Journal: Frontiers in Immunology

Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

doi: 10.3389/fimmu.2026.1833383

Figure Lengend Snippet: Overexpression of UPF1 in ccRCC inhibits M2 macrophage polarization and enhances the tumor suppressor function of CD8 + T cells, Caki-1 cell groups of sh-NC+PBS, sh-UPF1+PBS, or sh-UPF1 + 2-DG, and the conditioned medium was used to culture and polarize M0 macrophages. The polarized macrophages were cultured with activated CD8 + T cells. (A) CFSE assessed CD8 + T cell proliferative capacity. (B) Flow cytometry measured apoptosis rate of CD8 + T cells in different co-culture systems. (C) CD8 + T cells from culture systems were co-cultured with normal Caki-1 cells, with LDH assays determining cytotoxic effect of CD8 + T cells. *, p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; 2-DG, 2-Deoxy-D-glucose; CFSE, Carboxyfluorescein succinimidyl ester.

Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

Techniques: Over Expression, Cell Culture, Flow Cytometry, Co-culture Assay

Journal: Frontiers in Immunology

Article Title: UPF1-dependent glycolysis driven by LINC00887 facilitates macrophage M2 polarization in clear cell renal cell carcinoma

doi: 10.3389/fimmu.2026.1833383

Figure Lengend Snippet: LINC00887/UPF1-mediated aerobic glycolysis leads to lactic acid accumulation and facilitates immune evasion in ccRCC in vivo , A syngeneic tumor model was developed by injecting BALB/c mice with Renca cells expressing sh-NC or sh-LINC00887, followed by treatment with sh-NC or sh-UPF1 lentivirus. A-C: Assessment of tumor size (A) , volume (B) , and weight (C) . (D, E) qRT-PCR analyses for LINC00887 (D) and UPF1 (E) mRNA levels. (F) IHC analysis for UPF1, LDH-A, KI67 protein levels. (G) Kit assay for lactate levels in tumors. (H) Proportion of M2 macrophages (F4/80+CD206+) by flow cytometry. (I) Immunofluorescence assay for CD206+ macrophages. (J) Proportion of CD8 + T cells (CD3 + CD8 + ) in tumor tissues. ns, no significant difference. * p < 0.05. cRCC, Clear cell renal cell carcinoma; UPF1, Up-frameshift protein 1; LDH-A, Lactate dehydrogenase A; CD206, Mannose receptor C-type 1; CD3, Cluster of differentiation 3; CD8, Cluster of differentiation 8.

Article Snippet: Antibodies included: FITC Mouse monoclonal to CD68 (ab134351, abcam, UK), Pacific Blue anti-human CD86 Antibody (305417, yajimall, China), CD206 (MMR) Monoclonal Antibody (19.2)-PE (12-2069-42, ThermoFisher, USA), PerCP/Cyanine5.5 Anti-Mouse F4/80 Antibody (E-AB-F0995J, Elabscience, China), PE Anti-CD206 mouse antibody (E-AB-F1135D, Elabscience, China), APC Anti-mouse CD3 antibody (E-AB-F1013E, Elabscience, China), PE Anti-mouse CD8 alpha antibody (E-AB-F1104D, Elabscience, China).

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Analysis, Flow Cytometry, Immunofluorescence

Journal: Science Advances

Article Title: Harnessing skin-resident γδ T cells for immunotherapy in cutaneous squamous cell carcinoma

doi: 10.1126/sciadv.aec7215

Figure Lengend Snippet: ( A ) Expression of indicated chemokine receptors by in vitro expanded, live gated CD3 + Vδ1 + γδ T cells. Blood-derived αβ T cells were used as staining control. ( B ) As (A), graphical summary of the percentage of Vδ1 + γδ T cell expressing the chemokine receptors ( n = 5 skin donors). Error bars represent mean ± SD. ( C ) 10 × 10 6 human skin–derived T cells, containing approximately 7% of Vδ1 + γδ T cells, were injected intravenously (i.v.) into NSG mice carrying a xSCC of a volume ranging from 100 to 200 mm 3 [reached approximately 60 to 80 days (d) post–i.d. injection of SCC-13 cells]. Each mouse was injected intraperitoneally (i.p.) with recombinant IL-2 and IL-15 daily until the harvest day. Figure created in BioRender. I. Gratz (2026) https://BioRender.com/zmnd28b . ( D ) Representative plot of the percentage of Vδ1 + γδ T cells engrafted in the spleen, blood, xSCC and murine skin 2, 7, or 14 days posttransfer. ( E to G ) Bar graphs show the absolute numbers of ingoing Vδ1 + γδ T cells, Vδ1 + γδ T cells engrafting spleen and blood normalized to mouse weight (grams), and xSCC and murine skin normalized to tissue weight (grams). (E) n = 7 mice per group; pool of two independent experiments. (F) n = 12 mice per group; pool of four independent experiments; (G) n = 12 mice per group; pool of two independent experiments. Each symbol represents one skin donor. Error bars represent mean ± SD. Statistical significance was determined using the Kruskal-Wallis test with Dunn’s multiple comparisons test. All data points, including extreme values, are shown. ( H ) Representative immunofluorescent staining of colocalized TCRδ/DAPI in HD skin and xSCC 7 days after γδ transfer. Scale bars, 100 μm. Staining controls are shown in fig. S4.

