A potential target gene for the host-directed therapy of mycobacterial infection in murine macrophages
| Autor: | Ran Chen, Jianying Zhou, Xing Chen, Yake Yao, Pei Zhang, Zhang Bao, Yi-Lan Sun, Shan Lu, Xiaozheng Jin |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Collagen Type IV Male Vacuolar Proton-Translocating ATPases Cell Survival Cell phagosomal acidification Blotting Western Gene Expression macrophage Biology Cell Line 03 medical and health sciences 0302 clinical medicine RNA interference Gene expression Genetics medicine Macrophage Animals Gene Cells Cultured Phagosome Gene knockdown Microscopy Confocal Reverse Transcriptase Polymerase Chain Reaction Macrophages Cytoplasmic Vesicles General Medicine Articles Mycobacterium tuberculosis Col4a5 Hydrogen-Ion Concentration Molecular biology Mice Inbred C57BL 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis Host-Pathogen Interactions vacuolar-type H+-ATPase RNA Interference Intracellular |
| Zdroj: | International Journal of Molecular Medicine |
| ISSN: | 1791-244X 1107-3756 |
| Popis: | Mycobacterium tuberculosis (MTB), one of the major bacterial pathogens for lethal infectious diseases, is capable of surviving within the phagosomes of host alveolar macrophages; therefore, host genetic variations may alter the susceptibility to MTB. In this study, to identify host genes exploited by MTB during infection, genes were non-selectively inactivated using lentivirus-based antisense RNA methods in Raw264.7 macrophages, and the cells that survived virulent MTB infection were then screened. Following DNA sequencing of the surviving cell clones, 26 host genes affecting susceptibility to MTB were identified and their pathways were analyzed by bioinformatics analysis. In total, 9 of these genes were confirmed as positive regulators of collagen α-5(IV) chain (Col4a5) expression, a gene encoding a type IV collagen subunit present on the cell surface. The knockdown of Col4a5 consistently suppressed intracellular mycobacterial viability, promoting the survival of Raw264.7 macrophages following mycobacterial infection. Furthermore, Col4a5 deficiency lowered the pH levels of intracellular vesicles, including endosomes, lysosomes and phagosomes in the Raw264.7 cells. Finally, the knockdown of Col4a5 post-translationally increased microsomal vacuolar-type H+-ATPase activity in macrophages, leading to the acidification of intracellular vesicles. Our findings reveal a novel role for Col4a5 in the regulation of macrophage responses to mycobacterial infection and identify Col4a5 as a potential target for the host-directed anti-mycobacterial therapy. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|