title: Autophagy-targeted vaccine of LC3-LpqH DNA and its protective immunity in a murine model of tuberculosis.
abstract: The development of more effective antituberculosis vaccines would contribute to the control of the global problem of infection with Mycobacterium tuberculosis (MTB). Recently, the highlighted importance of autophagy in the host immune response against MTB has attracted the attention of researchers. However, the vaccines targeted at autophagy remain to be developed. In this study, we report on an autophagy-targeted vaccine of 19kDa MTB lipoprotein (LpqH) DNA that harbors another gene coding microtubule-associated protein light chain-3(LC3), which transports LpqH to autophagosomes and displays enhanced protective efficacy against MTB. After the transfection of pCMV-LpqH DNA, a significant increase LC3 II was detected in RAW264.7 cells, which was similar to that observed with treatment with rapamycin, a reagent used to induce autophagy. To target autophagy, the gene coding LC3, as a marked protein of autophagosome, was linked to the lpqH gene to express an LC3-LpqH fused protein. Interestingly, LC3-LpqH fused protein was determined to be transported to an autophagosome, which was demonstrated by the co-localization of GFP-LC3 with LC3-LpqH at autophagosomes. Notably, the mice immunized with LC3-LpqH/Ag85B displayed decreased mycobacterial loads in the lungs and spleen when challenged with virulent MTB by intravenous infection, which was consistent with increased IgG2a in serum and IFN-γ and IL-2 produced by splenocyte. In conclusion, our study demonstrates that an LC3-LpqH DNA vaccine could have autophagy as its target, which contributes to the enhancement of the Th1 immune response and vaccine protective efficacy.
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China. Electronic address: austhudong@126.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Hu
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China. Electronic address: wujing8008@126.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China. Electronic address: lory456@126.com.
authors.forename: Rongbo
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Chen
authors.affiliation: Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China.
authors.forename: Zhaoquan
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Medical Immunology, Medical School, Anhui University of Science and Technology, Huainan, China; Institute of infection and immunology, Anhui University of Science and Technology, Huainan, China.
authors.forename: Lifa
authors.initials: L
authors.lastname: Xu
authors.affiliation: School of Pharmacy, Wenzhou Medical College, Wenzhou, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Fengyu
authors.initials: F
authors.lastname: Hu
authors.affiliation: Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
authors.forename: Changyou
authors.initials: C
authors.lastname: Wu
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001343:Autophagy; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000276:immunology; D002460:Cell Line; D016207:Cytokines / Q000276:immunology; D005260:Female; D008074:Lipoproteins / Q000276:immunology; D008168:Lung / Q000382:microbiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008869:Microtubule-Associated Proteins / Q000276:immunology*; D010588:Phagosomes / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013154:Spleen / Q000382:microbiology; D014376:Tuberculosis / Q000517:prevention & control*; D032581:Tuberculosis Vaccines / Q000276:immunology*; D019444:Vaccines, DNA / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D016207:Cytokines; D008074:Lipoproteins; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA
keywords: Ag85B; Autophagy; LC3; LpqH; Mycobacterium tuberculosis; Protective immunity