Construction and virulence evaluation of nonreplicative vaccinia virus modified strain NTV-C7L
10.3760/cma.j.issn.1003-9279.2020.01.015
- VernacularTitle:非复制型痘苗病毒修饰株NTV-C7L构建及毒力初步评价
- Author:
Peng ZHANG
1
;
Ying ZHAO
;
Li ZHAO
;
Jiao REN
;
Wenjie TAN
;
Houwen TIAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Non-replicating vaccinia virus modified strain NTV-C7L;
Virulence;
Vaccinia virus vector
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(1):72-77
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the virulence of NTV modified strain NTV-C7L in vitro and in vivo.Methods The non-replicative vaccinia virus modified strain NTV-C7L was constructed by homologous recombination,in order to evaluate the virulence of NTV-C7L by cell diffusion ability and virus replication,intranasal challenge,and scratch of tail of mice.Results The homologous recombination non-replicating vaccinia virus modified strain NTV-C7L was selected by blue-white screening,the purity of NTV-C7L was identified by PCR and Western blot,the C7L gene was successfully inserted and expressed;In human embryonic lung fibroblasts (MRC-5) as well as mouse embryonic fibroblasts (BALB/C3T3),the replication ability of NTV-C7L was recovered compared with that of NTV,but was lower than that of VTT;the result of the intranasal route with 106 PFU vaccinia virus showed that the animals infected with TTV had slightly greater weight loss than NTV or NTV-C7L(t =6.56,P＜0.001;t =5.73,P＜0.001).Compared with the NTV-C7L group,the weight loss of the NTV group was not statistically significant (t =0.597,P =0.081);the intranasal route with 107pFU vaccinia virus showed that the weight loss of the NTV group and NTV-C7L group was statistically significant compared with the TTV group (t =12.86,P＜0.001;t =10.71,P＜ 0.001).Compared with the NTV-C7L group,the weight loss of the NTV group was statistically significant(t =4.616,P＜0.001);the result of skin scratch indicated that mice scarified with TTV (106 PFU) formed more obvious skin redness,swelling and injury on the post infected 5 days,whereas only redness was detected after scarification with a much higher dose (107) of NTV or NTV-C7L.Conclusions The intracellular and mouse virulence of NTV modified strain NTV-C7L was recovered compared with NTV,but was significantly lower than that of TTV,which provided reference for optimization and application of vaccinia virus vector.
