| 54 | 0 | 359 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 近年来，新型肠道病毒B75型(Enterovirus B75, EV-B75)在全球多地出现病例，可引起急性弛缓性麻痹等严重症状。但因病例稀少，其致病机制及引发严重疾病的机制尚未明确。本研究旨在通过构建EV-B75感染动物模型，明确其致病性和组织嗜性，为后续研究其致病机理等提供基础。方法 以2日龄I型干扰素受体缺陷型(Type I interferon receptor gene-knockout, IFNAR-/-)乳鼠为实验对象，经肌肉注射构建EV-B75感染模型，并通过体重检测、临床评分，生存率监测和测定各脏器组织中病毒载量来评估其致病性。采用HE染色观察组织病理变化，对肌肉组织进行RNA测序并通过GO、KEGG分析差异表达基因功能与通路。结果 感染乳鼠自第2天起出现生长迟滞，至第6天时实验组平均体重(1.7g)显著低于对照组(3.6g)。在感染后第4天，90%的乳鼠出现后肢麻痹症状，并于第6天全部死亡。脑和骨骼肌中病毒载量持续升高，并于第6天达到峰值(分别为105.5TCID50、10~7TCID50)。多脏器出现炎症损伤，以脑部水肿、骨骼肌纤维排列紊乱及断裂溶解最为突出。转录组测序分析共鉴定出1870个差异表达基因，显著富集于炎症小体激活、活性氧代谢、T细胞活化及JAK-STAT等免疫相关通路。结论EV-B75在IFNAR-/-乳鼠中表现出强神经侵袭性和高致病性，骨骼肌或为早期主要复制靶器官，病毒可经其扩散至脑等组织；差异表达基因在JAK-STAT等信号通路中显著富集，提示该通路异常激活及介导的过度炎症反应，可能是EV-B75感染后病毒复制增强、组织严重损伤的关键机制。本感染模型为探索EV-B75的致病机制及潜在抗病毒策略提供了重要实验平台。
Abstract：Enterovirus B75(EV-B75) has emerged in multiple regions worldwide in recent years and is capable of causing severe clinical manifestations, including acute flaccid paralysis. However, due to the scarcity of reported cases, its pathogenic mechanisms and determinants of severe disease remain largely undefined. In this study, we established an EV-B75 infection model in 2-day-old type I interferon receptor–deficient(IFNAR-/-) neonatal mice to characterize its pathogenicity and tissue tropism. Infection was induced via intramuscular inoculation, and pathogenicity was assessed through body-weight monitoring, clinical scoring, survival analysis, and quantification of viral loads across major organs. Histopathological changes were examined by hematoxylin-eosin staining, and RNA sequencing of skeletal muscle tissue was performed to identify differentially expressed genes and enriched functional pathways using GO and KEGG analyses. Infected pups developed growth retardation beginning on day 2 post-infection, and by day 6, the mean body weight of the infected group(1.7 g) was markedly lower than that of controls(3.6 g). By day 4, 90% of mice exhibited hindlimb paralysis, and all infected pups succumbed by day 6. Viral loads in the brain and skeletal muscle increased progressively, reaching peak titers on day 6(105.5 TCID50 and 107 TCID50, respectively). Multi-organ inflammatory injury was observed, most prominently characterized by cerebral edema and severe disruption and degeneration of skeletal muscle fibers. Transcriptome analysis identified 1,870 differentially expressed genes, significantly enriched in immune-related pathways including inflammasome activation, reactive oxygen species metabolism, T-cell activation, and JAK-STAT signaling. These findings demonstrate that EV-B75 exhibits strong neuroinvasiveness and high pathogenicity in IFNAR-/- neonatal mice. Skeletal muscle appears to serve as a primary early replication site, facilitating subsequent viral dissemination to the brain and other tissues. The marked enrichment of differentially expressed genes in the JAK-STAT pathway suggests that aberrant activation of this signaling axis and its associated excessive inflammatory responses may contribute to enhanced viral replication and severe tissue injury. This infection model provides a valuable platform for investigating the pathogenesis of EV-B75 and evaluating potential antiviral strategies.
[1]Simmonds P, Adriaenssens EM, Lefkowitz EJ, et al.Changes to virus taxonomy and the ICTV Statutes ratified by the International Committee on Taxonomy of Viruses(2024)[J]. Arch Virol, 2024, 169(11):236.DOI:10. 1007/s00705-024-06143-y.
[2]Xie Z, Khamrin P, Maneekarn N, et al. Epidemiology of enterovirus genotypes in association with human diseases[J]. Viruses, 2024, 16(7):1165. DOI:10. 3390/v16071165.
[3]Simmonds P, Gorbalenya AE, Harvala H, et al.Recommendations for the nomenclature of enteroviruses and rhinoviruses[J]. Arch Virol, 2020, 165(3):793-797. DOI:10. 1007/s00705-019-04520-6.
[4]曲艺璇，樊欢，邓斐，等. EV-D68感染所致急性呼吸道传染病的研究进展[J/OL].病毒学报，1-14[2026-01-15]. DOI:10. 13242/j. cnki. bingduxuebao. 250197.
[5]陶泽新，王海岩，徐爱强，等.人类肠道病毒B组73、75和97型山东地方株的鉴定及其基因特征分析[J].病毒学报，2010, 26(1):16-19. DOI:10. 13242/j.cnki. bingduxuebao. 002058.
[6]Simoes MP, Hodcroft EB, Simmonds P, et al.Epidemiological and clinical insights into the enterovirus D68 upsurge in Europe 2021-2022 and emergence of novel B3-derived lineages, ENPEN multicentre study[J]. J Infect Dis, 2024, 230(4):e917-e928. DOI:10. 1093/infdis/jiae154.
