sl032  (Genecopoeia)

Journal: Frontiers in Oncology

Article Title: Human Tumor Targeted Cytotoxic Mast Cells for Cancer Immunotherapy

doi: 10.3389/fonc.2022.871390

Figure Lengend Snippet: Breast cancer cell-induced BDMC mediator release. (A) Expression of HER2/ neu on BT474-1261 or SK-BR-3 cells using FACS. BDMC were sensitized with 1 μg/ml anti-HER/ neu IgE washed, and incubated with filtered BT474-1261 or SK-BR-3 cells and degranulation ( B ; left) and cytokine release ( B ; right) assessed. Data are from a single experiment representative of experiments performed on cells derived from three separate donors. Error bars represent ±SD. * p <0.05 compared with non-IgE (spontaneous) release.

Article Snippet: Luciferase/GFP dual-labeled SK-BR-3 cancer cell line was purchased from GeneCopoeia, Inc. Rockville, MD.

Techniques: Expressing, Incubation, Derivative Assay

Journal: Frontiers in Oncology

Article Title: Human Tumor Targeted Cytotoxic Mast Cells for Cancer Immunotherapy

doi: 10.3389/fonc.2022.871390

Figure Lengend Snippet: Mast cells bind to, penetrate, degranulate, and induce FcεRI-dependent apoptosis of BC cells. CellTracker™-Deep Red labelled ADMC (A, B) or BDMC (C) (7.5x10 4 ) were sensitized with 1 μg/ml non-specific IgE (A) or anti-HER2/ neu IgE (B, C) , washed, and incubated with MitoTracker™-Red HER2/ neu + BT474-1261 in culture medium containing a fluorescent apoptosis detecting dye (CellEvent Caspase 3/7; 1:100). Cells were monitored in a live cell chamber attached to a confocal microscope for up to 4 days. (D) . The amount of fluorescent intensity of the apoptosis dye was quantified using Image J of the average of ten fields of view at 63x magnification (±SD, *P < 0.05) (E) . Higher magnifications of several degranulating MCs is shown to illustrate the release and spreading of MC granules upon cancer cell HER2/ neu -induced activation of FcεRI. The red arrows indicate spreading MC granules within the BT474-1261 cancer cell mass. (F–H) . MitoTracker™-Red labelled MCs (7.5x10 4 ) were sensitized with 1 μg/ml non-specific IgE (F) or anti-HER2/ neu IgE (G, H) , washed, and incubated with HER2/ neu + BT474-1261 (F, G) or SK-BR-3 (H) cells and monitored in a live cell chamber for the indicated times. The red arrows indicate the dye-labelled ADMC, black arrows indicate cancer cell apoptotic bodies, and black circle indicates the outline of the ADMC within the tumor cell mass. Experiments are representative of 3 different donors.

Article Snippet: Luciferase/GFP dual-labeled SK-BR-3 cancer cell line was purchased from GeneCopoeia, Inc. Rockville, MD.

Techniques: Incubation, Microscopy, Activation Assay