Data_Sheet_1_Characterization of a Novel Tanay Virus Isolated From Anopheles sinensis Mosquitoes in Yunnan, China.docx

Globally, mosquitoes are known to be competent vectors to various arboviruses that cause serious and debilitating diseases to humans and animals. Conversely, mosquitoes harbor a wide array of insect specific viruses (ISVs) that are generally neglected. Extensive characterization of these ISVs is important in understanding their persistence infection effect on host behavior and arbovirus transmission. Herein, we report first time isolation of Tanay virus (TANAV) isolate YN15_103_01 in Anopheles sinensis mosquitoes from Yunnan Province, China. Phylogenetically, the isolate’s nucleotide identity had more than 14.47% variance compared to previous TANAV isolates, and it clustered into an independent branch within the genus Sandewavirus in the newly proposed taxon Negevirus. TANAV growth and high titers was attained in Aag2 cells (107 PFU/mL) but with no CPE observed up to 7 days.p.i. compared to C6/36 cells that exhibited extensive CPE at 48 h.p.i. with titers of 107 PFU/mL. Contrarywise, the viral isolate did not replicate in vertebrate cell lines. Electron microscopy analyses showed that its final maturation process takes place in the cell cytoplasm. Notably, the predicted viral proteins were verified to be corresponding to the obtained SDS-PAGE protein bands. Our findings advance forth new and vital knowledge important in understanding insect specific viruses, especially TANAV.

全球范围内，蚊虫被认为是多种导致人类和动物严重残疾疾病的虫媒病毒的有效传播媒介。反之，蚊虫携带着多种昆虫特异性病毒（ISV），这些病毒通常被忽视。对这些ISV的广泛表征对于理解其在宿主行为中的持久感染效应以及虫媒病毒的传播至关重要。本研究首次从中国云南省的中华按蚊中分离得到坦纳病毒（TANAV）分离株YN15_103_01。在系统发育学上，该分离株的核苷酸同源性相较于之前的TANAV分离株存在超过14.47%的变异，并聚类于新提出的分类单元Negevirus中沙德韦病毒属的一个独立分支。TANAV在Aag2细胞中生长并达到高滴度（107 PFU/mL），但相较于在48小时感染后出现广泛细胞病变效应（CPE）并达到107 PFU/mL滴度的C6/36细胞，Aag2细胞在感染后7天内未观察到CPE。相反，该病毒分离株在脊椎动物细胞系中不进行复制。电子显微镜分析显示，其最终成熟过程发生在细胞质中。值得注意的是，预测的病毒蛋白与获得的SDS-PAGE蛋白条带相吻合。我们的研究为理解昆虫特异性病毒，尤其是TANAV，提供了新的、至关重要的知识。