Journal: bioRxiv

Article Title: The metabolic reprogramming of T cells controls airway remodeling in severe asthma

doi: 10.64898/2026.03.19.712985

Figure Lengend Snippet: To model EA or MGA respectively, Balb/c or Il4ra -/- mice were given five intranasal (i.n.) challenges with 100ug house dust mite (HDM) extract every week (followed by two days resting) for three consecutive weeks. A final i.n. HDM challenge was given 24 hours prior to harvest. ( A ) Lung sections were stained and representative Masson trichrome (collagen), α-smooth muscle actin (αSMA), and hematoxylin and eosin (H&E) sections and accompanying quantifications are shown. ( B ) Airway resistance to increasing doses of methacholine exposure (6-8 mice per group). (C) Expression of mRNA for IL13 and IL17, assessed by quantitative PCR analyses of lung samples. mRNA expression was calculated relative to GAPDH for 5 mice per group. ( D ) Number of neutrophils or eosinophils in the left lung lobe of EA or MGA induced mice, as assessed by flow cytometry. ( E ) MGA-induced animals were treated with 50ug dexamethasone intratracheally one hour prior to each allergen challenge, beginning on day 7. Lung sections were stained and representative Masson trichrome and αSMA sections and accompanying quantifications are shown. ( F ) Number of neutrophils or eosinophils in the left lung lobe of MGA induced mice treated with or without dexamethasone, as assessed by flow cytometry. ( G ) MGA-induced animals were treated with 100ug anti-IL17A or matching isotype control i.p. beginning on day 7 and continuing every other day until harvest. Lung sections were stained, and representative H&E sections and accompanying quantifications are shown. Trichrome data show area of interest values covering full lung section of 6 individual mice per condition. αSMA data show mean values of all bronchi scored from 6 mice per condition. Data are representative of 3 independent experiments. *P < 0.05, **P < 0.005, ***P < 0.0005, ****P < 0.00005; HPF, high-power field; ns, nonsignificant.

Article Snippet: To block IL-17 signaling, 100ug anti-IL17A (17F3; BioXCell) or matching isotype control (MOPC-21; BioXCell) were given intraperitoneally (i.p.) to HDM induced Ilr4a -/- mice beginning on day 7 and continuing every other day for a total of 7 injections.

Techniques: Staining, Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Control

Journal: International journal of molecular sciences

Article Title: Inhibitory Effects of Extracellular Vesicles from iPS-Cell-Derived Mesenchymal Stem Cells on the Onset of Sialadenitis in Sjögren's Syndrome Are Mediated by Immunomodulatory Splenocytes and Improved by Inhibiting miR-125b.

doi: 10.3390/ijms24065258

Figure Lengend Snippet: Figure 3. Effects of iEVs on splenic Th17 cells. Four-month-old female NOD.B10.H2b mice were IV injected with PBS, PD15 iEVs, or PD45 iEVs twice a week for two weeks. Splenocytes were collected at two weeks after last injection. (A,B) Percentages of IL17+ cells in splenic CD4+ T cells were examined with flow cytometry. (C) The mRNA levels of Th17 markers (IL17a, IL21, and Rorc) and negative regulator IL1rn in spleen were examined with qRT-PCR. N = 5, ns: not significant, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001.

Article Snippet: These cells were stained with fluorescent-labeled antibodies against F4/80 (BD Pharmingen 123116, 123107, San Diego, CA, USA), CD3 (BioLegend 100236, San Diego, CA, USA), CD19 (BioLegend 152409), CD38 (BioLegend 102707), CD206 (BioLegend 141706), CD4 (BioLegend 100408), IL17a (130-112-009, Miltenyi Biotec, San Diego, CA, USA), or the corresponding isotype controls (BioLegend 400608, 400511, 400612, 400207, 1:100).

Techniques: Injection, Cytometry, Quantitative RT-PCR

Journal: International journal of molecular sciences

Article Title: Inhibitory Effects of Extracellular Vesicles from iPS-Cell-Derived Mesenchymal Stem Cells on the Onset of Sialadenitis in Sjögren's Syndrome Are Mediated by Immunomodulatory Splenocytes and Improved by Inhibiting miR-125b.

doi: 10.3390/ijms24065258

Figure Lengend Snippet: Figure 6. Effects of inhibiting miR-125b in aging iEVs on the polarization of splenic macrophages. Four-month-old female NOD.B10.H2b mice were IV injected with PBS, Ctrl EVs, or 125KD EVs twice a week for two weeks. Splenocytes were collected at two weeks after last injection. (A,B) Percentages of IL17+ cells in CD4+ splenocytes were examined with flow cytometry. (C) The mRNA levels of Th17 markers (IL17a, IL21, and Rorc) and negative regulator IL1rn in spleen were examined with qRT-PCR. N = 5, ns: not significant, *: p < 0.05, **: p < 0.01, ****: p < 0.0001.

