Journal: Heliyon

Article Title: Method validation of a bridging immunoassay in combination with acid-dissociation and bead treatment for detection of anti-drug antibody

doi: 10.1016/j.heliyon.2023.e13999

Figure Lengend Snippet: Target interference.

Article Snippet: The recombinant proteins FGL1 and LAG3 were supplied by Sino Biological, Beijing, China.

Techniques: Screening Assay, Confirmatory Assay, Inhibition

Journal: Journal for Immunotherapy of Cancer

Article Title: Optimizing anti-PI3Kδ and anti-LAG-3 immunotherapy dosing regimens in a mouse model of triple-negative breast cancer improves outcome by removing treatment-related adverse events

doi: 10.1136/jitc-2025-012157

Figure Lengend Snippet: LAG-3 ligand expression in mouse and human tumors. ( A ) Tumor expression of MHCII, Gal3 and FGL1. ( B ) Quantification of MHCII, Gal3 and FGL1 in the tumors at days 15, 21 and 28 post-tumor inoculation. ( C ) Quantification of LAG-3 expressing CD8 + T cells and FoxP3 + Tregs in direct contact with MHCII, Gal3 and FGL1 expressing cells within the tumor. ( D ) Representative expression of MHCII, Gal3 and FGL1 in the skin of vehicle or PI-3065 treated tumor bearing mice at day 28 postinoculation. ( E ) Representative expression of MHCII, Gal3 and FGL1 in human TNBC tumors. All data is displayed as the mean±SEM. Statistical significance was determined by a Mann-Whitney test (B) or Unpaired T test (C) (* p≤0.05, ** p≤0.01, *** p≤ 0.001).

Article Snippet: Tumor and skin sections were incubated with the primary abs to MHCII (anti-mouse–M5/114, Bio-Techne, anti-human–LGII-612.14, Cell Signaling Technology), Gal3 (anti-mouse/human–M3/38, BioLegend), FGL1 (anti-mouse–177R4, BioXcell, anti-human–E7C1Q, Cell Signaling Technology), LAG-3 (anti-mouse–ab209238, Abcam, anti-human–BLR028F, Fortis Life Sciences), CD8 (anti-mouse–4SM15, Invitrogen, anti-human–C8/144B, BioLegend), CD4 (anti-human–A19018, ABClonal), F4/80 (anti-mouse–D2S9R, Cell Signaling) or FoxP3 (anti-mouse–D6O8R, Cell Signaling Technology, anti-human–236A/E7, eBioscience) for 50 min at room temperature.

Techniques: Expressing, MANN-WHITNEY

Journal: Journal for Immunotherapy of Cancer

Article Title: Optimizing anti-PI3Kδ and anti-LAG-3 immunotherapy dosing regimens in a mouse model of triple-negative breast cancer improves outcome by removing treatment-related adverse events

doi: 10.1136/jitc-2025-012157

Figure Lengend Snippet: Combination anti-FGL1 antibody therapy promotes tumor control but does not reduce skin-associated irAE. ( A ) Schematic diagram shows the experimental protocol: Balb/C mice were treated daily for the duration of the study with either 75 mg/kg of PI-3065 or vehicle by oral gavage. 1 day later mice were inoculated with 4T1 tumor cells. Anti-FGL1 ab (100 ug) was given i.p. from day 10, two times per week until tumors were harvested at day 21–24. ( B ) Tumor growth curves of mice from each treatment arm (8 mice/group). ( C ) Representative images of skin condition of mice from each treatment group at day 24 post-tumor inoculation (8 mice/group). ( D ) Skin histology scores from mice at day 24 post-tumor inoculation (6–8 mice/group). All data are displayed as the mean±SEM. Statistical significance was determined by one-way ANOVA (***p≤0.001).

Article Snippet: Tumor and skin sections were incubated with the primary abs to MHCII (anti-mouse–M5/114, Bio-Techne, anti-human–LGII-612.14, Cell Signaling Technology), Gal3 (anti-mouse/human–M3/38, BioLegend), FGL1 (anti-mouse–177R4, BioXcell, anti-human–E7C1Q, Cell Signaling Technology), LAG-3 (anti-mouse–ab209238, Abcam, anti-human–BLR028F, Fortis Life Sciences), CD8 (anti-mouse–4SM15, Invitrogen, anti-human–C8/144B, BioLegend), CD4 (anti-human–A19018, ABClonal), F4/80 (anti-mouse–D2S9R, Cell Signaling) or FoxP3 (anti-mouse–D6O8R, Cell Signaling Technology, anti-human–236A/E7, eBioscience) for 50 min at room temperature.

Techniques: Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Optimizing anti-PI3Kδ and anti-LAG-3 immunotherapy dosing regimens in a mouse model of triple-negative breast cancer improves outcome by removing treatment-related adverse events

doi: 10.1136/jitc-2025-012157

Figure Lengend Snippet: Immune cell profiling of the skin. Balb/C mice were treated daily for the duration of the study with either 75 mg/kg of PI-3065 or vehicle by oral gavage. 1 day later mice were inoculated with 4T1 tumor cells. Anti-LAG-3 ab was given i.p. from day 10, three times per week, until tumors were harvested at day 24. Anti-CD8 depleting abs were administered i.p. at days 15, 18 and 24. ( A ) Tumor growth curves and ( B ) representative images of skin condition of treated mice (n=8 mice/group). ( C ) Representative skin sections from mice from each treatment group at day 24 post-tumor inoculation, stained with H&E (8 mice/group). Fluorescent ab multiplex staining depicting F4/80 + cells in the skin of vehicle-treated (i) and PI-3065+anti-LAG-3 ab-treated (ii) mice ( D ). Representative fluorescent ab multiplex staining depicting F4/80 + staining in the skin of PI-3065+anti-LAG-3 ab+anti-CD8 treated mice ( E ). All data are displayed as the mean±SEM. Statistical significance was determined by one-way ANOVA (A) (* p≤0.05, *** p≤0.001).

Article Snippet: Tumor and skin sections were incubated with the primary abs to MHCII (anti-mouse–M5/114, Bio-Techne, anti-human–LGII-612.14, Cell Signaling Technology), Gal3 (anti-mouse/human–M3/38, BioLegend), FGL1 (anti-mouse–177R4, BioXcell, anti-human–E7C1Q, Cell Signaling Technology), LAG-3 (anti-mouse–ab209238, Abcam, anti-human–BLR028F, Fortis Life Sciences), CD8 (anti-mouse–4SM15, Invitrogen, anti-human–C8/144B, BioLegend), CD4 (anti-human–A19018, ABClonal), F4/80 (anti-mouse–D2S9R, Cell Signaling) or FoxP3 (anti-mouse–D6O8R, Cell Signaling Technology, anti-human–236A/E7, eBioscience) for 50 min at room temperature.

Techniques: Staining, Multiplex Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Optimizing anti-PI3Kδ and anti-LAG-3 immunotherapy dosing regimens in a mouse model of triple-negative breast cancer improves outcome by removing treatment-related adverse events

doi: 10.1136/jitc-2025-012157

Figure Lengend Snippet: LAG-3 is a relevant marker in human breast cancer. ( A ) H&E stained whole section of a human TNBC paraffin-embedded specimen. (i) Immune cell low area, (ii) stroma rich area, (iii) immune cell high area. ( B ) Fluorescent antigen specific multiplex staining of the TNBC paraffin-embedded specimen depicted in ( A ). (i) whole tissue image, (ii) immune cell rich area, (iii) immune cell low area, and (iv) stroma rich area. Arrows indicate LAG-3 + CD4 + FoxP3 + T cells, asterisks indicate LAG-3 + CD8 + T cells. ( C ) Comparison of the percentage of CD8 + expressing LAG-3 + cells from nine matched TNBC and adjacent breast paraffin-embedded specimens. ( D ) Comparison of the percentage of FoxP3 + expressing LAG-3 + cells from nine matched TNBC and adjacent breast paraffin-embedded specimens (see also ). Statistical significance was determined by a Mann-Whitney test ( C and D ) (*p≤0.05, **p≤0.01).

Article Snippet: Tumor and skin sections were incubated with the primary abs to MHCII (anti-mouse–M5/114, Bio-Techne, anti-human–LGII-612.14, Cell Signaling Technology), Gal3 (anti-mouse/human–M3/38, BioLegend), FGL1 (anti-mouse–177R4, BioXcell, anti-human–E7C1Q, Cell Signaling Technology), LAG-3 (anti-mouse–ab209238, Abcam, anti-human–BLR028F, Fortis Life Sciences), CD8 (anti-mouse–4SM15, Invitrogen, anti-human–C8/144B, BioLegend), CD4 (anti-human–A19018, ABClonal), F4/80 (anti-mouse–D2S9R, Cell Signaling) or FoxP3 (anti-mouse–D6O8R, Cell Signaling Technology, anti-human–236A/E7, eBioscience) for 50 min at room temperature.

Techniques: Marker, Staining, Multiplex Assay, Comparison, Expressing, MANN-WHITNEY