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目的 禽网状内皮组织增殖病毒（Reticuloendotheliosis virus, REV）是一种经过鸡胚垂直传播的免疫抑制性病毒，感染REV的雏鸡胎粪以及粪便中可持续排出病毒造成水平传播，即使在一些母源抗体阳性鸡群也可以介导水平传播，本研究的目的便是验证雏鸡胎粪中存在REV感染相关外泌体并介导形成水平传播和免疫逃逸。方法 本研究将REV野毒株IBD-C1605株接种鸡胚后从出壳雏鸡检测为REV阳性的雏鸡胎粪中提取外泌体，进行纳米颗粒追踪分析、电镜观察、基因组测序和蛋白质组分析。本研究将REV阳性雏鸡胎粪外泌体经特异性REV抗体中和后接种DF-1细胞，以验证其逃避抗体中和的能力。本研究将REV游离病毒或REV阳性雏鸡胎粪外泌体分别接种REV母源抗体阳性雏鸡和阴性雏鸡，观察不同时间的雏鸡体重增长、器官发育指数、血液、泄殖腔和免疫器官中REV的复制情况等指标。结果 本研究证实了REV阳性雏鸡胎粪外泌体中包含REV完整基因组成分和3种主要蛋白。REV阳性雏鸡胎粪外泌体经特异性REV抗体中和后接种DF-1细胞，证实在细胞上检出REV高效感染，说明REV阳性雏鸡胎粪外泌体可介导REV逃避抗体中和形成体外感染。将REV游离病毒分别接种REV母源抗体阳性雏鸡和阴性雏鸡，观察不同时间的雏鸡体重增长、器官发育指数、血液、泄殖腔和免疫器官中REV的复制情况等指标，证实REV游离病毒接种母源抗体阴性雏鸡引起严重的体重增长迟缓、肝脏脾脏肿大，而接种母源抗体阳性雏鸡则因为中和作用而显著降低了对鸡的致病性。但REV阳性雏鸡胎粪外泌体按照上述操作接种REV母源抗体阳性雏鸡和阴性雏鸡后，两组鸡的相关致病指标无显著差异，证明母源抗体对外泌体介导的感染无法发挥有效抑制作用。结论 本研究解析了REV阳性胎粪外泌体并证实其可作为传染途径在体内外造成有效感染，其感染能力不受抗体的限制，推测胎粪外泌体是介导REV形成早期水平传播和突破抗体抑制的途径之一。
Abstract：Objective Reticuloendotheliosis virus(REV) is an immunosuppressive virus that can be vertically transmitted through chicken embryos. Chicks infected with REV continuously shed the virus in their meconium and feces, facilitating horizontal transmission, even in flocks with maternal antibodies. This study aimed to determine whether the REV infection-associated exosomes are present in chick meconium and to elucidate their role in mediating horizontal transmission and antibody-mediated immune evasion. Methods The wild-type REV strain IBD-C1605 was inoculated into chicken embryos. Exosomes were isolated from the meconium of hatched chicks confirmed to be REV-positive. The isolated exosomes were characterized by nanoparticle tracking analysis, transmission electron microscopy, genomic sequencing, and proteomic analysis. To assess their ability to evade antibody neutralization, REV-positive meconium exosomes were preincubated with REVspecific antibodies and subsequently inoculated into DF-1 cells. In parallel, free REV virions or REV-positive meconium exosomes were separately inoculated into chicks with positive or negative maternal REV antibodies. Body weight gain, organ development indices, and REV replication in blood, cloaca, and immune organs were monitored at multiple time points. Results REV-positive meconium exosomes were shown to contain the complete REV genome and three major viral proteins. Following neutralization with REV-specific antibodies, these exosomes were still able to mediate efficient REV infection in DF-1 cells, indicating that exosomeassociated REV can evade antibody neutralization and establish infection in vitro. In in vivo experiments inoculation with free REV caused severe growth retardation and hepatosplenomegaly in chicks lacking maternal antibodies, whereas pathogenic effects were significantly attenuated in chicks with maternal antibodies due to effective neutralization. In contrast, inoculation with REV-positive meconium exosomes resulted in comparable pathogenic outcomes in chicks with or without maternal antibodies, demonstrating that maternal antibodies failed to effectively inhibit exosome-mediated infection. Conclusion This study demonstrates that REVpositive meconium-derived exosomes serve as an effective vehicle for viral infection both in vitro and in vivo, and that their infectivity is not constrained by antibody-mediated neutralization. These findings suggest that meconium-derived exosomes represent an important pathway for early horizontal transmission of REV and contribute to viral immune evasion in the presence of maternal antibodies.
