il17a egfp  (Jackson Laboratory)

Journal: Molecular Medicine

Article Title: IL17F + naïve and IFNγ + memory CD8 T cells drive hepatic dysfunction in the cecal ligation and puncture model of sepsis

doi: 10.1186/s10020-025-01411-2

Figure Lengend Snippet: Effects of Immune Education, CLP, and Specific Cytokine Blockade on T cell Cytokine Production. A-C Immune Educated or Control IFNγ-eYFP reporter mice on a C57Bl/6 background were sacrificed prior to or 24 h. following CLP. IFNγ + T cells were assessed. Percent IFNγ + CD4 (Left) and CD8 (Right) T cells in the ( A ) spleen and ( B ) liver. C Histograms of IFNγ + (Red) and negative (Gray) splenic CD4 T cells in Educated mice following CLP. D-E Immune Educated or Control IL17A EGFP /IL17F mCherry reporter mice on a C57Bl/6 background were sacrificed prior to or 24 h. following CLP. IL17A + and IL17F + T cells were assessed. Percent IL17A + CD4 (Left), IL17A + CD8 (Middle Left), IL17F + CD4 (Middle Right) and IL17F + CD8 (Right) T cells in the ( D ) inguinal lymph node (ILN) and ( E ) liver. F Histograms of IL17A + (Orange) vs IL17F + (Green) ILN CD4 T cells in Educated mice following CLP. A-F * = p < 0.05 for comparisons between two groups, † = p < 0.05 for interaction effect, # = p < 0.05 for CLP effect, ^ = p < 0.05 for Education effect. N = 3–4/group. G Hepatic leukocytes were isolated from Educated or control C57Bl/6 mice prior to or 24 h. following CLP alone or with specific cytokine blockade and intracellular cytokine production was assessed following in vitro PMA/Ionomycin stimulation. Percent IFNγ + (Left) and IL17A + (Right) CD8 T cells. Analyzed by 2-way ANOVA. * = p < 0.05 for comparisons between two groups, # = p < 0.05 for specific cytokine effect, p values for interactions, effect of education or effect of blockade displayed. N = 3–5/group. Gating: FSC/SSC, Singlets, Live, CD90/CD4 or CD90/CD8, eYFP, eGFP, mCherry, IFNγ, IL17A, or TNF

Article Snippet: CD4 −/− (B6.129S2-CD4 tm1Mak /J), CD8 −/− (B6.129S2-CD8a tm1Mak /J), IFNγ −/− (B6.129S7-Ifng tm1Ts /J IFNγ), IFNγ-eYFP reporter mice (B6.129S4-Ifng tm3.1Lky /J), and IL17A-eGFP/IL17F-mCherry reporter mice. (C57Bl/6-Il17a tm1Bcgen Il17f em1Litt /J IL17A EGFP /IL17F mCherry ), all on a C57Bl/6 genetic backgound, were obtained from the Jackson Laboratory and bred and maintained in our barrier SPF animal facility.

Techniques: Control, Isolation, In Vitro

Journal: Nature Communications

Article Title: IL-17a induces age-related olfactory dysfunction by impairing regeneration and promoting respiratory metaplasia in mice

