猪源组织蛋白酶S抑制O型口蹄疫病毒在PK-15细胞复制

doi:10.11843/j.issn.0366-6964.2021.06.019

Abstract

Abstract: The previous research data showed that the expression level of cathepsin S (CTSS) in sow colostrum was significantly higher than that of mature milk and CTSS has the effect of inhibiting virus replication. This study aims to explore the effect of pig-derived CTSS on the replication of foot-and-mouth disease virus serotype O (FMDV-O) and the production of antiviral cytokines induced by FMDV-O.PK-15 cells were infected with FMDV-O. The effect of FMDV-O infection on the expression of endogenous CTSS was investigated by RT-qPCR and Western blot at the transcriptional and translational level, respectively, and the effect of FMDV-O infection on the enzyme activity of CTSS in vitro was determined by CTSS activity detection kit. According to the CTSS gene sequence (XM_021089893.1) published by GenBank, the eukaryotic expression plasmid of CTSS was constructed and transfected into PK-15 cells by liposome. The expression of CTSS was detected by Western blot. On this basis, the effect of overexpressed CTSS on FMDV-O replication and the level of antiviral cytokine mRNA induced by FMDV-O was detected by Western blot and RT-qPCR, and three pairs of siRNA specific to CTSS were designed and synthesized. Western blot and RT-qPCR were used to detect the changes of CTSS and FMDV-O. The results showed that PK-15 cells infected with FMDV-O could significantly up-regulate the expression of porcine CTSS and enhance the activity of CTSS enzyme; overexpression of CTSS could inhibit the replication of FMDV-O in PK-15 cells, which may be through promoting the production of antiviral cytokines induced by FMDV-O; the interference sequence siRNA-2947 down-regulated the expression of endogenous CTSS and promoted the replication of FMDV-O. Porcine CTSS promoting the production of host antiviral cytokines may be one of the reasons for inhibiting FMDV-O replication. This study provides a basis for further exploring the role and mechanism of host CTSS in anti-FMDV innate immune response.

Key words: cathepsin S, FMDV-O, PK-15 cell, antiviral function

CLC Number:

S852.659.6

SHI Xijuan, LIU Yuanzi, ZHANG Dajun, HOU Jing, SHEN Chaochao, YANG Bo, ZHANG Ting, YUAN Xingguo, REN Ruirui, DU Xiaohua, ZHANG Keshan, ZHENG Haixue, LIU Xiangtao. Porcine Cathepsin S Inhibits the Replication of Foot-and-Mouth Disease Virus Serotype O in PK-15 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(6): 1652-1661.

