Generation and evaluation of avian leukosis virus subgroup J envelope glycoprotein recombinant pseudovirions

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• 作者：Zhenjie Zhang^a ; Lina Cui^b ; Liang Wang^c ; Zhikun Yang^d ; Zhizhong Cui^a ; Weishan Chang^a ; ^{wschang@sdau.edu.cn" class="auth_mail}
• 关键词：Avian leukosis virus subgroup J ; Pseudovirions ; Cell-tropism ; Micro-neutralization assay
• 刊名：Journal of Virological Methods
• 出版年：15 June, 2014
• 年：2014
• 卷：202
• 期：Complete
• 页码：1-7
• 全文大小：842 K

文摘

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.