easysep human cd4 t cell isolation kit 17952  (STEMCELL Technologies Inc)

Journal: Nature Cardiovascular Research

Article Title: Low-dose interleukin-2 induces clonal expansion of BACH2-repressed effector regulatory T cells following acute coronary syndrome

doi: 10.1038/s44161-025-00652-y

Figure Lengend Snippet: a , LILACS study sample collection and workflow schematic. Three treatment groups from Part B of LILACS are evaluated in this study: placebo ( n = 4), 1.5 MIU d −1 IL-2 ( n = 6) and 2.5 MIU d −1 IL-2 ( n = 6). b , Uniform manifold approximation and projection (UMAP) visualization of unsupervised clustering revealed 8 distinct T cell populations ( n = 41,050 cells). c , Heat map with the average expression of canonical T cell function-associated genes. The histogram on the right of the heat map shows the number of cells within each cluster. d , Single-cell TCR workflow schematic. e , Gene segment usage and gene–gene pairing landscapes are illustrated graphically using four vertical stacks (one for each V and J segment) connected by curved segments with thickness proportional to the number of TCRs with the respective gene pairing. Each stack has genes with the highest frequencies stacked on top with subsequent genes in descending order. f , Plot showing the frequency of clonotype sizes across all T cells, CD4 + T cells or CD8 + T cells. g , Network analysis showing larger ( n ≥ 15) clonotypes. Each number is a clonotype identification number, each individual color represents a single patient and the size of the dots represents the number of cells. Linked dots are clonotypes that are closely related. Panels a and d created with BioRender.com .

Article Snippet: Primary human T reg cells were isolated from fresh (collected within 12 h) apheresis cones using a Lymphoprep gradient and a subsequent EasySep Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit (Stemcell) or Human CD4 + CD25 + CD127 dim/− Regulatory T Cell Isolation Kit II (Miltenyi).

Techniques: Expressing, Cell Function Assay

Journal: Nature Cardiovascular Research

Article Title: Low-dose interleukin-2 induces clonal expansion of BACH2-repressed effector regulatory T cells following acute coronary syndrome

doi: 10.1038/s44161-025-00652-y

Figure Lengend Snippet: a , scRNA-seq data showing the effect of IL-2 versus placebo on T cell subsets. There was a significant increase in T reg cell numbers after 1.5 MIU d −1 and 2.5 MIU d −1 of IL-2 treatment compared with pooled pretreatment counts: * P = 0.034; **P = 0.0072 by two-tailed t -test. b , Histogram showing the distribution of clonotype size across 5 bins: 1, 2–5, 6–10, 11–20 and 21+. The bars represent the proportion of that clonotype size in each treatment allocation. *** P < 0.001 (by chi-squared (one-tailed) test), indicating significant difference in clonotype size proportions between 1.5 MIU d − 1 or 2.5 MIU d −1 doses and placebo. c , Histograms showing the effect of the treatment allocations on TCR diversity using Shannon entropy and D50. d , The left panel shows a UMAP with the CD4 + T cell subsets overlaid. The other panels show the effect of either placebo or IL-2 treatment on CD4 + T cell clonotype size. The shades of red or blue represent the size of clonotypes on a scale from 0 to >5. Clonotype size is defined as the number of CD4 + T cells that share highly similar TCR sequences .

Article Snippet: Primary human T reg cells were isolated from fresh (collected within 12 h) apheresis cones using a Lymphoprep gradient and a subsequent EasySep Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit (Stemcell) or Human CD4 + CD25 + CD127 dim/− Regulatory T Cell Isolation Kit II (Miltenyi).

