猪繁殖与呼吸综合征病毒M蛋白的ITIM基序对IFN-β产生的正调节作用

doi:10.11843/j.issn.0366-6964.2018.12.017

Abstract

Abstract:

To study the activation of ITIM motif of PRRSV M protein to IFN-β production, the full-length ORF6 gene fragment and the full-length ORF6 gene fragment without ITIM motif were amplified using the PRRSV Hn-1/06 cDNA as template by PCR method, respectively. The two gene fragments were respectively inserted into the eukaryotic expression vector pcDNA3.0 to yield recombinant plasmids, designated by pcDNA3.0-GP6 and pcDNA3.0-GP6ΔITIM (lacking of ITIM motif). Then, the two recombinant plasmids were used to transfected the Marc-145 cells, respectively. The TRIF, MyD88, TRAF6, IRF3, IRF7, NF-κB and IFN-β mRNA levels of the transfected Marc-145 cells were detected by qRT-PCR. The results showed that the TRIF, IRF3 and IRF7 mRNA levels of the Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were up-regulated significantly (This is the general trend,which may not be completely consistent with individual time points.The same as below), while the MyD88 and TRAF6 mRNA levels of the Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were down-regulated significantly, when compared with the Marc-145 cells transfected with pcDNA3.0-GP6ΔITIM recombinant plasmid. The two recombinant plasmids were used to transfected the Marc-145 cells transfected with Poly (I:C), respectively. The TRIF, IRF3, IRF7, NF-κB and IFN-β mRNA levels of the transfected Marc-145 cells were detected by qRT-PCR. The results showed that the TRIF, IRF3 and IRF7 mRNA levels of the Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were up-regulated significantly, while the NF-κB and IFN-β mRNA levels of the Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were up-regulated significantly at 36 h, when compared with the Marc-145 cells transfected with pcDNA3.0-GP6ΔITIM recombinant plasmid. The recombinant plasmids pcDNA3.0-GP6 and pcDNA3.0-GP6△ITIM were respectively transfected into the Marc-145 cells silenced with SHP-1-siRNA-781 or SHP-2-siRNA-1335, then the TRIF, IRF3, IRF7, NF-κB and IFN-β mRNA levels of the transfected Marc-145 cells were detected by qRT-PCR. The TRIF, IRF3, IRF7,NF-κB and IFN-β mRNA levels of the SHP-1 silenced Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were down-regulated significantly at any timepoints, compared with the SHP-1 silenced Marc-145 cells transfected with pcDNA3.0-GP6ΔITIM recombinant plasmid, the TRIF, IRF3, IRF7, NF-κB and IFN-β mRNA levels of the SHP-2 silenced Marc-145 cells transfected with pcDNA3.0-GP6 recombinant plasmid were down-regulated significantly at early phase, then recovery to normal levels at later phase, when compared with the SHP-2 silenced Marc-145 cells transfected with pcDNA3.0-GP6ΔITIM recombinant plasmid. In summary, the ITIM motif of PRRSV M protein may induce the IFN-β production by interacting with SHP-1 in TLR3-TRIF-dependent signaling pathway, however the relationship between ITIM motif and SHP-2 needs to further study. This study provides an important reference for understandin the role of ITIM motif.

CLC Number:

S852.659.6

WEI Feng-ling, ZHANG Ying-ying, XU Rui-qin, LI Wen, LI Xiang-tong, SUN Yang-yang, ZHANG Liu-jun, YANG Guo-yu, XIA Ping-an, ZHANG Gai-ping. Positive Regulation of ITIM of PRRSV M Protein to IFN-β Production[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2680-2689.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.xmsyxb.com/EN/10.11843/j.issn.0366-6964.2018.12.017

