IL-33 gene recombination and expression does not affect the phenotypic characteristics of rabies virus in vitro.
- Author:
Ting GAO
1
;
Zhizhong MI
2
;
Ming SUN
2
;
Ximin TANG
2
;
Yong WANG
2
;
Yingying LI
3
Author Information
1. Basic Medical College of Dali University, Dali Nursing Vocational College, Dali 671000, China.
2. Basic Medical College of Dali University, Dali 671000, China.
3. Basic Medical College of Dali University, Dali 671000, China. *Corresponding author, E-mail: liyingying@dali.edu.cn.
- Publication Type:Journal Article
- MeSH:
Animals;
Cricetinae;
Mice;
Cell Line;
Interleukin-33/genetics*;
Rabies virus/genetics*;
Phenotype
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(7):586-591
- CountryChina
- Language:Chinese
-
Abstract:
Objective To create a recombinant rabies virus overexpressing IL-33 and to clarify the effect of IL-33 overexpression on the phenotypic characteristics of recombinant virus in vitro. Methods The IL-33 gene was obtained and amplified from the brain of a highly virulent strain of rabies infected mouse. It was then inserted between the G and L genes of the parental virus LBNSE genome by reversing genetic manipulation and rescuing a recombinant virus overexpressing IL-33. BSR cells or mouse NA cells were infected with recombinant rabies virus (rLBNSE-IL33) and the parental strain LBNSE. Sequencing and fluorescent antibody virus neutralization assay was employed to detect the stability of recombinant virus at multiplicity of infection=0.01. Viral titres focal forming units (FFU) were detected to plot multi-step growth curves (multiplicity of infection=0.01). Cytotoxicity assay kit was used to detect cellular activity. ELISA was adopted to identify the IL-33 in the supernatant of infected cells of different multiplicity of infection. Results Rescued rLBNSE-IL33 overexpressing IL-33 remained stable for at least 10 consecutive generations and had virus titers of approximately 108 FFU/mL. rLBNSE-IL33 was able to express IL-33 at high levels in a dose-dependent manner, but no high expression of IL-33 was detected in the supernatant of cells infected by LBNSE. Examination of the titers of rLBNSE-IL33 and the parental strain LBNSE in BSR and NA cells over 5 days showed no significant differences and similar kinetic properties in growth. Overexpression of IL-33 had no significant effect on the proliferation and activity of infected cells. Conclusion Overexpression of IL-33 does not significantly affect the phenotypic characteristics of recombinant rabies virus in vitro.