doi: 10.1038/s41467-025-67786-2

Figure Lengend Snippet: a UMAP visualization of scRNA-seq data, colored by annotated T cell subtypes. b Compositional bar plot showing the proportional abundance of each T cell subtype. c Feature plots demonstrating expression patterns of canonical markers of representative cell types. d Dot plot showing the expression levels of key marker genes across T cell subtypes. Dot size reflects the percentage of cells expressing the gene; color intensity indicates average expression level. e List of marker genes used to identify CD4 + Th cell subsets. f Violin plots showing expression patterns of representative marker genes across CD4 + Th cell subsets. g Differentiation index scores for CD4 + Th cell subsets, calculated based on marker genes listed in ( e ). The horizontal solid lines represent the median, and the horizontal dashed lines represent the first/third quartile. h Proportion of IL-17a-expressing cells within each T cell subset. i Concentrations of IL-17a and IL-23 in plasma and OE tissues from young and aged mice ( n = 3 biologically independent experiments). j Western blot analysis of IL-17a protein expression in OE tissues from young and aged mice. k qPCR analysis showing Il17a mRNA expression in OE tissues of young and aged mice ( n = 5 biologically independent experiments). l Violin plot showing Il17a expression levels in OE tissues from young versus aged mice based on scRNA-seq data. m Confocal images of OE tissue sections immunostained for CD3, CD4 and IL-17a in young and aged mice. n Quantification of IL-17a + cells from scRNA-seq data in young and aged OE samples. o Correlation analysis between Il17a mRNA expression level and time taken to locate hidden cookies in young and aged mice ( n = 14 individual mice). The association was evaluated by simple linear regression. p Olfactory behavioral test measuring the time required to locate cookies in young and aged mice ( n = 20 biologically independent samples). Data are presented as mean ± SD ( g , i , k , l , p ). Statistical analyses are performed using the Wilcoxon test ( g, l ) or an unpaired two-sided t-test ( i , k , p ). Confocal microscopy images are representative of at least three biologically independent experiments with consistent results. All nuclei are counterstained with Hoechst. Scale bars, 50 μm. Source data are provided as a Source Data file.

Article Snippet: C57BL/6 JCya-Tg (Lck-Cre), C57BL/6JCya- Il17a em1flox , C57BL/6JCya- Il17a tm2(EGFP) mice were purchased from Cyagen Biologics (Suzhou, China).

Techniques: Expressing, Marker, Clinical Proteomics, Western Blot, Olfactory, Confocal Microscopy

Journal: Nature Communications

Article Title: IL-17a induces age-related olfactory dysfunction by impairing regeneration and promoting respiratory metaplasia in mice

doi: 10.1038/s41467-025-67786-2

Figure Lengend Snippet: a Experimental workflow for scRNA-seq analysis of OE tissues from Il17a CKO and Flox mice at 7 and 14 dpi. Created in BioRender. Shenglei, W. (2025) https://BioRender.com/o9l4wkk . b UMAP visualization of OE cells, color-coded by annotated cell types as shown in the legend. c Dot plot displaying representative marker gene expression for each cell type. Dot size reflects the percentage of cells expressing the gene; color intensity represents average expression level. d Bar graph showing the proportional distribution of cell types across samples, with color coding corresponding to panel ( b ). e Feature plots illustrating expression of key marker genes: Krt5 (HBC), Neurod1 (GBC), Tubb3 (iOSN), Omp (mOSN), Trpm5 (MV), Plp1 (OEG), Rgs5 (SUS), Krt19 (Res BC), Sox2 (SUS/GBC), Ctla4 (CD4 + Th), Adgre1 (Mac), and Il17a (Th17 cells). Gradient from gray to blue indicates low to high expression. f Violin plots showing expression levels of Omp , Tubb3 , Ascl1 and Il17a in relevant cell subsets. g Latency time for locating hidden cookies in behavioral tests performed on Flox and CKO mice at 1, 3, 7, and 14 dpi ( n = 7 biologically independent replicates). h Olfactory habituation / dishabituation tests using serial dilutions of geraniol and citronitrile in mineral oil ( n = 4 biologically independent replicates). i, j Representative immunofluorescence images ( i ) and statistical analysis ( j ) of OMP + and Tuj1 + cells in the OE of Flox and CKO mice at 7 and 14 dpi ( n = 9 biologically independent samples). k Dot plot from CellChat analysis showing differential signaling interactions between T cells and OE cell types in Flox and CKO mice. Data are presented as mean ± SD ( g , j ). Statistical analyses are performed using one-way ANOVA with Sidak’s multiple comparison test ( j ) or two-way ANOVA with Sidak’s multiple comparison test ( g , h ). Confocal microscopy images are representative of at least three biologically independent experiments with consistent results. All nuclei are counterstained with Hoechst. Scale bars, 50 μm. Source data are provided in the Source Data file.

Article Snippet: C57BL/6 JCya-Tg (Lck-Cre), C57BL/6JCya- Il17a em1flox , C57BL/6JCya- Il17a tm2(EGFP) mice were purchased from Cyagen Biologics (Suzhou, China).

