title: Generation and evaluation of avian leukosis virus subgroup J envelope glycoprotein recombinant pseudovirions.
abstract: Retroviral and lentiviral vector pseudotypes (based on human immunodeficiency virus type 1, HIV-1) have been used for stable and safe gene transfer because of their broad host ranges and high mechanical strength. In the present study, a recombinant avian leukosis virus subgroup J (ALV-J) polypeptide pseudotyped with lentivirus membrane glycoproteins gp85 and gp37, HIV/env-ALV, was generated, characterized in vitro and evaluated for its ability to infect natural host cells. We optimized the newly developed micro-neutralization (MN) assay using recombinant pseudovirion HIV/env-ALV expressing enhanced green fluorescent protein and well-characterized sera from chickens with confirmed ALV-J disease or virus-free controls. HIV/env-ALV could infect CEF and DF-1 but not pk15, 293FT, MDCK or VERO E6 cells, therefore demonstrating a cellular tropism similar to the wild-type ALV-J. The MN assay indicated that the IC50 values of positive sera offered a considerable advantage in both speed and accuracy. These results suggest that this pseudotyped lentivirus is a good model for studying the functions of ALV-J env and that the MN assay is a reliable serological method for assessing antibody levels in investigating the actual status of the current ALV-J epidemic. These recombinant pseudovirions may prove to be useful for studying ALV-J biology in lower biosafety level laboratory environments, and also for the detection and quantification of neutralizing antibodies to ALV-J in a manner akin to ELISA assays, but that would also be applicable to other viruses.
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, Shandong, China.
authors.forename: Zhenjie
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Cui
authors.affiliation: China Animal Husbandry and Veterinary Bureau of Yiyuan, Zibo 256100, Shandong, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institute of Veterinary Drug Quality Inspection of Shandong Province, Jinan 250022, Shandong, China.
authors.forename: Zhikun
authors.initials: Z
authors.lastname: Yang
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, Shandong, China.
authors.forename: Zhizhong
authors.initials: Z
authors.lastname: Cui
authors.affiliation: College of Veterinary Medicine, Shandong Agricultural University, Taian 271018, Shandong, China. Electronic address: wschang@sdau.edu.cn.
authors.forename: Weishan
authors.initials: W
authors.lastname: Chang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D001354:Avian Leukosis Virus / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D002645:Chickens; D016086:Lentivirus / Q000235:genetics* / Q000502:physiology; D008562:Membrane Glycoproteins / Q000235:genetics*; D009500:Neutralization Tests / Q000379:methods; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*; D056189:Viral Tropism; D014771:Virion / Q000235:genetics*; D019065:Virus Assembly
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins
keywords: Avian leukosis virus subgroup J; Cell-tropism; Micro-neutralization assay; Pseudovirions