Fig. 2.

The effect of artificial bioorthogonal targeting over cell surface on CAR-T cell recognition, migration, and antitumor response. (a) The interaction between CAR-T and Raji cell was analyzed by confocal imaging. The white circles indicated cell clusters of T and Raji cells. (b) the number of cell cluster in the co-culture of CAR-T and Raji cells with or without treatment by Ac₄ManN-BCN (20 μM) and Ac₄GalNAz (50 μM) respectively, at early 20 min. (c) Transwell assay of CAR-T cell migration with or without bioorthogonal targeting using flow cytometry. (d) Fluorescence imaging analysis of CAR-T cell infiltration into tumor spheroids.(e-f)CAR-T cell infiltration was quantified by flow cytometry using anti-Myc tag antibodies. (g–i) The release of IL-2, IFN-γ and TNF-α was determined using ELISA assay. *p < 0.05, **p < 0.01.