[7]Oberste MS, Michele SM, Maher K, et al. Molecular identification and characterization of two proposed new enterovirus serotypes, EV74 and EV75[J]. J Gen Virol, 2004, 85(Pt 11):3205-3212. DOI:10. 1099/vir. 0. 80148-0.
[8]Tao Z, Cui N, Lin X, et al. Complete genome sequence analysis of an enterovirus 75 isolate from China[J]. Arch Virol, 2014, 159(10):2809-2813. DOI:10. 1007/s00705-014-2014-6.
[9]Zhang K, Hong M, Zhang Y, et al. Molecular epidemiological, serological, and pathogenic analysis of EV-B75 associated with acute flaccid paralysis cases in Tibet, China[J]. Front Microbiol, 2021, 11:632552.DOI:10. 3389/fmicb. 2020. 632552.
[10]Adil MN, Abbas MA, Alam MM, et al. Molecular detection of non-polio enteroviruses and emergence of novel enterovirus D111 in stool samples from children reported with acute flaccid paralysis symptoms during2021 in Pakistan[J]. J Med Virol, 2025, 97(9):e70566. DOI:10. 1002/jmv. 70566.
[11]Maan HS, Dhole TN, Chowdhary R. Identification and characterization of nonpolio enterovirus associated with nonpolio-acute flaccid paralysis in polio endemic state of Uttar Pradesh, Northern India[J]. PLoS One, 2019, 14(1):e0208902. DOI:10. 1371/journal. pone. 0208902.
[12]Faleye TOC, Adewumi MO, Japhet MO, et al. Nonpolio enteroviruses in faeces of children diagnosed with acute flaccid paralysis in Nigeria[J]. Virol J, 2017, 14(1):175. DOI:10. 1186/s12985-017-0846-x.
[13]Bero DM, da Silva EE, de Sousa Júnior IP, et al.Enterovirus detection in stool samples from Mozambican children with acute gastroenteritis[J]. Acta Trop,2023, 238:106755. DOI:10. 1016/j.actatropica. 2022. 106755.
[14]Cabrerizo M, Echevarria JE, González I, et al.Molecular epidemiological study of HEV-B enteroviruses involved in the increase in meningitis cases occurred in Spain during 2006[J]. J Med Virol, 2008,80(6):1018-1024. DOI:10. 1002/jmv. 21197.
[15]林小嫚，潘倩瑜，何雅青，等. RNF126介导IKKγ的K63位泛素化促进EV71感染炎症反应的机制研究[J].病毒学报，2025, 41(1):71-78. DOI:10. 13242/j. cnki. bingduxuebao. 004628.
[16]Forcic D, Kosuti?-Gulija T, Santak M, et al.Comparisons of mumps virus potency estimates obtained by 50%cell culture infective dose assay and plaque assay[J]. Vaccine, 2010, 28(7):1887-1892. DOI:10. 1016/j. vaccine. 2009. 11. 049.
[17]张卓，刘学武，陈相池，等.不同浓度柯萨奇病毒B组3型感染幼龄大鼠动物模型的研究[J].中国实验动物学报，2025, 33(7):1032-1042. DOI:10. 3969/j.issn. 1005-4847. 2025. 07. 010.
[18]刘社兰，林君芬，张静. WHO手足口病临床管理与公共卫生应对指南第四部分EV71的致病机制[J].疾病监测，2013, 28(3):251-252. DOI:CNKI:SUN:JBJC. 0. 2013-03-031.
[19]陈亚坤，孙婧，蒋亚君，等.Ⅰ型干扰素受体（Ifnar）基因敲除小鼠的繁育及基因型鉴定[J].实验动物科学，2019, 36(4):35-38, 43. DOI:10. 3969/j. issn. 1006-6179. 2019. 04. 009.
[20]Huang HI, Chio CC, Lin JY, et al. EV-A71 induced IL-1β production in THP-1 macrophages is dependent on NLRP3, RIG-I, and TLR3[J]. Sci Rep, 2022, 12(1):21425. DOI:10. 1038/s41598-022-25458-x.
[21]Kra?un D, G?rlach A, Snedeker JG, et al. Reactive oxygen species in tendon injury and repair[J]. Redox Biol, 2025, 81:103568. DOI:10. 1016/j.redox. 2025. 103568.
[22]Hu X, Li J, Fu M, et al. The JAK/STAT signaling pathway:from bench to clinic[J]. Signal Transduct Target Ther, 2021, 6(1):402. DOI:10. 1038/s41392-021-00791-1.
[23]Zhao P, Sun T, Lyu C, et al. Cell mediated ECMdegradation as an emerging tool for anti-fibrotic strategy[J]. Cell Regen, 2023, 12(1):29. DOI:10. 1186/s13619-023-00172-9.
[24]Mahjoor M, Mahmoudvand G, Farokhi S, et al. Double-edged sword of JAK/STAT signaling pathway in viral infections:novel insights into virotherapy[J]. Cell Commun Signal, 2023, 21(1):272. DOI:10. 1186/s12964-023-01240-y.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250324
中图分类号:R373.2
引用信息:
[1]韩晓梅,张珂艺,路环环,等.肠道病毒B75型感染2日龄IFNAR~(-/-)乳鼠的特征研究[J].病毒学报,2026,42(01):60-70.DOI:10.13242/j.cnki.bingduxuebao.250324.
基金信息:
国家疾病预防控制局公共卫生人才培养支持项目（项目号：GJJKJ-2024-ZY）; 传染病溯源预警与智能决策全国重点实验室（NITFID）资助项目（项目号：ZDGWNLJS25-36）,题目：未知及新发病原体监测及预测预警技术研究; 国家重点研发计划项目（项目号：2021YFC2302003）,题目：病毒监测网络数据标准及数据平台建设~~