Construction of a mavs-inactivated MDCK cell line for facilitating the propagation of canine distemper virus (CDV)

被引:3
作者
Cai, Cong [1 ,2 ]
Wang, Xiaoping [1 ,2 ]
Zhao, Ya [1 ,2 ]
Yi, Chenyang [1 ,2 ]
Jin, Zehua [1 ,2 ]
Zhang, Anding [1 ,2 ]
Han, Li [1 ,2 ]
机构
[1] Huazhong Agr Univ, Coll Vet Med, State Key Lab Agr Microbiol, Wuhan 430070, Hubei, Peoples R China
[2] Minist Sci & Technol, Int Res Ctr Anim Dis, Key Lab Prevent Vet Med Hubei Prov, Wuhan 430070, Hubei, Peoples R China
关键词
Canine distemper virus (CDV); CRISPR/Cas9; technology; Mays (mitochondrial antiviral signaling); Madin-Darby Canine Kidney Epithelial (MDCK); Interferon response; GENE-EXPRESSION; INTERFERON; PATHOGENESIS; VIRULENT; STRAINS; MORBILLIVIRUSES; CULTURES; PROTEIN; GROWTH; STAT2;
D O I
10.1016/j.molimm.2019.06.013
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Canine distemper is a highly contagious disease of wild and domestic carnivores. Obtaining of a suitable cell line for canine distemper virus (CDV) propagation is very important for field CDV isolation and vaccine antigen preparation. However, the cell line currently developed cell lines for CDV propagation are a marmoset lymphoid cell line (B95a), which could cause the virus to potentially infect human cells, and canine SLAM-expressing Vero cells, which may cause the virus to lose virulence, Therefore, a canine cell line constructed for efficient CDV propagation would be attractive. In the present study, a Madin-Darby Canine Kidney Epithelial (MDCK) cell line with mays (mitochondrial antiviral signaling) inactivation was constructed by CRISPR/Cas9 technology. The interferon-I response induced by poly(I:C), an analogue of viral RNA, was significantly blocked in the constructed cell line, designated MDCK-KOmays. Moreover, the propagation of a filed CDV strain was approximately 100 times higher in MDCK-KOmays cells than in wild-type MDCK cells. Therefore, in the present study, a canine cell line facilitating CDV propagation was successfully constructed, and the results suggested that the constructed canine cell line was more efficient than the wild-type cell line for the isolation of field CDVs. In addition, the rapid propagation of CDVs to high titers in the constructed MDCK-KOmays cell line indicated that this cell line could also be an alternative cell line for the preparation of vaccine antigens.
引用
收藏
页码:133 / 138
页数:6
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