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Article(id=1259888469254464102, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250892, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1764604800000, receivedDateStr=2025-12-02, revisedDate=null, revisedDateStr=null, acceptedDate=1767542400000, acceptedDateStr=2026-01-05, onlineDate=1778310418666, onlineDateStr=2026-05-09, pubDate=1777824000000, pubDateStr=2026-05-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1778310418666, onlineIssueDateStr=2026-05-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1778310418666, creator=13701087609, updateTime=1778310418666, updator=13701087609, issue=Issue{id=1259888457367806489, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='5', pageStart='2031', pageEnd='2556', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=0, articleOrder=1, issueType=-1, specialIssue=null, createTime=1778310415832, creator=13701087609, updateTime=1778320153326, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1259929299465921482, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1259929299465921483, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1259888457367806489, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2430, endPage=2443, ext={EN=ArticleExt(id=1259888472383414919, articleId=1259888469254464102, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Establishment of a Senecavirus A infection model in C57BL/6J IFNAR-/-mice and evaluation of its infection characteristics, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a mouse model that effectively simulates the key clinical features of porcine Senecavirus A (SVA) infection, providing a crucial experimental tool for elucidating its pathogenesis and evaluating prevention and control products. Methods Five-week-old SPF C57BL/6J wild-type (WT) mice and type I interferon receptor-deficient (C57BL/6J IFNR-/- ) mice were inoculated via intraperitoneal, subcutaneous, and intramuscular routes. Blood and tissue samples were collected on days 1, 3, and 5 post-infection (dpi) for analysis of gross pathology, histopathology, viral load, and dynamic determination of inflammatory cytokines at the mRNA level. Results Compared with the mock-infected control group, both mouse strains developed gross lesions (e.g., swollen inguinal lymph nodes, yellowish livers, splenomegaly) and histopathological lesions (e.g., cortical disintegration of lymph nodes, hepatocellular necrosis, atrophy of splenic white pulp, and renal tubular necrosis). However, these lesions were more severe in C57BL/6J IFNR-/- mice. Viral RNA was widely distributed in tissues of both groups but was significantly higher in the C57BL/6J IFNR-/- group. Notably, viremia was undetectable in WT mice, whereas in C57BL/6J IFNR-/- mice, the virus was detected in whole blood as early as 1 dpi, peaked at 3 dpi, and then declined rapidly. Inflammatory cytokine analysis revealed significantly higher mRNA levels and protein levels of IL-1β and IL-6 in C57BL/6J IFNR-/- mice than in WT mice. Conclusion The C57BL/6J IFNR-/- mouse model successfully simulates, for the first time, the transient viremia characteristic of porcine SVA infection. It comprehensively replicates key features, including the multi-organ viral distribution, high viral load, and self-limiting recovery, providing a more effective animal model for delving into the pathogenic mechanism of SVA and evaluating vaccines and antiviral drugs.

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E-mail: JIA Meiyu: ;
WANG Yu’e:
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These authors contributed equally to this work.

, authorsList=Lingqiao YOU, Qing ZHANG, Shengbin GAO, Lizhi FU, Meiyu JIA, Yu’e WANG), CN=ArticleExt(id=1259888489710084995, articleId=1259888469254464102, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=A型塞内卡病毒C57BL/6J IFNR-/- 小鼠感染模型建立及感染特性评价, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

目的 构建能有效模拟猪A型塞内卡病毒(Senecavirus A, SVA)关键临床特征的小鼠模型，为阐明其致病机制及开展防控产品评价提供关键实验工具。 方法 通过腹腔、皮下、肌内注射3种途径分别接种5周龄SPF级C57BL/6J野生型(WT)小鼠和I型干扰素受体缺失(C57BL/6J IFNR-/- )小鼠，于感染后第1、3、5天采集血液及组织样本，进行宏观病理、组织病理、病毒载量及炎性因子mRNA动态检测。 结果 与未接毒对照组相比，2种小鼠均出现腹股沟淋巴结肿大、肝脏土黄色变及脾肿大等宏观病变和显微病变(淋巴结皮质崩解、肝细胞坏死、脾白髓萎缩、肾小管坏死)，但C57BL/6J IFNR-/- 小鼠病变更为严重；病毒RNA在2种小鼠组织中广泛分布，但C57BL/6J IFNR-/- 组显著高于WT组。此外，WT小鼠未出现病毒血症，C57BL/6J IFNR-/- 小鼠在感染后1 dpi全血中即可检出病毒，3 dpi达到峰值后快速下降。炎症因子检测显示，C57BL/6J IFNR-/- 小鼠IL-1β和IL-6炎性因子mRNA水平和蛋白水平均显著高于WT小鼠。 结论 C57BL/6J IFNR-/- 小鼠首次模拟了猪SVA短暂病毒血症特征，可全面复现病毒多器官分布、高病毒载量及自限性恢复等特征，为深入研究其致病机制及评价疫苗与抗病毒药物提供了更为有效的动物模型。

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作者贡献声明

游灵巧：实验进行，稿件撰写与修改；张卿：文献检索与整理，稿件撰写与修改；高晟斌：数据收集与分析；付利芝：格式修改与校对，对文献查漏补缺；贾梅玉：文章框架构思与设计，项目支持，语言润色，监督管理；王玉娥：文章框架构思与设计，提供资源，稿件审阅与投稿，监督管理。

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A: Inguinal lymph node; B: Liver; C: Spleen; D: Brain; E: Kidney; F: Heart. From left to right: intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group., figureFileSmall=GuLxCbEC2blyddjLOOzg4Q==, figureFileBig=WG0c+7I07kdae6BrDxlfXw==, tableContent=null), ArticleFig(id=1259928465168523484, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图1, caption=C57BL/6J WT小鼠病理剖检结果, figureFileSmall=GuLxCbEC2blyddjLOOzg4Q==, figureFileBig=WG0c+7I07kdae6BrDxlfXw==, tableContent=null), ArticleFig(id=1259928468536549628, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 2, caption=SVA infection-induced parenchymal injury in C57BL/6J WT mice (400×). A-D: Inguinal lymph nodes of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right); E-H: Liver of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right); I-L: Spleen of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right)., figureFileSmall=MlZzvSw+MtbLNZvbziwd4g==, figureFileBig=SvXvtKG9vWH1qRR0kaxYxA==, tableContent=null), ArticleFig(id=1259928470142968063, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图2, caption=SVA感染引起C57BL/6J WT小鼠出现实质性损伤(400×), figureFileSmall=MlZzvSw+MtbLNZvbziwd4g==, figureFileBig=SvXvtKG9vWH1qRR0kaxYxA==, tableContent=null), ArticleFig(id=1259928471078297867, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 3, caption=Viral load of SVA in tissues of C57BL/6J WT mice. A: Intraperitoneal injection group; B: Subcutaneous injection group; C: Intramuscular injection group. ns: P>0.05; *: P<0.05; **: P<0.01; ***: P<0.001; ****: P<0.000 1., figureFileSmall=oz8WVe86WHecdoZztXKOZA==, figureFileBig=9gUr46KStXhhRS97dycTeQ==, tableContent=null), ArticleFig(id=1259928472697299225, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图3, caption=SVA在C57BL/6J WT小鼠组织中的病毒载量, figureFileSmall=oz8WVe86WHecdoZztXKOZA==, figureFileBig=9gUr46KStXhhRS97dycTeQ==, tableContent=null), ArticleFig(id=1259928474014310692, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 4, caption=Viremia in C57BL/6J IFNR-/- mice. A: Intraperitoneal injection group; B: Subcutaneous injection group; C: Intramuscular injection group; D: Negative control. ns: P>0.05; *: P<0.05; ***: P<0.001; ****: P<0.000 1., figureFileSmall=lIe78FpahEygD/6dwFNPUw==, figureFileBig=GEdz5+4LcpTsDt0OkJIjsg==, tableContent=null), ArticleFig(id=1259928475155161388, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图4, caption=C57BL/6J IFNR-/- 小鼠出现病毒血症, figureFileSmall=lIe78FpahEygD/6dwFNPUw==, figureFileBig=GEdz5+4LcpTsDt0OkJIjsg==, tableContent=null), ArticleFig(id=1259928476484755773, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 5, caption=Pathological autopsy findings in C57BL/6J IFNR-/- mice. A: Inguinal lymph node; B: Liver; C: Spleen; D: Brain; E: Kidney; F: Heart. From left to right: intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group., figureFileSmall=BxI1k/oARo+4kXs873UV3A==, figureFileBig=ByHJjR5JvKFNnfHeRbAPzA==, tableContent=null), ArticleFig(id=1259928480599368004, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图5, caption=C57BL/6J IFNR-/- 小鼠病理剖检结果, figureFileSmall=BxI1k/oARo+4kXs873UV3A==, figureFileBig=ByHJjR5JvKFNnfHeRbAPzA==, tableContent=null), ArticleFig(id=1259928483157893457, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 6, caption=SVA infection causes more severe histopathological damage in C57BL/6J IFNR-/- mice (400×). A-D: Inguinal lymph nodes of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right); E-H: Liver of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right); I-L: Spleen of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right); M-P: Kidney of mice from the intraperitoneal injection group, subcutaneous injection group, intramuscular injection group, and negative control group (from left to right)., figureFileSmall=pcfW3mJif4cP4rfSmhCOGA==, figureFileBig=BkyqQVkGqbV7VcGA3C1qfQ==, tableContent=null), ArticleFig(id=1259928484030308700, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图6, caption=SVA感染C57BL/6J IFNR-/- 小鼠出现更为严重的组织病理损伤(400×), figureFileSmall=pcfW3mJif4cP4rfSmhCOGA==, figureFileBig=BkyqQVkGqbV7VcGA3C1qfQ==, tableContent=null), ArticleFig(id=1259928485225685348, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 7, caption=Viral load of SVA in tissues of C57BL/6J IFNR-/- mice. A: Intraperitoneal injection group; B: Subcutaneous injection group; C: Intramuscular injection group. ns: P>0.05; *: P<0.05; **: P<0.01; ***: P<0.001; ****: P<0.000 1., figureFileSmall=Qb/HG28nq2/I0jYtSw4OkA==, figureFileBig=HvsjvMa89AncYYGikjXIug==, tableContent=null), ArticleFig(id=1259928487821959537, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图7, caption=SVA在C57BL/6J IFNR-/- 小鼠组织中的病毒载量, figureFileSmall=Qb/HG28nq2/I0jYtSw4OkA==, figureFileBig=HvsjvMa89AncYYGikjXIug==, tableContent=null), ArticleFig(id=1259928490254655869, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Figure 8, caption=The expression levels of TNF-α, IL-6, IL-1β in the inguinal lymph nodes and serum of SVA-infected C57BL/6J WT and C57BL/6J IFNR-/- mice. A: TNF-α mRNA levels in inguinal lymph nodes; B: IL-6 mRNA levels in inguinal lymph nodes; C: IL-1β mRNA levels in inguinal lymph nodes; D: TNF-α protein levels in serum; E: IL-6 protein levels in serum; F: IL-1β protein levels in serum. ns: P>0.05; *: P<0.05; **: P<0.01; ***: P<0.001; ****: P<0.000 1., figureFileSmall=wP3yfxsf6860LGu4pImrzw==, figureFileBig=I4pemBL1Q+i+5+J6lh7jUQ==, tableContent=null), ArticleFig(id=1259928491940766087, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=图8, caption=SVA感染C57BL/6J WT和C57BL/6J IFNR-/- 小鼠腹股沟淋巴结和血清中TNF-α、IL-6、IL-1β的表达水平, figureFileSmall=wP3yfxsf6860LGu4pImrzw==, figureFileBig=I4pemBL1Q+i+5+J6lh7jUQ==, tableContent=null), ArticleFig(id=1259928493207445906, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=EN, label=Table 1, caption=

Primer used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene namesForward primers (5′→3′)Reverse primers (5′→3′)
SVA 3DGCGTCGCATCAAGATTACCGAGGTCAATGCCAGAGCAGTC
IL-6AGTTGCCTTCTTGGGACTGACAGAATTGCCATTGCACAAC
IL-1βGCAACTGTTCCTGAACTCAACTATCTTTTGGGGTCCGTCAACT
TNF-αCGATGAGGTCAATCTGCCCACCAGGTCACTGTCCCAGCATC
GAPDHAAATGGTGAAGGTCGGTGTGAACCAACAATCTCCACTTTGCCACTG
), ArticleFig(id=1259928494025335196, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1259888469254464102, language=CN, label=表1, caption=

本研究所用引物

, figureFileSmall=null, figureFileBig=null, tableContent=
Gene namesForward primers (5′→3′)Reverse primers (5′→3′)
SVA 3DGCGTCGCATCAAGATTACCGAGGTCAATGCCAGAGCAGTC
IL-6AGTTGCCTTCTTGGGACTGACAGAATTGCCATTGCACAAC
IL-1βGCAACTGTTCCTGAACTCAACTATCTTTTGGGGTCCGTCAACT
TNF-αCGATGAGGTCAATCTGCCCACCAGGTCACTGTCCCAGCATC
GAPDHAAATGGTGAAGGTCGGTGTGAACCAACAATCTCCACTTTGCCACTG
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A型塞内卡病毒C57BL/6J IFNR-/- 小鼠感染模型建立及感染特性评价
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游灵巧 1 , 张卿 1 , 高晟斌 2 , 付利芝 3, 4, 5 , 贾梅玉 1 , 王玉娥 1, 3
微生物学报 | 研究报告 2026,66(5): 2430-2443
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微生物学报 | 研究报告 2026, 66(5): 2430-2443
A型塞内卡病毒C57BL/6J IFNR-/- 小鼠感染模型建立及感染特性评价
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游灵巧1, 张卿1, 高晟斌2, 付利芝3, 4, 5, 贾梅玉1 , 王玉娥1, 3
作者信息
  • 1.西南大学 动物医学院，重庆
  • 2.中国动物卫生与流行病学中心，山东 青岛
  • 3.国家生猪技术创新中心，重庆
  • 4.重庆市畜牧科学院，重庆
  • 5.农业农村部动物疫病荣昌野外科学观测研究站，重庆
Establishment of a Senecavirus A infection model in C57BL/6J IFNAR-/-mice and evaluation of its infection characteristics
Lingqiao YOU1, Qing ZHANG1, Shengbin GAO2, Lizhi FU3, 4, 5, Meiyu JIA1 , Yu’e WANG1, 3
Affiliations
  • 1.College of Veterinary Medicine, Southwest University, Chongqing, China
  • 2.China Animal Health and Epidemiology Center, Qingdao, Shandong, China
  • 3.National Center of Technology Innovation for Pigs, Chongqing, China
  • 4.Chongqing Academy of Animal Sciences, Chongqing, China
  • 5.Rongchang Field Scientific Observation and Research Station for Animal Diseases, Ministry of Agriculture and Rural Affairs, Chongqing, China
出版时间: 2026-05-04 doi: 10.13343/j.cnki.wsxb.20250892
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目的 构建能有效模拟猪A型塞内卡病毒(Senecavirus A, SVA)关键临床特征的小鼠模型，为阐明其致病机制及开展防控产品评价提供关键实验工具。 方法 通过腹腔、皮下、肌内注射3种途径分别接种5周龄SPF级C57BL/6J野生型(WT)小鼠和I型干扰素受体缺失(C57BL/6J IFNR-/- )小鼠，于感染后第1、3、5天采集血液及组织样本，进行宏观病理、组织病理、病毒载量及炎性因子mRNA动态检测。 结果 与未接毒对照组相比，2种小鼠均出现腹股沟淋巴结肿大、肝脏土黄色变及脾肿大等宏观病变和显微病变(淋巴结皮质崩解、肝细胞坏死、脾白髓萎缩、肾小管坏死)，但C57BL/6J IFNR-/- 小鼠病变更为严重；病毒RNA在2种小鼠组织中广泛分布，但C57BL/6J IFNR-/- 组显著高于WT组。此外，WT小鼠未出现病毒血症，C57BL/6J IFNR-/- 小鼠在感染后1 dpi全血中即可检出病毒，3 dpi达到峰值后快速下降。炎症因子检测显示，C57BL/6J IFNR-/- 小鼠IL-1β和IL-6炎性因子mRNA水平和蛋白水平均显著高于WT小鼠。 结论 C57BL/6J IFNR-/- 小鼠首次模拟了猪SVA短暂病毒血症特征，可全面复现病毒多器官分布、高病毒载量及自限性恢复等特征，为深入研究其致病机制及评价疫苗与抗病毒药物提供了更为有效的动物模型。

A型塞内卡病毒  /  动物模型  /  C57BL/6J野生型小鼠  /  C57BL/6J IFNR-/- 小鼠  /  感染特性

Objective To establish a mouse model that effectively simulates the key clinical features of porcine Senecavirus A (SVA) infection, providing a crucial experimental tool for elucidating its pathogenesis and evaluating prevention and control products. Methods Five-week-old SPF C57BL/6J wild-type (WT) mice and type I interferon receptor-deficient (C57BL/6J IFNR-/- ) mice were inoculated via intraperitoneal, subcutaneous, and intramuscular routes. Blood and tissue samples were collected on days 1, 3, and 5 post-infection (dpi) for analysis of gross pathology, histopathology, viral load, and dynamic determination of inflammatory cytokines at the mRNA level. Results Compared with the mock-infected control group, both mouse strains developed gross lesions (e.g., swollen inguinal lymph nodes, yellowish livers, splenomegaly) and histopathological lesions (e.g., cortical disintegration of lymph nodes, hepatocellular necrosis, atrophy of splenic white pulp, and renal tubular necrosis). However, these lesions were more severe in C57BL/6J IFNR-/- mice. Viral RNA was widely distributed in tissues of both groups but was significantly higher in the C57BL/6J IFNR-/- group. Notably, viremia was undetectable in WT mice, whereas in C57BL/6J IFNR-/- mice, the virus was detected in whole blood as early as 1 dpi, peaked at 3 dpi, and then declined rapidly. Inflammatory cytokine analysis revealed significantly higher mRNA levels and protein levels of IL-1β and IL-6 in C57BL/6J IFNR-/- mice than in WT mice. Conclusion The C57BL/6J IFNR-/- mouse model successfully simulates, for the first time, the transient viremia characteristic of porcine SVA infection. It comprehensively replicates key features, including the multi-organ viral distribution, high viral load, and self-limiting recovery, providing a more effective animal model for delving into the pathogenic mechanism of SVA and evaluating vaccines and antiviral drugs.

Senecavirus A  /  animal model  /  C57BL/6J wild-type mice  /  C57BL/6J IFNR-/- mice  /  infection characteristics
游灵巧, 张卿, 高晟斌, 付利芝, 贾梅玉, 王玉娥. A型塞内卡病毒C57BL/6J IFNR-/- 小鼠感染模型建立及感染特性评价. 微生物学报, 2026 , 66 (5) : 2430 -2443 . DOI: 10.13343/j.cnki.wsxb.20250892
Lingqiao YOU, Qing ZHANG, Shengbin GAO, Lizhi FU, Meiyu JIA, Yu’e WANG. Establishment of a Senecavirus A infection model in C57BL/6J IFNAR-/-mice and evaluation of its infection characteristics[J]. Acta Microbiologica Sinica, 2026 , 66 (5) : 2430 -2443 . DOI: 10.13343/j.cnki.wsxb.20250892
正文
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A型塞内卡病毒(Senecavirus A, SVA)属于小RNA病毒科塞内卡病毒属，是一种无囊膜的单股正链RNA病毒[1]。其基因组长度约7 300 bp，呈典型的L-4-3-4结构，即包含L前体蛋白、结构蛋白(VP4、VP2、VP3和VP1)的P1区、非结构蛋白(2A、2B和2C)的P2区、非结构蛋白(3A、3B、3C和3D)的P3区，编码的结构与非结构蛋白共同完成病毒的复制与组装。猪感染该病毒后会表现出类似口蹄疫的临床症状。自2007年加拿大首次报道以来[2]，SVA疫情已蔓延至全球多个主要养猪国家，如美国、巴西、中国、哥伦比亚、泰国等，成为一种不容忽视的新发病毒性传染病[3]。2015年3月，我国广东省首次报道了SVA疫情，之后疫情由南往北迅速蔓延开来，目前我国已有多地的猪场感染SVA，湖南、广西、福建、河南、湖北、江西、新疆、黑龙江等省(自治区)相继发生疫情[4]。Li等[5]统计2021年前的数据发现，中国西南地区SVA个体阳性率达11.70%，群体阳性率高达50%；Preis等[6]对2022年美国近百个猪场的监测显示，SVA血清阳性率约17.3%，提示SVA的流行形势依然严峻。
SVA是猪特发性水疱病(porcine idiopathic vesicular disease, PIVD)和流行性短暂新生仔猪死亡(epidemic transient neonatal losses, ETNL)的主要病原，感染者表现为嗜睡、跛行及口鼻部、蹄部出现水疱性病变[7-8]。尽管SVA的总体发病率较高，但其致死率因猪群日龄不同而有所差异，1-4日龄新生仔猪感染后死亡率可高达30%-70%[9]。该病毒具有广泛的组织嗜性，可在扁桃体、淋巴结、脾脏、肝脏、肾脏等多种组织中复制，并诱导宿主产生短暂的病毒血症[10-11]；其早期产生的中和抗体与病毒清除密切相关[12-13]。
然而，SVA的流行病学和致病机制等诸多方面仍不清楚。猪虽被视为SVA的自然宿主，但血清学及病毒分离证据表明，该病毒可能存在跨物种传播风险。国外研究者在牛、鼠等动物血清中检出SVA抗体[2,14]；国内研究者于2021年从广东省的水牛体内首次分离到SVA，进一步证实了这一风险[15]。此外，研究表明自然感染的小鼠可携带具有感染性的病毒颗粒，家蝇和库蠓等昆虫也被认为是潜在传播媒介，提示SVA的传播链复杂，对养猪业构成持续威胁[16-18]。
目前，针对SVA的致病机制和传播规律仍有许多未明之处，亟需建立可靠的动物模型以支撑相关研究。虽然猪是理想的自然感染模型，但其成本高、周期长、操作不便等限制了大通量研究的开展。小鼠因具备遗传背景清晰、繁殖快、成本低廉等优势，被广泛应用于病毒机制、疫苗免疫评价及药物筛选研究[19-20]。然而，常规C57BL/6J野生型(wild-type, WT)小鼠模型感染SVA后不引起病毒血症，无法模拟猪只急性期的关键病理过程。因此，本研究系统比较了C57BL/6J WT与其I型干扰素受体基因敲除(C57BL/6J IFNR-/- )小鼠在不同攻毒途径下对SVA的感染反应，通过分析病毒复制动态、组织病理损伤、免疫应答特征等指标，构建可重现SVA病毒血症与多器官病变的小鼠模型，以期为深入揭示SVA致病机制及开展防控产品评价提供关键实验工具。
1 材料与方法
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1.1 病毒和实验动物
SVA毒株由本实验室分离并保存。实验动物选用5周龄雌性无特定病原体(specific pathogen free, SPF) C57BL/6J WT小鼠，购自重庆恩斯维尔实验动物销售有限公司；C57BL/6J IFNR-/- 小鼠(I型干扰素受体基因敲除)由中国农业科学院哈尔滨兽医研究所翁长江与黄丽研究员惠赠。本研究所有动物实验均获得重庆市畜牧科学院实验动物伦理委员会批准，编号为XKY-202402B01。
1.2 主要试剂和仪器
TRIzol试剂和ChamQ Universal SYBR qPCR Master Mix，南京诺唯赞生物科技股份有限公司；PrimeScript RT Master Mix试剂盒，TaKaRa公司；乙醚，上海凌峰化学试剂有限公司；肝素钠，上海源叶生物科技有限公司；4%中性福尔马林溶液，上海麦克林生化科技股份有限公司；小鼠IL-6、TNF-α及IL-1β ELISA试剂盒，杭州华安生物技术有限公司。
冷冻研磨仪，上海净信实业发展有限公司；低温高速离心机，上海卢湘仪离心机仪器有限公司；PCR仪，Bio-Rad公司；实时荧光定量PCR仪，西安天隆科技有限公司；酶标仪，赛默飞世尔科技公司。
1.3 小鼠攻毒试验
将48只C57BL/6J WT小鼠随机分成4组(n=12)，包括3个感染组(腹腔注射、皮下注射、肌内注射)和1个对照组。小鼠适应饲养3 d后，感染组每只小鼠经相应途径接种200 µL SVA病毒液(滴度为2×107 TCID50/只，原液浓度为107 TCID50/0.1 mL)；对照组小鼠不做任何处理。攻毒当天记为0 d，每日观察并记录小鼠临床症状，分别于感染后第1、3、5天(days post infection, dpi)采集各组小鼠抗凝全血和血清。C57BL/6J IFNR-/- 小鼠的实验分组、病毒接种剂量与时间点均与WT小鼠保持一致。
1.4 病理剖检
在1、3、5 dpi，每组随机选取4只小鼠，采用乙醚麻醉后实施安乐死，立即进行系统剖检。重点观察并记录肝、脾、肺、肾、腹股沟淋巴结等组织器官的病变情况，同时拍照存档。在无菌条件下采集上述组织，每个样本均分为2份：一份置于-80 ℃冰箱保存，用于病原检测；另一份置于4%福尔马林溶液中固定，用于后续组织病理学分析。
1.5 病原检测
将采集的病料组织和抗凝全血加入TRIzol试剂，经冷冻研磨仪充分匀浆。4 ℃、12 000 r/min离心10 min后收集上清液，进行总RNA抽提。使用PrimeScript RT Master Mix试剂盒将RNA反转录成cDNA。以cDNA为模板，使用ChamQ Universal SYBR qPCR Master Mix在实时荧光定量PCR仪上对SVA 3D基因及各类细胞因子mRNA进行扩增检测。以GAPDH为内参，采用ΔΔCt法对炎症因子等靶基因进行相对定量分析；采用绝对定量PCR技术检测病毒在动物体组织脏器中的载量。所用引物序列如表1所示。
1.6 苏木素-伊红(H.E.)染色
福尔马林溶液固定后的组织经常规包埋，制作石蜡切片。切片经脱蜡、水化后，进行苏木素-伊红染色，具体步骤如下：苏木精染色5 min，经水洗-分化-水洗-返蓝-水洗处理后，再依次用85%、95%的乙醇梯度脱水5 min，伊红复染各5 min，最后用中性树胶封片，在光学显微镜下观察并评估组织病理变化。
1.7 酶联免疫吸附试验(enzyme-linked immunosorbent assay, ELISA)
将标准品和采集的血清样本各10 µL，加入50 µL分析缓冲液，再加入50 µL检测抗体，用封板膜覆盖后在微孔板振荡器上室温孵育1 h。随后弃去液体，每孔加300 µL洗涤液，静置后弃去并拍干，重复4次。每孔加入100 µL链霉亲和素-HRP，用封板膜覆盖后在微孔板振荡器上室温孵育30 min。随后弃去液体，每孔加300 µL洗涤液，静置后弃去并拍干，重复4次。每孔加入100 µL TMB底物溶液，室温避光孵育15 min。每孔加入100 µL终止液，用酶标仪在450 nm波长下读取吸光值。根据绘制的标准曲线计算样品浓度。
1.8 统计学分析与作图
采用GraphPad Prism 8.0软件对数据进行处理及统计学分析。数据以平均值±标准差(mean±SD)表示。采用t检验进行组间差异性比较。P<0.05表示差异具有统计学意义。
2 结果与分析
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2.1 C57BL/6J WT小鼠病理剖检结果
剖检结果显示，SVA感染组C57BL/6J WT小鼠呈现一致性的系统性病变：腹股沟淋巴结均明显肿胀，质地变硬(图1A)；肝脏颜色变浅，呈土黄色(图1B)；通过腹腔和皮下注射途径感染的小鼠脾脏表现出轻度至中度肿大(图1C)。脑、肾、心脏等器官外观未见异常(图1D-1F)。上述结果表明，SVA感染可导致C57BL/6J WT小鼠产生明显的病理损伤。
2.2 C57BL/6J WT小鼠组织病理学观察
由于剖检仅发现腹股沟淋巴结、肝脏和脾脏出现异常，因此对这3类脏器进行组织病理学检查。与对照组相比，感染组小鼠腹股沟淋巴结出现网状内皮细胞增生、炎性细胞浸润(图2A-2D)；肝脏表现为肝细胞空泡变性、核碎裂或消失，并见散在的局灶性坏死(图2E-2H)；脾脏则出现浆液渗出、淋巴细胞数量减少(图2I-2L)。这些结果进一步证实SVA在WT小鼠体内感染可引起实质性损伤。
2.3 SVA在C57BL/6J WT小鼠组织中的病毒载量
通过实时荧光定量PCR系统检测感染后1、3、5 d小鼠组织中的病毒RNA。结果显示(图3)，经不同途径感染的小鼠在肝脏、脾脏、肺、腹股沟淋巴结等组织中均能检测到病毒RNA，但全血始终为阴性，提示未形成病毒血症。病毒含量呈明显的组织偏好性：腹股沟淋巴结病毒载量最高，约为1×105-3×105 copies/mg；肝脏和脾脏次之，载量分别为3×104-2×105 copies/mg和1×103-1×104 copies/mg；心脏、肺、肾、脑等脏器中病毒载量水平约为1×102 copies/mg。腹腔注射组在大脑、胸腺、心脏、肺、肾、空肠及肠系膜淋巴结中的病毒载量均高于皮下和肌内注射组(图3A-3C)，表明腹腔注射途径更易造成全身性感染。此外，病毒于1 dpi可在各个组织中检出，随着时间延长，病毒载量显著降低，提示WT小鼠可通过天然免疫逐步控制SVA复制。
2.4 C57BL/6J IFNR-/- 小鼠出现病毒血症
已有研究表明，猪感染SVA早期伴随短暂且高效的病毒血症，这是临床关键特征之一[12]。由于C57BL/6J WT小鼠未出现病毒血症，难以完整模拟SVA感染过程。因此，本研究进一步在C57BL/6J IFNR-/- 小鼠中评估病毒血症的发生与消长规律。实时荧光定量PCR结果显示(图4)，3种攻毒途径均可在1 dpi的全血中检出SVA RNA，峰值可达1×106 copies/mL；皮下和肌内注射组病毒载量水平在3 dpi时进一步升高，随后快速下降，至5 dpi降至4×103 copies/mL左右。病毒血症的动态曲线与猪只早期报道一致，提示C57BL/6J IFNR-/- 小鼠能够更精准地模拟SVA感染情况。
2.5 C57BL/6J IFNR-/- 小鼠病理剖检结果
与对照组及WT感染组相比，C57BL/6J IFNR-/- 小鼠呈现更为显著的系统性病变(图5)。腹股沟淋巴结体积明显增大、质地变硬(图5A)；肝脏色泽普遍变浅，呈土黄色改变(图5B)；各攻毒组脾脏均可见中度至重度肿大，边缘钝圆(图5C)；脑、肾、心脏等脏器外观未见异常(图5D-5F)。结果提示，I型干扰素信号缺失可显著放大SVA引起的系统性病变。
2.6 C57BL/6J IFNR-/- 小鼠组织病理学观察
由于SVA对C57BL/6J IFNR-/- 小鼠的宏观病变显著重于WT组，本研究对腹股沟淋巴结、肝、脾和肾组织进行了组织病理学评估。H.E.染色结果显示，腹股沟淋巴结皮质区结构崩解，淋巴细胞显著减少(图6A-6D)；肝脏出现肝细胞空泡样变性及灶性坏死，肝小叶之间界限模糊不清(图6E-6H)；脾脏白髓萎缩，淋巴细胞数量明显减少(图6I-6L)；肾脏则表现为肾小管上皮细胞肿胀、核固缩，管腔内可见细胞坏死及脱落(图6M-6P)。上述多器官实质性损伤证实I型干扰素信号缺失显著加剧了SVA的嗜组织性与致病力。
2.7 C57BL/6J IFNR-/- 小鼠各组织中具有更高的病毒载量
定量PCR显示，SVA在C57BL/6J IFNR-/- 小鼠体内分布更为广泛且能够高效增殖(图7)。除肝脏、脾脏、肺和腹股沟淋巴结外，小脑、心脏、肾脏、空肠及肠系膜淋巴结中均能检出病毒RNA。腹股沟淋巴结仍是病毒载量最高的组织，腹腔与皮下注射组可达1×107-2×107 copies/mg，肌内注射组约为7×105 copies/mg；肝脏次之，病毒载量约为3×105-1×107 copies/mg，腹腔和皮下注射组显著高于肌内注射组；其他脏器中病毒载量峰值均维持在3×102-5×105 copies/mg。病毒RNA在1 dpi即可在各组织中检出；至3 dpi，多数组织的病毒载量迅速升高至峰值，较1 dpi上升1-3 log10，且显著高于同期WT小鼠(P<0.01)，表明SVA在C57BL/6J IFNR-/- 小鼠体内活跃复制；到5 dpi时病毒载量普遍下降2-5 log10，提示病毒复制已得到宿主有效控制。
2.8 C57BL/6J WT和C57BL/6J IFNR-/- 小鼠炎性因子表达动态
由于C57BL/6J IFNR-/- 小鼠感染SVA后在临床表现、组织损伤以及病毒载量等方面均较WT小鼠更为显著，本研究进一步探究了2组小鼠腹股沟淋巴结和血清中关键炎性因子的动态变化。如图8所示，与WT小鼠相比，C57BL/6J IFNR-/- 小鼠呈现更强且动态变化的炎症谱型：TNF-α与IL-6 mRNA于1 dpi显著升高，3 dpi即明显回落；对应血清蛋白TNF-α和IL-6也分别由24 pg/mL和6 pg/mL降至16 pg/mL和3 pg/mL；IL-1β mRNA则在3 dpi显著升高，但蛋白水平无明显变化且仅约为0.6 pg/mL，提示第二波炎症放大。相比之下，WT小鼠TNF-α变化趋势与C57BL/6J IFNR-/- 小鼠相似(图8A、8D)，IL-6 mRNA水平仅在3 dpi上调，血清中IL-6含量无明显变化(图8B、8E)，而IL-1β在WT小鼠则无明显变化(图8C、8F)。综上所述，SVA在C57BL/6J IFNR-/- 小鼠诱发的炎症反应更为强烈，与临床上猪自然感染SVA早期的症状更相符。
3 讨论与结论
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建立能够准确模拟病原体关键致病特征的动物模型是解析SVA致病机制与评价防控产品的首要前提。猪作为SVA的自然宿主，其高成本、长周期以及伦理限制等因素使其难以满足高通量研究的需求。现有小鼠模型虽可用于评估SVA疫苗的免疫原性初评[21-23]和药物筛选[24]，但因缺乏病毒血症而难以模拟急性期全身扩散的特征。本研究系统比较WT与C57BL/6J IFNR-/- 小鼠在3种接种途径下的感染动力学，证实SVA在两系小鼠中呈广谱组织嗜性，病毒RNA高效分布于肝、脾、肺、腹股沟淋巴结、大脑、心脏、肾、空肠及肠系膜淋巴结等组织，且以淋巴结、肝脏、脾脏的病毒载量最高，这与猪感染后以扁桃体-淋巴结-脾脏-肺-肝为主要靶器官的分布规律高度一致(小鼠无扁桃体)。皮下接种在可操作性、临床相似性与病毒载量之间取得了最佳平衡，因此被推荐为标准攻毒途径。
病毒血症是猪SVA感染早期最显著的标志性事件[12-13]，但此前的小鼠模型难以复制这一过程。本研究在C57BL/6J IFNR-/- 小鼠中成功模拟了这一关键过程：血液病毒载量在1 dpi即达1×106 copies/mL，并于3 dpi达到峰值后迅速下降，重现了“短暂而高效”的宿主内传播模式。更重要的是，绝对定量PCR显示，病毒在多组织中呈现高度一致的“检出-指数扩增-清除”复制动力学曲线(1-3 dpi病毒载量上升1-3 log10)，这与病毒血症动态完全吻合，构成了病毒成功建立系统性感染的坚实证据。因此，尽管抗原原位定位受限于现有抗体工具未能实现，但本研究所呈现的完整定量动力学数据，已充分满足了验证一个新感染模型的核心要求。
从病理组织学变化来看，C57BL/6J IFNR-/- 小鼠呈现出“剂量-时间”依赖的显著病变：宏观可见淋巴结肿大、肝土黄色变、脾肿大；镜下呈现淋巴结皮质崩解、肝脏小叶坏死、脾白髓萎缩及肾小管变性，损伤程度较WT组显著加重，却未触发IL-6、TNF-α、IL-1β等风暴式升高，炎性因子仅在早期短暂上调后即回落，且蛋白水平均较低，因强烈的免疫反应易造成机体严重损伤[25-26]，这些特征与猪群“自限性恢复”的临床特征相符[12,27]，提示SVA致病机制以直接细胞损伤为主，而非过度炎症。此外，干扰素系统是宿主抵御病毒感染的第一道防线[28]。I型干扰素受体缺失使C57BL/6J IFNR-/- 小鼠无法启动经典的IFN-α/β-JAK-STAT抗病毒信号通路，导致病毒复制失控，病毒载量较WT组整体高1-3 log10，直接证实干扰素系统在抑制SVA早期扩散中发挥核心作用，也为后续研究SVA与宿主先天免疫互作、筛选干扰素佐剂或拮抗剂提供了直观平台。
综上所述，本研究成功建立并表征了C57BL/6J WT与C57BL/6J IFNR-/- 小鼠的SVA感染模型，明确了最佳感染途径(皮下注射)、病毒组织分布规律及免疫应答特征。其中，C57BL/6J IFNR-/- 小鼠模型因能模拟猪的病毒血症特征，适用于病毒-宿主互作、抗病毒药物和疫苗效力评价；WT小鼠成本低、操作简便，可用于病毒毒株毒力比较、中和抗体初筛及早期免疫机制探索。这2种模型互为补充，为SVA致病机制研究、防控产品快速评价及免疫策略优化提供了可靠、高效且经济的动物平台。
基金
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  • 国家自然科学基金(32402877)
  • 重庆市自然科学基金(CSTB2024NSCQ-MSX2567)
文献
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2026年第66卷第5期
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doi: 10.13343/j.cnki.wsxb.20250892
  • 接收时间：2025-12-02
  • 首发时间：2026-05-09
  • 出版时间：2026-05-04
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出版历史
  • 收稿日期：2025-12-02
  • 录用日期：2026-01-05
基金
The National Natural Science Foundation of China(32402877)
国家自然科学基金(32402877)
The Natural Science Foundation of Chongqing(CSTB2024NSCQ-MSX2567)
重庆市自然科学基金(CSTB2024NSCQ-MSX2567)
作者信息
    1.西南大学 动物医学院，重庆
    2.中国动物卫生与流行病学中心，山东 青岛
    3.国家生猪技术创新中心，重庆
    4.重庆市畜牧科学院，重庆
    5.农业农村部动物疫病荣昌野外科学观测研究站，重庆
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https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20250892
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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