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Classical swine fever virus triggers RIG-I and MDA5-dependent signaling pathway to IRF-3 and NF-κB activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages.
Dong, Xiao-Ying; Liu, Wen-Jun; Zhao, Ming-Qiu; Wang, Jia-Ying; Pei, Jing-Jing; Luo, Yong-Wen; Ju, Chun-Mei; Chen, Jin-Ding.
Afiliação
  • Dong XY; College of Veterinary Medicine, South China Agricultural University, No,483 Wu Shan Road, Tian He District, Guangzhou 510642, China. jdchen@scau.edu.cn.
Virol J ; 10: 286, 2013 Sep 13.
Article em En | MEDLINE | ID: mdl-24034559
ABSTRACT

BACKGROUND:

Classical swine fever (CSF) caused by CSF virus (CSFV) is a highly contagious disease of pigs. The RNA helicases retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA-5) are differentially involved in the detection of various RNA viruses. In present study, we investigated the roles of RIG-I and MDA-5 in eliciting antiviral and inflammatory responses to CSFV shimen strain in Porcine alveolar macrophages (PAMs).

METHODS:

CSFV Shimen strain was used as challenge virus in this study and PAMs were cultured in vitro. Interferon regulatory factor (IRF)-3 and nuclear factor-kappa B (NF-κB) translocation was detected using immunofluorescent staining; RIG-I, MDA5, interferon promoter-stimulating factor 1 (IPS-1), IRF-3 and NF-κB expression was measured by Western Blotting; Interferon beta (IFN-ß), IFN-α, interleukin-1beta (IL-1ß), IL-6 and tumor necrosis factor (TNF-α) expression was tested by Enzyme-linked immunosorbent assays (ELISA) and shRNA-mediated knockdown of MDA5 or RIG-I was performed.

RESULTS:

The findings suggested that the initial response to CSFV infection resulted in the higher expression of RIG-I and MDA5 leading to the activation of IPS-1, IRF-3 and NF-κB in a dose-dependent manner. Evaluation of IFN-α, IFN-ß, IL-1ß, IL-6 or TNF-α expressed by PAMs showed significant differences between infected and uninfected cells. CSFV infected cells induced to express high levels of IFN-α, IFN-ß, IL-1ß, IL-6 and TNF-α in a dose-dependent way within 24 h post-infection (hpi). At the same time, CSFV improved the nuclear translocation of IRF-3 and NF-κB. We also directly compared and assessed the roles of RIG-I and MDA5 in triggering innate immune actions during CSFV infection through shRNA-mediated knockdown of MDA5 or RIG-I. We found that, compared to the control, the production of IFN-α, IFN-ß, IL-1ß, IL-6 and TNF-α in response to CSFV infection was heavily reduced in RIG-I knockdown cells while it was moderately decreased in MDA5 knockdown cells. PAMs derived from knockdown of both RIG-I and MDA5 almost failed to produce IFNs and inflammatory cytokines.

CONCLUSIONS:

It indicates that CSFV can be recognized by both RIG-I and MDA5 to initiate the RIG-I signaling pathway to trigger innate defenses against infection.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transdução de Sinais / Citocinas / Macrófagos Alveolares / RNA Helicases DEAD-box / Interações Hospedeiro-Patógeno / Vírus da Febre Suína Clássica Limite: Animals Idioma: En Revista: Virol J Ano de publicação: 2013 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Transdução de Sinais / Citocinas / Macrófagos Alveolares / RNA Helicases DEAD-box / Interações Hospedeiro-Patógeno / Vírus da Febre Suína Clássica Limite: Animals Idioma: En Revista: Virol J Ano de publicação: 2013 Tipo de documento: Article