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. 2021 Oct 16;24(11):103295. doi: 10.1016/j.isci.2021.103295

Figure 2.

Figure 2

Different CAR-Ts possess cytolytic activity that mediated the killing of RBD, S1-loaded, and S1-expressing target cells

(A) Different CAR-Ts kill targets loaded with the RBD peptide. (Left panel) 293-ACE2 cells were loaded with the RBD peptide followed by washing and incubation with CAR-Ts for 4–5 h prior to a measure of killing. (Right panel) The presence of RBD peptides on loaded 293-ACE2 cells was detected with anti-S1 and anti-RBD antibodies. Data are shown as mean ± SD of triplicate samples. ∗∗∗∗p < 0.0001 (compared with CR3014-28Z by two-way ANOVA, n = 4).

(B) CR3022 antibody blocks CR3022-8a-28Z and CR3022-IgG4-28Z kill RBD-coated targets. 293-ACE2-RBD cells were incubated with CR3022, or the isotype control, at 37°C for 30 min prior to the incubation with CAR-Ts. Data are shown as mean ± SD of triplicate samples. ∗∗∗∗p < 0.0001 (compared with isotype control groups by paired Student’s t test, two tailed, n = 2).

(C) Different CAR-Ts kill targets coated with S1 peptide. (Left panel) 293-ACE2 cells were loaded with the S1 peptide followed by washing and incubation with CAR-Ts for 4–5 h prior to a measure of killing. (Right panel) The presence of S1 peptides on loaded 293-ACE2 cells was detected with anti-S1 and anti-RBD antibodies. Data are shown as mean ± SD of triplicate samples. ∗∗∗∗p < 0.0001 (compared with CR3014-28Z by two-way ANOVA, n = 3).

(D) Different CAR-Ts kill NIH/3T3 cells expressing the S1 peptide. (Lower panel) The S1 overexpressing NIH/3T3 cells were sorted by FACS staining of S1. These cells showed high levels of anti-S1 staining, whereas the staining with the CR3022 antibody was lower. (Upper panel) S1-expressing NIH/3T3 cells were co-cultured with CAR-Ts for 4–5 h prior to a measure of killing. Data are shown as mean ± SD of triplicate samples. ∗∗∗∗p < 0.0001 (compared with CR3014-28Z by two-way ANOVA, n = 3) (also see Figure S6).