mouse anti yfp  (TaKaRa)

Journal: European Journal of Immunology

Article Title: Tissue‐Resident Regulatory T Cells Expressing CD83 Maintain Local Homeostasis and Restrict Th2 Responses in Asthma

doi: 10.1002/eji.202451525

Figure Lengend Snippet: CD83 mitigates activation status of lung tissue‐resident Tregs and restrains effector T cell activation. FACS analysis of Tregs from cervical lymph node (cLN) and lungs of 8–12‐week‐old CD83cKO and control mice. (A) Frequencies of FoxP3 + Tregs among all living CD45 + in the respective organs. (B, C) Frequency of ST2 + and KLGR1 + cells among Tregs. (D) Surface receptor expression of ST2, KLRG1, and GITR on Tregs. (Ctrl n = 8–13, CD83cKO n = 8–13, pool of three independent experiments). (E–I) Lung and cervical lymph node (cLNs) derived lymphocytes from 8 to 12‐week‐old mice were stimulated for 48 h with anti‐CD3/anti‐CD28 and cytokine and chemokine secretion was measured (Ctrl n = 20–21, CD83cKO n = 20–21, pool of six independent experiments). In all graphs, data are represented as mean ± SEM and a two‐tailed Mann–Whitney U ‐test was used to analyze the data.

Article Snippet: Sorted CD3 + CD4 + YFP + Tregs (1 × 10 5 ) were pipetted into a 96‐well U‐bottom plate in 100 μL final volume and stimulated overnight with IL‐4 (10 ng/mL, Milteny Biotec) in R10 medium.

Techniques: Activation Assay, Control, Expressing, Derivative Assay, Two Tailed Test, MANN-WHITNEY

Journal: European Journal of Immunology

Article Title: Tissue‐Resident Regulatory T Cells Expressing CD83 Maintain Local Homeostasis and Restrict Th2 Responses in Asthma

doi: 10.1002/eji.202451525

Figure Lengend Snippet: Asthmatic lungs derived from CD83cKO mice show enhanced Th2 effector T‐cell responses and a highly activated phenotype. (A–F) Flow cytometric analysis of lung‐resident CD4 + T cells and Tregs from asthmatic and control mice: (A) CD4 + T‐cell frequencies among all CD45 + cells. (B) Percentage of GITR + cells among CD4 + T cells and Tregs. (C) Th2 effector cells (Foxp3 − GATA3 + ) frequencies in the CD4 + T cell population. (D) Representative graphs of ST2 and FoxP3 populations among CD4 + cells. (E) Analysis of ST2 + Foxp3 − effector cells and ST2 + Foxp3 + Tregs among CD4 + T cells. (F) Percentages of St2 − Foxp3 + Tregs among lung‐resident T cells, and median expression of FoxP3 on tissue‐resident Tregs. (G) Cytokine and (H) chemokine analysis of lung lymphocytes re‐stimulated with OVA for 24 h. In all graphs, data are represented as mean ± SEM and a two‐tailed Mann–Whitney U ‐test was used to analyze the data (Ctrl n = 4–5, CD83cKO n = 4–5, one representative experiment out of two).

Article Snippet: Sorted CD3 + CD4 + YFP + Tregs (1 × 10 5 ) were pipetted into a 96‐well U‐bottom plate in 100 μL final volume and stimulated overnight with IL‐4 (10 ng/mL, Milteny Biotec) in R10 medium.

Techniques: Derivative Assay, Control, Expressing, Two Tailed Test, MANN-WHITNEY

Journal: European Journal of Immunology

Article Title: Tissue‐Resident Regulatory T Cells Expressing CD83 Maintain Local Homeostasis and Restrict Th2 Responses in Asthma

doi: 10.1002/eji.202451525

Figure Lengend Snippet: IL‐4 dependent destabilization of CD83cKO Tregs leads to insufficient control of Th2 cell differentiation. (A) Schematic overview of DC/T cell coculture: LPS‐stimulated DCs are co‐cultured with CellTrace Violet (CTV)‐labeled CD4 + T cells from wild‐type or CD83cKO mice in the presence of stimuli (anti‐CD3/anti‐IFNγ/IL‐4) for 4 days. (B) Representative histogram showing the proliferation of CD83CKO and control cells and the evaluation by the replication index. (C) Representative plots of GATA3 expression in CD4 + T cells under different cultivation conditions. (D) Frequency of GATA3 + T cells amongst CD4 + T cells. (E) Assessment of IL‐13 production in supernatants of co‐cultures ( n = 9, pool of three independent experiments). (F) Representative dot plots of CD4 + T cells (upper panel) and (lower panel) from the suppression assay. Percentages of (G) CD25 + /FoxP3 + cells and (H) ex‐Tregs among CTV − cells ( n = 5, pooled from two independent experiments). (I) Gene expression analysis of Il4ra in sorted Tregs (CD3 + CD4 + YFP + ) from spleens of Ctrl and CD83cKO mice. (J) Median fluorescence of CD25 and GITR on Foxp3 + Tregs after stimulation with IL‐4 in relation to nontreated control (NTC) ( n = 6, pool of three independent experiments). Data are represented as mean ± SEM and a two‐tailed Mann–Whitney U ‐test was used for analysis.

Article Snippet: Sorted CD3 + CD4 + YFP + Tregs (1 × 10 5 ) were pipetted into a 96‐well U‐bottom plate in 100 μL final volume and stimulated overnight with IL‐4 (10 ng/mL, Milteny Biotec) in R10 medium.

Techniques: Control, Cell Differentiation, Cell Culture, Labeling, Expressing, Suppression Assay, Gene Expression, Fluorescence, Two Tailed Test, MANN-WHITNEY