flu influenza peptides  (Miltenyi Biotec)

Journal: Cell Death & Disease

Article Title: Insulin enables acquisition of the IL7R + memory phenotype in PD1 + T cells in RA tissues

doi: 10.1038/s41419-026-08916-6

Figure Lengend Snippet: A Glucose metabolism. Studied metabolic pathways are in bold. B Box plots of standardized expression sum (SES), by RNA-seq, of genes within metabolic pathways in CD4 + cells of RA ( n = 11), healthy controls (HC, n = 57), and of CD4 + cells split by mean expression of BIRC5 gene in high (B5 hi ) and low (B5 lo ) ( n = 24). Boxes indicate IQR and whiskers indicate the minimum and maximum values. GO-terms used for pathway annotation: Glycolysis (GO:0006096); oxidative phosphorylation (OxPhos, GO:0006119 and GO:0022900); pentose phosphate pathway (PPP, GO:0006098); tricarboxylic acid cycle (TCA, GO:0006099). P -values are calculated by Wilcoxon unpaired test. C Heatmap of Spearman’s correlation between HAT, HDAC and metabolic pathways as above. GO-terms used for annotation HAT (GO:0000123) and HDAC (GO:0000118). Asterisks indicate p -values. * < 0.05, ** < 0.01, *** <0.001. D Heatmap of Spearman’s correlation between HAT and HDAC and insulin signaling (IS). Asterisks indicate p -values * < 0.05, ** < 0.01, *** <0.001. E Scatter plot of Spearman’s correlation between plasma insulin levels and SES of insulin signaling. F Uniform manifold approximation and projection (UMAP) map of scaled expression intensity of BIRC5 , IFNG , and TNF in T cells of RA synovial tissue (ST), by single cell transcriptome. BIRC5 Hi CD4 + clusters are indicated. G Heatmap of scaled expression intensity of metabolic pathway (annotated as above) in peripheral blood (PB), synovial fluid (SF), and ST of RA patients, by scRNA-seq. H Violin plot of insulin signaling by SES of INSR, IRS1, IRS2 and IGF1R genes in BIRC5 Hi CD4 + clusters. P-value was calculated by chi-square test. I Violin plot of metabolic pathways SES in cells with high and low IS in BIRC5 Hi CD4 + clusters. P -values are calculated by Wilcoxon unpaired test. Asterisks indicate * < 0.05, ** < 0.01, *** <0.001, **** <0.0001 J . Box plot of BIRC5, IFNG , and TNF expression in Tph cells with high and low IS. Boxes indicate IQR and whiskers indicate the minimum and maximum values. P -values are calculated by Wilcoxon unpaired test. K Heatmap of expression for IL7R-signaling targets in Tph cells high and low IS. Genes regulated by survivin-H3K27ac are in bold. P -values are calculated by Wilcoxon unpaired test. Asterisks indicate * < 0.05, ** < 0.01, *** <0.001, **** <0.0001.

Article Snippet: Cells were stimulated with concanavalin A (ConA, 0.625 μg/mL, MP Biomedicals), and lipopolysaccharide (LPS, 5 μg/mL, Sigma-Aldrich) for 48 h. For immunophenotyping, the cultures were supplemented with insulin (Humalog 100 U/mL, Eli Lilly, Indianapolis, IN, USA) 25 nM and/or the class I HDAC inhibitor (HDACi) valproate, 50 μg/mL (stock 200 mg/mL, Ergenyl, Sanofi, Paris, France) after 24 h. Supernatants were collected for cytokine measures and cells were analyzed by flow cytometry.

Techniques: Expressing, RNA Sequencing, Phospho-proteomics, Clinical Proteomics, Single Cell

Journal: Cell Death & Disease

Article Title: Insulin enables acquisition of the IL7R + memory phenotype in PD1 + T cells in RA tissues

doi: 10.1038/s41419-026-08916-6

Figure Lengend Snippet: A Confocal microscopy images of THP1 cells stained with antibodies to H3K27ac (red) in nucleus (blue) after stimulation with insulin (top), and HDAC-inhibitor (bottom) for 24 h. Images are acquired with 40× magnification with additional digital magnification 1.5×. B Scatter plot of nuclear H3K27ac staining intensity after stimulation with insulin (top) and HDAC-inhibitor (bottom). P -values are calculated by Wilcoxon unpaired test. Asterisks indicate * < 0.05, ** < 0.01, *** <0.001, **** <0.0001. C Histogram of phosphorylated serine 473 (p)AKT1 mean fluorescence intensity in THP1 cells stimulated with 10 nM insulin, by flow cytometry. D Histogram of H3K27ac mean fluorescence intensity in THP1 cells stimulated with increasing concentrations of insulin for 24 h, by flow cytometry. E Histogram of pAKT1 mean fluorescence intensity in human lymphocytes stimulated with increasing concentrations of insulin for 30 min, by flow cytometry.

Article Snippet: Cells were stimulated with concanavalin A (ConA, 0.625 μg/mL, MP Biomedicals), and lipopolysaccharide (LPS, 5 μg/mL, Sigma-Aldrich) for 48 h. For immunophenotyping, the cultures were supplemented with insulin (Humalog 100 U/mL, Eli Lilly, Indianapolis, IN, USA) 25 nM and/or the class I HDAC inhibitor (HDACi) valproate, 50 μg/mL (stock 200 mg/mL, Ergenyl, Sanofi, Paris, France) after 24 h. Supernatants were collected for cytokine measures and cells were analyzed by flow cytometry.

