Construction of NTV-ΔF1L-C7L modified strain of non-replication vaccinia virus NTV and evaluation of its immunological effects
10.3760/cma.j.cn112866-20231122-00061
- VernacularTitle:非复制型痘苗病毒NTV修饰株NTV-ΔF1L- C7L的构建及免疫效果评价
- Author:
Jiao REN
1
;
Hang YUAN
;
Li ZHAO
;
Yamei DOU
;
Shiyuan LIU
;
Xin MENG
;
Houwen TIAN
;
Wenling WANG
;
Wenjie TAN
Author Information
1. 中国疾病预防控制中心病毒病预防控制所，北京 102206
- Keywords:
Vaccinia virus;
Recombinant virus;
Immunogenicity
- From:
Chinese Journal of Experimental and Clinical Virology
2024;38(2):181-187
- CountryChina
- Language:Chinese
-
Abstract:
Objective:We genetically modified our non-replicating vaccinia virus NTV to improve its immunogenicity.Methods:We constructed NTV-modified strain NTV-ΔF1L-C7L by homologous recombination of vaccinia virus based on CRISPR-Cas9 technology by inserting the C7L gene while deleting the F1L gene. The recombinant virus NTV-ΔF1L-C7L was then immunized with 10 7 PFU in BALB/c mice, and the levels of humoral and cellular immunity induced by NTV-ΔF1L-C7L were detected by ELISA and ELISpot method, respectively, and the levels of neutralizing antibodies were determined by the phage-reduced neutralization assay. Results:The PCR and western- blot identification proved that the F1L gene of the constructed NTV-modified strain NTV-ΔF1L-C7L was missing, while the C7L gene was inserted back in the region, and the C7L gene could be expressed normally, indicating that the recombinant virus was constructed correctly. After immunization of mice with NTV-ΔF1L-C7L, ELISA result showed that the recombinant virus NTV-ΔF1L-C7L induced a higher level of IgG antibody than NTV; ELISpot result also showed that the recombinant virus was able to induce a higher level of IFN-γ; and the result of plaque reduction neutralization test showed that the recombinant virus was able to induce a higher level of IFN-γ antibody than that of NTV.Conclusions:We correctly constructed the NTV gene-modified strain NTV-ΔF1L-C7L, which induced stronger humoral and cellular immunity compared with NTV, and provided reference data for the research and development of replacement products for smallpox or monkeypox vaccines.
