表达猪繁殖与呼吸综合征病毒GP5的重组伪狂犬病毒的构建与鉴定

Zhi-Jun Tian, Hua-Ji Qiu, Jian-Qiang Ni, Yan-Jun Zhou, Xue-Hui Cai, Guo-Hui Zhou, Yun-Feng Wang, Guang-Zhi Tong
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摘要

短句来源将猪繁殖与呼吸综合征病毒(PRRSV)的GP5基因整合到伪狂犬病毒(PRV)疫苗株Bartha-K61的TK基因位点上,得到了一株含有GP5基因的TK和gE阴性重组PRV,命名为rPRV-GP5。采用单步生长法、Western blot法和间接免疫荧光法检测rprv -GP5感染细胞对GP5的体外表达。结果表明,GP5基因可以在rprv -GP5感染细胞的细胞质中真实表达。与亲本病毒相比,在几种细胞培养中,rPRV-GP5在病毒复制和细胞致病作用方面没有明显差异。用10(6.0)PFU rPRV-GP5肌内免疫的4只PRV阴性羊完全免受高毒性猪源PRV S株10(3)LD50的攻击。间接免疫荧光试验、酶联免疫吸附试验和病毒中和试验显示,接种后第63天,10头PRV和PRRSV阴性仔猪经鼻灌胃10(7.0)PFU的rPRV-GP5,并经鼻灌胃10(5.0)TCID50的强毒PRRSV CH-1a株,在灌胃后第3、5、14天产生了抗PRRSV抗体。结果提示,rPRV-GP5能够诱导接种动物对PRRS产生遗忘性免疫反应。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Construction and characterization of a recombinant pseudorabies virus expressing porcine reproductive and respiratory syndrome virus GP5.

The GP5 gene of porcine reproductive and respiratory syndrome virus (PRRSV) was integrated into the TK gene locus of pseudorabies virus (PRV) vaccine strain Bartha-K61, resulting in a TK- and gE- negative recombinant PRV harboring GP5 gene, designated as rPRV-GP5. The in vitro expression of the GP5 by rPRV-GP5-infected cells was analyzed by single-step growth analysis,Western blot,and indirect immunofluorescence test. It was shown that GP5 gene can be expressed authentically in the cytoplasm of rPRV-GP5-infected cells. Compared to its parental virus, rPRV-GP5 showed no obvious difference regarding viral replication and cytopathogenic effects in several cell cultures. Four PRV-negative sheep immunized intramuscularly with 10(6.0) PFU of rPRV-GP5 were fully protected from challenge with 10(3) LD50 of highly virulent PRV S strain of porcine origin. Ten PRV- and PRRSV-negative piglets given intranasally with 10(7.0) PFU of rPRV-GP5 and challenged intranasally with 10(5.0) TCID50 of virulent PRRSV CH-1a strain at day 63 post-inoculation developed antibodies against PRRSV 3, 5, 14 days post-challenge, as revealed by indirect immunofluorescence test, enzyme-linked immunosorbent assay and virus neutralization test. The results suggest that rPRV-GP5 is capable of inducing anamnestic immune response to PRRS in inoculated animals.

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