Journal: Cancer Cell International

Article Title: Adipose stem cell exosomes, stimulated by pro-inflammatory factors, enhance immune evasion in triple-negative breast cancer by modulating the HDAC6/STAT3/PD-L1 pathway through the transporter UCHL1

doi: 10.1186/s12935-024-03557-1

Figure Lengend Snippet: ADSCs-Exos IFN−γ+TNF−α promoted immune escape of TNBC cells by transporting UCHL1. ADSCs were treated with 10 ng/mL IFN-γ and TNF-α, and transfected with si-NC or si-UCHL1. The exosomes were extracted from the supernatant. TNBC cells were treated with 10 μg/mL of the ADSCs-CM and different exosomes for 24 h, and then co-cultured with THP-1 cells for 48 h or with T cells for 16 h. A UCHL1 protein levels in TNBC cells in the indicated groups. B Viability of TNBC cells in the indicated groups. C Representative images showing Ki-67 expression in TNBC cells in the indicated groups. Scale bar = 100/25 μm. D Representative images showing migration of TNBC cells in the indicated groups. Scale bar = 100 μm. * P < 0.05 vs . MDA-MB-231 or MDA-MB-468. # P < 0.05 vs . Exos + MDA-MB-231 or Exos + MDA-MB-468. & P < 0.05 vs . Exos IFN−γ+TNF−α+con + MDA-MB-231 or Exos IFN−γ+TNF−α+con + MDA-MB-468. E The proportion of CD68 + CD206 + THP-1 cells in the indicated groups. F Arg-1, CD206, TNF-α, and iNOS expression in THP-1 cells. * P < 0.05 vs . THP-1. # P < 0.05 vs . MDA-MB-231 + THP-1 or MDA-MB-468 + THP-1. & P < 0.05 vs . Exos + THP-1. @ P < 0.05 vs . Exos IFN−γ+TNF−α+con + THP-1. G The proportion of CD3 + CD8 + T cells in the indicated groups. H Perforin and GZMB levels in T cells. * P < 0.05 vs . T cells. # P < 0.05 vs . MDA-MB-231 + T cells or MDA-MB-468 + T cells. & P < 0.05 vs . Exos + T cells. @ P < 0.05 vs . Exos IFN−γ+TNF−α+con + T cells

Article Snippet: Granular enzyme B (GZMB) and perforin levels were analyzed using specific GZMB (CSB-E08718h, CUSABIO, China) and perforin (CSB-E09313h, CUSABIO, China) ELISA kits according to the instructions.

Techniques: Transfection, Cell Culture, Expressing, Migration

Journal: Cancer Cell International

Article Title: Adipose stem cell exosomes, stimulated by pro-inflammatory factors, enhance immune evasion in triple-negative breast cancer by modulating the HDAC6/STAT3/PD-L1 pathway through the transporter UCHL1

doi: 10.1186/s12935-024-03557-1

Figure Lengend Snippet: ADSCs-Exos IFN−γ+TNF−α promoted tumor growth in vivo. BALB/c mice were subcutaneously injected with 4T1 cells (2 × 10 6 ) and treated with 100 μg of the different exosomes every 3 days. A Tumor weight and volume in the indicated groups. B In-situ expression of Ki-67 in the tumor tissues of indicated groups. Scale bar = 100/25 μm. C The F4/80 + /CD206 + cells in the tumor tissues of indicated groups. Scale bar = 25 μm. D The proportion of CD3 + CD8 + T cell in the tumor tissues of indicated groups. E Perforin and GZMB levels in the tumor tissues of indicated groups. F and G CD8 + /GZMB + cells and CD8 + /perforin + cells in the tumor tissues of indicated groups. (H) HDAC6, UCHL1, PD-L1, p-STAT3 and STAT3 levels in the tumors of indicated groups. * P < 0.05 vs . 4T1. # P < 0.05 vs . Exos + 4T1. & P < 0.05 vs . Exos IFN−γ+TNF−α+con + 4T1

Article Snippet: Granular enzyme B (GZMB) and perforin levels were analyzed using specific GZMB (CSB-E08718h, CUSABIO, China) and perforin (CSB-E09313h, CUSABIO, China) ELISA kits according to the instructions.

