Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Nature Biotechnology

Article Title: Protection of cell therapeutics from antibody-mediated killing by CD64 overexpression

doi: 10.1038/s41587-022-01540-7

Figure Lengend Snippet: a , b , Human thyroid epiCs TPO and epiCs TPO,CD64t were challenged in impedance NK cell ADCC ( a ) and CDC ( b ) assays with different concentrations of an anti-TPO IgG1 antibody (mean ± s.d.; three independent replicates per group and timepoint). c – e , Human thyroid epiCs TPO and epiCs TPO,CD64t were incubated with serum from Hashimoto’s patients 1 ( c ), 2 ( d ) and 3 ( e ) in impedance CDC assays (mean ± s.d.; three independent replicates per group and timepoint). ULN, upper limit of normal. f , 5 × 10 4 human thyroid epiCs TPO or epiCs TPO,CD64t were injected subcutaneously into NSG mice with 10 6 human NK cells. Both groups received three subcutaneous doses of anti-TPO IgG1 1 mg on days 0, 1 and 2. g , h , BLI signals of thyroid epiCs TPO ( g ) and epiCs TPO,CD64t ( h ) were followed (all individual mice were plotted, and BLI pictures of one representative mouse per group are shown). s.c., subcutaneous.

Article Snippet: A 24-well plate was coated with gelatin, and 5 × 10 4 iPSC-derived beta cells and beta cells CD64t (Y10108, Takara Bio) per well were seeded in 500 μl of Cellartis hiPS beta cell media and incubated for 24 hours at 37 °C in 5% CO 2 .

Techniques: Incubation, Injection

Journal: Nature Biotechnology

Article Title: Protection of cell therapeutics from antibody-mediated killing by CD64 overexpression

doi: 10.1038/s41587-022-01540-7

Figure Lengend Snippet: a , Flow cytometry histograms for anti-CD19 scFv and CD64t expression on human CAR T and CAR T CD64t cells (representative graphs of two independent experiments). b , Flow cytometry histograms for the binding of free IgG1 F c (anti-TPO IgG1; representative graph of two independent experiments). c , The kinetics of Nalm6 target cell killing by T cells, CAR T cells and CAR T CD64t cells is expressed as hours that it takes for the cell index to drop from 1 to 0.5. Different T cell-to-Nalm6 ratios are shown (mean ± s.d.; three independent replicates per group and timepoint). d , The kinetics of Nalm6 target cell killing by CAR T CD64t in the presence and absence of 1 μg ml −1 of anti-CD52 is expressed as hours that it takes for the cell index to drop from 1 to 0.5. Different CAR T cell-to-Nalm6 ratios are shown (mean ± s.d.; three independent replicates per group and timepoint). e , f , Human CAR T and CAR T CD64t cells were challenged in impedance NK cell ADCC ( e ) and CDC ( f ) assays with antibodies against HLA (HLA-A2), non-HLA (CD52 and CD3), rhesus blood type antigen D (Rh(D)) and the CAR (anti-CD19 scFv) at 1 μg ml −1 (mean ± s.d.; three independent replicates per group and timepoint).

Article Snippet: A 24-well plate was coated with gelatin, and 5 × 10 4 iPSC-derived beta cells and beta cells CD64t (Y10108, Takara Bio) per well were seeded in 500 μl of Cellartis hiPS beta cell media and incubated for 24 hours at 37 °C in 5% CO 2 .

Techniques: Flow Cytometry, Expressing, Binding Assay