HomeHome

Respiratory syncytial virus prolifically infects N2a neuronal cells, leading to TLR4 and nucleolin protein modulations and RSV F protein co-localization with TLR4 and nucleolin

Xiaoling Yuan; Tao Hu; Hanwen He; Huan Qiu; Xuan Wu; Jingxian Chen; Minmin Wang; Cheng Chen; Shenghai Huang

Title:
Respiratory syncytial virus prolifically infects N2a neuronal cells, leading to TLR4 and nucleolin protein modulations and RSV F protein co-localization with TLR4 and nucleolin
Author(s):
Yuan, Xiaoling
Hu, Tao
He, Hanwen
Qiu, Huan
Wu, Xuan
Chen, Jingxian
Wang, Minmin
Chen, Cheng
Huang, Shenghai
Date:
Type:
Articles
Department(s):
Pathology and Cell Biology
Volume:
25
Persistent URL:
Book/Journal Title:
Journal of Biomedical Science
Notes:
Respiratory syncytial virus (RSV), Toll-like receptor 4 (TLR4), Nucleolin (C23), Virus infection, N2a cells
Abstract:
Background Respiratory syncytial virus (RSV) infects the central nervous system, resulting in neurological symptoms. However, the precise underlying pathogenic mechanisms have not been elucidated. In the present study, the infectivity of RSV on N2a neuronal cells and the possible roles of Toll-like receptor 4 (TLR4) and nucleolin (C23) during RSV infection were investigated. Methods We compared two experimental groups (infected and non-infected) and monitored the RSV viral titers in the culture supernatant by a viral plaque assay. We also inspected the morphology of the nucleus in infected N2a cells. We measured the level of RSV F protein and studied its co-localization with TLR4 and nucleolin using immunofluorescence assays and laser confocal microscopy. The potential interaction of RSV F protein with TLR4 and nucleolin was examined by coimmunoprecipitation. The expression changes of TLR4, nucleolin, TLR3 and TLR7 proteins in N2a cells and IL-6 and TNF-α in the culture supernatant were investigated by Western Blot analysis and ELISA assay. Changes in neuronal cell apoptosis status was examined by flow cytometry. Results The results demonstrated prolific RSV infection of N2a cells, which triggered a decrease of NeuN protein expression, coinciding with an increase of nuclear lesions, F protein expression, RSV viral titers, and late apoptotic levels of N2a cells. RSV infection induced co-localization of RSV F protein with TLR4 and nucleolin, which could potentially lead to a direct interaction. Furthermore, it was found that TLR4 and nucleolin levels increased early after infection and decreased subsequently, whereas TLR3 and TLR7 expression increased throughout RSV infection. Conclusion The RSV Long strain can prolifically infect N2a neuronal cells, modulating the expression of TLR4 and nucleolin, as well as TLR3, TLR7 and their downstream inflammatory factors, and inducing the co-localization of the RSV F protein with TLR4 and nucleolin.
Subject(s):
Respiratory syncytial virus
Virus diseases
Medicine
Biology
Pathology
Publisher DOI:
https://doi.org/10.1186/s12929-018-0416-6
Item views
18
Metadata:
text | xml
Suggested Citation:
Xiaoling Yuan, Tao Hu, Hanwen He, Huan Qiu, Xuan Wu, Jingxian Chen, Minmin Wang, Cheng Chen, Shenghai Huang, , Respiratory syncytial virus prolifically infects N2a neuronal cells, leading to TLR4 and nucleolin protein modulations and RSV F protein co-localization with TLR4 and nucleolin, Columbia University Academic Commons, .

Columbia University Libraries | Policies | FAQ