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黑鲤鱼的 STAT1a 和 STAT1b 在先天免疫防御 GCRV 中发挥重要作用。

STAT1a and STAT1b of black carp play important roles in the innate immune defense against GCRV.

机构信息

State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, 410081, China.

State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, 410081, China.

出版信息

Fish Shellfish Immunol. 2019 Apr;87:386-394. doi: 10.1016/j.fsi.2019.01.037. Epub 2019 Jan 28.

Abstract

Signal transducer and activator of transcription 1 (STAT1) plays an important role in the Janus kinase (JAK)-STAT signaling of human and mammals; however, the mechanism of STAT1 in innate immune activation of teleost fishes remains largely unknown. In this study, two STAT1 homologues (bcSTAT1a and bcSTAT1b) of black carp (Mylopharyngodon piceus) have been cloned and characterized. Both bcSTAT1a and bcSTAT1b transcription in host cells was obviously increased in response to the stimulation of poly (I:C), lipopolysaccharide (LPS), grass carp reovirus (GCRV) and interferon (IFN); however, the increase rate of bcSTAT1b transcription post stimulation was obviously higher than that of bcSTAT1a. bcSTAT1a and bcSTAT1b were distributed in both cytoplasm and nucleus in the immunofluorescence staining assay. Self-association of bcSTAT1a and bcSTAT1b, and the interaction between bcSTAT1a and bcSTAT1b have been detected through co-immunoprecipitation (co-IP) assay; and the data of native polyacrylamide gel electrophoresis (PAGE) implied that bcSTAT1a and bcSTAT1b might form homodimer and heterodimer in vivo like their mammalian counterparts. Both bcSTAT1a and bcSTAT1b presented IFN-inducing ability in report assay, and both bcSTAT1a and bcSTAT1b showed antiviral activities against GCRV in EPC cells. Our data support the conclusion that both bcSTAT1a and bcSTAT1b play important roles in host antiviral innate immune activation initiated by GCRV.

摘要

信号转导子和转录激活子 1(STAT1)在人类和哺乳动物的 Janus 激酶(JAK)-STAT 信号转导中发挥重要作用;然而,STAT1 在硬骨鱼类先天免疫激活中的机制在很大程度上仍然未知。在本研究中,克隆并表征了草鱼(Mylopharyngodon piceus)的两种 STAT1 同源物(bcSTAT1a 和 bcSTAT1b)。bcSTAT1a 和 bcSTAT1b 的转录在宿主细胞中明显增加,以响应多聚(I:C)、脂多糖(LPS)、草鱼虹彩病毒(GCRV)和干扰素(IFN)的刺激;然而,bcSTAT1b 转录后的增加率明显高于 bcSTAT1a。在免疫荧光染色测定中,bcSTAT1a 和 bcSTAT1b 分布在细胞质和细胞核中。通过共免疫沉淀(co-IP)测定检测到 bcSTAT1a 和 bcSTAT1b 的自我缔合以及 bcSTAT1a 和 bcSTAT1b 之间的相互作用;并且天然聚丙烯酰胺凝胶电泳(PAGE)的数据表明,bcSTAT1a 和 bcSTAT1b 可能像它们的哺乳动物对应物一样在体内形成同源二聚体和异源二聚体。bcSTAT1a 和 bcSTAT1b 在报告测定中均具有 IFN 诱导能力,并且 bcSTAT1a 和 bcSTAT1b 在 EPC 细胞中均显示出对 GCRV 的抗病毒活性。我们的数据支持以下结论:bcSTAT1a 和 bcSTAT1b 均在 GCRV 引发的宿主抗病毒先天免疫激活中发挥重要作用。

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