Journal: iScience

Article Title: Ambient temperature regulates CD4 + T cell tonic T cell receptor signaling and responsiveness

doi: 10.1016/j.isci.2026.115801

Figure Lengend Snippet: Thermoneutrality augments TCR signaling strength (A–J) 8-to-10-week-old Nur77 GFP C57BL/6J male mice were housed in Tn (30 °C)- or Ts (22 °C)-conditions. After 2 weeks, splenic T cells (CD45 + TCRβ + TCRγδ − NK1.1 - ) were characterized via flow cytometry (n = 4–10 biological replicates per group). (A) Schematic overview. (B) Representative flow cytometric plot of Nur77-GFP expression in splenic CD4 + (left) and CD8 + (right) T cells. (C, D) Frequency of Nur77-GFP + CD4 + T cells (C) and CD8 + T cells (D). (E, F) Relative mean fluorescence intensity (MFI) of Nur77-GFP in CD4 + T cells (E) and CD8 + T cells (F). (G, H) Relative MFI of CD69 in CD4 + T cells (G) and CD8 + T cells (H). (I, J) Frequency of CD5 hi Ly6C − CD4 + T cells (I) and CD8 + T cells (J). (K, L) 8-to-10-week-old WT C57BL/6J male mice were housed in Tn (30 °C)- or Ts (22 °C)-conditions. After 2 weeks, splenic CD4 + T cells were negatively selected via magnetic separation and stimulated ex vivo with anti-CD3 and anti-CD28 for the indicated times. Levels of phosphorylated ZAP70 (p-ZAP70) and total ZAP70 were quantified via western blot analysis ( n = 4 biological replicates per group). (K) Representative western blot. (L) Quantification of relative phosphorylated-to-total ZAP70 ratio. Statistical analysis was performed between each time point, in addition to the total area under the curve (AUC), for each group. (E-H) MFI of each data point was divided by the average MFI in the control (Ts-housed) group per experiment to yield relative MFI. (L) Relative phospho-to-total ZAP70 ratios of each timepoint were normalized to that of the 0 min timepoint of the Ts replicate per membrane. (C-L) Mann-Whitney U test. ∗: p < 0.05, ∗∗: p < 0.01, and ∗∗∗: p < 0.001 (Ts vs. Tn). #: p < 0.05 (Ts AUC vs. Tn AUC). For bar graphs, data represent mean ± SEM. For the line graph, data represent mean ± SEM, and the shaded region under the lines represents the area used to calculate AUC.

Article Snippet: Nur77 GFP mice , Jackson Laboratory , Strain #: 016617; RRID: IMSR_JAX:016617.

Techniques: Flow Cytometry, Expressing, Fluorescence, Ex Vivo, Western Blot, Control, Membrane, MANN-WHITNEY

Journal: iScience

Article Title: Ambient temperature regulates CD4 + T cell tonic T cell receptor signaling and responsiveness

doi: 10.1016/j.isci.2026.115801

Figure Lengend Snippet: Thermoneutrality alters CD4 + T cell transcriptome upon tonic TCR signaling activation 8-to-10-week-old Nur77 GFP C57BL/6J male mice were housed in Tn- or Ts-conditions. After 2 weeks, splenic Nur77-GFP + and Nur77-GFP - CD4 + T cells (CD45 + TCRβ + TCRγδ − NK1.1 − CD4 + ) from each condition were isolated via fluorescence-activated cell sorting. Transcriptomic analysis was then performed via bulk RNA-sequencing (n = 3–4 biological replicates per group). (A) Schematic overview. (B) Principal component analysis. Orange: Nur77-GFP - CD4 + T cells from Tn-housed mice. Red: Nur77-GFP + CD4 + T cells from Tn-housed mice. Light blue: Nur77-GFP - CD4 + T cells from Ts-housed mice. Dark blue: Nur77-GFP + CD4 + T cells from Ts-housed mice. Ovals represent a visual enclosing shape. (C) Volcano plot of gene expression levels between Nur77-GFP + CD4 + T cells from Tn- or Ts-housed mice. Red dots represent genes significantly upregulated (FC > 1.5, p -value <0.05) in Tn-housed mice. Blue dots represent genes significantly upregulated (FC < −1.5, p -value <0.05) in Ts-housed mice. (D) ToppGene functional enrichment analysis of genes significantly upregulated in Ts-housed mice. (E) ToppGene functional enrichment analysis of genes significantly upregulated in Tn-housed mice.

Article Snippet: Nur77 GFP mice , Jackson Laboratory , Strain #: 016617; RRID: IMSR_JAX:016617.