Article Snippet: Human skin–derived T lymphocytes were labeled with Cell Proliferation Dye eFluor 450 (10 μM, Thermo Fisher Scientific, catalog no. 65- 0842-85), washed with PBS, and subsequently stimulated for 6 days with rhuIL-2 (100 IU/ml; BioLegend, catalog no. 589108), rhuIL-15 (20 ng/ml; BioLegend, catalog no. 570603), in addition with mouse anti-human CD3 monoclonal antibody (1 μg/ml; Miltenyi Biotec, catalog no. 130-093-387, RRID:AB_1036144) alone or with rhuIL-1α (9 ng/ml; BioLegend, catalog no. 570004) and rhuIL-18 (9 ng/ml, BioLegend, catalog no. 592102) applied individually or in combination in cTexMacs medium.

Techniques: Expressing, In Vitro, Derivative Assay, Staining, Control, Injection, Recombinant

Journal: Science Advances

Article Title: Harnessing skin-resident γδ T cells for immunotherapy in cutaneous squamous cell carcinoma

doi: 10.1126/sciadv.aec7215

Figure Lengend Snippet: ( A ) Levels of cytokines (pg/mg tissue) produced by HD skin, huSCC, ES, and xSCC. Heatmap bars represent the mean of n = 5 HD skin and huSCC donors, and mean n = 5 of xenograft mice. ( B ) Representative gating strategy and bar graphs of the human skin–derived and ex vivo expanded Vδ1 + γδ T cells expressing IL-1RAcP and IL-18Rα. Peripheral blood αβ T cells and ex vivo expanded skin-derived αβ T cells were used as staining controls. Mean of n = 5 skin donors. ( C ) eFluor450-labeled γδ T cells were cultured under basal conditions [unstimulated or with IL-2 (100 IU/ml) and IL-15 (20 ng/ml)] or stimulated with anti-CD3 (1 μg/ml) and/or IL-1α and IL-18 (9 ng/ml) for 6 days. Proliferation was assessed by the median fluorescence intensity (MFI) of eFluor450 in Vδ1 + by flow cytometry. The representative histograms show the eFluor450 dilution in Vδ1 + T cells in the different conditions. Cell counts were normalized to unit area. ( D ) Bar graphs show the fold change of efluor450 MFI of Vδ1 + treated with anti-CD3, IL-1α, and IL-18 relative to IL-2 and IL-15. Mean of n = 6 skin donors. Statistical analysis was performed using a Friedman test followed by Dunn’s multiple comparisons test. Data in bar graphs (B) and (D) are shown as mean ± SD.

Article Snippet: Human skin–derived T lymphocytes were labeled with Cell Proliferation Dye eFluor 450 (10 μM, Thermo Fisher Scientific, catalog no. 65- 0842-85), washed with PBS, and subsequently stimulated for 6 days with rhuIL-2 (100 IU/ml; BioLegend, catalog no. 589108), rhuIL-15 (20 ng/ml; BioLegend, catalog no. 570603), in addition with mouse anti-human CD3 monoclonal antibody (1 μg/ml; Miltenyi Biotec, catalog no. 130-093-387, RRID:AB_1036144) alone or with rhuIL-1α (9 ng/ml; BioLegend, catalog no. 570004) and rhuIL-18 (9 ng/ml, BioLegend, catalog no. 592102) applied individually or in combination in cTexMacs medium.

Techniques: Produced, Derivative Assay, Ex Vivo, Expressing, Staining, Labeling, Cell Culture, Fluorescence, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Engineered Salmonella -mediated c-di-AMP delivery activates STING to remodel the tumor microenvironment

doi: 10.1016/j.omton.2026.201185

Figure Lengend Snippet: Effect of SL disA therapy on T cells and macrophage from tumor samples (A, B) Flow Cytometry analysis of CD3 and CD8 surface markers(A) and CD3 + CD8 + cells statistical graph (B). (C, D) Flow cytometry analysis of CD3 and CD4 surface markers (C) and CD3 + CD4 + cells statistical graph (D). (E, F) Flow cytometry analysis of F4/80 and CD86 surface markers (E) and F4/80 + CD86 + cells statistical graph (F). Data are expressed as mean ± SEM, n = 3. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, one-way ANOVA with Tukey’s multiple comparisons tests.

Article Snippet: The following antibodies were used: FITC anti-mouse F4/80 (clone CI: A3-1), APC anti-mouse CD86 (clone GL-1), FITC anti-mouse CD3 (clone 17A2), APC anti-mouse CD4 (clone GK1.5), and APC anti-mouse CD8 (clone YTS-169), all purchased from Elabscience Biotechnology Co., Ltd.

Techniques: Flow Cytometry