title: Viperin inhibits rabies virus replication via reduced cholesterol and sphingomyelin and is regulated upstream by TLR4.
abstract: Viperin (virus inhibitory protein, endoplasmic reticulum-associated, IFN-inducible) is an interferon-inducible protein that mediates antiviral activity. Generally, rabies virus (RABV) multiplies extremely well in susceptible cells, leading to high virus titres. In this study, we found that viperin was significantly up-regulated in macrophage RAW264.7 cells but not in NA, BHK-21 or BSR cells. Transient viperin overexpression in BSR cells and stable expression in BHK-21 cells could inhibit RABV replication, including both attenuated and street RABV. Furthermore, the inhibitory function of viperin was related to reduce cholesterol/sphingomyelin on the membranes of RAW264.7 cells. We explored the up-stream regulation pathway of viperin in macrophage RAW264.7 cells in the context of RABV infection. An experiment confirmed that a specific Toll-like receptor 4 (TLR4) inhibitor, TAK-242, could inhibit viperin expression in RABV-infected RAW264.7 cells. These results support a regulatory role for TLR4. Geldanamycin, a specific inhibitor of interferon regulatory factor 3 (IRF3) (by inhibiting heat-shock protein 90 (Hsp90) of the IRF3 phosphorylation chaperone), significantly delayed and reduced viperin expression, indicating that IRF3 is involved in viperin induction in RAW264.7 cells. Taken together, our data support the therapeutic potential for viperin to inhibit RABV replication, which appears to involve upstream regulation by TLR4.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, Guangxi, China.|Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Hai-Bo
authors.initials: HB
authors.lastname: Tang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, Guangxi, China.|Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Zhuan-Ling
authors.initials: ZL
authors.lastname: Lu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, Guangxi, China.|Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Xian-Kai
authors.initials: XK
authors.lastname: Wei
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Tao-Zhen
authors.initials: TZ
authors.lastname: Zhong
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Yi-Zhi
authors.initials: YZ
authors.lastname: Zhong
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Ling-Xuan
authors.initials: LX
authors.lastname: Ouyang
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Xing-Wei
authors.initials: XW
authors.lastname: Xing
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Liao
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Ke-Ke
authors.initials: KK
authors.lastname: Peng
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Chao-Qian
authors.initials: CQ
authors.lastname: Deng
authors.affiliation: Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Minamoto
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, Guangxi, China.|Laboratory of Animal Infectious Diseases, College of Animal Sciences and Veterinary Medicine, Guangxi University, Nanning 530004, Guangxi, China.
authors.forename: Ting Rong
authors.initials: TR
authors.lastname: Luo
mesh_terms: D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D006224:Cricetinae; D004285:Dogs; D051379:Mice; D011506:Proteins / Q000378:metabolism*; D000067996:RAW 264.7 Cells; D011820:Rabies virus / Q000502:physiology*; D013109:Sphingomyelins / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D014779:Virus Replication; D000090123:Viperin Protein
chemical_list: D002784:Cholesterol; D011506:Proteins; D013109:Sphingomyelins; D051197:Toll-Like Receptor 4; C493487:Tlr4 protein, mouse; C417529:Rsad2 protein, mouse; D000090123:Viperin Protein