Techniques: In Vivo, Injection, In Situ, Expressing

Journal: Frontiers in Immunology

Article Title: NK cell function down regulated by HMGB2 through ANGPT1/PI3K/AKT pathway and its effect on esophageal squamous carcinoma cells

doi: 10.3389/fimmu.2025.1666199

Figure Lengend Snippet: Down-regulation of HMGB2 enhances the function of NK cells. (a) qRT-PCR analysis of HMGB2 mRNA levels in NK-92 cells transfected with siNC, siHMGB2 #1, siHMGB2 #2, and siHMGB2 #3. (b) Western blot analysis of HMGB2 protein levels in NK-92 cells transfected with siNC or siHMGB2. (c) The secretion levels of perforin-1 and granzyme B were measured by ELISA in NK-92 cells after HMGB2 knockdown. (d) Cytokine release (TNF-α and IFN-γ) from NK-92 cells with HMGB2 knock down were showed in histogram. (e) Cytotoxicity of NK-92 cells with HMGB2 knockdown against various ESCC cell lines (EC109, KYSE70, KYSE150, KYSE450) was assessed at 24h and 48h using a cytotoxicity assay. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test for normally distributed data, or Kruskal–Wallis tests followed by Dunn’s post hoc tests for non-normally distributed data. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent the mean ± SD.

Article Snippet: After centrifugation, the culture supernatant was collected, and the concentrations of granzyme B and perforin 1 were measured using the Human Granzyme B ELISA Kit (E-EL-H1617c, Elabscience, Beijing, China) and Human Perforin 1 ELISA Kit (E-EL-H1123c, Elabscience), respectively, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Knockdown, Cytotoxicity Assay

Journal: Frontiers in Immunology

Article Title: NK cell function down regulated by HMGB2 through ANGPT1/PI3K/AKT pathway and its effect on esophageal squamous carcinoma cells

doi: 10.3389/fimmu.2025.1666199

Figure Lengend Snippet: Overexpression of HMGB2 inhibits the function of NK-92 cells. (a) qRT-PCR analysis of HMGB2 mRNA levels in NK-92 cells transfected with pcDNA3.1-Vector or pcDNA3.1-HMGB2. (b) Western blot analysis of HMGB2 protein expression in NK-92 cells transfected with pcDNA3.1-Vector or pcDNA3.1-HMGB2. (c) The secretion levels of perforin-1 and granzyme B were measured by ELISA in NK-92 cells after HMGB2 overexpression. (d) Cytokine release (TNF-α and IFN-γ) from NK-92 cells with HMGB2 overexpression was showed in histogram. (e) Cytotoxicity of NK-92 cells with HMGB2 overexpression against various ESCC cell lines (EC109, KYSE70, KYSE150, KYSE450) at 24h and 48h was assessed using a cytotoxicity assay. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test for normally distributed data, or Kruskal–Wallis tests followed by Dunn’s post hoc tests for non-normally distributed data. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent the mean ± SD.

Article Snippet: After centrifugation, the culture supernatant was collected, and the concentrations of granzyme B and perforin 1 were measured using the Human Granzyme B ELISA Kit (E-EL-H1617c, Elabscience, Beijing, China) and Human Perforin 1 ELISA Kit (E-EL-H1123c, Elabscience), respectively, according to the manufacturer’s instructions.

Techniques: Over Expression, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay

Journal: Frontiers in Immunology

Article Title: NK cell function down regulated by HMGB2 through ANGPT1/PI3K/AKT pathway and its effect on esophageal squamous carcinoma cells

doi: 10.3389/fimmu.2025.1666199

Figure Lengend Snippet: Confirmation of HMGB2 knockout and functional assessment of KO-HMGB2 NK-92 cells. (a) Sanger sequencing chromatograms showing two distinct allelic edits in the KO-HMGB2 NK-92 cells. (b) Western blot analysis of HMGB2 protein levels in wild-type (WT) and KO-HMGB2 NK-92 cells. (c) ELISA analysis of perforin-1 and granzyme B secretion in supernatants collected from KO-HMGB2 and WT NK-92 cells co-cultured with KYSE450 cells for 12 hours. (d) qRT-PCR analysis of activating receptors (NKG2D, NKp44, NKp46, and CD107a) in KO-HMGB2 and WT NK-92 cells. (e) Cytotoxicity of NK-92 cells with HMGB2 knock out against various ESCC cell lines (EC109, KYSE70, KYSE150, KYSE450) at 24h and 48h was assessed using a cytotoxicity assay. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test for normally distributed data, or Kruskal–Wallis tests followed by Dunn’s post hoc tests for non-normally distributed data. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent the mean ± SD.

Article Snippet: After centrifugation, the culture supernatant was collected, and the concentrations of granzyme B and perforin 1 were measured using the Human Granzyme B ELISA Kit (E-EL-H1617c, Elabscience, Beijing, China) and Human Perforin 1 ELISA Kit (E-EL-H1123c, Elabscience), respectively, according to the manufacturer’s instructions.

Techniques: Knock-Out, Functional Assay, Sequencing, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, Quantitative RT-PCR, Cytotoxicity Assay