Journal: Frontiers in Immunology

Article Title: Structural and functional characterization of IgG- and non-IgG-based T-cell-engaging bispecific antibodies

doi: 10.3389/fimmu.2024.1376096

Figure Lengend Snippet: Granzyme/perforin activity and cytokine production by anti-EGFR/CD3 DVD-Ig and BiTE (A) Representative anti-granzyme B immunoblots using the WCL from co-culture of tumor cells (CaOV3 or MDA-MB-231) and T cells exposed to DVD-Ig or BiTE for 2 days. First lane represents tumors cell only. Actin was used as loading control. (B, C) Granzyme B and perforin measurement in co-culture supernatants was performed using ELISA kit. This experiment was performed in biological triplicates, and data is presented as mean ± SD. **p < 0.01 (D) Detection of human cytokines in co-culture supernatant of CaOV3 was performed using array kit. Green boxes (and associated green cytokine coordinates) represent increased dot intensity under BiTE treatment condition; blue boxes (and associated blue cytokine coordinates) represent increased array dot intensity under both DVD-Ig and BiTE treatment conditions.

Article Snippet: Quantitative measurement of Granzyme B was performed using Human Granzyme B Quantikine ELISA Kit (cat# DGZB00, R and D Systems, Minneapolis, MN) and perforin was measured using Perforin (PRF1) Human ELISA Kit (cat# ab46068, Abcam, Waltham, MA).

Techniques: Activity Assay, Western Blot, Co-Culture Assay, Control, Enzyme-linked Immunosorbent Assay

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