cd8a microbeads  (Miltenyi Biotec)

Journal: International Journal of Biological Sciences

Article Title: GZMK⁺CD8⁺ T Cells Induce Acinar Cell Senescence and Salivary Secretion Impairment in Sjögren's Syndrome via Mitochondrial Dysfunction and TMEM16A Chloride Channel Disruption

doi: 10.7150/ijbs.134478

Figure Lengend Snippet: Senescence-associated GZMK⁺CD8⁺ T cells exhibit significant interactions with SS acinar cells. (A) UMAP plot showing the re-annotation and subclassification of T/NK cell populations into 14 distinct subsets from labial gland samples. (B) Bar graph displaying the proportional composition of T cell subsets in young and elderly SS patients and age- and sex-matched HCs, highlighting the increased proportion of GZMK⁺CD8⁺ T cells in SS patients. (C) The key markers associated with different T cell subsets in SS and HC groups. (D) Boxplot showing the proportion of GZMK⁺CD8⁺ T cells in the young and elderly SS groups compared to young and elderly HC groups, with the highest abundance observed in the elderly SS group. (E) Volcano plot of DEGs in GZMK⁺CD8⁺ T cells from SS patients compared to HCs, showing significantly upregulated functional genes in SS. (F) Boxplots showing the expression levels of GZMK in GZMK⁺CD8⁺ T cells from young and elderly SS patients and HCs, with significantly higher expression in SS groups, especially in the elderly SS group. (G) CellChat analysis of intercellular communication between GZMK⁺CD8⁺ T cells and various resident tissue cell types in labial glands, highlighting significant interactions with acinar cells. (H) Top 15 ligand-receptor pairs involved in the interaction between GZMK⁺CD8⁺ T cells and acinar cells, demonstrating the close communication between these cell types.

Article Snippet: CD8+ T cells were purified by positive selection using rat CD8a MicroBeads (130-090-318, Miltenyi Biotec, Germany) according to the manufacturer's instructions.

Techniques: Functional Assay, Expressing, Analysis, Tissue

Journal: Cancer Discovery

Article Title: Senescent-Like Neutrophils Shape Angiogenic Immunosuppressive Niches in Colorectal Cancer Liver Metastasis

doi: 10.1158/2159-8290.CD-25-0820

Figure Lengend Snippet: Proangiogenic and immunosuppression phenotype of TANs in the CRLM microenvironment. A, Heatmap showing signature scores for angiogenesis, myeloid cell recruitment, neutrophil cytotoxicity, MDSC, and NET across each TAN and PMN subset. B, Comparison of angiogenesis, myeloid cell recruitment, MDSC, and NET signature scores among TAN and PMN subsets. C, Dot plot illustrating expression levels of major cytokines and chemokines in each TAN and PMN subset. D, Representative IHC images of CD31 staining in CRLM tissues. Scale bars, 50 µm. E, Correlation analysis between CD31 and TAN1 density using Spearman’s rank correlation ( n = 102). F, Representative images and quantification of tube formation in HUVECs when cocultured with conditioned medium from human LGALS3 − neutrophils or LGALS3 + neutrophils ( n = 6). G, UMAP plot showing six monocyte/macrophage subsets across colorectal cancer, adjacent liver tissues, CRLM, and PB samples. H, Shell-like plot depicting ligand–receptor interaction strength between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. I, Selected ligand–receptor interactions between TAN1_LGASL3 and neutrophil/monocyte/macrophage subset in CRLM. J, Representative mIHC images of CD68, CCL3L1, CD66b, CCR1, and BHLHE40 in CRLM. Scale bars, 50 µm. K, Representative images of macrophage migration assays and effects of conditioned medium from human LGALS3 + neutrophils on migration of macrophages (induced from THP-1 cells) in the presence of CCR1 inhibitor (BX471) or anti-CCL3 antibody ( n = 6). L, Comparison of immunosuppressive macrophage signature scores between colorectal cancer and CRLM. M, Schematic of the CRLM mouse model. C57BL/6 mice received intrasplenic injections of mouse colorectal cancer cells (MC38). Mice were subsequently treated with IgG control (IgG; n = 6), anti-Ly6G antibody (anti-Ly6G; n = 6), or anti-Ly6G plus anti-CD8a antibodies (anti-Ly6G + anti-CD8a; n = 6). Livers were harvested on day 14. N, Representative images of liver metastases from IgG, anti-Ly6G, and anti-Ly6G + anti-CD8a groups. O, Comparison of liver weights among the three groups. P values were calculated by the Wilcoxon rank-sum test ( B and L ) and unpaired Student t test ( F , K , and O ). *, P < 0.05; **, P < 0.01; ***, P < 0.001. CRC, colorectal cancer; Ctrl, control.

Article Snippet: For CD8 + T-cell depletion, 200 μg anti-mouse CD8a (Bio X Cell, BE0061, RRID: AB_1125541) was injected intraperitoneally twice a week.

Techniques: Comparison, Expressing, Staining, Analysis, Migration, Control