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. 2022 Mar 31;10(3):e003939. doi: 10.1136/jitc-2021-003939

Figure 1.

Figure 1

rBCG-S.FimH exhibited stronger bladder adhesion and internalization ability in vitro and in vivo. (A) Schematic representation for the construction of plasmids expressing FimH, EGFP or FimH-EGFP fused to the membrane protein Mbt19. (B) Only recombinant BCGs expressing FimH can bind to mannose residues in the HRP protein (BCG: wild-type BCG; S.FimH: rBCG-S.FimH; S.EGFP: rBCG-S.EGFP; S.FimH-EGFP: rBCG-S.FimH-EGFP; M: 100 µM D-mannose). (C–D) Mouse bladder cancer cell line MB49-luc was incubated with rBCG-S.EGFP and rBCG-S.FimH-EGFP for 4 hours, fluorescence images were taken by confocal (C) and fluorescence intensity quantitated using image J (D). Scale bar: 100 µm. (E–F) rBCG-S.FimH-EGFP showed higher adhesion to the urothelium in normal mouse bladder. C57BL/6 mice were deprived of water for 8 hours, followed by intravesical injection with PBS, rBCG-S.EGFP or rBCG-S.FimH-EGFP (5×105 CFU/mL, 50 µL/mice), the bladders were collected 4 hours later and frozen sections were subjected to immunofluorescence staining and confocal microscopy (E), fluorescence intensity were quantitated using image J (F). Scale bars: 250 µm and 40 µm. ****P<0.0001. HRP, horseradish peroxidase; PBS, phosphate buffered saline.