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Updated: Apr 28, 2026
An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis
Published on: August 16, 2021
[Preparation of an autophagy-targeted antituberculosis DNA vaccine encoding LpqH fused with LC3]
Dong Hu1, Xiaokang Yang1, Jing Wu1
1Institute of Infection and Immunology, Department of Immunology, Medical School, Anhui University of Science and Technology, Huainan 232001, China.
Objective:
To construct an autophagy-targeted vaccine harboring the genes encoding lipoprotein antigen precursor LpqH from Mycobacterium tuberculosis and microtubule-associated protein light chain-3(LC3), and to investigate its efficacy of inducing and targeting autophagy.
Methods:
The expressions of LC-3 and LC3-LpqH in RAW264.7 cells were detected by Western blotting after transfected with pCMV-LpqH and pCMV-LC3-LpqH plasmids respectively. The pCMV-LC3-LpqH or pCMV-LpqH plasmids were transfected into GFP-LC3-RAW264.7 cells to analyze the localization of LC3-LpqH by immunofluorescence staining.
Results:
After transfected with pCMV-LpqH DNA, RAW264.7 cells showed a significant increase of LC-3 amount. The LC3-LpqH fusion protein was also detected in RAW264.7 cells after pCMV-LC3-LpqH transfection and in a dose-dependent manner. Interestingly, LpqH was found to be transported to autophagosomes through the fusion protein, which was demonstrated by the co-localization of GFP-LC3 and LC3-LpqH on autophagosomes.
Conclusion:
The recombinant plasmid encoding pCMV-LC3-LpqH could enhance the autophagy in vitro, and facilitate the localization of LpqH on autophagosomes. Our study provides a new practical strategy for the development of improved vaccines against Mycobacterium tuberculosis.
