title: Intracellular expression of an anti-idiotypic antibody single-chain variable fragment reduces porcine reproductive and respiratory syndrome virus infection in MARC-145 cells.
abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent of porcine reproductive and respiratory syndrome; it is one of the most economically important viral diseases affecting the swine industry worldwide. At present, neither live-attenuated nor inactivated PRRSV vaccines can provide sustainable disease control. Our previous studies have demonstrated that PRRSV infection can produce the auto-anti-idiotypic antibodies (aAb2s) specific to the idiotypic antibodies against PRRSV GP5, which plays an important role in the host immune responses to PRRSV infection. In the present study, a single-chain variable antibody fragment (scFv) from the monoclonal anti-idiotypic antibody specific for the idiotypic antibody against GP5 was expressed in MARC-145 cells and its effect on virus infection in vitro was evaluated.
  
  
  
  An scFv was constructed from the anti-idiotypic antibody (Mab2-5G2) and was named 5G2scFv. The lentiviral vector system was used as a vehicle to deliver 5G2scFv into MARC-145 cells. The effect of 5G2scFv expression in MARC-145 was analysed by determining the PRRSV N protein level and the virus titre in the supernatant. Virus attachment and the level of type I interferon (IFN) were determined to elucidate the mechanism of the scFv effect.
  
  
  
  5G2scFv was delivered in MARC-145 cells using the lentiviral vector system as confirmed by the western blot and indirect immunofluorescence assays. The PRRSV challenge experiments demonstrated that expressed 5G2scFv in MARC-145 strongly reduced PRRSV infection and replication by inhibiting protein synthesis and progeny virus production. This effect was not due to the change of viability or virus binding, but increased IFN-α at messenger RNA and protein levels.
  
  
  
  The expression of the anti-idiotypic antibody 5G2scFv in MARC-145 cells has the interferential effect on PRRSV infection in the cells by induction of IFN-α, which provides a novel therapeutic approach for PRRSV infection.
authors.affiliation: Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, China.
authors.forename: Qiongyi
authors.initials: Q
authors.lastname: Li
authors.forename: Xiangpeng
authors.initials: X
authors.lastname: Wang
authors.forename: Chengbao
authors.initials: C
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Yu
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Jiming
authors.initials: J
authors.lastname: Gao
authors.forename: Hongliang
authors.initials: H
authors.lastname: Liu
authors.forename: Huali
authors.initials: H
authors.lastname: Xie
authors.forename: Baicheng
authors.initials: B
authors.lastname: Huang
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Li
authors.forename: Ning
authors.initials: N
authors.lastname: Kong
authors.forename: Gaiping
authors.initials: G
authors.lastname: Zhang
authors.forename: Walter H
authors.initials: WH
authors.lastname: Hsu
authors.forename: En-Min
authors.initials: EM
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival; D005786:Gene Expression Regulation; D006801:Humans; D007372:Interferons; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D053585:Virus Attachment; D014779:Virus Replication
chemical_list: D000888:Antibodies, Anti-Idiotypic; D012333:RNA, Messenger; D007372:Interferons