SOCS3负调控流感病毒诱导干扰素表达机理的初步研究

彭本群; 胡精蕴; 毛亚楠; 王淑霖; 王佳俊; 陈梦颖; 尤冬雪; 王松

SOCS3负调控流感病毒诱导干扰素表达机理的初步研究

彭本群^{1, 2, 3,},
胡精蕴^{1, 2, 3},
毛亚楠^{1, 2},
王淑霖^{1, 2},
王佳俊^{1, 3},
陈梦颖^{1, 3},
尤冬雪^{1, 3},
王松^{1, 2, 3, ,}

1.
福建农林大学动物科学学院，福建　福州　350002
2.
福建省畜禽病原感染与免疫学重点实验室，福建　福州　350002
3.
闽台动物病原生物学福建省高校重点实验室，福建　福州　350002

基金项目: 国家自然科学基金项目（U23A20235）；福建省自然科学基金项目（2020J06016）；福建农林大学科技创新专项基金（KFb22063XA）

详细信息

作者简介:
彭本群（1999 —），男，硕士研究生，主要从事预防兽医学研究，E-mail：enjamin_Pong@outlook.com

通讯作者:
王松（1983 —），男，博士，教授，主要从事动物病原微生物与免疫学研究，E-mail：wscookie@163.com

中图分类号: S852.65
计量
- 文章访问数: 43
- HTML全文浏览量: 33
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2024-01-03
- 修回日期: 2024-02-29
- 网络出版日期: 2024-03-28

Preliminary Research on the Mechanism of SOCS3 Negatively Regulating the Expression of Interferons Induced by Influenza Virus

PENG Benqun^{1, 2, 3
,},
HU Jingyun^{1, 2, 3},
MAO Yanan^{1, 2},
WANG Shulin^{1, 2},
WANG Jiajun^{1, 3},
CHEN Mengying^{1, 3},
YOU Dongxue^{1, 3},
WANG Song^{1, 2, 3
, ,}

1.
College of Animal Sciences, Fujian Agriculture and Forestry University, Fuzhou, , Fujian　350002, China
2.
Key Laboratory of Animal Pathogen Infection and Immunology of Fujian Province, Fuzhou, Fujian　350002, China
3.
Key Laboratory of Fujian-Taiwan Animal Pathogen Biology, Fuzhou, Fujian　350002, China

摘要

摘要: 目的探究细胞因子信号转导抑制因子3（Suppressor of cytokine signaling 3, SOCS3）在流感病毒（Influenza virus）感染过程中对干扰素（Interferon, IFN）信号通路的调控作用。方法通过慢病毒感染的方式在人非小细胞肺癌细胞（A549细胞）中过表达SOCS3，利用siRNA技术在A549细胞中敲低SOCS3的表达，然后使用流感病毒感染SOCS3过表达或敲低的细胞系以及对照细胞系，于不同时间点收取RNA或蛋白样品，采用RT-PCR和Western blot技术检测干扰素信号通路中关键节点分子的表达或活化情况。结果检测发现细胞中过表达SOCS3后I型干扰素IFN-β和III型干扰素IL-28（Interleukin-28）、IL-29（Interleukin-29）表达水平下降，而敲低SOCS3表达后IFN-β及IL-28、IL-29的表达水平升高。进一步研究发现，过表达SOCS3对识别流感病毒RNA的模式识别受体视黄酸诱导基因-I（Retinoic acid-inducible gene 1, RIG-I）、黑色素瘤分化相关蛋白5（Melanoma differentiation-associated protein 5, MDA5）、Toll样受体3（Toll-like receptor 3, TLR3）以及干扰素调节因子7（Interferon regulatory factor 7, IRF7）的mRNA表达均有抑制作用。同时SOCS3还能影响干扰素下游信号转导与转录激活因子 1（Signal transducer and activator of transcription 1, STAT1）的活化，细胞中过表达SOCS3抑制流感病毒诱导的STAT1的磷酸化，而敲低SOCS3表达则使STAT1的磷酸化水平升高。结论流感病毒感染后，SOCS3能够在转录水平下调模式识别受体及干扰素调节因子的表达，抑制Ⅰ型和Ⅲ型干扰素的产生，同时还影响STAT1的活化从而阻断干扰素信号的传递。
- 流感病毒 /
- SOCS3 /
- 干扰素 /
- 天然免疫 /
- STAT1
Abstract: Objective The regulatory impact of SOCS3 on the interferon signaling pathway during influenza virus infection was studied. Method The A549 cell lines with SOCS3 overexpression and knockdown were constructed using lentivirus infection and siRNA technology, respectively. Subsequently, these cell lines, along with control cell lines, were infected with influenza virus. Samples were collected at different time points to assess the expression and activation of crucial molecules within the interferon signaling pathway using RT-PCR and Western blot analysis. Result The findings demonstrated a decrease in expression levels of type I interferon IFN-β and type III interferons IL-28 and IL-29 following the overexpression of SOCS3 in cells. Conversely, the expression levels of IFN-β, IL-28, and IL-29 were observed to increase upon the knockdown of SOCS3 expression. Subsequent investigations indicated that the overexpression of SOCS3 suppressed the expression of interferon regulatory factor IRF7, as well as pattern recognition receptors RIG-I, MDA5, and TLR3 responsible for detecting influenza virus RNA. Additionally, the phosphorylation level of STAT1 after virus infection was detected, and the results showed that overexpression of SOCS3 in cells inhibited the phosphorylation of STAT1 induced by influenza virus, while knocking down the expression of SOCS3 increased the phosphorylation level of STAT1. Conclusion Upon influenza virus infection, SOCS3 down-regulated the mRNA expression of pattern recognition receptors and interferon regulatory factor, inhibited the production of type I and III interferons, as well as the activation of STAT1. This ultimately led to the blocking of interferon signal transmission.
- influenza virus /
- SOCS3 /
- interferon /
- innate immunity /
- STAT1

