Deficiency of cathepsin B suppresses Brucella intracellular infection
Objective To investigate the role of cathepsin B(CTSB)inhuman cervical cancer HeLa cellswith Brucella infections.Methods The ctsb knockout(KO)HeLa cell line was constructed using CRISPR/Cas9.The effects of ctsbKO on cell proliferation and survival of intracellular bacteria in case of Brucella infection were detected by CCK-8 assay and intracellular bacteria count while the molecular mechanism of ctsb regulation on Brucella infection was revealed by Western blotting.Results The ctsbKO cell line was constructed.CCK-8 results showed that KO of ctsb had no impact on cell proliferationwith or without Brucella infection.The results of intracellular bacteria count showed a significant decrease in intracellular Brucella four days after infection in the KO cell line.Western blotting assay suggested that the expression levels of autophagy-related proteins were significantly down-regulated in the KO cell line with long-term Brucella infection.Conclusion KO of ctsb inhibits the intracellular survival of Brucella in long-term infections,providing a potential target for the treatment of chronic brucellosis.
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