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. 2021 Dec 23;12:763460. doi: 10.3389/fimmu.2021.763460

Figure 3.

Figure 3

H84T-BanLec CAR-NK cells decrease S-pseudotyped viral infection. (A) Schema of SARS-CoV-2 pseudovirus infection of hACE2.293T. Pseudoviral particles contain plasmids encoding ffLuc. Following viral entry, cells emit bioluminescence (BL) after D-Luciferin metabolism. (B) Measurement of target cell BL emission following transduction with S-protein pseudotyped virus carrying ffLuc reporter gene. Assay performed in triplicate. (C) Schematic representation of the BanLec-CAR NK cells blocking hACE2.293T infection. (D) NK cells plated with target cells (hACE2.293T) and pseudovirus at indicated effector to target (E:T) ratios. Percent bioluminescence (BL) reduction from maximal measured on day 2. The condition with 293T.ACE2 and pseudovirus alone (100%) represents the positive control (E:T = 0.4, p = 0.034; E:T = 1, p = 0.054, n = 6, 2 separate experiments using 3 independent NK cell donors, each experiment performed in triplicate). (E) Quantification (pg/mL) of IFNγ and TNFα present in culture media of NK cells at baseline and in co-culture with S-pseudotyped virus infected hACE2.293T (black: no target, red: co-culture; n=3 donors; Mean value+/- SEM; baseline vs. co-culture *p < 0.05, **p < 0.01, ***p < 0.001, not significant if not indicated).