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微生物学通报

猪圆环病毒3型衣壳蛋白(Cap)抑制宿主细胞天然免疫应答
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国家重点研发计划(2016YFD0500702-2);中国农业科学院前沿探索项目(1610312017003)


Inhibition effect of PCV3 capsid protein on innate immune response of host cells
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    摘要:

    【背景】猪圆环病毒是引起猪圆环病毒病的病原,能够引起严重的免疫抑制和临床症状,给养猪业造成严重的经济损失。【目的】研究猪圆环病毒3型(porcine circovirus 3,PCV3)衣壳蛋白(Capsid protein,Cap)对宿主天然免疫应答的调控作用,解析其免疫抑制机制对阐明猪圆环病毒致病机制具有重要意义。【方法】通过构建Cap真核表达质粒,利用Western blotting进行表达验证,实时荧光定量(RT-PCR)、双荧光素酶基因报告系统和ELISA探究Cap对I型干扰素通路活化的影响,通过免疫共沉淀探索其作用机制。【结果】真核表达质粒成功表达并且证实Cap可以抑制DNA模拟物poly(dA:dT)诱导的I型干扰素通路的活化;Cap可以与天然免疫通路节点分子MITA相互作用。【结论】研究发现PCV3病毒Cap蛋白与干扰素通路节点分子MITA相互作用发挥免疫抑制作用,这为阐明PCV3免疫抑制机制提供了理论依据。

    Abstract:

    [Background] Porcine circovirus causes porcine circovirus associated disease, severe immunosuppression and clinical symptoms in pigs, causing substantial economic losses. [Objective] To study the role of porcine circovirus 3 (PCV3) capsid protein (Cap) in regulating host innate immune response, we constructed a eukaryotic plasmid expressing PCV3 protein Cap. [Methods] The expression of Cap was verified by Western blotting. The real-time quantification PCR (RT-PCR), dual luciferase gene reporting system and ELISA assay were used to study the effect of Cap on type I interferon pathway activation, and the Co-immunoprecipitation was done to study the involved antagonistic mechanism. [Results] The expression of Cap was verified, suggesting that Cap was highly expressed in the transfected cells, and Cap inhibited the activation of type I interferon signaling pathway stimulated by a synthetic DNA, poly(dA:dT). The interaction of Cap with the adaptor protein MITA of innate immune signaling pathway was identified. [Conclusion] These results indicate that Cap plays an important role in inhibiting the innate immune response of host cells by interacting with MITA. Our study provides a theoretical basis for clarification of the molecular mechanisms for PCV3-mediated immune suppression.

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赵振翔,朱紫祥,杨帆,曹伟军,李莎莎,康慧,杨孝朴,郑海学. 猪圆环病毒3型衣壳蛋白(Cap)抑制宿主细胞天然免疫应答[J]. 微生物学通报, 2019, 46(7): 1761-1771

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  • 在线发布日期: 2019-07-02
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