1. [LRG47/EBP50 recombinant lentivirus-targeted vector vaccine enhances anti-tuberculosis immunity of RAW264.7 macrophages and its mechanism].
- Author
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Guo Y, Zhou J, Le F, Wang Y, Fu P, Su R, Huang Z, Luo Q, and Li J
- Subjects
- Humans, Lentivirus genetics, Lentivirus metabolism, Macrophages metabolism, Nitric Oxide metabolism, Phosphoproteins genetics, Sodium-Hydrogen Exchangers metabolism, Tuberculosis, Vaccines
- Abstract
Objective To investigate the effect of the gene vaccine in anti-tuberculosis immunity by constructing immunity-related p47 GTPase/ezrin-radixin-moesin-binding phosphoprotein 50 (LRG47/EBP50) gene co-expression recombinant lentivirus targeting vector. Methods Recombinant lentiviral plasmid vector pLenti-EBP50-LRG47 was established by using molecular cloning and packaged as lentivirus LV-EBP50-LRG47 and H37Rv infect macrophages. Then their bactericidal ability was tested by colony-forming units while the cellular autophagy and apoptosis was detected by flow cytometry. iNOS protein was detected by Western blotting and the expression level of nitric oxide (NO) was detected by ultraviolet spectrophotometer. Results The recombinant lentivirus LV-EBP50-LRG47 successfully up-regulated the expression of EBP50 and LRG47 after infecting macrophages. Compared with the control group, LV-EBP50-LRG47 can significantly inhibit the growth of intracellular H37Rv. The autophagy and apoptosis levels of LV-EBP50-LRG47 infected macrophages increased significantly, and the expression levels of iNOS and NO were significantly up-regulated. Conclusion LRG47/EBP50 gene co-expression enhances macrophages autophagy and apoptosis, and increases generation of iNOS and NO, which significantly inhibites the growth of intracellular H37Rv.
- Published
- 2022