Techniques: Two Tailed Test, One-tailed Test

Journal: Nature Cardiovascular Research

Article Title: Low-dose interleukin-2 induces clonal expansion of BACH2-repressed effector regulatory T cells following acute coronary syndrome

doi: 10.1038/s44161-025-00652-y

Figure Lengend Snippet: a , Venn diagrams showing the overlap of CD4 + TCR clonotypes (and associated T cells) that match between the pre- and post-dose timepoints in the various treatment allocations. *** P < 0.001 (by chi-squared (one-tailed) test), indicating a significant difference. b , Schematic showing the concept of TCR tracking. We use the TCR as a barcode to track cells over two timepoints before and after dosing. By then pairing scTCR-seq and scRNA-seq, we can phenotype these tracked cells. c , Using TCR tracking, we show the effect of acute MI (treated with placebo) on matched TCRs and their cell phenotype over two timepoints. Connected lines from the pre- and post-dose timepoints represent CD4 + T cells that have the same TCR clonotype. The labels on the sides describe the scRNA-seq phenotype of the cell. Only those clonotypes tracked onto T reg cells at either the pre- or post-dose timepoints are shown. The numbers of cells underlying this figure are provided in Supplementary Table . d , Results of transition matrix calculation within each treatment group followed by outgoing flow visualization using CellRank 2. Panel b created with BioRender.com .

Article Snippet: Primary human T reg cells were isolated from fresh (collected within 12 h) apheresis cones using a Lymphoprep gradient and a subsequent EasySep Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit (Stemcell) or Human CD4 + CD25 + CD127 dim/− Regulatory T Cell Isolation Kit II (Miltenyi).

Techniques: One-tailed Test

Journal: Nature Cardiovascular Research

Article Title: Low-dose interleukin-2 induces clonal expansion of BACH2-repressed effector regulatory T cells following acute coronary syndrome

doi: 10.1038/s44161-025-00652-y

Figure Lengend Snippet: A , Weblogo plot showing differences in aligned CDR3 composition in clonally expanded CD4+ non-Tregs. The height of the letters represent the relative frequency with decreasing frequency stacked on top. B , Volcano plot showing differentially expressed genes in expanded CD4+ non-Tregs vs. nonexpanded CD4+ non-Tregs (enriched in the expanded (red) or non-expanded (blue).

Article Snippet: Primary human T reg cells were isolated from fresh (collected within 12 h) apheresis cones using a Lymphoprep gradient and a subsequent EasySep Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit (Stemcell) or Human CD4 + CD25 + CD127 dim/− Regulatory T Cell Isolation Kit II (Miltenyi).

Techniques:

Journal: Nature Cardiovascular Research

Article Title: Low-dose interleukin-2 induces clonal expansion of BACH2-repressed effector regulatory T cells following acute coronary syndrome

doi: 10.1038/s44161-025-00652-y

Figure Lengend Snippet: a , Network plot from regulon analysis of expanded versus non-expanded T reg cells using SCENIC. b , Expression of BACH2 and encompassing regulons in expanded versus non-expanded T reg cells from the LILACS dataset. c , Representative data from flow cytometry analysis ( n = 3) of pSTAT5 levels in primary human T reg cells after 48 h of stimulation and an 8-h rest in serum-free media ± BACH2 inhibitor (inh.) (RGFP966), followed by 15 min of IL-2 treatment. Numeric values indicate percentages. SSC, side scatter; MFI, mean fluorescence intensity; FC norm, normalized fold change. d , pSTAT5 and CD25 levels assessed via flow cytometry ( n = 3). e , RT-qPCR analysis of IL-32 ( n = 5) and IL-10 ( n = 5) after 24 h in primary human T reg cells treated with BACH2 inhibitor and/or IL-2 (error bars represent s.e.m.; P values were determined by two-tailed independent t -test; n = number of independent experiments).

Article Snippet: Primary human T reg cells were isolated from fresh (collected within 12 h) apheresis cones using a Lymphoprep gradient and a subsequent EasySep Human CD4 + CD127 low CD25 + Regulatory T Cell Isolation Kit (Stemcell) or Human CD4 + CD25 + CD127 dim/− Regulatory T Cell Isolation Kit II (Miltenyi).

Techniques: Expressing, Flow Cytometry, Fluorescence, Quantitative RT-PCR, Two Tailed Test