Hepatitis E virus infects human testicular tissue and Sertoli cells

Globally, hepatitis E virus (HEV) infections are prevalent. The finding of high viral loads and persistent viral shedding in ejaculate suggests that HEV replicates within the human male genital tract, but its target organ is unknown and appropriate models are lacking. We aimed to determine the HEV tropism in the human testis and its potential influence on male reproductive health. We conducted an ex vivo culture of human testis explants and in vitro culture of primary human Sertoli cells. Clinically derived HEV genotype 1 (HEV1) and HEV3 virions, as well as rat-derived HEV-C1, were used for inoculation. Transcriptomic analysis was performed on testis tissues collected from tacrolimus-treated rabbits with chronic HEV3 infection. Our findings reveal that HEV3, but not HEV1 or HEV-C1, can replicate in human testis explants and primary human Sertoli cells. Tacrolimus treatment significantly enhanced the replication efficiency of HEV3 in testis explants and enabled successful HEV1 infection in Sertoli cells. HEV3 infection disrupted the secretion of several soluble factors and altered the cytokine microenvironment within primary human Sertoli cells. Finally, intratesticular transcriptomic analysis of immunocompromised rabbits with chronic HEV infection indicated downregulation of genes associated with spermatogenesis. HEV can infect the human testicular tissues and Sertoli cells, with increased replication efficiency when exposed to tacrolimus treatment. These findings shed light on how HEV may persist in the ejaculate of patients with chronic hepatitis E and provide valuable ex vivo tools for studying countermeasures.戊型肝炎病毒（HEV）感染在全球范围内均呈流行态势。研究人员在精液中检测到高病毒载量与持续性病毒排毒现象，这提示HEV可在人类男性生殖道内复制，但目前该病毒的靶器官尚不明确，且缺乏合适的研究模型。本研究旨在明确HEV在人类睾丸中的组织嗜性，及其对男性生殖健康的潜在影响。我们采用人类睾丸组织外植体离体培养与原代人类塞尔托利细胞（Sertoli cell）体外培养的实验方法，所用病毒粒子包括临床分离的1型HEV（HEV1）、3型HEV（HEV3）以及大鼠来源的HEV-C1，用于病毒接种。我们对经他克莫司处理且慢性感染HEV3的家兔睾丸组织进行了转录组学分析。研究结果显示，仅HEV3可在人类睾丸外植体与原代塞尔托利细胞中复制，HEV1与HEV-C1均无此能力。他克莫司处理可显著提升HEV3在睾丸外植体中的复制效率，同时使HEV1得以在塞尔托利细胞中成功感染。HEV3感染会干扰原代人类塞尔托利细胞中多种可溶性因子的分泌，并改变其细胞因子微环境。最后，对慢性HEV感染的免疫缺陷家兔进行睾丸内转录组学分析后发现，与精子发生相关的基因呈现表达下调态势。本研究证实HEV可感染人类睾丸组织与塞尔托利细胞，且经他克莫司处理后其复制效率会得到提升。上述研究结果阐明了HEV可在慢性戊型肝炎患者精液中持续存在的潜在机制，并为抗病毒防治策略的研究提供了极具价值的离体实验工具。