Techniques: Marker, Gene Expression, Expressing, Olfactory, Immunofluorescence, Comparison, Confocal Microscopy

Journal: bioRxiv

Article Title: ITK signaling regulates a switch between T helper 17 and T regulatory cell lineages via a calcium-mediated pathway

doi: 10.1101/2023.04.01.535229

Figure Lengend Snippet: ( a ) Naïve WT, Itk -/- or ITK as IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions (anti-CD3/28, IL6 and TGFβ) in presence of ITK as inhibitor 2MBPP1 or DMSO control as indicated, followed by flow cytometric analysis for percentage of GFP + /IL17 + cells and RFP + /Foxp3 + Treg-like cells. ( b ) Top: Naïve WT IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions in presence of ITK inhibitor CPI-818 or DMSO control, followed by quantification of percentage of GFP + /IL17 + cells and RFP + /Foxp3 + Treg-like cells. Bottom: Naïve WT (left) or ITK as (right) IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions in presence of ITK inhibitor CPI-818 (left) ITK as inhibitor 3MBPP1 (right) or DMSO control, followed by quantification of percentage of GFP + /IL17 + cells and RFP + /Foxp3 + Treg-like cell. Mean ± SEM, one-way ANOVA was performed for statistical significance where * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001 and **** p ≤ 0.0001, 3 independent experiments. ( c ) Switched Foxp3 + Treg-like cells generated under Th17 differentiation conditions in presence of ITK inhibitor, or in vitro generated Th17 cells, were then further reactivated under Th17 differentiation conditions in presence of ITK inhibitor CPI-818 or DMSO control for a duration of 5 days (Switched Foxp3 + Treg-like cells and Th17 cells) or 10 days (Th17 cells). Mean ± SEM, Student’s T test was performed for statistical significance where * p ≤ 0.05, ** p ≤ 0.005, 3 independent experiments.

Article Snippet: WT, Itk -/- and ITK as IL17-GFP/Foxp3-RFP dual reporter strains were generated by crossing IL17-GFP (B-IL17A-EGFP tm1 , Biocytogen, Worchester, MA) with Foxp3-RFP (C57BL/6- Foxp3 tm1Flv /J, Jackson Laboratory, Bar Harbor, ME) strain ( ) as previously reported ( ).

Techniques: Generated, In Vitro

Journal: bioRxiv

Article Title: ITK signaling regulates a switch between T helper 17 and T regulatory cell lineages via a calcium-mediated pathway

doi: 10.1101/2023.04.01.535229

Figure Lengend Snippet: ( a ) Naïve WT IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions followed by addition of ITK inhibitor CPI-818 after 1, 2, 3 or 4 days of culture, followed by analysis on day 5. ( b ) Naïve WT IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions in the presence of the ITK inhibitor CPI-818, followed by removal of inhibitor after 1, 2, 3 or 4 days of culture, with analysis on day 5. Mean ± SEM, one-way ANOVA was performed for statistical significance where * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001 and **** p ≤ 0.0001, 2 independent experiments.

Article Snippet: WT, Itk -/- and ITK as IL17-GFP/Foxp3-RFP dual reporter strains were generated by crossing IL17-GFP (B-IL17A-EGFP tm1 , Biocytogen, Worchester, MA) with Foxp3-RFP (C57BL/6- Foxp3 tm1Flv /J, Jackson Laboratory, Bar Harbor, ME) strain ( ) as previously reported ( ).

Techniques:

Journal: bioRxiv

Article Title: ITK signaling regulates a switch between T helper 17 and T regulatory cell lineages via a calcium-mediated pathway

doi: 10.1101/2023.04.01.535229

Figure Lengend Snippet: ( a ) Foxp3 + Treg-like cells that are generated from WT IL17A-GFP/Foxp3-RFP CD4 + T cells activated under Th17 differentiation conditions in presence of ITK inhibitor CPI-818, and compared to iTregs, pTregs and nTregs for expression of select Treg markers. Expression represented as mean fluorescence intensity. ( b ) CD45.2 switched Foxp3 + Treg-like cells (generated in absence of ITK activity under Th17 conditions) and iTregs were sort purified, followed by co-culture with CFSE labelled CD45.1 naïve CD4 + T cell responders. Representative CFSE plots (top). Percentage suppression of proliferation of naïve CD4 + T cell responders by Foxp3 + Treg-like cells and iTregs was quantified (bottom). Mean ± SEM for percentage of cells undergoing division. One-way ANOVA performed for statistical significance where * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001 and **** p ≤ 0.0001, 3 independent experiments.