HTML全文

图 1 SOCS3抑制流感病毒感染后干扰素的产生

Figure 1. SOCS3 inhibits the production of interferons during influenza virus infection

下载: 全尺寸图片幻灯片

图 2 SOCS3抑制模式识别受体的表达

Figure 2. SOCS3 inhibits the expression of pattern recognition receptors

下载: 全尺寸图片幻灯片

图 3 SOCS3抑制IRF7的表达

Figure 3. SOCS3 inhibits the expression of IRF7

下载: 全尺寸图片幻灯片

图 4 SOCS3抑制流感病毒诱导的STAT1磷酸化

Figure 4. SOCS3 inhibits the phosphorylation of STAT1 induced by influenza virus infection

下载: 全尺寸图片幻灯片

表 1 RT-PCR引物序列

Table 1. Sequences of primers for RT-PCR

引物 Primer	上游引物 5′-3′ Forward primer 5′-3′	下游引物 5′-3′ Reverse primer 5′-3′
GAPDH	TGGGTGTGAACCATGAGAAGT	AAGGCCATGCCAGTGAGCTT
NP (Influenza virus)	TCAAACGTGGGATCAATG	GTGCAGACCGTGCTAAAA
SOCS3	CAGTACGATGCCCCGCTTTA	GAAGCTGGAGACTCAGGTGG
IFN-β	GCACTGGCTGGAATGAGACTA	ACTCTGACTATGGTCCAGGCA
IL-28	CCTGGTGGACGTCTTGGAC	GCGGAAGAGGTTGAAGGTGA
IL-29	TGGTGACTTTGGTGCTAGGC	GGCCTTCTTGAAGCTCGCTA
RIG-I	GAAGATCCAGAATGCCAGAATC	CTGGGATCTGATTCGCAAAA
MDA5	CTGCTGCAGAAAACAATGGA	TGGCTGAACTGTGGTTGAAA
TLR3	AGTGCCGTCTATTTGCCACA	GCATCCCAAAGGGCAAAAGG
IRF3	GGGGAGTGATGAGCTACGTG	CTACAATGAAGGGCCCCAGG
IRF7	GAGCTGTGCTGGCGAGAAG	CTCTCCAGGAGCCTTGGTTG

下载: 导出CSV

参考文献(19)

