mouse triple negative 4t1 breast cancer cell line  (ATCC)

Journal: Journal of Translational Medicine

Article Title: Tumor necrosis factor receptor 2 promotes endothelial cell-mediated suppression of CD8+ T cells through tuning glycolysis in chemoresistance of breast cancer

doi: 10.1186/s12967-024-05472-5

Figure Lengend Snippet: Endothelial PD-L1 mediates suppression of CD8+ T cell within tumor endothelium. A Orthotopic 4T1-bearing murine model. B Tumor growth curves in both groups; the yellow arrows indicate PTX or PBS administration. C Representative images of tumor samples (day 26). Immunofluorescence staining of CD31 (green), CD8 (red), and DAPI (blue); yellow arrows indicate cell–cell interactions between CD8+ T cells (CD8) and ECs (CD31). Scale bars: 1 mm (left) and 200 µm (right). D Representative histograms and quantification of PD-L1 expression on ECs from 4T1-bearing mice (day 26). Fluorescence Minus One (FMO) served as a negative control. E The splenocytes from OT-I mice were incubated with IFNγ-stimulated sEND.1 at a ratio of 500:1 with or without pre-administration of anti-PD-L1 antibody (10 μg/ml) 1 h prior to co-incubation. 72 h later, the proliferation of CD8+ T cells was detected by flow cytometry using the FITC channel. F The quantitation of CD8+ T cell prefoliation. n = 3; G immunofluorescence staining of CD31 (green) and CD8 (red) in tumor samples. The number of samples: PBS-day12, n = 7; PTX-day 12, n = 6; PTX-day 16, n = 7; PTX-day 26, n = 6. Scale bars: 200 µm. H Values of M1 (CD8) and M2 (CD31) over time corresponding to G . *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant

Article Snippet: Human umbilical vein endothelial cell (HUVEC; RRID: CVCL_2959) and the mouse triple-negative breast cancer cell line 4T1 (RRID: CVCL_0125) were purchased from ATCC (Manassas, VA).

Techniques: Immunofluorescence, Staining, Expressing, Fluorescence, Negative Control, Incubation, Flow Cytometry, Quantitation Assay

Journal: Journal of Translational Medicine

Article Title: Tumor necrosis factor receptor 2 promotes endothelial cell-mediated suppression of CD8+ T cells through tuning glycolysis in chemoresistance of breast cancer

doi: 10.1186/s12967-024-05472-5

Figure Lengend Snippet: TNFR2 blockade and PTX combination therapy. A Schematic representation of 4T1-transplanted murine model and therapeutic procedure. B Tumor growth curves. C Primary tumors harvested from mice. D Representative histogram (left) and quantification (right) of PD-L1 expression on tumor ECs (CD45−CD31+PD-L1+) isolated from mice. E , F Gating scheme for ( E ) CD45+CD3+CD8+ T cells and ( F ) PD-1+ T cells (CD45+CD3+CD8+PD-1+) from tumor-bearing mice. G Quantification of the proportion of PD-1+CD8+ T cells. H Schematic illustration showing immune features of CD8+ T cells and ECs from responders and non-responders following PTX therapy. The expression of the ligands PD-L1, PVR, and MHC class II molecules on ECs and the corresponding receptors PD-1 and LAG3 on CD8+ T cells increase in non-responders but decrease in responders. Endothelial TNF–TNFR2 signaling enables inhibition of CD8+ T cell through PD-L1/PD-1 axis via regulating cellular NF-κB–glycolysis pathway on the basis of IFNγ-mediated immune feedback. Schematic illustration was created on the website of “Home for Researchers”. *P < 0.05, ***P < 0.001, ****P < 0.0001

Article Snippet: Human umbilical vein endothelial cell (HUVEC; RRID: CVCL_2959) and the mouse triple-negative breast cancer cell line 4T1 (RRID: CVCL_0125) were purchased from ATCC (Manassas, VA).

Techniques: Expressing, Isolation, Inhibition