Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity

Figure 5

The luciferase IVT RNA-based assay efficiently assesses mAb-induced ADCC and CDC of tumour cell lines. ADCC assay using (a) KATO-III and (b) NUGC-4 cells. KATO-III and NUGC-4 cells endogenously expressing hCLDN18.2 were transfected with 7 μg luc2 IVT RNA and seeded into 96-well plates independently. 4 h later, IMAB 362 at different concentrations and human PBMCs (E : T ratio = 40 : 1) from 6 different donors were added to the target cells and incubated for 24 h. ADCC was determined 40 and 45 min after addition of D-luciferin substrate to the KATO-III and NUGC-4 cells, respectively. (c) CDC assay. CHO-K1 cells stably expressing hCLDN18.2 were transfected with 7 μg luc2 IVT RNA and seeded into 96-well plates. 24 h later, cells were incubated for 80 min with IMAB 362 diluted in human serum (final concentration of 20%) from 6 different healthy donors. CDC was determined 45 min after addition of D-luciferin substrate. Results are the mean ± SD ().

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