References 42

[1]	ALEXANDERSEN S, MOWAT N. Foot-and-mouth disease:host range and pathogenesis[M]//MAHY B W J. Foot-and-Mouth Disease Virus. Berlin, Heidelberg:Springer, 2005:9-42.
[2]	AL AMIN M, ALI M R, ISLAM M R, et al. Development and serology based efficacy assessment of a trivalent foot-and-mouth disease vaccine[J]. Vaccine, 2020, 38(32):4970-4978.
[3]	CROFT S, AEGERTER J N, MASSEI G, et al. The risk of foot-and-mouth disease becoming endemic in a wildlife host is driven by spatial extent rather than density[J]. PLoS One, 2019, 14(6):e0218898.
[4]	YANG F, ZHU Z X, CAO W J, et al. Genetic determinants of altered virulence of type O foot-and-mouth disease virus[J]. J Virol, 2020, 94(7):e01657-19.
[5]	ZHAO F R, XIE Y L, LIU Z Z, et al. Lithium chloride inhibits early stages of foot-and-mouth disease virus (FMDV) replication in vitro[J]. J Med Virol, 2017, 89(11):2041-2046.
[6]	MASON P W, GRUBMAN M J, BAXT B. Molecular basis of pathogenesis of FMDV[J]. Virus Res, 2003, 91(1):9-32.
[7]	SUN C, LIU M M, CHANG J T, et al. Heterogeneous nuclear ribonucleoprotein L negatively regulates foot-and-mouth disease virus replication through inhibition of viral RNA synthesis by interacting with the internal ribosome entry site in the 5' untranslated region[J]. J Virol, 2020, 94(10):e00282-20.
[8]	BUSCH G K, TATE E W, GAFFNEY P R J, et al. Specific N-terminal protein labelling:use of FMDV 3Cpro protease and native chemical ligation[J]. Chem Commun (Camb), 2008, (29):3369-3371.
[9]	LIN Y, HUNG C Y, BHATTACHARYA C, et al. An effective way of producing fully assembled antibody in transgenic tobacco plants by linking heavy and light chains via a self-cleaving 2A peptide[J]. Front Plant Sci, 2018, 9:1379.
[10]	DUAN X, SUN P, LAN Y, et al. 1IFN-α modulates memory Tfh cells and memory B cells in mice, following recombinant FMDV adenoviral challenge[J]. Front Immunol, 2020, 11:701.
[11]	WANG D, FANG L R, LUO R, et al. Foot-and-mouth disease virus leader proteinase inhibits dsRNA-induced type I interferon transcription by decreasing interferon regulatory factor 3/7 in protein levels[J]. Biochem Biophys Res Commun, 2010, 399(1):72-78.
[12]	LI D, LEI C Q, XU Z S, et al. Foot-and-mouth disease virus non-structural protein 3A inhibits the interferon-β signaling pathway[J]. Sci Rep, 2016, 6(1):21888.
[13]	NGUYEN T A, PANG K C, MASTERS S L. Intercellular communication for innate immunity[J]. Mol Immunol, 2017, 86:16-22.
[14]	GLADUE D P, O'DONNELL V, BAKER-BRANSETTER R, et al. Interaction of foot-and-mouth disease virus nonstructural protein 3A with host protein DCTN3 is important for viral virulence in cattle[J]. J Virol, 2014, 88(5):2737-2747.
[15]	BARARIA D, HILDEBRAND J A, STOLZ S, et al. Cathepsin S alterations induce a tumor-promoting immune microenvironment in follicular lymphoma[J]. Cell Rep, 2020, 31(5):107522.
[16]	ZHANG J, SHAN Y, LI Y, et al. Palmitate impairs angiogenesis via suppression of cathepsin activity[J]. Mol Med Rep, 2017, 15(6):3644-3650.
[17]	NI H E, XU S, CHEN H W, et al. Nicotine modulates CTSS (Cathepsin S) synthesis and secretion through regulating the autophagy-lysosomal machinery in atherosclerosis[J]. Arterioscler, Thromb, Vasc Biol, 2020, 40(9):2054-2069.
[18]	WIENER J J M, WICKBOLDT JR A T, WIENER D K, et al. Discovery and SAR of novel pyrazole-based thioethers as cathepsin S inhibitors. Part 2:modification of P3, P4, and P5 regions[J]. Bioorg Med Chem Lett, 2010, 20(7):2375-2378.
[19]	Cathepsin S protease mediates T-cell response in non-hodgkin lymphoma[J]. Cancer Discovery, 2020, 10(6):761.
[20]	HSIEH M J, LIN C W, CHEN M K, et al. Inhibition of cathepsin S confers sensitivity to methyl protodioscin in oral cancer cells via activation of p38 MAPK/JNK signaling pathways[J]. Scientific Reports, 2017, 7(1):45039.
[21]	CHANG Z Y, ZHAO G Z, ZHAO Y, et al. BAF60a deficiency in vascular smooth muscle cells prevents abdominal aortic aneurysm by reducing inflammation and extracellular matrix degradation[J]. Arterioscler, Thromb, Vasc Biol, 2020, 40(10):2494-2507.
[22]	RIESE R J, WOLF P R, BRÖMME D, et al. Essential role for cathepsin S in MHC class II-associated invariant chain processing and peptide loading[J]. Immunity, 1996, 4(4):357-366.