https://www.xmsyxb.com/EN/Y2018/V49/I12/2680

References

[1] 陈溥言.兽医传染病学[M].5版.北京:中国农业出版社,2006.
CHEN P Y. Veterinary lemology[M]. 5th ed.Beijing:China Agriculture Press,2006.(in Chinese)
[2] DEA S, GAGNON C A, MARDASSI H, et al. Current knowledge on the structural proteins of porcine reproductive and respiratory syndrome (PRRS) virus:Comparison of the North American and European isolates[J]. Arch Virol, 2000, 145(4):659-688.
[3] SNIJDER E J, KIKKERT M, FANG Y. Arterivirus molecular biology and pathogenesis[J]. J Gen Virol, 2013, 94(Pt 10):2141-2163.
[4] YOO D, WOOTTON S K, LI G, et al.Colocalization and interaction of the porcine arterivirus nucleocapsid protein With the small nucleolar RNA-associated protein fibrillarin[J]. J Virol, 2003, 77(22):12173-12183.
[5] KIMMAN T G, CORNELISSEN L A, MOORMANN R J, et al. Challenges for porcine reproductive and respiratory syndrome virus (PRRSV) vaccinology[J].Vaccine, 2009, 27(28):3704-3718.
[6] BEURA L K, SARKAR S N, KWON B, et al. Porcine reproductive and respiratory syndrome virus nonstructural protein 1β modulates host innate immune response by antagonizing IRF3 activation[J]. J Virol, 2010, 84(3):1574-1584.
[7] LUO R,XIAO S B,JIANG Y B, et al. Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses interferon-β production by interfering with the RIG-I signaling pathway[J]. Mol Immunol, 2008, 45(10):2839-2846.
[8] GAO J M, JI P C, ZHANG M D, et al. GP5 expression in Marc-145 cells inhibits porcine reproductive and respiratory syndrome virus infection by inducing beta interferon activity[J].Vet Microbiol, 2014,174(3-4):409-418.
[9] STEBBING R E, IRVIN S C, RIVERA-SERRANO E E, et al. An ITAM in a nonenveloped virus regulates activation of NF-κB, induction of beta interferon, and viral spread[J].J Virol, 2014, 88(5):2572-2583.
[10] ALOULOU M, BEN MKADDEM S, BIARNES-PELICOT M, et al. IgG1 and IVIg induce inhibitory ITAM signaling through FcγRⅢ controlling inflammatory responses[J]. Blood, 2012, 119(13):3084-3096.
[11] FUCHS A, CELLA M, KONDO T, et al. Paradoxic inhibition of human natural interferon-producing cells by the activating receptor NKp44[J].Blood, 2005, 106(6):2076-2082.
[12] CHRISTOPHI G P, PANOS M, HUDSON C A, et al. Interferon-β treatment in multiple sclerosis attenuates Inflammatory gene expression through inducible activity of the phosphatase SHP-1[J]. Clin Immunol, 2009, 133(1):27-44.
[13] AN H Z, HOU J, ZHOU J, et al. Phosphatase SHP-1 promotes TLR-and RIG-I-Activated production of type I interferon by inhibiting the kinase IRAK1[J].Nat Immunol, 2008, 9(5):542-550.
[14] KIM E J, LEE S M, SUK K, et al.CD300a and CD300f differentially regulate the MyD88 and TRIF-mediated TLR signalling pathways through activation of SHP-1 and/or SHP-2 in human monocytic cell lines[J]. Immunology, 2012, 135(3):226-235.
[15] AN H Z, ZHAO W, HOU J, et al. SHP-2 Phosphatase negatively regulates the TRIF adaptor protein-dependent type I interferon and proinflammatory cytokine production[J].Immunity, 2006, 25(6):919-928.
[16] HUANG C, ZHANG Q, GUO X K, et al. Porcine reproductive and respiratory syndrome virus nonstructural protein 4 antagonizes beta interferon expression by targeting the NF-κB essential modulator[J]. J Virol, 2014, 88(18):10934-10945.
[17] ZHANG Y N, ZHOU Y H, YANG Q Y, et al. Ligation of Fc gamma receptor ⅡB enhances levels of antiviral cytokine in response to PRRSV infection in vitro[J].Vet Microbiol, 2012, 160(3-4):473-480.
[18] RAHIM M M A, TAI L H, TROKE A D, et al.Ly49Q positively regulates type I IFN production by plasmacytoid dendritic cells in an immunoreceptor tyrosine-based inhibitory motif-dependent manner[J]. J Immunol, 2013, 190(8):3994-4004.
[19] KASAMATSU J,DENG M Y,AZUMA M,et al.Double-stranded RNA analog and type I interferon regulate expression of Trem paired receptors in murine myeloid cells[J].BMC Immunol,2016,17(1):9.
[20] BRUNI D, SEBASTIA J, DUNNE S, et al. A novel IRAK1-IKKε signaling axis limits the activation of TAK1-IKKβ downstream of TLR3[J]. J Immunol, 2013, 190(6):2844-2856.
[21] CHONG Z Z, MAIESE K. The Src homology 2 domain tyrosine phosphatases SHP-1 and SHP-2:Diversified control of cell growth, inflammation, and injury[J].Histol Histopathol, 2007, 22(11):1251-1267.

