Virology
Influence of cellular autophagy on replication dynamics of human T cell leukemia virus typeⅠ(HTLV-1) in HeLa cells
Qingsong Huang, Ziqiang Ding, Zhiguo Niu, Tingting Wang, Xiaomei Hou, Lushuang Mao, Ruohan Lyu, Ze Li, Xinxiang Huang, Hui Wang
Published 2017-03-31
Cite as Chin J Microbiol Immunol, 2017, 37(3): 208-212. DOI: 10.3760/cma.j.issn.0254-5101.2017.03.008
Abstract
ObjectiveTo investigate the replication dynamics and cellular states of host cells with human T cell leukemia virus type 1 (HTLV-1) infection.
MethodsHTLV-1 positive cell line MT2 was co-cultured with HeLa cells for 4 to 36 hours to acquire the virus early infection model. Immunoblotting was used to detect HTLV-1 core protein p19, apoptosis-related proteins caspase-3/9 and autophagy-related protein LC3 at different time points. Real-time PCR was used to measure viral loads. Luciferase reporter gene assay was used to detect the enzymatic activities of caspase-3/7/9 and the transcriptional activities of 3′-LTR at different time points after co-culturing. Autophagosomes were analyzed by immunofluorescence after co-culturing. The HTLV-1 long terminal repeat reporter gene (pHTLV-1-LTR-luc) was transfected into HeLa cells by lipidosome 2000-mediated transfection and the luciferase activities at different time points were detected.
ResultsThe HTLV-1 infection model was successfully constructed by co-culturing MT2 cells with HeLa cells in vitro. Viral load and transcriptional activity of the 3′-LTR gradually increased at 4 to 16 h after virus infection (P<0.01) and the viral load reached a steady state at 24 h. The expression of core protein p19 could be detected at 4 h after infection, decreased at 8 h and increased at 16 h, followed by reaching a steady state at 24 h. The enzymatic activity and cleaved protein of caspase-3/7/9 had no significant change at different time points after infection (P>0.05), but the proportion of LC3B/LC3A showed a gradual increasing trend with the persistent increase of autophagosomes (P<0.01).
ConclusionViral replication dynamics of HTLV-1 is closely related to autophagy in host cells.
Key words:
Human T cell leukemia virus type 1; Replication dynamics; Apoptosis; Autophagy
Contributor Information
Qingsong Huang
School of Medicine, Jiangsu University, Zhenjiang 212013, China
Ziqiang Ding
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Zhiguo Niu
Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine in Henan Province, Xinxiang 453003, China
Tingting Wang
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Xiaomei Hou
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Lushuang Mao
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Ruohan Lyu
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Ze Li
Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China
Xinxiang Huang
School of Medicine, Jiangsu University, Zhenjiang 212013, China
Hui Wang
Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine in Henan Province, Xinxiang 453003, China