Techniques: Confocal Microscopy, Staining, Fluorescence, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Insulin enables acquisition of the IL7R + memory phenotype in PD1 + T cells in RA tissues

doi: 10.1038/s41419-026-08916-6

Figure Lengend Snippet: A Analysis strategy of survivin and H3K27ac deposition, by ChIP-seq, in cis-regulatory elements ( cis -RE) and connected genes in human CD4 + T cells. B Venn diagram of insulin-responsive (InsResp) genes connected to cis -RE with survivin-H3K27ac co-deposition. Genes are identified by DESeq2 analysis of CD4 + cells transcriptome after a) direct insulin stimulation (InsStim) and b) in regression to corresponding plasma insulin levels, by RNA-seq. C Scatter plot of pathway enrichment of insulin-responsive genes, by GSEA GO-terms. Circle size indicates the number of genes in the pathway, and color intensity indicates p -value. D Venn diagram of insulin-responsive (InsResp) genes connected to cis -RE with H3K27ac deposition. Genes are identified by DESeq2 analysis of CD4 + cells transcriptome after a) direct insulin stimulation (InsStim) and b) HDAC inhibition. E Scatter plot of protein subunits with survivin-H3K27ac in HAT (E1) and HDAC (E3) complexes, annotated by STRING database. Circle size represents number of cis- RE with survivin-H3K27ac connected to the gene. Circle color represents beta-coefficient of expression difference by DESeq2 analysis of CD4 + cell transcriptome in regression to corresponding plasma insulin levels. (E2 and E4) Heatmap of transcription difference, by log2FoldChange (FC), in CD4 + cells in regression to plasma insulin levels (InsReg), and after HDAC inhibitor treatment in vitro. F . Bar plot of percentage of H3K27ac and survivin-H3K27ac connected genes in each metabolic pathway. GO-terms used for pathway annotation as in Fig. . G Scatter plot of expression difference of survivin-H3K27ac connected genes in transcriptome of CD4 + cells in regression to plasma insulin levels and after HDACi treatment, by RNA-seq. Expression difference is calculated by DESeq2 analysis. Colour fill corresponds to metabolic pathway annotation of the gene.

Article Snippet: Cells were stimulated with concanavalin A (ConA, 0.625 μg/mL, MP Biomedicals), and lipopolysaccharide (LPS, 5 μg/mL, Sigma-Aldrich) for 48 h. For immunophenotyping, the cultures were supplemented with insulin (Humalog 100 U/mL, Eli Lilly, Indianapolis, IN, USA) 25 nM and/or the class I HDAC inhibitor (HDACi) valproate, 50 μg/mL (stock 200 mg/mL, Ergenyl, Sanofi, Paris, France) after 24 h. Supernatants were collected for cytokine measures and cells were analyzed by flow cytometry.

Techniques: ChIP-sequencing, Clinical Proteomics, RNA Sequencing, Inhibition, Expressing, In Vitro

Journal: Cell Death & Disease

Article Title: Insulin enables acquisition of the IL7R + memory phenotype in PD1 + T cells in RA tissues

doi: 10.1038/s41419-026-08916-6

Figure Lengend Snippet: A Transition model of PD1 hi Tph cells to the IL7R + T memory cells, influenced by insulin and HDAC inhibition. B Box plot of protein cytokine levels in supernatants of CD4 + cells ( n = 12) stimulated with insulin (25 nM) and HDACi (50 µg/mL) for 24 h, by ELISA. Boxes indicate IQR and whiskers indicate the minimum and maximum values. P -values are calculated by Wilcoxon Sign rank test. C Gating strategy to CD4 + T cell subset analysis by flow cytometry. D Box plot of PD1 and CD27 expression in CD4 + cells ( n = 8) stimulated by insulin and HDACi, by mean fluorescence intensity (MFI). Boxes indicate IQR and whiskers indicate the minimum and maximum values. P -values are calculated by Wilcoxon Sign rank paired test. E Heatmap of expression difference of IL7R-related signaling molecules in CD4 + cells after in vitro stimulation with insulin (25 nM), HDACi (50 µg/mL), IFNγ (50 ng/ml), and survivin inhibitor YM155 (10 μM), by RNA-seq. Genes connected to cis -RE containing survivin-H3K27ac co-deposition are indicated bold. Asterisks indicate RNA-seq p -values * < 0.05, ** < 0.01, *** <0.001, **** <0.0001. F Heatmap showing expression difference, by log2FC, of cluster markers of Tph cluster after insulin stimulation, HDAC inhibition. In bold are the genes supervised by a cis-RE containing survivin-H3K27ac deposition. P -values are calculated by DESeq2 analysis. Asterisks indicate nominal p -values * < 0.05, ** < 0.01, *** <0.001, **** <0.0001.

Article Snippet: Cells were stimulated with concanavalin A (ConA, 0.625 μg/mL, MP Biomedicals), and lipopolysaccharide (LPS, 5 μg/mL, Sigma-Aldrich) for 48 h. For immunophenotyping, the cultures were supplemented with insulin (Humalog 100 U/mL, Eli Lilly, Indianapolis, IN, USA) 25 nM and/or the class I HDAC inhibitor (HDACi) valproate, 50 μg/mL (stock 200 mg/mL, Ergenyl, Sanofi, Paris, France) after 24 h. Supernatants were collected for cytokine measures and cells were analyzed by flow cytometry.

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Fluorescence, In Vitro, RNA Sequencing