Techniques: Activation Assay, Isolation, Fluorescence, FACS, RNA Sequencing, Gene Expression, Functional Assay

Journal: iScience

Article Title: Ambient temperature regulates CD4 + T cell tonic T cell receptor signaling and responsiveness

doi: 10.1016/j.isci.2026.115801

Figure Lengend Snippet: Thermoneutrality-driven augmentation of CD4 + T cell TCR signaling strength is mediated by MHC class II presentation of self-peptide (A and B) 8-to-10-week-old OT-II transgenic (Tg) C57BL/6J male mice were housed in Tn- or Ts-conditions. After 2 weeks, splenic CD4 + T cells (CD45 + TCRβ + TCRγδ − NK1.1 − CD4 + ) were characterized via flow cytometry (n = 5–9 biological replicates per group). (A) Schematic overview. (B) Relative MFI of Nur77. (C–F) 8-to-10-week-old WT C57BL/6J male mice were housed in Tn- or Ts-conditions. Mice were treated with anti-MHCII antibodies (250 μg/mouse) or saline control via intraperitoneal injections every three days for 2 weeks. After 2 weeks, splenic immune cells (CD45 + ) were characterized via flow cytometry ( n = 3 biological replicates per group). (C) Schematic overview. (D) Cell counts of MHCII + B cells (B220 + CD11c − ). (E) Cell counts of MHCII + dendritic cells (DCs; CD11c + ). (F) Relative MFI of Nur77 in CD4 + T cells. (G and H) 8-to-10-week-old WT C57BL/6J male mice were housed in Tn- or Ts-conditions. After 2 weeks, splenic immune cells (CD45 + ) were characterized via flow cytometry ( n = 3 biological replicates per group). Relative MFI of CD80 and CD86 in B cells (G) and DCs (H). (B, F-H) MFI of each data point was divided by the average MFI in the control (Ts-housed) group per experiment to yield relative MFI. (B, D-F) Permutation test with Bonferroni correction for multiple comparisons. (G-H) Mann-Whitney U test. ∗: p < 0.05, ∗∗: p < 0.01, ∗∗∗: p < 0.001, and ∗∗∗∗: p < 0.0001. For bar graphs, data represent mean ± SEM.

Article Snippet: Nur77 GFP mice , Jackson Laboratory , Strain #: 016617; RRID: IMSR_JAX:016617.

Techniques: Transgenic Assay, Flow Cytometry, Saline, Control, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Targeting T cell metabolism and polarization to modulate post-stroke immune responses and improve outcomes

doi: 10.3389/fimmu.2026.1703552

Figure Lengend Snippet: Study design. 12–14 week old male Nur77 GFP mice underwent tMCAO, transient middle cerebral artery occlusion. At the timepoint of reperfusion, mice were injected intraperitoneally with LPS (40 µg/kg) or vehicle (PBS). Two hours after tMCAO and daily in the morning, mice were injected intraperitoneally with Soraphen A (SorA, 2.5 µmol/kg) or vehicle (PEG buffer). Magnetic resonance imaging (MRI) was conducted 16 hours and 7 days after tMCAO to verify infarct induction and calculate lesion volumes. Mice were scored daily and a set of behavioral tests (corner tests, cylinder test and inclined plane test) was conducted before as well as 2 and 6 days after tMCAO. At each experimental time point (16 h, 2 d, 3 d, 7 d after MCAO) brain, lungs, spleen, blood and ingiunal lymph nodes (ILN) were harvested and prepared for flow cytometric analysis of T cell activation and polarization, differentiating between antigen-specific and -unspecific T cell activation via GFP expression. To evaluate the impact of SorA treatment on stroke pathology under post-stroke and post-stroke inflammatory conditions associated with infection, we compared treatment groups to assess: the effect of LPS alone (vehicle/vehicle vs. LPS/vehicle), the effect of SorA alone (vehicle/vehicle vs. vehicle/SorA), and the effect of SorA under proinflammatory conditions (LPS/vehicle vs. LPS/SorA) in MCAO mice. Image created in BioRender. Vogelgesang, A. (2026) https://BioRender.com/pr62lu3 .

Article Snippet: The Nur77 GFP mouse model was generated by Kristin Hogquist (C57BL/6-Tg(Nr4a1-EGFP/cre)820Khog/J mice, Stock No: 016617, The Jackson Laboratory) ( ) and express eGFP under the control of the Nr4a1 promoter/enhancer regions.

Techniques: Injection, Magnetic Resonance Imaging, Activation Assay, Expressing, Infection