Deficiency of cathepsin B suppresses Brucella intracellular infection

VernacularTitle:组织蛋白酶B基因敲除抑制布鲁菌胞内感染
Author: Jingjing LU ¹ ; Heling XU ; Qi ZHANG ; Fang HUANG ; Shanhu LI
Author Information

1. 军事科学院军事医学研究院,北京 100850
Keywords: Brucella; cathepsin B; autophagy; CRISPR/Cas9 gene editing
From: Military Medical Sciences 2024;48(5):362-368
CountryChina
Language:Chinese
Abstract: 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.