cd8 t cell isolation beads  (MedChemExpress)

Journal: Cancer Research

Article Title: Tetraspanin 13 Enhances Immune Evasion in Breast Cancer by Promoting MHC-I Degradation

doi: 10.1158/0008-5472.CAN-25-1223

Figure Lengend Snippet: TSPAN13 as a negative regulator of CD8 + T-cell infiltration in breast cancer. A, Workflow for identifying negative regulators of activated CD8 + T cells and effector memory CD8 + T cells. B, The Venn diagram illustrates the intersection of differentially expressed genes between the TCGA-BRCA and GSE20685 datasets. C–F, Correlation between TSPAN13 levels (top 25% vs. bottom 25% samples) and CD8A expression across different breast cancer subtypes ( C, basal-like, n = 89 per group; D, HER2 + , n = 42 per group; E, luminal A, n = 255 per group; F, luminal B, n = 101 per group). G and H, Kaplan–Meier survival curves (log-rank test) were used to compare the survival of patients with high and low TSPAN13 expression in HER2 + ( n = 36 per group; G ) and basal-like ( n = 84; H ) breast cancer subtypes. I and J, Prognostic predictive value of TSPAN13 expression in HER2 + ( I ) and basal-like ( J ) breast cancer subtypes. K, IHC staining was used to analyze the expression pattern of TSPAN13 in the TME of both human samples and the MMTV-PyMT mouse model (low magnification scale bar, 100 μm; high magnification scale bar, 50 μm). L, Left, representative IHC staining images of TSPAN13 and CD8α in human breast cancer tissues (low magnification scale bar, 100 μm; high magnification scale bar, 50 μm). Right, CD8 + T-cell density (cells/mm 2 ) in the TME was compared between samples with low and high TSPAN13 expression ( n = 60 per group, Wilcoxon rank-sum test). M and N, The effect of Tspan13 knockdown on the growth of orthotopically implanted 4T1 breast tumors in BALB/c nude mice. Tumor volume ( M ) and tumor images and tumor weight ( N ) are shown. n = 5 per group, mean ± SD; two-way ANOVA ( M ); two-tailed t test ( N ). O and P, Evaluation of the effect of CD8 + T-cell blockade on tumor growth in orthotopic breast tumor models using 4T1 cells with different TSPAN13 expression levels O, tumor volume, n = 5; two-way ANOVA; P, tumor size and tumor weight, n = 5 per group, two-tailed t test, mean ± SD. n.s., not significant. A, Created in BioRender. Liu, Y. (2025). https://BioRender.com/ydc9cdo .

Article Snippet: Primary CD8 + T cells were isolated from the spleens of OT-1 mice using CD8 + T-cell isolation beads (Miltenyi Biotec, cat. #130-104-453).

Techniques: Expressing, Immunohistochemistry, Knockdown, Two Tailed Test

Journal: Cancer Research

Article Title: Tetraspanin 13 Enhances Immune Evasion in Breast Cancer by Promoting MHC-I Degradation

doi: 10.1158/0008-5472.CAN-25-1223

Figure Lengend Snippet: TSPAN13 restricts CD8 + T-cell activation and effector functions by impairing antigen presentation. A–C, Flow cytometry was used to assess the surface expression levels of MHC-I in MDA-MB-468 ( A ), 4T1 ( B ), and E0771 ( C ) cells under different TSPAN13 expression levels, with or without IFNγ stimulation. n = 3 per group; mean ± SD; one-way ANOVA, multiple comparisons with the Tukey method. D, Flow cytometry was used to assess the surface expression levels of H-2Kb SIINFEKL in E0771-OVA cells under different TSPAN13 expression levels, with or without IFNγ stimulation. n = 3 per group; mean ± SD; two-tailed t test, one-way ANOVA, multiple comparisons with Tukey method. E, Schematic of the coculture setup between OT-1 CD8 + T cells and OVA-expressing mouse tumor cells. F–J, Quantification of CD69 + ( F ), IFNγ + ( G ), TNFα + ( H ), and GZMB + ( I ) CD8 + T cells by flow cytometry. Naive OT-I T cells were cocultured with sh-NC/sh- Tspan13 –expressing E0771-OVA cells at a ratio of 2.5:1 (T cells: E0771-OVA cells) for 24 hours. n = 3 per group; mean ± SD; two-tailed t test ( J ). K, ELISPOT assay was used to evaluate the ability of CD8 + T cells, isolated from the spleens of mice on day 18 after transplantation of sh-NC or sh- Tspan13 E0771-OVA cells into immunocompetent mice, to produce IFNγ in response to SIINFEKL stimulation. n = 3 per group; mean ± SD; two-tailed t test. L, In vitro cytotoxicity assay to evaluate the killing effect of activated OT-I T cells on Tspan13 knockdown or control E0771-OVA cells. n = 3 per group; two-way ANOVA. E:T, effector-to-target ratio. M, Naïve OT-1 CD8 + T cells were cocultured with E0771-OVA cells in the presence of H-2Kb-SIINFEKL antibodies or IgG control antibodies for 48 hours. OT-1 cell proliferation was assessed using the CellTrace Violet Cell Proliferation Kit. n = 3 per group; mean ± SD; one-way ANOVA, multiple comparisons with Tukey method. E, Created in BioRender. Liu, Y. (2025). https://BioRender.com/kmvwmfr .

Article Snippet: Primary CD8 + T cells were isolated from the spleens of OT-1 mice using CD8 + T-cell isolation beads (Miltenyi Biotec, cat. #130-104-453).