[1]	HUTCHINSON E C. Influenza virus [J]. Trends in Microbiology, 2018, 26(9): 809−810. doi: 10.1016/j.tim.2018.05.013
[2]	UYEKI T M, HUI D S, ZAMBON M, et al. Influenza [J]. Lancet, 2022, 400(10353): 693−706. doi: 10.1016/S0140-6736(22)00982-5
[3]	GORAYA M U, ZAIGHUM F, SAJJAD N, et al. Web of interferon stimulated antiviral factors to control the influenza A viruses replication [J]. Microbial Pathogenesis, 2020, 139: 103919. doi: 10.1016/j.micpath.2019.103919
[4]	WONG P T, GOFF P H, SUN R J, et al. Combined intranasal nanoemulsion and RIG-I activating RNA adjuvants enhance mucosal, humoral, and cellular immunity to influenza virus [J]. Molecular Pharmaceutics, 2021, 18(2): 679−698. doi: 10.1021/acs.molpharmaceut.0c00315
[5]	MA W, HUANG G, WANG Z, et al. IRF7: Role and regulation in immunity and autoimmunity [J]. Frontiers in Immunology, 2023, 14: 1236923. doi: 10.3389/fimmu.2023.1236923
[6]	YAO D D, BAO L L, LI F D, et al. H1N1 influenza virus dose dependent induction of dysregulated innate immune responses and STAT1/3 activation are associated with pulmonary immunopathological damage [J]. Virulence, 2022, 13(1): 1558−1572. doi: 10.1080/21505594.2022.2120951
[7]	HALLER O, KOCHS G. Mx genes: Host determinants controlling influenza virus infection and trans-species transmission [J]. Human Genetics, 2020, 139(6): 695−705.
[8]	周斌, 万少兵, 王瑛, 等. SOCS3通过调控JAK2/STAT3信号通路改善急性肺损伤 [J]. 山西医科大学学报, 2023, 54(6):778−784. ZHOU B, WAN S B, WANG Y, et al. SOCS3 improves acute lung injury by regulating JAK2/STAT3 signaling pathway [J]. Journal of Shanxi Medical University, 2023, 54(6): 778−784. (in Chinese)
[9]	SIMS N A. The JAK1/STAT3/SOCS3 axis in bone development, physiology, and pathology [J]. Experimental & Molecular Medicine, 2020, 52: 1185−1197.
[10]	LIU S S, YAN R X, CHEN B, et al. Influenza virus-induced robust expression of SOCS3 contributes to excessive production of IL-6 [J]. Frontiers in Immunology, 2019, 10: 1843. doi: 10.3389/fimmu.2019.01843
[11]	SUN Y P, JIANG J W, PO T E, et al. IFN-λ: A new spotlight in innate immunity against influenza virus infection [J]. Protein & Cell, 2018, 9(10): 832−837.
[12]	BIONDO C, LENTINI G, BENINATI C, et al. The dual role of innate immunity during influenza [J]. Biomedical Journal, 2019, 42(1): 8−18. doi: 10.1016/j.bj.2018.12.009
[13]	HUANG B Z, CHEN H P, ZHENG Y B. MiR-103/miR-107 inhibits enterovirus 71 replication and facilitates type I interferon response by regulating SOCS3/STAT3 pathway [J]. Biotechnology Letters, 2021, 43(7): 1357−1369. doi: 10.1007/s10529-021-03115-z
[14]	YAKASS M B, FRANCO D, QUAYE O. Suppressors of cytokine signaling and protein inhibitors of activated signal transducer and activator of transcriptions As therapeutic targets in flavivirus infections [J]. Journal of Interferon & Cytokine Research, 2020, 40(1): 1−18.
[15]	LI L, WU H Y, LI Q M, et al. SOCS3-deficient lung epithelial cells uptaking neutrophil-derived SOCS3 worsens lung influenza infection [J]. Molecular Immunology, 2020, 125: 51−62. doi: 10.1016/j.molimm.2020.06.022
[16]	YANG H, DONG Y R, BIAN Y, et al. The influenza virus PB2 protein evades antiviral innate immunity by inhibiting JAK1/STAT signalling [J]. Nature Communications, 2022, 13: 6288. doi: 10.1038/s41467-022-33909-2
[17]	ZHANG Y L, XU L L, ZHANG Z, et al. Enterovirus D68 infection upregulates SOCS3 expression to inhibit JAK-STAT3 signaling and antagonize the innate interferon response of the host [J]. Virologica Sinica, 2023, 38(5): 755−766. doi: 10.1016/j.virs.2023.08.007
[18]	WANG X W, JIA Y Q, REN J, et al. Newcastle disease virus nonstructural V protein upregulates SOCS3 expression to facilitate viral replication depending on the MEK/ERK pathway [J]. Frontiers in Cellular and Infection Microbiology, 2019, 9: 317. doi: 10.3389/fcimb.2019.00317
[19]	XIE J Y, WANG M S, CHENG A C, et al. DHAV-1 inhibits type I interferon signaling to assist viral adaption by increasing the expression of SOCS3 [J]. Frontiers in Immunology, 2019, 10: 731. doi: 10.3389/fimmu.2019.00731