Ramjag Anushka, Cutrone Sergej, Lu Kai, Crasto Christine, Jin Jing, Bakkour Sonia, Carrington Christine V F, Simmons Graham
Department of Preclinical Sciences, The University of the West Indies, St. Augustine Campus, St. Augustine, Trinidad and Tobago.
Vitalant Research Institute, 270 Masonic Avenue, San Francisco, CA, 94118, USA.
Virol J. 2022 Oct 29;19(1):170. doi: 10.1186/s12985-022-01906-y.
Several studies have demonstrated neutralizing antibodies to be highly effective against alphavirus infection in animal models, both prophylactically and remedially. In most studies, neutralizing antibodies have been evaluated for their ability to block viral entry in vitro but recent evidence suggests that antibody inhibition through other mechanisms, including viral budding/release, significantly contributes to viral control in vivo for a number of alphaviruses.
We describe a BSL-2, cell-based, high-throughput screening system that specifically screens for inhibitors of alphavirus egress using chikungunya virus (CHIKV) and Mayaro virus (MAYV) novel replication competent nano-luciferase (nLuc) reporter viruses. Screening of both polyclonal sera and memory B-cell clones from CHIKV immune individuals using the optimized assay detected several antibodies that display potent anti-budding activity.
We describe an "anti-budding assay" to specifically screen for inhibitors of viral egress using novel CHIKV and MAYV nLuc reporter viruses. This BSL-2 safe, high-throughput system can be utilized to explore neutralizing "anti-budding" antibodies to yield potent candidates for CHIKV and MAYV therapeutics and prophylaxis.
多项研究已证明，在动物模型中，中和抗体在预防和治疗方面对甲病毒感染均具有高效性。在大多数研究中，已对中和抗体在体外阻断病毒进入的能力进行了评估，但最近有证据表明，通过包括病毒出芽/释放在内的其他机制产生的抗体抑制作用，对多种甲病毒在体内的病毒控制有显著贡献。
我们描述了一种基于细胞的二级生物安全水平（BSL-2）高通量筛选系统，该系统使用基孔肯雅病毒（CHIKV）和马亚罗病毒（MAYV）新型具有复制能力的纳米荧光素酶（nLuc）报告病毒，专门筛选甲病毒出芽的抑制剂。使用优化后的检测方法，对来自CHIKV免疫个体的多克隆血清和记忆B细胞克隆进行筛选，检测到了几种具有强大抗出芽活性的抗体。
我们描述了一种“抗出芽检测法”，使用新型CHIKV和MAYV nLuc报告病毒专门筛选病毒出芽的抑制剂。这种二级生物安全水平（BSL-2）的安全高通量系统可用于探索中和性“抗出芽”抗体，以产生用于CHIKV和MAYV治疗及预防的有效候选物。
