A Zoonotic Strain of Rocahepevirus ratti Hepatitis E Virus Does Not Replicate Efficiently within Human Placental JEG-3 Cells

Kush K. Yadav, Jake Hofstetter, Scott P. Kenney
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Abstract

Paslahepevirus balayani and Rocahepevirus ratti are genetically diverse species of hepatitis E virus [HEV]. Previously, only members of the Paslahepevirus genus were known to infect humans but recently some Rocahepevirus members have been found to be infectious to both immunocompromised and immunocompetent humans. Paslahepevirus balayani genotypes (gt) 1, 2, and 4 are known for their detrimental effects during pregnancy, causing pregnancy-related disorders. Recent findings have demonstrated the ability of Paslahepevirus balayani gt3 to replicate within placental cell lines, suggesting a direct effect on the placenta and fetus. To study whether zoonotic rat HEV strains possess a similar human-host placental tropism, we utilized JEG-3 cells to understand the replicative ability of an infectious clone of a recently reported strain of Rocahepevirus ratti, the LCK-3110 strain. Infectious cDNA clones of Pasla-, Avi-, and Rocahepevirus were transcribed and then, transduced into JEG-3 cells. Cells were harvested, and cell lysates were used for testing infectivity. Five days post-transfection or after inoculation onto naive HepG2/C3A cells, the cells were analyzed for infection. Replication in transduced JEG-3 cells and the infection potential in HepG2/C3A cells were assessed via an indirect immunofluorescence assay and a flow-cytometry assay. We found that the Rocahepevirus ratti LCK-3110 strain did not have efficient replication in JEG-3 cell cultures.
人畜共患的 Rocahepevirus ratti 戊型肝炎病毒株在人胎盘 JEG-3 细胞内不能有效复制
Paslahepevirus balayani 和 Rocahepevirus ratti 是戊型肝炎病毒 [HEV] 的不同基因种类。以前只知道 Paslahepevirus 属的成员会感染人类,但最近发现 Rocahepevirus 属的一些成员既可感染免疫力低下的人类,也可感染免疫力正常的人类。众所周知,巴拉伊病毒基因型(gt)1、2和4在怀孕期间会产生有害影响,导致与妊娠有关的疾病。最近的研究结果表明,巴拉耶病毒 gt3 能够在胎盘细胞系中复制,这表明它对胎盘和胎儿有直接影响。为了研究人畜共患病大鼠 HEV 株系是否具有类似的人-宿主胎盘趋向性,我们利用 JEG-3 细胞了解了最近报道的 Rocahepevirus ratti 株系(LCK-3110 株系)的感染性克隆的复制能力。转录 Pasla、Avi 和 Rocahepevirus 的感染性 cDNA 克隆,然后将其转导到 JEG-3 细胞中。收获细胞,用细胞裂解液检测感染性。转染后五天或接种到天真的 HepG2/C3A 细胞后,分析细胞的感染情况。通过间接免疫荧光检测法和流式细胞仪检测法评估了转导的 JEG-3 细胞的复制情况和 HepG2/C3A 细胞的感染潜力。我们发现,Rocahepevirus ratti LCK-3110 株在 JEG-3 细胞培养物中不能有效复制。
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