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