乙型脑炎重组痘苗病毒疫苗安全性评价研究
发布时间：2018-07-16 08:48
【摘要】：近年来，乙型脑炎病毒（JEV）在我国甚至亚洲地区广泛流行，威胁着人和家畜的安全，经过这些年我国政府及相关部门的努力，患乙型脑炎的人的发病率及死亡率不断下降，但是对于猪的乙型脑炎的预防还没有猪专用的乙型脑炎病毒疫苗，目前一般采用弱毒疫苗SA14-14-2对猪进行免疫。因此，根据当前我国养猪行业的形势，需要一种猪专用的乙型脑炎病毒疫苗，本实验室研制的含乙型脑炎E抗原基因重组痘苗病毒疫苗rVTT-JEVE具有良好的免疫效果，但是目前尚不能证明该疫苗的安全性。 本研究对本实验室研制的重组痘苗病毒疫苗rVTT-JEVE的安全性进行评价，首先对重组痘苗病毒rVTT-JEVE的理化特性进行研究，通过将其进行温度、酸碱度、乙醚、胰蛋白酶、氯仿、超声波及紫外线等理化因子的适当的不同变量的处理，检测其经处理后的病毒滴度的变化，证明其对高温敏感，对酸碱有一定的耐受性，对氯仿、胰蛋白酶、超声波、紫外线均有一定程度的敏感性，完成了理化特性的研究。 首先应用小鼠进行了免疫相关研究，且对重组痘苗病毒疫苗株rVTT-JEVE的安全性进行了初步评价，免疫小鼠后产生的特异性抗体水平显著高于弱毒疫苗组和对照组（P0.05），经RT-qPCR方法检测重组痘苗病毒在小鼠体内残留发现，重组痘苗病毒在小鼠体内5-7d后拷贝数明显减少，通过分析其体液免疫及病毒残留，证明该候选疫苗具有良好的体液免疫且病毒在小鼠体内残留仅5-7d。通过猪体免疫，分析其免疫特性、病毒残留和环境释放，，证实了该重组痘苗病毒候选疫苗株rVTT-JEVE有良好的免疫特性，重组痘苗病毒在猪体内残留仅3d且不会向环境中释放，表明该候选疫苗具有良好安全性。
[Abstract]:In recent years, Japanese encephalitis virus (JEV) has been widely prevalent in China and even in Asia, threatening the safety of human beings and livestock. Through the efforts of our government and related departments, the morbidity and mortality of people suffering from encephalitis B have been decreasing. However, for the prevention of swine encephalitis B, there is no swine specific vaccine against Japanese encephalitis virus. Currently, attenuated vaccine SA14-14-2 is generally used to immunize pigs. Therefore, according to the current situation of pig industry in our country, we need a kind of special type B encephalitis virus vaccine. The recombinant vaccinia virus vaccine rVTT-JEVE containing E antigen gene of Japanese encephalitis developed by our laboratory has good immune effect. But it is not yet possible to prove the vaccine's safety. In this study, the safety of recombinant vaccinia virus vaccine rVTT-JEVE was evaluated. Firstly, the physical and chemical properties of recombinant vaccinia virus rVTT-JEVE were studied. The proper treatment of different variables of physical and chemical factors, such as ultrasonic wave and ultraviolet ray, to detect the changes of virus titer after treatment, proved that it is sensitive to high temperature, has certain tolerance to acid and alkali, and is resistant to chloroform, trypsin, ultrasonic wave, and so on. UV rays were sensitive to a certain degree and the physicochemical properties were studied. First of all, immune-related studies were carried out in mice, and the safety of recombinant vaccinia virus vaccine strain rVTT-JEVE was preliminarily evaluated. The specific antibody level of immunized mice was significantly higher than that of attenuated vaccine group and control group (P0.05). The recombinant vaccinia virus remained in mice by RT-qPCR, and the copy number of recombinant vaccinia virus in mice was significantly decreased after 5-7 days. By analyzing its humoral immunity and virus residues, it was proved that the candidate vaccine had good humoral immunity and the virus remained only 5-7 days in mice. The immune characteristics, virus residues and environmental release of the recombinant vaccinia virus candidate strain rVTT-JEVE were analyzed. The recombinant vaccinia virus remained in pigs for only 3 days and could not be released into the environment, the results showed that the recombinant vaccinia virus candidate strain rVTT-JEVE had good immune properties. The result shows that the candidate vaccine is safe.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.79
[Abstract]:In recent years, Japanese encephalitis virus (JEV) has been widely prevalent in China and even in Asia, threatening the safety of human beings and livestock. Through the efforts of our government and related departments, the morbidity and mortality of people suffering from encephalitis B have been decreasing. However, for the prevention of swine encephalitis B, there is no swine specific vaccine against Japanese encephalitis virus. Currently, attenuated vaccine SA14-14-2 is generally used to immunize pigs. Therefore, according to the current situation of pig industry in our country, we need a kind of special type B encephalitis virus vaccine. The recombinant vaccinia virus vaccine rVTT-JEVE containing E antigen gene of Japanese encephalitis developed by our laboratory has good immune effect. But it is not yet possible to prove the vaccine's safety. In this study, the safety of recombinant vaccinia virus vaccine rVTT-JEVE was evaluated. Firstly, the physical and chemical properties of recombinant vaccinia virus rVTT-JEVE were studied. The proper treatment of different variables of physical and chemical factors, such as ultrasonic wave and ultraviolet ray, to detect the changes of virus titer after treatment, proved that it is sensitive to high temperature, has certain tolerance to acid and alkali, and is resistant to chloroform, trypsin, ultrasonic wave, and so on. UV rays were sensitive to a certain degree and the physicochemical properties were studied. First of all, immune-related studies were carried out in mice, and the safety of recombinant vaccinia virus vaccine strain rVTT-JEVE was preliminarily evaluated. The specific antibody level of immunized mice was significantly higher than that of attenuated vaccine group and control group (P0.05). The recombinant vaccinia virus remained in mice by RT-qPCR, and the copy number of recombinant vaccinia virus in mice was significantly decreased after 5-7 days. By analyzing its humoral immunity and virus residues, it was proved that the candidate vaccine had good humoral immunity and the virus remained only 5-7 days in mice. The immune characteristics, virus residues and environmental release of the recombinant vaccinia virus candidate strain rVTT-JEVE were analyzed. The recombinant vaccinia virus remained in pigs for only 3 days and could not be released into the environment, the results showed that the recombinant vaccinia virus candidate strain rVTT-JEVE had good immune properties. The result shows that the candidate vaccine is safe.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S859.79
【共引文献】
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