[1]Campbell WF, Baxter-Gabbard KL, Levine AS. Avian reticuloendotheliosis virus(strain T). I. Virological characterization[J]. Avian Dis, 1971, 15(4):837-849.
[2]BAXTER-GABBARD K L. Reticuloendotheliosis virus and related viruses.[J]. Diseases of poultry. 9th ed.1991, 439-456.
[3]吉荣，崔治中，王锡乐，等.分子克隆化禽网状内皮组织增生症病毒传染性及其前病毒全基因组序列研究[J].病毒学报，2005, 21(6):448-455. DOI:10. 13242/j. cnki. bingduxuebao. 001583.
[4]Mussman HC, Twiehaus MJ. Pathogenesis of reticuloendothelial virus disease in chicks—an acute runting syndrome[J]. Avian Dis, 1971, 15(3):483-502.
[5]Paul PS, Pomeroy KA, Muscoplat CC, et al.Characteristics of two new reticuloendotheliosis virus isolates of turkeys[J]. Am J Vet Res, 1977, 38(3):311-315.
[6]Dren CN, Nemeth I, Sari I, et al. Isolation of a reticuloendotheliosis-like virus from naturally occurring lymphoreticular tumours of domestic goose[J]. Avian Pathol, 1988, 17(2):259-277. DOI:10. 1080/03079458808436446.
[7]Sarma PS, Jain DK, Mishra NK, et al. Isolation and characterization of viruses from natural outbreaks of reticuloendotheliosis in turkeys[J]. J Natl Cancer Inst,1975, 54(6):1355-1359. DOI:10. 1093/jnci/54. 6. 1355.
[8]Jiang L, Qi X, Gao Y, et al. Molecular characterization and phylogenetic analysis of the reticuloendotheliosis virus isolated from wild birds in Northeast China[J].Vet Microbiol, 2013, 166(1-2):68-75. DOI:10. 1016/j. vetmic. 2013. 05. 008.
[9]Marbán E. The secret life of exosomes:what bees can teach us about next-generation therapeutics[J]. J Am Coll Cardiol, 2018, 71(2):193-200. DOI:10. 1016/j.jacc. 2017. 11. 013.
[10]Gallo A, Vella S, Miele M, et al. Global profiling of viral and cellular non-coding RNAs in Epstein-Barr virusinduced lymphoblastoid cell lines and released exosome cargos[J]. Cancer Lett, 2017, 388:334-343. DOI:10. 1016/j. canlet. 2016. 12. 003.
[11]Li M, Zeringer E, Barta T, et al. Analysis of the RNA content of the exosomes derived from blood serum and urine and its potential as biomarkers[J]. Philos Trans R Soc Lond B Biol Sci, 2014, 369(1652):20130502.DOI:10. 1098/rstb. 2013. 0502.
[12]Madison MN, Welch JL, Okeoma CM. Isolation of exosomes from semen for in vitro uptake and HIV-1infection assays[J]. Bio Protoc, 2017, 7(7):e2216.DOI:10. 21769/BioProtoc. 2216.
[13]Yang C, Chen J, Zhao Y, et al. Salivary exosomes exacerbate colitis by bridging the oral cavity and intestine[J]. iScience, 2024, 27(11):111061. DOI:10. 1016/j. isci. 2024. 111061.
