抗体免疫压力下猪繁殖与呼吸综合征病毒ORF3和 ORF5基因变异研究

Abstract

Abstract: To study the mutants of a highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV), HPPRRSV-JXA1 strain was continuously passed in Marc-145 cell cultures with or without anti-JXA1 pig sera for 30 passagesand named JXA1-Abf30 and JXA1-f30 respectively. Infected cells were collected for extraction of viral RNA every 10 passages,ORF3 and ORF5 genes were amplified,cloned and sequenced. Sequence analysis indicated that numbers of nucleotide mutations on ORF3 and ORF5 genes in JXA1-Abf30 and JXA1-f30 strain were 18 and 9, numbers of amino acids mutations were 15 and 3 respectively. The ratio of nucleotide mutative speed between the two strains was 186% , The ratio of amino acid mutative speed of two strains was 500%, non-synonymous VS synonymous mutation ratios (NS/S) of ORF3 and ORF5 genes in two strains were 5.0 and 0.5. Besides,there were 4 stable NS mutations in ORF5 gene and two of them (related to AA31 and AA190) were related to the known epitopes during passages with anti-PRRSV sera,which were not detected in passages without specific antibody. It indicated that mutations in PRRSV under antibody immune selection pressure were significantly faster than PRRSV without antibody immune selection pressure, anti-PRRSV sera had strong immune selective pressures on viral evolution in its ORF5 genes, but for ORF3 gene, mutation influence was less than ORF5 genes.

Key words: PRRSV; ORF3; ORF5; immune selection pressure; genetic mutations

CLC Number:

HU Jing, JIANG Ai-xiang, NIU Xiao-yun, SU Dan-ping, CHEN Rui-ai, HE Dong-sheng. Mutations of ORF3 and ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus under Antibody Selective Pressure[J]. , 2012, 39(11): 13-17.

References

1 王剑非,孙建全,张金强,等.PRRSV分离株Nsp2基因和ORF5基因的遗传变异分析[J]. 家畜生态学报, 2011,32(1):48~53.
2 孙建富,郝建伟,荣福龙,等. 猪繁殖与呼吸综合征病毒的分离与鉴定及 ORF5基因的序列分析[J].中国畜牧兽医, 2011,38(9):138~142.
3 赵鹏,马诚太,崔治中,等.猪繁殖与呼吸综合征病毒在抗体免疫选择压下的变异[J].中国科学, 2010,40(10):952~962.
4 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997.
5 郭宝清,陈章水,刘文兴,等.从疑似PRRS流产胎儿分离PRRSV的研究[J].中国畜禽传染病,1996,87(2):1~4.
6 崔乃忠. PRRSV BJ-10株主要结构蛋白质基因序列分析.长春:吉林农业大学,2001.
7 童光志,周艳君,郝晓芳,等.高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,29(5):323~327.
8 蔡雪辉.PRRSV国内分离毒株病毒结构蛋白抗原表位鉴别及其ORF5的RFLP分析.哈尔滨:东北农业大学,2002.
9 Benn S, Rutleddge R, Folks T, et al. Genomic heterogeneity of AIDS retroviral isolates from North America and Zaire[J]. Science, 1985,230(4728):949~951.
10 Conzelmann K K, Visser N, Van Woensel P,et al. Molecular characterization of porcine reproductive and respiratory syndrome virus, a member of the arterivirus group[J].Virology, 1993,193:329~339.
11 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, 2000,145:659~688.
12 Hwang S S, Boyle T J, Lyerly H K, et al. Identification of the envelope V3 loop as the primary determinant of cell tropism in HIV1[J].Science, 1991,253:71~74.
13 Key K F, Haqshenas G, Guenette D K, et al. Genetic variation and phylogenetic analyses of the ORF5 gene of acute porcine reproductive and respiratory syndrome virus isolates[J].Vet Microbiol, 2001,83:249~263.
14 Li B, Xiao S, Wang Y, et al. Immunogenicity of the highly pathogenic porcine reproductive and respiratory syndrome virus GP5 protein encoded by a synthetic ORF5 gene[J].Vaccine, 2009,27:1957~1963.
15 Plana D J, Climent I, Sarraseca J, et al. Baculovirus expression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/91. Involvement of ORF3 and ORF5 proteins in protection[J].Virus Genes, 1997,14:19~29.
16 Rodriguez M J, Sarraseca J, Fominaya J, et al. Identification of an immunodominant epitope in the C terminus of glycoprotein 5 of porcine reproductive and respiratory syndrome virus[J].J Gen Virol, 2001,82(5):995~999.
17 Shioda T, Levy L A, Chen M C. Small amino acid changes in the V3 hypervariable region of gp120 can affect the T-cell-line and macrophage tropism of human immunodeficiency virus typeⅠ[J].Proc Natl Acad Sci USA, 1992,89:9434~9438.
18 Tian K, Yu X, Zhao T, et al. Emergence of fatal PRRSV variants: Unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unike hallmark[J].PLoS One, 2007,2:e526.
19 Vu H L, Kwon B.Immune evasion of porcine reproductive and respiratory syndrome virus through glycan shielding involves both glycoprotein 5 as well as glycoprotein 3[J].J Virol, 2011, 85(11):5555~5564.
20 Woelk C H, Jin L, Holmes E C, et al. Immune and artificial selection in the hemagglutinin(H) glycoprotein of measles virus[J].J GenVirol, 2001,82:2463~2474.