Article Snippet: These cells were stained with fluorescent-labeled antibodies against F4/80 (BD Pharmingen 123116, 123107, San Diego, CA, USA), CD3 (BioLegend 100236, San Diego, CA, USA), CD19 (BioLegend 152409), CD38 (BioLegend 102707), CD206 (BioLegend 141706), CD4 (BioLegend 100408), IL17a (130-112-009, Miltenyi Biotec, San Diego, CA, USA), or the corresponding isotype controls (BioLegend 400608, 400511, 400612, 400207, 1:100).

Techniques: Injection, Cytometry, Quantitative RT-PCR

Journal: eLife

Article Title: Short-range interactions between fibrocytes and CD8 + T cells in COPD bronchial inflammatory response

doi: 10.7554/eLife.85875

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-IL-17-PE-Cy7 (mouse monoclonal) , Miltenyi Biotec , Cat. #:130-120-413, RRID: AB_2752086 , FC (1:20).

Techniques: Blocking Assay, Recombinant

Journal: Frontiers in Genetics

Article Title: Exosomes containing long non-coding RNA AGAP2-AS1 promote the differentiation of CD4 + T cells through the miR-424-5p/SGK1 axis in psoriasis

doi: 10.3389/fgene.2025.1521470

Figure Lengend Snippet: ASAP2-AS1 promotes the differentiation of Th1 and Th17 in CD4 + T cells and serves as a sponge for miR-424-5p. (A) Fluorescence microscopy was used to detect the subcellular localization of AGAP2-AS1 in CD4 + T cells. AGAP2-AS1 was labeled with Cy3, cell nuclei were counterstained with DAPI (blue). (B) The expression of AGAP2-AS1 in CD4 + T cells after co-incubation of TNF-α-treated HaCaTs-derived exosomes. (C) CD4 + T cells were transfected with LV-AGAP2-AS1 or LV-NC. The percentage of Th1 and Th17 cells was detected by flow cytometry. (D) The protein levels of IFN-γ and IL-17A are measured by ELISA. (E) Prediction of the binding sequence between AGAP2-AS1 and miR-424-5p and construction of the AGAP2-AS1 MUT sequence. (F) Dual-luciferase reporter gene assay confirmed the binding relationship between AGAP2-AS1 and miR-424-5p in HEK-293T cells. (G) AGAP2-AS1 expression level in CD4 + T cells transfected with miR-424-5p mimics or NC mimics and miR-424-5p inhibitor or NC inhibitor. WT wild type, MUT mutant type. Th, T helper. Experiments were repeated three times. *P < 0.05, **P < 0.01, ***P < 0.001. ns: no significance.

Article Snippet: For intracellular cytokine staining, cells were fixed with fixation buffer (E-CK-A109, Elabscience) and then stained intracellularly with antibodies against human IFN-γ (E-AB-F1196C, Elabscience) and IL-17A (E-AB-F1173D, Elabscience).

Techniques: Fluorescence, Microscopy, Labeling, Expressing, Incubation, Derivative Assay, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Binding Assay, Sequencing, Luciferase, Reporter Gene Assay, Mutagenesis

Journal: Frontiers in Genetics

Article Title: Exosomes containing long non-coding RNA AGAP2-AS1 promote the differentiation of CD4 + T cells through the miR-424-5p/SGK1 axis in psoriasis

doi: 10.3389/fgene.2025.1521470

Figure Lengend Snippet: AGAP2-AS1/miR-424-5p regulated CD4 + T cell differentiation via the SGK1 signaling pathway. (A) The relative expression levels of SGK1 in 293T cells that were transfected with siRNA targeting SGK1. (B) SGK1 expression level in CD4 + T cells transfected with LV-AGAP2-AS1 or LV-NC and si-AGAP2-AS1 or si-NC. (C) Western blot confirmed that transfection of AGAP2-AS1 in CD4 + T cells promoted the expression of SGK1, and the miR-424-5p mimics reversed this process. (D) Flow cytometry analysis of the percentages of Th1 and Th17 cells in CD4 + T cells transfected with LV-AGAP2-AS1 and miR-424-5p mimics or NC mimics. (E) The protein levels of IFN-γ and IL-17A are measured by ELISA. (F) Flow cytometry analysis of the percentages of Th1 and Th17 cells in CD4 + T cells transfected with LV-AGAP2-AS1 and SGK1 siRNA or NC siRNA. (G) The protein levels of IFN-γ and IL-17A are measured by ELISA. The experiments were repeated three times. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: For intracellular cytokine staining, cells were fixed with fixation buffer (E-CK-A109, Elabscience) and then stained intracellularly with antibodies against human IFN-γ (E-AB-F1196C, Elabscience) and IL-17A (E-AB-F1173D, Elabscience).

Techniques: Cell Differentiation, Expressing, Transfection, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay