| B型流感病毒冷适应株B/Yamagata/16/88反向遗传操作平台的搭建及BALB/c小鼠感染模型的建立 |
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| 引用本文： | 孙伟洋,于志君,李雪,陈强,高晓龙,国娇,李元果,张坤,王铁成,杨松涛,黄耕,赵永坤,高玉伟,夏咸柱.B型流感病毒冷适应株B/Yamagata/16/88反向遗传操作平台的搭建及BALB/c小鼠感染模型的建立[J].实验动物与比较医学,2015,23(1). |
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| 作者姓名： | 孙伟洋 于志君 李雪 陈强 高晓龙 国娇 李元果 张坤 王铁成 杨松涛 黄耕 赵永坤 高玉伟 夏咸柱 |
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| 作者单位： | 中国人民解放军军事医学科学院军事兽医研究所,北京协和医学院和中国医学科学院医学实验动物研究所,长春生物制品研究所有限责任公司,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所,中国人民解放军军事医学科学院军事兽医研究所 |
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| 基金项目： | 国家高技术研究发展计划（863计划） |
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| 摘 要： | 【摘要】 目的 本研究利用基因工程技术全基因合成B型流感病毒冷适应株B/Yamagata/16/88的8个基因节段，并利用反向遗传技术从体外拯救B型流感病毒冷适应株B/Yamagata/16/88，同时建立BALB/c小鼠感染模型，为下一步研究B型流感病毒致病机制、传播机制以及开发新型疫苗奠定基础。方法 通过基因合成和反向遗传技术体外拯救B型流感病毒冷适应株B/Yamagata/16/88。全基因组测序验证拯救病毒基因组序列与Genbank序列的一致性。将拯救病毒以105EID50的攻毒剂量人工感染BALB/c小鼠，通过体重变化、生存率、肺脏病毒复制等方面进行致病性分析，建立小鼠感染模型。结果 成功从体外拯救出B型流感病毒冷适应株B/Yamagata/16/88，命名为B-S9。全基因组测序结果表明，B-S9基因组序列与Genbank公布序列一致。B-S9能够人工感染BALB/c小鼠，但不致死，对BALB/c小鼠呈现低致病性； 攻毒后第3天，B-S9感染小鼠体重出现下降，攻毒后第8天，小鼠体重开始回升；攻毒后第3天和第6天，B-S9感染小鼠的肺脏内均能检测到病毒复制，且攻毒后第3天的小鼠肺脏病毒滴度比攻毒后第6天的小鼠肺脏滴度高132倍。结论 成功搭建B型流感病毒冷适应株B/Yamagata/16/88反向遗传操作平台，并建立BALB/c小鼠感染模型。目前国内外对B型流感病毒的研究还较少，该反向遗传操作平台的建立为B型流感病毒致病机制和传播机制的研究奠定了基础，同时也为包括B型流感病毒减毒活疫苗在内的新型疫苗的研制开辟了新途径。
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| 关 键 词： | B型流感病毒 反向遗传操作 致病性 BALB/c小鼠 |
| 收稿时间： | 2014/8/25 0:00:00 |
| 修稿时间： | 2014/9/3 0:00:00 |
The establishment of a reverse genetics system of cold-adaptated influenza B virus B/Yamagata/16/88 and a model of influenza B virus infection in BALB/c mice |
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| Abstract: | Abstract] Objective The whole genome of the cold-adapted influenza B virus B/Yamagata/16/88 were synthesized by using the genetic engineering technology in this study. To build a foundation for studying the pathogenesis, the mechanism of transmission, and developing of the new vaccines of the influenza B virus, the cold-adapted influenza B virus was rescued in vitro by using the reverse genetic technology, and the model of BALB/c mice, which can be infected with this virus, were established. Methods Using genetic synthesis and the reverse genetic technology, the cold-adapted influenza B virus was rescued in vitro. We utilized the whole genome sequencing approach to validate the identity between the rescued viral genome sequences and Genbank sequences. To establish the model of BALB/c mice infection in influenza B virus, the BALB/c mice were infected with 105 EID50 dose of the rescued virus, and the weight change, survival rate, and viral replication in the lungs were analyzed. Results We successfully rescued the cold-adapted influenza B virus B/Yamagata/16/88 in vitro, and this virus was named B-S9. The genome sequencing results showed that Genbank sequence was consistent with the genome sequence of B-S9. BALB/c mice can be artificially infected with B-S9, but no dead mice were found due to infection in this study. The above results indicated B-S9 is low pathogenicity to the BALB/c mice. The mice that were infected with B-S9 showed weight decline in 3 days post inoculation (dpi) but restore in 8 dpi. The virus titers can be detected in the lungs of mice infected with B-S9 in 3 and 6 dpi, respectively. Furthermore, the virus titers in the lungs of the BALB/c mice in 3 dpi were 132 times higher than that in 6 dpi. Conclusions A reverse genetic system of cold-adapted influenza B virus B/Yamagata/16/88 were successfully established, and the model of the influenza B virus infection in BALB/c mice was also established. To date, the study of influenza B virus was less at home and abroad. The establishment of this reverse genetic system provides not only a platform for studying the pathogenesis and the mechanism of transmission of influenza B virus, but also a way to develop a new live-attenuated influenza B virus vaccine. |
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| Keywords: | Influenza B virus Reverse genetic system Pathogenicity BALB/c mice |
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