Positive Regulation of ITIM of PRRSV M Protein to IFN-β Production

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	HUANG Jin, LI Siyuan, MAO Li, CAI Xuhang, XIE Lingling, WANG Fu, ZHOU Hua, LI Jizong, LI Bin. Eukaryotic Expression of Bovine Coronavirus S1 Protein and Establishment and Application of Indirect ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2050-2060.
[2]	MA Rumeng, ZHAO Yuliang, MA Mingshuang, GUO Guihai, LIU Xinzi, LI Jiaxuan, CUI Wen, JIANG Yanping, SHAN Zhifu, ZHOU Han, WANG Li, QIAO Xinyuan, TANG Lijie, WANG Xiaona, LI Yijing. Comparative Study on the Immune Response Induced by the Different Porcine Receptor Bacteria with Expressing the Protective Antigen S1 of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2090-2099.
[3]	XU Hong, SHANG Hongqi, ZHANG Xue, QIAN Jiali, WANG Chuanhong, SONG Xu, BAO Meiying, LIU Shiyu, ZHANG Gege, GUO Rongli, ZHAO Yongxiang, FAN Baochao, LI Bin. Inhibition Effect of C8orf4 Gene Encoding Protein on in vitro Replication of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2100-2108.
[4]	XIONG Ting, HE Xianming, ZHAO Xiya, ZHUANG Tingting, HUANG Meizhen, LIANG Shijin, YU Chuanzhao, LIANG Xuejing, CHEN Ruiai. Whole Genome Analysis of Three Predominant Epidemic Strains of Chicken Infectious Bronchitis Virus and Their Pathogenicity [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2109-2122.
[5]	WANG Jing, ZHANG Shujuan, HU Xia, LIU Xiangyang, ZHANG Xingcui, SONG Zhenhui. CD44 Regulates Na⁺/H⁺ Exchanger 3 Activity by Influencing Porcine Epidemic Diarrhea Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2176-2185.
[6]	LIN Lili, ZHANG Mengdi, ZHU Linlin, MA Hailong, SUN Qi, HE Qigai, ZHANG Mengjia, LI Wentao. Establishment of Neutralizing Antibody Detection Method based on Recombinant Fluorescent Virus of Porcine Epidemic Diarrhea Virus GⅡb Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1649-1660.
[7]	JING Yang, WANG Yumiao, LI Yang, CHANG Hui, MA Zhiqian, LI Zhiwei, XIAO Shuqi. Construction of MARC-145^ORF6 Cell Line Stably Overexpressing PRRSV M Protein and Its Effect on PRRSV Proliferation [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1159-1169.
[8]	YU Qisheng, ZHU Qing, ZHOU Qun, SONG Xin, ZHANG Jiaqi, CHEN Taoyun, XU Lin, ZHANG Chaohui, ZHANG Bin. Expression of BCoV Spike Protein by Baculovirus Expression System and Its Immunogenicity in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 640-648.
[9]	ZHANG Jinpeng, CHEN Cuiteng, LIN Lin, FU Huanru, LI Zhaolong, JIANG Bin, HUANG Yu, WAN Chunhe. Establishment of a Real-time Fluorescent RT-PCR Assay for Detection of Pigeon Megrivirus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 860-866.
[10]	SONG Wenyan, ZHANG Hanwen, WU Aodi, ZHANG Liyan, LIU Zhao, YE Tongtong, CHEN Chuangfu, SHENG Jinliang. Screening of Nanobodies against Porcine Reproductive and Respiratory Syndrome Virus GP5 Protein and Exploration of Their Inhibitory Effect on Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 258-270.
[11]	WANG Ying, LI Jiakang, ZENG Yue, SHI Kaituo, WEI Liting, PENG Jiajia, LI Qiuyan, CAO Longlong, ZHOU Dengyuan. Development of Monoclonal Antibodies against Feline Calicivirus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 401-405.
[12]	LI Huihui, DONG Keer, LIU Xinbo, ZHANG Chunxiao, MA Chao, CHEN Liping, ZHONG Qi, YAO Gang, MA Xuelian. Establishment and Application of Rapid Detection Method for Bovine Norovirus and Bovine Rotavirus Dual RAA-LFD [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 406-412.
[13]	ZHOU Jianhao, WANG Dongfang, LIU Ying, WANG Shujuan, MA Zhenyuan, XIE Caihua, ZHAO Xueli, YANG Haibo, FENG Guidan, KANG Taisheng, HU Yufeng, LI Bowen, YAN Ruoqian. Establishment and Preliminary Application of a Quantitative Droplet Digital PCR Assay for Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 413-418.
[14]	ZHANG Jiaqi, JIA Xining, ZHOU Qun, SONG Xin, ZHU Chenxi, LI Gege, ZHANG Bin. Construction of BVDV-1 Virus-like Particles based on E2 Protein and Evaluation of Its Immune Efficacy in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 5143-5153.
[15]	ZHOU Likun, DONG Xinyuan, LI Jiahui, CUI Zhiying, ZHAO Shijie, XU Jie, CHEN Jing, ZHANG Yina, XIA Ping’an. Regulatory Role of MID2 on Porcine Reproductive and Respiratory Syndrome Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(11): 4735-4744.