Article Snippet: WT, Itk -/- and ITK as IL17-GFP/Foxp3-RFP dual reporter strains were generated by crossing IL17-GFP (B-IL17A-EGFP tm1 , Biocytogen, Worchester, MA) with Foxp3-RFP (C57BL/6- Foxp3 tm1Flv /J, Jackson Laboratory, Bar Harbor, ME) strain ( ) as previously reported ( ).

Techniques: Generated, Expressing, Fluorescence, Activity Assay, Purification, Co-Culture Assay

Journal: bioRxiv

Article Title: ITK signaling regulates a switch between T helper 17 and T regulatory cell lineages via a calcium-mediated pathway

doi: 10.1101/2023.04.01.535229

Figure Lengend Snippet: Switched Foxp3 + Treg-like cells were compared with in vitro generated Th17 and iTregs by ATAC-Seq analysis. The chromatin accessibility profiles were compared via ( a ) PCA analysis, ( b ) heatmap of global differential peaks in chromatin accessibility, and fold changes of global differential peaks in chromatin accessibility for switched Foxp3 cells compared to ( c ) Th17 cells and iTregs. ( d ) Tracks indicate chromatin areas chromatin accessibility for Foxp3, RORC, IL17A and IL17F in Foxp3 + Treg like cells, compared to Th17 cells and iTreg cells.

Article Snippet: WT, Itk -/- and ITK as IL17-GFP/Foxp3-RFP dual reporter strains were generated by crossing IL17-GFP (B-IL17A-EGFP tm1 , Biocytogen, Worchester, MA) with Foxp3-RFP (C57BL/6- Foxp3 tm1Flv /J, Jackson Laboratory, Bar Harbor, ME) strain ( ) as previously reported ( ).

Techniques: In Vitro, Generated

Journal: bioRxiv

Article Title: ITK signaling regulates a switch between T helper 17 and T regulatory cell lineages via a calcium-mediated pathway

doi: 10.1101/2023.04.01.535229

Figure Lengend Snippet: ( a ) Naïve Itk -/- IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions in presence of ionomycin or DMSO control, followed by analysis of percentage of GFP + /IL17 + cells and percentage of RFP + /Foxp3 + Treg-like cells. Representative flow plots (top), Quantified (bottom). ( b ) Naïve WT IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under Th17 differentiation conditions in presence of ionomycin or DMSO control, with or without ITK inhibitor CPI-818, followed by analysis of percentage of GFP + /IL17 + cells and percentage of RFP + /Foxp3 + Treg-like cells. Representative flow plots (top), Quantified (bottom). ( c ) Naïve Itk -/- IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under iTreg differentiation conditions in presence of ionomycin or DMSO control, followed by analysis of percentage of GFP + /IL17 + cells and percentage of RFP + /Foxp3 + Treg-like cells. Representative flow plots (top), Quantified (bottom). ( d ) Naïve WT IL17A-GFP/Foxp3-RFP CD4 + T cells were activated under iTreg differentiation conditions in presence of ionomycin or DMSO control, with or without ITK inhibitor CPI-818, followed by analysis of percentage of GFP + /IL17 + cells and percentage of RFP + /Foxp3 + Treg-like cells. Representative flow plots (top), Quantified (bottom). Mean ± SEM, T test was performed for statistical significance where * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.001 and **** p ≤ 0.0001, 3 independent experiments.

Article Snippet: WT, Itk -/- and ITK as IL17-GFP/Foxp3-RFP dual reporter strains were generated by crossing IL17-GFP (B-IL17A-EGFP tm1 , Biocytogen, Worchester, MA) with Foxp3-RFP (C57BL/6- Foxp3 tm1Flv /J, Jackson Laboratory, Bar Harbor, ME) strain ( ) as previously reported ( ).

Techniques: