Journal: Cancer research

Article Title: Focused Ultrasound Delivers Targeted Immune Cells to Metastatic Brain Tumors

doi: 10.1158/0008-5472.CAN-12-2609

Figure Lengend Snippet: Histological quantification of HER2-specific NK-92 cells accumulating at the tumor site. Effector cells were co-localized with CD45 IHC (upper panel) and Prussian blue histochemistry (lower panel) in the three experimental groups, A. HER2-specific NK-92 cells reaching the tumor were quantitatively assessed (mean ± SEM), B. When NK-92 cells were injected prior to BBBD, the number reaching the tumor was significantly higher than if they were injected following or without BBBD (group 3 vs groups 1 and 2: 0.95 ± 0.23 vs 0.09 ± 0.11, 0.21 ± 0.15, p < 0.01). There was no statistical difference between groups 1 and 2. These results are in agreement with the iron-sensitive MR imaging in Figure 4.

Article Snippet: Expression of CD45 was evaluated on a cell smear of targeted cells using mouse anti-human CD45 (RnD systems, Minneapolis, MN USA) and the procedures described above.

Techniques: Injection, Imaging

Journal: Cancer research

Article Title: Focused Ultrasound Delivers Targeted Immune Cells to Metastatic Brain Tumors

doi: 10.1158/0008-5472.CAN-12-2609

Figure Lengend Snippet: FUS causes the translocation of HER2-specific NK-92 cells from the vasculature into the brain and tumor when they are present in the circulation at the time of BBBD. A, CD45 IHC depicting a vessel from which a large number of cells have extravasated and appear to track to the tumor (indicated by the star). B, the corresponding Prussian blue stained section is shown, colocalizing the HER2-specific NK-92 cells. C, a normal capillary adjacent the tumor but within the sonicated region, shows HER2-specific NK-92 cells forced to the adluminal surface of the vessel. FUS results in HER2-specific NK-92 cells circumventing both the BBB and BTB. D, the corresponding Prussian blue section. These cell distributions were seen exclusively in group 3 animals.

Article Snippet: Expression of CD45 was evaluated on a cell smear of targeted cells using mouse anti-human CD45 (RnD systems, Minneapolis, MN USA) and the procedures described above.

Techniques: Translocation Assay, Staining, Sonication

Journal: bioRxiv

Article Title: Oseltamivir (Tamiflu), a Commonly Prescribed Antiviral Drug, Mitigates Hearing Loss in Mice

doi: 10.1101/2024.05.06.592815

Figure Lengend Snippet: (A) Noise exposure and treatment schedule. Mice were exposed to 100 dB noise for 2 hours (8-16 kHz) and treated with oseltamivir for 3 days starting 24 hours following noise exposure. Cochleae were collected 4 days post noise insult. (B) Representative confocal images of cochlear cryosections stained with CD45 (red) and DAPI (blue). (C) Quantification of CD45 positive cells per cochlear section. Four treatment groups are carrier alone (black), oseltamivir alone (yellow), noise + carrier (red), and oseltamivir + noise (blue). Data shown as means ± SEM, **P<0.01 compared to noise alone by one-way ANOVA with Bonferroni post hoc test. n=3-6 mice.

Article Snippet: Tissues were stained overnight at 4°C with mouse CD45 antibody (1:50; Af114, R&D Systems).

Techniques: Staining

Journal: Cancer immunology research

Article Title: Combined anti-VEGF and anti–CTLA-4 therapy elicits humoral immunity to galectin-1 which is associated with favorable clinical outcomes

doi: 10.1158/2326-6066.CIR-16-0385

Figure Lengend Snippet: Anti-Gal-1 antibodies isolated from an Ipi-Bev-treated patient abrogate Gal-1 binding to CD45. Anti-Gal-1 antibodies were affinity purified from plasma. Biotinylated HAS-Gal-1 (250 ng/ml) was incubated with a commercial anti-Gal-1 polyclonal antibody (Gal-1 Ab) or control antibody (10 μg/ml), enriched endogenous Gal-1 antibodies (Gal-1 Ig) or normal human IgG (1.98 μg/ml) prior to incubation with coated CD45. The binding of HAS-Gal-1 to CD45 was detected with streptavidin-HRP. Sucrose and lactose were added to the reaction at 5 mM. Data are presented as Mean ± SD of 3 experiments.

Article Snippet: Recombinant human CD45 (R&D Systems) was coated onto 96-well plates (25 ng/well) at 4°C overnight.

Techniques: Isolation, Binding Assay, Affinity Purification, Clinical Proteomics, Incubation, Control

Journal: Journal of Nanobiotechnology

Article Title: A near infrared light-triggerable modular formulation for the delivery of small biomolecules

doi: 10.1186/s12951-019-0530-y

Figure Lengend Snippet: Intracellular trafficking of RA1-CB[6] @AuNRs as evaluated by immunocytochemistry. a Schematic representation of the experiment. b Confocal images of leukemic cells incubated with RA1-CB[6] @AuNR-PEG-TRITC. Cells were incubated for 4 h with AuNR (50 µg/mL), washed to remove the non-internalised AuNRs, added new cell culture media, irradiated for 2 min with a 780 nm laser (power: 2 W/cm 2 ) and finally fixed with PFA (4%). The cell membrane was immunostained for CD45, while endosomes were immunostained for EEA1. White scale bar corresponds to 30 µm. c Intracellular localization of RA1-CB[6] @AuNRs before NIR light irradiation was confirmed by a Z-stack scan. Scale bar corresponds to 10 µm. d The co-localization between RA1-CB[6] @AuNR-PEG-TRITC and early endosomes before and after irradiation was quantified by image analyses. Results are average ± SEM ( n = 3). Statistical analysis was performed using a t-test. ***Denotes statistical significance (P < 0.001)

Article Snippet: Cell membrane was stained with monoclonal mouse anti-human CD45 antibody, diluted 1:50 (R&D Systems, MAB1430) and using Alexa Fluor-488 goat anti-mouse igG as secondary antibody, diluted 1:1000 (Life technologies, A11001).

Techniques: Immunocytochemistry, Incubation, Cell Culture, Irradiation, Membrane