[1]	KANG Pu, REN Zhaowen, CAI Rujian, LYU Zongji, HUANG Yuhao, WANG Xiaohu. Variation Regularity and Transmission of PRRSV-2 Lineage 8 Strains in Guangdong and Surrounding Provinces [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2001-2011.
[2]	CHEN Tianbao, WANG Yani, LIU Baoling, WANG Gang, WANG Xiaohu, LIU Xuhui, CAI Rujian, HE Dongsheng. Research Progress on the Anti-Porcine Reproductive and Respiratory Syndrome Virus Effect and Mechanism of Traditional Chinese Medicine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2114-2129.
[3]	LI Huawei, GUO Kewei, WANG Xuying, ZHANG Xiaoyu, LU Ziwei, LI Xinfeng, MA Hui, HOU Wenjing. Eukaryotic Expression and Subcellular Localization of Nonstructural Protein 7 of Porcine Reproductive and Respiratory Syndrome Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1160-1168.
[4]	LIU Qingqing, WU Peng, LI Peidong, ZHANG Jiangwei, CHEN Chuangfu, XIAO Chencheng. Construction of Recombinant Adenovirus Expressing Porcine Reproductive and Respiratory Syndrome Virus N Protein and Its Immunogenicity Analysis [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1024-1031.
[5]	WANG Zhaoli, YANG Siyu, WU Yanmei, SONG Xiaoxiao, MU Xiang, ZHANG Tao. Effects of PRRSV-LPS Treatment on Neutrophil Adhesion to Porcine Pulmonary Microvascular Endothelial Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 471-479.
[6]	SUN Lu, ZHANG Jing, YIN Miao, CHENG Yaodie, DAI Kun, WU Qian, ZHAO Hong, CHEN Xiwen. Study on the Antibacterial and Anti-PRRSV Activities of Four Heat Clearing and Detoxifying Compound Chinese Herbal Medicine in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 746-754.
[7]	CHU Pinpin, JIANG Zhiyong, LI Chunling, GOU Hongchao, LI Yan, CAI Rujian. Antiviral Effect of Silver Nanoparticles on Porcine Reproductive and Respiratory Syndrome Virus in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4356-4363.
[8]	JIAO Hanwei, ZHOU Zhixiong, LI Mengjuan, XIAO Yu, LI Bowen, GUO Xiaoyi, ZENG Hui, GU Guojing, SHUAI Xuehong. Identification of lncRNA-mRNA Regulatory Network of Riboflavin Metabolism in HepG2 Cells Affected by Swine Hepatitis E Virus ORF3 Protein [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4618-4627.
[9]	DENG Jiping, CHEN Zhilong, GUO Songyangnian, ZHANG Kun, XU Daojun, YANG Qing, YUAN Anwen. Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolated from Hunan Province [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3795-3802.
[10]	LIU Hanyang, ZHANG Yi, XIAO Ran, CHEN Bin, WU Yongsheng, SU Zhipeng, LI Yupeng, PEI Chaoxin, XU Zhenying, ZHU Jiawen. Molecular Epidemiological Investigation on ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus in Sichuan in 2018 [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1172-1182.
[11]	ZHOU Shumian, CAO Mixia, HU Tingjun. A Study on Effect of Anti-PRRSV of n-butanol Part of Flavonoids from Polygonum hydropiper L. in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 605-611.
[12]	ZHANG Xiyi, WEN Guilan, GUAN Guodan, CHEN Guang, TIAN Lang, YANG Baiqi. Cloning and Genetic Variation Analysis of NSP2 Gene of Porcine Reproductive and Respiratory Syndrome Virus Guizhou Strain GZ-R [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3659-3666.
[13]	DENG Tongwei, XUE Yanan, LU Jianzhou, XIA Yanxun, CHEN Lulu, GUO Yiwen, PENG Liming, JIANG Zenghai, QIAO Hongxing, XU Yaohui, ZHANG Xiaozhan. Diagnosis and Pathogen Analysis of Mixed Infection of Porcine Reproductive and Respiratory Syndrome Virus,Porcine Circovirus Type 2 and Haemophilus parasuis [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(10): 3401-3409.
[14]	ZHANG Hailong, CHEN Shengnan, XIE Bo, LI Li, HUANG Liangzong. Diagnosis of Mixed Infection of Porcine Reproductive and Respiratory Syndrome Virus and Haemophilus parasuis [J]. , 2019, 46(6): 1856-1864.
[15]	LIU Shengli, LIU Lingling, LYU Yujin, WU Fengsun, LI Wengang. Research on Multiplex PCR Combined with DNA Microarray for Detecting Five Kinds of Reproductive Disease Virus [J]. , 2019, 46(5): 1532-1540.

Mutations of ORF3 and ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus under Antibody Selective Pressure

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments