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[Preparation of an autophagy-targeted antituberculosis DNA vaccine encoding LpqH fused with LC3].
- Source :
-
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2014 Jun; Vol. 30 (6), pp. 581-4. - Publication Year :
- 2014
-
Abstract
- 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.<br />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.<br />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.<br />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.
- Subjects :
- Animals
Antigens, Bacterial genetics
Antigens, Bacterial metabolism
Antitubercular Agents administration & dosage
Antitubercular Agents immunology
Blotting, Western
Cell Line
Fluorescent Antibody Technique
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Lipoproteins genetics
Lipoproteins metabolism
Macrophages cytology
Macrophages immunology
Macrophages metabolism
Mice
Mice, Inbred BALB C
Microtubule-Associated Proteins genetics
Microtubule-Associated Proteins metabolism
Mycobacterium tuberculosis genetics
Mycobacterium tuberculosis immunology
Mycobacterium tuberculosis metabolism
Phagosomes immunology
Phagosomes metabolism
Plasmids genetics
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Transfection
Tuberculosis immunology
Tuberculosis microbiology
Tuberculosis prevention & control
Tuberculosis Vaccines administration & dosage
Tuberculosis Vaccines genetics
Vaccines, DNA administration & dosage
Vaccines, DNA genetics
Vaccines, DNA immunology
Antigens, Bacterial immunology
Autophagy immunology
Lipoproteins immunology
Microtubule-Associated Proteins immunology
Recombinant Fusion Proteins immunology
Tuberculosis Vaccines immunology
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1007-8738
- Volume :
- 30
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
- Publication Type :
- Academic Journal
- Accession number :
- 24909277