Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Pearl River Fishery Research Institute, Chinese Academy of Fishery Sciences, Liwan District, Guangzhou, Guangdong, China; Institute of Military Veterinary, Academy of Military Medical Science, Changchun 130062, China.
Institute of Military Veterinary, Academy of Military Medical Science, Changchun 130062, China.
Virus Res. 2014 May 12;184:7-13. doi: 10.1016/j.virusres.2014.01.021. Epub 2014 Jan 30.
Rabies is a fatal central nervous system (CNS) disease caused by the neurotropic rabies virus (RABV). The therapeutic management of RABV infections is still problematic, and novel antiviral strategies are urgently required. We established the RVG-BHK-21 cell line, which expresses RABV glycoprotein on the cell surface, to select aptamers. Through 28 iterative rounds of selection, single-stranded DNA (ssDNA) aptamers were generated by exponential enrichment (SELEX). A virus titer assay and a real-time quantitative reverse transcription PCR (qRT-PCR) assay revealed that four aptamers could inhibit the replication of RABV in cultured baby hamster kidney (BHK)-21 cells. However, the aptamers did not inhibit the replication of other virus, e.g., canine distemper virus (CDV) and canine parvovirus (CPV). In addition, the GE54 aptamer was found to effectively protect mice against lethal RABV challenge. After inoculation with aptamers for 24h or 48h, followed by inoculation with CVS-11, approximately 25-33% of the mice survived. In summary, we selected aptamers that could significantly protect from a lethal dose of RABV in vitro and in vivo.
狂犬病是一种致命的中枢神经系统（CNS）疾病，由神经嗜性狂犬病病毒（RABV）引起。RABV 感染的治疗管理仍然存在问题，迫切需要新的抗病毒策略。我们建立了 RVG-BHK-21 细胞系，该细胞系在细胞表面表达 RABV 糖蛋白，以筛选适体。通过 28 轮的选择，通过指数富集（SELEX）产生了单链 DNA（ssDNA）适体。病毒滴度测定和实时定量逆转录 PCR（qRT-PCR）检测表明，四种适体可以抑制培养的仓鼠肾细胞（BHK-21）中 RABV 的复制。然而，适体并没有抑制其他病毒（如犬瘟热病毒（CDV）和犬细小病毒（CPV））的复制。此外，发现 GE54 适体可有效保护小鼠免受致死性 RABV 攻击。在用适体接种 24 小时或 48 小时后，再接种 CVS-11，大约 25-33%的小鼠存活下来。总之，我们选择了可以在体外和体内显著保护免受致死剂量 RABV 感染的适体。
