoxP3 (clone 150D, 32008, BioLegend) for 1 h at 4°C in the dark.

Techniques: Migration, Positive Control, Comparison

Journal: PLOS ONE

Article Title: Anti-inflammatory effect of the combined treatment of LMT-28 and kaempferol in a collagen-induced arthritis mouse model

doi: 10.1371/journal.pone.0302119

Figure Lengend Snippet: (A) Arthritis scores of mice were measured every 3 days starting at 21 days after the first immunization. (B) DBA1/J mice showed deformation and swelling of the toe and ankle joints. (C) Paraffin-embedded sections of CIA mice joint were stained with hematoxylin and eosin. (D) Histological analysis was performed based on the analysis criteria: inflammation, synovial hyperplasia, pannus formation, and erosion of cartilage and bone. (E) The protein levels of IL-6, IL-1β and IL-17 in CIA mouse serum were measured using ELISA. Data are presented as mean ± SEM. # p <0.05 vs. normal group, * p <0.05, ** p <0.01, and *** p <0.005 vs. CIA control group.

Article Snippet: The cells were incubated with anti-IL-17A-PE antibody (Miltenyi Biotec) or anti-mouse FOXP3 antibody (Invitrogen, Waltham, MA, USA) diluted in 1× permeabilization buffer (eBioscience) for 30 min at 4°C in the dark.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Control

Journal: Annals of Translational Medicine

Article Title: Mesenchymal stromal cells ameliorate acute lung injury induced by LPS mainly through stanniocalcin-2 mediating macrophage polarization

doi: 10.21037/atm.2020.02.105

Figure Lengend Snippet: The secretion of IL-10 (left) and TSG6 (right) from STC2 silenced MSCs or control MSCs detected by ELISA. IL-10, interleukin 10; TSG6, tumor necrosis factor α stimulated gene 6. Data are shown as mean ± SD (n=3). *, P<0.05. STC2, stanniocalcin-2; MSC, mesenchymal stem cell; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Protein levels of interleukin-17A and TNF-α in the BALF were quantified by specific enzyme-linked immunosorbent assay (ELISA) kits (Boster Biological Technology Co., Ltd., China) according to standard protocols.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Annals of Translational Medicine

Article Title: Mesenchymal stromal cells ameliorate acute lung injury induced by LPS mainly through stanniocalcin-2 mediating macrophage polarization

doi: 10.21037/atm.2020.02.105

Figure Lengend Snippet: STC2 contributed to MSCs-mediated ameliorating effects on LPS-induced ALI. Male adult BALB/c mice (8 to 10 weeks old) were randomly divided into four different groups: PBS + PBS, LPS + PBS, LPS + MSCshNTC, and LPS + MSCshSTC2. The ALI model was established by LPS (5 mg/kg) installation intranasally. The control group received the same volume of PBS. The mice were killed to obtain lung tissues 24 h after LPS treatment or were used to collect BALF. (A) The degree of lung injury was assessed with H&E staining; (B) histopathological mean lung injury scores; (C) neutrophil count in BALF; (D) the pulmonary permeability index was measured by Evans blue dye. TNF-α (E) and IL-17A (F) levels in BALF were detected using ELISA assay. n=10 each group. *, P<0.05; **, P<0.01; ***, P<0.001. STC2, stanniocalcin-2; MSC, mesenchymal stem cell; ALI, acute lung injury; BALF, bronchoalveolar lavage fluid; ELISA, enzyme-linked immunosorbent assay.

Article Snippet: Protein levels of interleukin-17A and TNF-α in the BALF were quantified by specific enzyme-linked immunosorbent assay (ELISA) kits (Boster Biological Technology Co., Ltd., China) according to standard protocols.

Techniques: Control, Staining, Permeability, Enzyme-linked Immunosorbent Assay