[23]	KIM S J, SCHÄTZLE S, AHMED S S, et al. Increased cathepsin S in Prdm1-/- dendritic cells alters the T cell repertoire and contributes to lupus[J]. Nat Immunol, 2017, 18(9):1016-1024.
[24]	FIGUEIREDO J L, AIKAWA M, ZHENG C Y, et al. Selective cathepsin S inhibition attenuates atherosclerosis in apolipoprotein E-deficient mice with chronic renal disease[J]. Am J Pathol, 2015, 185(4):1156-1166.
[25]	KIM S Y, JIN H, SEO H R, et al. Regulating BRCA1 protein stability by cathepsin S-mediated ubiquitin degradation[J]. Cell Death Differ, 2019, 26(5):812-825.
[26]	IMUS J K, LEHMKUHL H D, WOODS L W. Resistance of colostrum-deprived domestic lambs to infection with deer adenovirus[J]. J Vet Diagn, 2019, 31(1):78-82.
[27]	CONUS S, SIMON H U. Cathepsins and their involvement in immune responses[J]. Swiss Med Wkly, 2010, 140:w13042.
[28]	STEIMLE A, GRONBACH K, BEIFUSS B, et al. Symbiotic gut commensal bacteria act as host cathepsin S activity regulators[J]. J Autoimmun, 2016, 75:82-95.
[29]	GLADUE D P, O'DONNELL V, BAKER-BRANSTETTER R, et al. Foot-and-mouth disease virus nonstructural protein 2C interacts with beclin1, modulating virus replication[J]. J Virol, 2012, 86(22):12080-12090.
[30]	ZHANG W, YANG F, ZHU Z X, et al. Cellular DNAJA3, a novel VP1-interacting protein, inhibits foot-and-mouth disease virus replication by inducing lysosomal degradation of VP1 and attenuating its antagonistic role in the beta interferon signaling pathway[J]. J Virol, 2019, 93(13):e00588-19.
[31]	FENG H H, ZHU Z X, CAO W J, et al. Foot-and-mouth disease virus induces lysosomal degradation of NME1 to impair p53-regulated interferon-inducible antiviral genes expression[J]. Cell Death Dis, 2018, 9(9):885.
[32]	ZHU Z X, WANG G Q, YANG F, et al. Foot-and-mouth disease virus viroporin 2B antagonizes RIG-I-mediated antiviral effects by inhibition of its protein expression[J]. J Virol, 2016, 90(24):11106-11121.
[33]	FU R Z, GUO H, JANGA S, et al. Cathepsin S activation contributes to elevated CX3CL1(fractalkine) levels in tears of a Sj gren's syndrome murine model[J]. Sci Rep, 2020, 10(1):1455.
[34]	KLINNGAM W, FU R Z, JANGA S R, et al. Cathepsin S alters the expression of pro-inflammatory cytokines and MMP-9, partially through protease-Activated receptor-2, in human corneal epithelial cells[J]. Int J Mol Sci, 2018, 19(11):3530.
[35]	PAN L L, LI Y L, JIA L X, et al. Cathepsin S deficiency results in abnormal accumulation of autophagosomes in macrophages and enhances Ang II-induced cardiac inflammation[J]. PLoS One, 2012, 7(4):e35315.
[36]	DE MINGO PULIDO Á, DE GREGORIO E, CHANDRA S, et al. Differential role of cathepsins S and B in hepatic APC-mediated NKT cell activation and cytokine secretion[J]. Front Immunol, 2018, 9:391.
[37]	ZHANG K S, YAN M H, HAO J H, et al. Foot-and-mouth disease virus structural protein VP1 destroys the stability of TPL2 trimer by degradation TPL2 to evade host antiviral immunity[J]. J Virol, 2021, 95(6):e02149-20.
[38]	LI D, WEI J, YANG F, et al. Foot-and-mouth disease virus structural protein VP3 degrades Janus kinase 1 to inhibit IFN-γ signal transduction pathways[J]. Cell Cycle, 2016, 15(6):850-860.
[39]	DE LOS SANTOS T, DE AVILA BOTTON S, WEIBLEN R, et al. The leader proteinase of foot-and-mouth disease virus inhibits the induction of beta interferon mRNA and blocks the host innate immune response[J]. J Virol, 2006, 80(4):1906-1914.
[40]	VISSER L J, ALOISE C, SWATEK K N, et al. Dissecting distinct proteolytic activities of FMDV Lpro implicates cleavage and degradation of RLR signaling proteins, not its deISGylase/DUB activity, in type I interferon suppression[J]. PLoS Pathog, 2020, 16(7):e1008702.
[41]	MEDINA G N, KNUDSEN G M, GRENINGER A L, et al. Interaction between FMDV Lpro and transcription factor ADNP is required for optimal viral replication[J]. Virology, 2017, 505:12-22.
[42]	LI D, ZHANG J, YANG W P, et al. Poly (rC) binding protein 2 interacts with VP0 and increases the replication of the foot-and-mouth disease virus[J]. Cell Death Dis, 2019, 10(7):516.

Porcine Cathepsin S Inhibits the Replication of Foot-and-Mouth Disease Virus Serotype O in PK-15 Cells

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