非洲猪瘟病毒多基因家族MGF360-13L基因功能的初步研究

doi:10.11843/j.issn.0366-6964.2022.12.027

Abstract

Abstract: The aim of this study was to investigate the function of MGF360-13L gene of African swine fever virus (ASFV) multigene family. The MGF360-13L gene was analyzed by bioinformatics software. The eukaryotic expressing plasmid pVAX1-MGF360-13L was constructed and transfected into 293T cells for expression. The MGF360-13L gene deleted strain ASFV-ΔMGF360-13L was constructed by homologous recombination. After infecting the gene deleted virus strain and parent virus strain (ASFV CN/GS/2018) in porcine alveolar macrophages (PAMs) with the same multiplicity of infection (MOI=0.01), the virus titer was calculated by erythrocyte adsorption test (HAD) and the in vitro growth curve was drawn; ASFV-ΔMGF360-13L and ASFV CN/GS/2018 were infected with porcine bone marrow macrophages (BMDM) with MOI=1. The expression levels of inflammatory factors IL-1β, IL-6 and TNF-α were determined by qPCR and ELISA. The results showed that, MGF360-13L gene was relatively conservative in ASFV gene type II. The coding protein belongs to non secretory type, without transmembrane region and with good hydrophilicity; The eukaryotic expression plasmid pVAX1-MGF360-13L was expressed by 293T cells; The MGF360-13L deleted strain ASFV-ΔMGF360-13L was successfully constructed. There was no significant difference between ASFV-ΔMGF360-13L and ASFV CN/GS/2018 in in vitro replication. After ASFV-ΔMGF360-13L infected BMDM, the levels of transcription and secretion of inflammatory cytokines IL-1β, IL-6 and TNF-α were significantly higher than those of ASFV CN/GS/2018. In this study, the MGF360-13L gene deleted strain of ASFV were successfully constructed, and it was confirmed as a non-essential gene for ASFV replication and have the function of inhibiting the expression of inflammatory factors. This study provides clues for further annotation of ASFV MGF360-13L gene function.

Key words: ASFV, MGF360-13L, inflammatory cytokines, growth curve measurement

CLC Number:

S852.651

CHEN Danian, MA Xusheng, DAI Junfei, WANG Yang, LI Qian, BAI Heng, MAO Tiantian, LIU Yongsheng, DING Long, CHEN Haohan, CHEN Siyan, RAO Yufei, JIA Ning, ZHANG Jie, ZHENG Haixue, LIU Xiangtao. Preliminary Study on the Function of MGF360-13L Gene of African Swine Fever Virus Multigene Family[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4419-4428.

References 23

[1]	DIXON L K,CHAPMAN D A G,NETHERTON C L,et al.African swine fever virus replication and genomics[J].Virus Res,2013,173(1):3-14.
[2]	MALOGOLOVKIN A,KOLBASOV D.Genetic and antigenic diversity of African swine fever virus[J].Virus Res,2019,271:197673.
[3]	GE S Q,LI J M,FAN X X,et al.Molecular characterization of African swine fever virus,China,2018[J].Emerg Infect Dis,2018,24(11):2131-2133.
[4]	CACKETT G,MATELSKA D,SÝKORA M,et al.The African swine fever virus transcriptome[J].J Virol,2020,94(9):e00119-20.
[5]	ZHU Z Z,CHEN H T,LIU L,et al.Classification and characterization of multigene family proteins of African swine fever viruses[J].Brief Bioinform,2021,22(4):bbaa380.
[6]	FARLOW J,DONDUASHVILI M,KOKHREIDZE M,et al.Intra-epidemic genome variation in highly pathogenic African swine fever virus (ASFV) from the country of Georgia[J].Virol J,2018,15(1):190.
[7]	DIXON L K,ISLAM M,NASH R,et al.African swine fever virus evasion of host defences[J].Virus Res,2019,266:25-33.
[8]	O’DONNELL V,HOLINKA L G,GLADUE D P,et al.African swine fever virus Georgia isolate harboring deletions of MGF360 and MGF505 genes is attenuated in swine and confers protection against challenge with virulent parental virus[J].J Virol,2015,89(11):6048-6056.
[9]	郭泽恒.非洲猪瘟病毒MGF360-13L蛋白通过cGAS-STING信号通路抑制Ⅰ型干扰素的产生[D].武汉:华中农业大学,2021.GUO Z H.African swine fever virus MGF360-13L inhibited type I interferon production through cGAS-STING pathway[D].Wuhan:Huazhong Agricultural University,2021.(in Chinese)
[10]	KUPRASH D V,NEDOSPASOV S A.Molecular and cellular mechanisms of inflammation[J].Biochemistry (Mosc),2016,81(11):1237-1239.
[11]	CORREIA S,VENTURA S,PARKHOUSE R M.Identification and utility of innate immune system evasion mechanisms of ASFV[J].Virus Res,2013,173(1):87-100.
[12]	LI D,YANG W P,LI L L,et al.African swine fever virus MGF-505-7R negatively regulates cGAS–STING-mediated signaling pathway[J].J Immunol,2021,206(8):1844-1857.
[13]	LI D,LIU Y G,QI X L,et al.African swine fever virus MGF-110-9L-deficient mutant has attenuated virulence in pigs[J].Virol Sin,2021,36(2):187-195.
[14]	潘力,罗瑞,王涛,等.非洲猪瘟病毒研究进展:组学视角[J].病毒学报,2022,38(2):246-256.PAN L,LUO R,WANG T,et al.Advances in African swine fever virus research:An omics view[J].Chinese Journal of Virology,2022,38(2):246-256.(in Chinese)
[15]	URBANO A C,FERREIRA F.Role of the DNA-binding protein pA104R in ASFV genome packaging and as a novel target for vaccine and drug development[J].Vaccines (Basel),2020,8(4):585.
[16]	O’DONNELL V,RISATTI G R,HOLINKA L G,et al.Simultaneous deletion of the 9GL and UK genes from the African swine fever virus Georgia 2007 isolate offers increased safety and protection against homologous challenge[J].J Virol,2016,91(1):e01760-16.
[17]	BORCA M V,RAMIREZ-MEDINA E,SILVA E,et al.Development of a highly effective African swine fever virus vaccine by deletion of the I177L gene results in sterile immunity against the current epidemic Eurasia strain[J].J Virol,2020,94(7):e02017-19.
[18]	RAMIREZ-MEDINA E,VUONO E A,RAI A,et al.Evaluation in swine of a recombinant African swine fever virus lacking the MGF-360-1L gene[J].Viruses,2020,12(10):1193.
[19]	RAMIREZ-MEDINA E,VUONO E,PRUITT S,et al.X69R is a non-essential gene that,when deleted from African swine fever,does not affect virulence in swine[J].Viruses,2020,12(9):918.
[20]	GALINDO I,ALONSO C.African swine fever virus:A review[J].Viruses,2017,9(5):103.
[21]	黄瑶,赵军,曾喻兵,等.非洲猪瘟病毒调控宿主细胞因子表达水平的研究进展[J].病毒学报,2021,37(6):1484-1490.HUANG Y,ZHAO J,ZENG Y B,et al.Research advances in regulation of expression of cytokines in hosts by the African swine fever virus[J].Chinese Journal of Virology,2021,37(6):1484-1490.(in Chinese)
[22]	HAMIDZADEH K,CHRISTENSEN S M,DALBY E,et al.Macrophages and the recovery from acute and chronic inflammation[J].Annu Rev Physiol,2017,79:567-592.
[23]	BORCA M V,O’DONNELL V,HOLINKA L G,et al.The L83L ORF of African swine fever virus strain Georgia encodes for a non-essential gene that interacts with the host protein IL-1β[J].Virus Res,2018,249:116-123.