Techniques: Activation Assay, Immunopeptidomics, Flow Cytometry, Expressing, Two Tailed Test, Enzyme-linked Immunospot, Isolation, Transplantation Assay, In Vitro, Cytotoxicity Assay, Knockdown, Control

Journal: Cancer Research

Article Title: Tetraspanin 13 Enhances Immune Evasion in Breast Cancer by Promoting MHC-I Degradation

doi: 10.1158/0008-5472.CAN-25-1223

Figure Lengend Snippet: Targeting TSPAN13 enhances antigen presentation in tumor cells and boosts antitumor immunity. A, Flow cytometry was used to analyze the surface H-2Kb levels on tumor cells in 4T1 mouse models with varying Tspan13 expression levels. n = 5 per group; mean ± SD; two-tailed t test. B, Flow cytometry was used to analyze the proportions of CD44 + CD62L − , CD44 − CD62L + , and CD44 + CD62 + subsets among tumor-infiltrating T lymphocytes in the 4T1 tumor-bearing mouse model with different levels of TSPAN13 expression. n = 3 per group; two-tailed t test; mean ± SD. C and D, Immunofluorescence ( C ) and flow cytometry ( D ) were used to assess the levels of CD8 + T-cell infiltration in tumor tissues from 4T1 mouse models with different Tspan13 expression levels. n = 5 per group; mean ± SD; two-tailed t test. E–G, Flow cytometry was used to analyze the proportions of IFNγ + ( E ), TNFα + ( F ), and GZMB + ( G ) CD8 + T cells within tumor-infiltrating lymphocytes from 4T1 mouse models following restimulation. n = 5 per group; mean ± SD; two-tailed t test. H–J, mRNA coexpression analysis of IFN γ ( H ), TNF α ( I ), and GZMB ( J ), with TSPAN13 in BRCA . Data were obtained from the TCGA database and analyzed by GEPIA2. K, Schematic representation of the 4T1 mouse model. A total of 1 × 10 5 cells were implanted into the right fourth mammary fat pad of each BALB/c mouse. On day 3 after model establishment, IgG or anti–H-2K mAb was administered via i.p. injection. L–N, Effect of H-2 blockade on tumor growth in orthotopic breast tumor models with varying TSPAN13 expression levels. Tumor volume ( L ), tumor size ( M ), and tumor weight ( N ). n = 4 per group; mean ± SD; two-way ANOVA ( L ); two-tailed t test ( N ). K, Created in BioRender. Liu, Y. (2025). https://BioRender.com/58a7i0v .

Article Snippet: Primary CD8 + T cells were isolated from the spleens of OT-1 mice using CD8 + T-cell isolation beads (Miltenyi Biotec, cat. #130-104-453).

Techniques: Immunopeptidomics, Flow Cytometry, Expressing, Two Tailed Test, Immunofluorescence, Injection

Journal: Cancer Research

Article Title: Tetraspanin 13 Enhances Immune Evasion in Breast Cancer by Promoting MHC-I Degradation

doi: 10.1158/0008-5472.CAN-25-1223

Figure Lengend Snippet: TSPAN13 ablation enhances immunotherapy efficacy. A, Schematic illustration of the adoptive transfer experiment of OT-1 CD8 + T cells in the E0771-OVA nude mouse model. B, Tumor growth curves of mice receiving adoptive transfer of OT-1 CD8 + T cells. n = 4 per group; two-way ANOVA. C, Representative tumor-bearing mice after treatment. D–G, Association analysis between TSPAN13 expression levels and predicted response to ICIs based on the TCIA cohort. No immunotherapy (PD-1 − /CTLA4 − ; D ), anti–PD-1 therapy (PD-1 + /CTLA4 − ; E ), anti–CTLA4 therapy (PD-1 − /CTLA4 + ; F ), and combination therapy (PD-1 + /CTLA4 + ; G ). n = 546 per group; Wilcoxon rank-sum test. ips, immunophenoscores. H, Schematic representation of the 4T1 mouse model. A total of 1 × 10 5 cells were implanted into the right fourth mammary fat pad of each BALB/c mouse. On day 8 after model establishment, IgG or anti–PD-L1 mAb was administered via i.p. injection. The experiment concluded on day 90 after model establishment. I, Tumor growth curves of mice treated with either anti–PD-L1 or IgG control. n = 7 per group; two-way ANOVA. J, Tumor size in mice on the 28th day after treatment with either anti–PD-L1 or IgG control. K, Survival curves of mice treated with either anti–PD-L1 or IgG control. n = 7 per group; log-rank Mantel–Cox test. L, Immunofluorescence staining was performed to evaluate tumor-infiltrating CD8 + T cells in mice treated with either anti–PD-L1 or IgG control. CD8a was used to label CD8 + T cells, EGFP to identify tumor cells, and DAPI to visualize nuclei. Scale bars, 50 μm. n = 5 mice per group; one-way ANOVA, multiple comparisons with Tukey method. M, Tspan13 expression in breast tumor tissues under different therapeutic regimens. Control group, n = 18; each treatment group, n = 6; Wilcoxon rank-sum test. A, Created in BioRender. Liu, Y. (2025). https://BioRender.com/zs6yq0t ; H, Created in BioRender. Liu, Y. (2025). https://BioRender.com/ojwnb3n .

Article Snippet: Primary CD8 + T cells were isolated from the spleens of OT-1 mice using CD8 + T-cell isolation beads (Miltenyi Biotec, cat. #130-104-453).

Techniques: Adoptive Transfer Assay, Expressing, Injection, Control, Immunofluorescence, Staining