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用于增强猪瘟病毒复制的PK15细胞的克隆与鉴定

Cloning and Identification of PK15 Cells for Enhanced Replication of Classical Swine Fever Virus.

作者信息

Yin Mei, Hu Dongfang, Li Peng, Kong Lingyun, Ning Hongmei, Yue Feng, Jiang Jinqing, Wang Xuannian

机构信息

College of Animal Science and Technology, Henan Institute of Science and Technology, Xinxiang, Henan, 453003, China.

College of Life Science and Technology, Xinxiang University, Xinxiang, Henan, 453003, China.

出版信息

J Vet Res. 2020 Mar 24;64(1):9-14. doi: 10.2478/jvetres-2020-0020. eCollection 2020 Mar.

Abstract

INTRODUCTION

Classical swine fever virus (CSFV) causes an economically important and highly contagious disease of pigs, leading to economic losses around the world. Attenuated live vaccines with CSFV antigens have played an important role in the prevention and control of the disease. Porcine kidney 15 (PK15) cells have been widely used for the propagation of CSFV, but this cell line is not efficient or homogeneously susceptible to viral infection.

MATERIAL AND METHODS

To achieve a homogeneous PK15 cell line which enabled high titre replication of CSFV, we used the limiting dilution cell cloning method.

RESULTS

We developed two cell clones, PK15-1A6 and PK15-3B1, which respectively have high- and low-permissive phenotypes to CSFV infection. The PK15-1A6, PK15-3B1, and PK15 parent cells showed different characteristics in cell proliferation rate, susceptibility to CSFV infection, and CSFV production. The mean virus titres per millilitre reflected by TCID values in PK15-1A6, PK15-3B1, and PK15 parent cells were 106.85, 103.63, and 104.74, respectively.

CONCLUSION

The PK15-1A6 cell clone is more permissive to CSFV infection than the PK15 parent cells. The screened high-permissive cells will be useful for CSFV propagation and vaccine development , and facilitate research on the pathogenicity of CSFV.

摘要

引言

经典猪瘟病毒(CSFV)引发一种对猪来说具有重要经济意义且传染性极强的疾病,在全球范围内造成经济损失。含有CSFV抗原的减毒活疫苗在该疾病的预防和控制中发挥了重要作用。猪肾15(PK15)细胞已被广泛用于CSFV的增殖,但该细胞系对病毒感染的效率不高且易感性不均一。

材料与方法

为获得能够高效复制CSFV的均一PK15细胞系,我们采用了有限稀释细胞克隆法。

结果

我们培育出两个细胞克隆,PK15 - 1A6和PK15 - 3B1,它们对CSFV感染分别具有高允许和低允许表型。PK15 - 1A6、PK15 - 3B1和PK15亲本细胞在细胞增殖速率、对CSFV感染的易感性以及CSFV产生方面表现出不同特征。PK15 - 1A6、PK15 - 3B1和PK15亲本细胞中通过TCID值反映的每毫升平均病毒滴度分别为106.85、103.63和104.74。

结论

PK15 - 1A6细胞克隆比PK15亲本细胞对CSFV感染更敏感。筛选出的高允许细胞将有助于CSFV的增殖和疫苗开发,并促进对CSFV致病性的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9ff9/7105985/f794654d8d3c/jvetres-64-009-g001.jpg

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