Journal: Pharmaceuticals

Article Title: Click-to-Release for Controlled Immune Cell Activation: Tumor-Targeted Unmasking of an IL12 Prodrug

doi: 10.3390/ph18091380

Figure Lengend Snippet: In vivo characterization of diabody-Tz 12 and IL12-TCO-PEG 2 10 . ( A ) Blood kinetics of I-125 labelled IL12 and IL12-TCO-PEG 2 (FG 11), administered i.v. in tumor-free mice (10 µg/mouse), followed by detection of radioactivity in blood samples collected at various timepoints (2, 30, 60 min, 3, 6, 24, and 72 h post injection). ( B ) Blood kinetics of I-125-labelled diabody-Tz 12 , administered i.v. in tumor-free mice (40 µg/mouse), followed by detection of radioactivity in blood samples collected at various timepoints (2, 30, 60 min, 3, 6, 24, and 72 h post injection). ( C ) Biodistribution of I-125-labelled diabody-Tz 12 , administered i.v. in LS174T-tumor bearing mice (150 µg/mouse), followed by detection of radioactivity in various tissues at 48 h post-injection. ( D ) For selected groups, the tumor from a representative mouse was isolated and snap-frozen in OCT matrix, and the presence of reactive Tz in the tumor was detected by TCO-PEG 4 -biotin on 5 μm tissue slides. ( E ) I-125 Autoradiography of the same tumor slices as panel ( D ) with arrows indicating areas where clusters of Tz are visible. Data represent the mean percentage injected dose (% ID) or injected dose per gram (% ID/g) with s.d. ( n = 4).

Article Snippet: The human colon cancer LS174T cell line was obtained from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 medium (Gibco/Thermo Fisher Scientific (Waltham, MA, USA)) supplemented with 2 mM glutamine (Gibco) and 10% fetal calf serum (FCS; Sigma Aldrich (St. Louis, MA, USA)).

Techniques: In Vivo, Radioactivity, Injection, Isolation, Autoradiography

Journal: Pharmaceuticals

Article Title: Click-to-Release for Controlled Immune Cell Activation: Tumor-Targeted Unmasking of an IL12 Prodrug

doi: 10.3390/ph18091380

Figure Lengend Snippet: In vivo IL12 detection. ( A ) Schematic workflow to detect (unmasked) IL12 in tumors and plasma after trigger pretargeting. Diabody-Tz was injected i.v. in mice bearing LS174T tumors (40 µg in 100 µL PBS per mouse; n = 4). Twenty-four hours later, when the diabody was virtually cleared from blood, TCO-masked IL12 was injected i.v. (FG 11; 10 µg in 100 µL PBS). Twenty-four hours later, tumors and blood were isolated and processed with protease inhibitor cocktail present. Before the ELISA, some samples received an excess of Tz 8 to release all remaining masks. ( B ) ELISA analysis showing no IL12 unmasking in plasma in vivo due to low blood level of Tz at the time of IL12-TCO-PEG injection (left panel). Right panel shows that pre-localized Tz in the tumor is able to unmask the IL12 construct locally. Data represent mean OD450nm values with SEM ( n = 4). Significance: n.s. indicates p > 0.05; *** indicates p ≤ 0.001.

Article Snippet: The human colon cancer LS174T cell line was obtained from ATCC (Manassas, VA, USA) and cultured in RPMI-1640 medium (Gibco/Thermo Fisher Scientific (Waltham, MA, USA)) supplemented with 2 mM glutamine (Gibco) and 10% fetal calf serum (FCS; Sigma Aldrich (St. Louis, MA, USA)).

Techniques: In Vivo, Clinical Proteomics, Injection, Isolation, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Construct

Journal: Microbiome Research Reports

Article Title: The biofunction of Akkermansia muciniphila in intestinal-related diseases

doi: 10.20517/mrr.2024.12

Figure Lengend Snippet: The mechanism of Akk and its effective components in intestinal-related diseases

Article Snippet: Colon cancer cell line LS174T , ATCC BAA-835 , Cell adhesion , Bind with normal colon cells and degrade Muc2 secreted by colon cancer cells, as well as promoting the mutual adhesion of colon cancer cells with high Muc2 expression , [ ] .

Techniques: Expressing, Gene Expression, Phospho-proteomics