Bell Mikaela G, Parry Rhys H, Lee Tricia S E, Habarugira Gervais, McMahon Isabella E, Thompson Madeline A, Modhiran Naphak, Watterson Daniel, Batovska Jana, Lynch Stacey E, Hall Roy A, Harrison Jessica J, Hobson-Peters Jody
School of Chemistry and Molecular Biosciences, The University of Queensland St Lucia, Brisbane, Australia.
Australian Infectious Diseases Research Centre, Brisbane, Australia.
Npj Viruses. 2024 Sep 30;2(1):45. doi: 10.1038/s44298-024-00052-2.
Few insect-specific alphaviruses (ISA) have been discovered, with even fewer culturable to facilitate full characterisation. Here, we report the recovery of an infectious clone of Yada Yada virus (YYV)-a virus previously only detected by metagenomic sequencing of mosquito homogenates. Using the infectious clone, we confirmed the inability of YYV to replicate in vertebrate cells in vitro, with replication limited to only Aedes mosquito-derived cell lines. We further produced and characterised the first monoclonal antibodies (mAbs) to ISAs. Through successful replacement of the structural proteins of YYV with those of other ISAs, Eilat virus, Agua Salud (ASALV), Taï Forest (TALV) and Mwinilunga alphaviruses (MWAV), we established that a replication block for in vitro culture of TALV and MWAV in mosquito cells does not exist at virus entry. Unexpectedly, ASALV structural proteins were recognised by cross-reactive mAbs made to chikungunya (CHIKV) and Ross River viruses (RRV), suggesting a potential antigenic link between ASALV and pathogenic alphaviruses. The YYV genetic backbone was also investigated to generate chimeras displaying the structural proteins of various pathogenic vertebrate-infecting alphaviruses including CHIKV, RRV, Barmah Forest, Sindbis and Mayaro viruses. These chimeras retained the antigenic properties of the parental viruses and did not replicate in vertebrate cells, demonstrating the potential of the YYV platform for vaccine and diagnostic antigen production.
已发现的昆虫特异性甲病毒（ISA）很少，可培养以进行全面表征的更少。在此，我们报告了Yada Yada病毒（YYV）感染性克隆的获得，该病毒此前仅通过蚊子匀浆的宏基因组测序检测到。利用该感染性克隆，我们证实YYV无法在脊椎动物细胞中体外复制，其复制仅限于源自伊蚊的细胞系。我们还制备并表征了首批针对ISA的单克隆抗体（mAb）。通过成功将YYV的结构蛋白替换为其他ISA（埃拉特病毒、阿瓜萨尔杜病毒（ASALV）、塔伊森林病毒（TALV）和姆维尼伦加甲病毒（MWAV））的结构蛋白，我们确定TALV和MWAV在蚊子细胞中体外培养的复制障碍在病毒进入时不存在。出乎意料的是，ASALV结构蛋白被针对基孔肯雅病毒（CHIKV）和罗斯河病毒（RRV）制备的交叉反应性mAb识别，这表明ASALV与致病性甲病毒之间存在潜在的抗原联系。我们还研究了YYV的遗传骨架，以产生展示包括CHIKV、RRV、巴马森林病毒、辛德毕斯病毒和马亚罗病毒在内的各种致病性脊椎动物感染性甲病毒结构蛋白的嵌合体。这些嵌合体保留了亲本病毒的抗原特性，且不在脊椎动物细胞中复制，证明了YYV平台在疫苗和诊断抗原生产方面的潜力。