Preliminary Study on the Function of MGF360-13L Gene of African Swine Fever Virus Multigene Family

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 23

Related Articles 9

Recommended Articles

Metrics

Comments

[1]	WANG Hui, FENG Baoliang, WU Dan, XIANG Guangming, WANG Nan, MU Yulian, LI Kui, LIU Zhiguo. Research Progress of CD163 Gene and Disease-Resistant Breeding on Porcine Reproductive and Respiratory Syndrome [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3127-3138.
[2]	QIU Yingwu, CHANG Hao, PENG Jie, YI Heyou, WANG Qiumei, GUO Yanchen, WU Qianwen, CAO Xuezhen, LIN Limiao, LI Wei, ZHOU Qingfeng, ZHANG Guihong, LI Qunhui, GONG Lang. A Real-time PCR Method for the Rapid Detection of African Swine Fever Virus I177L Gene Deletion Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2521-2527.
[3]	LIU Yankun, LUO Runbo, LIN Yan, ZHU Weiyun. Effects of Phage Cocktail on Growth Performance, Blood Parameters and Fecal Microbiota of Weaned Piglets [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1555-1567.
[4]	CHANG Hao, QIU Yingwu, PENG Jie, GAO Qi, SONG Zebu, CHEN Yang, LI Wei, LIN Limiao, CAO Xuezhen, ZHOU Qingfeng, ZHANG Guihong, LI Qunhui, ZHENG Zezhong. Establishment of a Dual Real-time Fluorescence Quantitative PCR Assay for Genotype I ASFV [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4428-4432.
[5]	XU Lingyu, CAO Chenfu, LI Zhonsheng, CHEN Junhong, LIU Tengfei, WANG Xinkai, JIA Weixin. Preparation of Monoclonal Antibody against African Swine Fever Virus p54 Protein and Development of Blocking ELISA [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 813-821.
[6]	WANG Lijuan, SHI Zhengwang, YANG Bo, MA Yuan, LUO Juncong, WAN Ying, SONG Rui, TIAN Hong, ZHENG Haixue. Construction, Expression and Activity Identification of Anti-ASFV Single-Chain Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(5): 1328-1336.
[7]	XU Zhaokun, WANG Jianhong, LI Wu, WANG Yujiong. The effects of the Recombinant Mycobacterium Tuberculosis CFP10 Protein on the TLR Signal-mediated Inflammatory Responses in A549 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(10): 2509-2517.
[8]	HE Huan，CHEN Xin-nuo，ZENG Ze，REN Yu-peng，TANG Cheng，ZHANG Bin，YUE Hua. The Initial Research of OmpP2 in Haemophilus parasuis Induces Pro-inflammatory Cytokine mRNA Transcription and Inflammatory Mechanism in Porcine Alveolar Macrophages [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1428-1434.
[9]	SHAO Bin; ZHANG Caixia;ZHAO Ruqian;ZHANG Shuxia . Effect of Transportation on the Antioxidative Capacity and Proinflammatory Cytokines in Piglets [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2009, 40(10): 0-1549.