[14]鲁丹，方一泰，罗德炎，等.外泌体参与新冠病毒感染的机制及其应用的研究进展[J].病毒学报，2024, 40(1):160-168. DOI:10. 13242/j. cnki.bingduxuebao. 004454.
[15]Mathivanan S, Simpson RJ. ExoCarta:a compendium of exosomal proteins and RNA[J]. Proteomics, 2009, 9(21):4997-5000. DOI:10. 1002/pmic. 200900351.
[16]Liao L, Chen W, Zhang X, et al. Semen extracellular vesicles mediate vertical transmission of subgroup J avian leukosis virus[J]. Virol Sin, 2022, 37(2):284-294. DOI:10. 1016/j. virs. 2022. 01. 026.
[17]任志浩，李阳，苏红芹，等.齐多夫定和单克隆抗体联合应用去除疫苗毒种中REV的污染[J].病毒学报，2017, 33(4):605-609. DOI:10. 13242/j. cnki.bingduxuebao. 003189.
[18]崔帅. IBDV疫苗毒种中REV的分离鉴定及对SPF雏鸡的致病性研究[D].泰安：山东农业大学，2018.
[19]Su Q, Zhang Y, Cui Z, et al. Semen-derived exosomes mediate immune escape and transmission of reticuloendotheliosis virus[J]. Front Immunol, 2021,12:735280. DOI:10. 3389/fimmu. 2021. 735280.
[20]Bose HR. Reticuloendotheliosis virus and disturbance in immune regulation[J]. Microbiol Sci, 1984, 1(5):107-112.
[21]Gould SJ, Booth AM, Hildreth JEK. The Trojan exosome hypothesis[J]. Proc Natl Acad Sci U S A,2003, 100(19):10592-10597. DOI:10. 1073/pnas. 1831413100.
[22]Bao X, Zhuang T, Xu Y, et al. Exosomes secreted by CSFV-infected cells evade neutralizing antibody to activate innate immune responses and establish productive infection in recipient cells[J]. Vet Microbiol,2024, 292:110062. DOI:10. 1016/j.vetmic. 2024. 110062.
[23]Patters BJ, Kumar S. The role of exosomal transport of viral agents in persistent HIV pathogenesis[J].Retrovirology, 2018, 15(1):79. DOI:10. 1186/s12977-018-0462-x.
[24]Fu Y, Xiong S. Exosomes mediate Coxsackievirus B3transmission and expand the viral tropism[J]. PLoS Pathog, 2023, 19(1):e1011090. DOI:10. 1371/journal. ppat. 1011090.
[25]Wang Z, He M, He H, et al. Nonenveloped avian reoviruses released with small extracellular vesicles are highly infectious[J]. Viruses, 2023, 15(7):1610.DOI:10. 3390/v15071610.
[26]Deng L, Jiang W, Wang X, et al. Syntenin regulates hepatitis C virus sensitivity to neutralizing antibody by promoting E2 secretion through exosomes[J]. J Hepatol, 2019, 71(1):52-61. DOI:10. 1016/j.jhep. 2019. 03. 006.
[27]Wang Z, Cui H, Zhang Y, et al. DF-1-Derived exosomes mediate transmission of reticuloendotheliosis virus and resist REV-specific antibodies[J]. Virol J,2024, 21(1):177. DOI:10. 1186/s12985-024-02445-4.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250364
中图分类号:S852.65
引用信息:
[1]傅子豪,耿瑞强,任志浩,等.禽网状内皮组织增殖病毒IBD-C1605株经胎粪外泌体介导感染并逃逸抗体抑制[J].病毒学报,2026,42(01):253-268.DOI:10.13242/j.cnki.bingduxuebao.250364.
基金信息:
国家自然科学基金面上项目(项目号:31972663)题目:精液外泌体介导的禽网状内皮增生病毒感染与逃逸机制~~