SARSCoV灭活疫苗对恒河猴的免疫原性、保护性与安全性研究
发布时间：2018-08-03 15:32
【摘要】：目的: 建立SARS相关冠状病毒感染恒河猴动物模型,并用该模型评价一种实验性SARS冠状病毒灭活疫苗免疫原性、保护性与安全性,同时也为该疫苗进一步完善及投入临床试验提供理论和实验依据。 方法: 动物免疫分3组,分别为肌肉注射0.5μg、5μg、50μg疫苗组及磷酸盐缓冲液(PBS)对照组,同时设超大剂量(5000μg)观察疫苗安全性。于初次免疫后7天再次以同等剂量加强免疫,0.5μg、5μg、50μg剂量组和阳性对照组于免疫后22天接受SARS相关冠状病毒的攻击,并在攻毒后15天处死所有动物。观察免疫和攻毒前后动物临床症状和生化指标的改变。ELISA检测IgG和IgA抗体,观察不同动物组IgG的变化和鼻咽部IgA分泌情况,了解动物体液免疫和粘膜免疫的情况。双抗体夹心ELISA试剂盒定量分析恒河猴血清IL-4和IFN-γ水平。采用病毒分离、免疫荧光、光镜及RT-PCR方法对实验动物不同时间、不同组织或分泌物进行检测。病理切片了解免疫和攻毒前后动物脏器的改变。 结果: 1.血清IgG抗体反应:大部分免疫猴在初次免疫7天后产生较高IgG滴度(100),高剂量组呈现高水平抗体滴度(≥1600),随疫苗剂量的升高,免疫应答亦增强。 2.粘膜免疫:肌肉接种引起的粘膜抗体反应,12只免疫动物中有5只(41.7%)表达SARS-CoV特异性分泌型IgA抗体。 3.细胞因子评价:初次免疫后,5μg,50μg疫苗组动物血清中IFN-γ显著增加,明显高于免疫前水平(P0.05和0.01),再次免疫后继续增加。0.5μg剂量组动物仅在再次免疫后IFN-γ显著升高(P0.05),而PBS组无明显变化。三个实验组及对照组IL-4水平在免疫前后均无显著变化。 4.血清中和抗体效价:在初次免疫后14天,三个免疫组出现不同水平针对SARS-CoV的中和抗体,同时随着疫苗剂量的增加,中和抗体滴度也升高。 5.组织病理学检查:各免疫组及阴性对照动物及肺部、肝脏、肾脏病理检查均未发现异常,而阳性对照动物肺部及肝脏病理学检查均发现异常变化。
[Abstract]:Objective: to establish an animal model of SARS associated coronavirus infection in rhesus monkeys and to evaluate the immunogenicity of an experimental SARS coronavirus inactivated vaccine. Protection and safety also provide theoretical and experimental basis for further improvement and application of the vaccine in clinical trials. Methods: animal immunizations were divided into 3 groups, which were intramuscularly injected with 0.5 渭 g of 5 渭 g or 50 渭 g of (PBS) phosphate buffer, respectively. The safety of the vaccine was observed at a high dose (5000 渭 g). On the 7th day after the first immunization, the same dose of 0.5 渭 g ~ 5 渭 g ~ (5 渭 g) ~ (50 渭 g) and the positive control group were given SARS related coronavirus attack again, and all the animals were killed 15 days after the initial immunization. To observe the changes of clinical symptoms and biochemical indexes before and after immunization. Elisa was used to detect IgG and IgA antibodies, to observe the changes of IgG and the secretion of nasopharyngeal IgA in different groups, and to understand the humoral immunity and mucosal immunity of animals. Serum levels of IL-4 and IFN- 纬 in rhesus monkeys were determined by double antibody sandwich ELISA kit. Virus isolation, immunofluorescence, light microscopy and RT-PCR were used to detect different tissues and secretions of experimental animals at different time. Pathological sections were used to understand the changes of animal organs before and after immunization. Results: 1. Serum IgG antibody response: most of the immunized monkeys produced high IgG titer (100) 7 days after the first immunization, and the high level antibody titer (鈮，
本文编号：2162180
[Abstract]:Objective: to establish an animal model of SARS associated coronavirus infection in rhesus monkeys and to evaluate the immunogenicity of an experimental SARS coronavirus inactivated vaccine. Protection and safety also provide theoretical and experimental basis for further improvement and application of the vaccine in clinical trials. Methods: animal immunizations were divided into 3 groups, which were intramuscularly injected with 0.5 渭 g of 5 渭 g or 50 渭 g of (PBS) phosphate buffer, respectively. The safety of the vaccine was observed at a high dose (5000 渭 g). On the 7th day after the first immunization, the same dose of 0.5 渭 g ~ 5 渭 g ~ (5 渭 g) ~ (50 渭 g) and the positive control group were given SARS related coronavirus attack again, and all the animals were killed 15 days after the initial immunization. To observe the changes of clinical symptoms and biochemical indexes before and after immunization. Elisa was used to detect IgG and IgA antibodies, to observe the changes of IgG and the secretion of nasopharyngeal IgA in different groups, and to understand the humoral immunity and mucosal immunity of animals. Serum levels of IL-4 and IFN- 纬 in rhesus monkeys were determined by double antibody sandwich ELISA kit. Virus isolation, immunofluorescence, light microscopy and RT-PCR were used to detect different tissues and secretions of experimental animals at different time. Pathological sections were used to understand the changes of animal organs before and after immunization. Results: 1. Serum IgG antibody response: most of the immunized monkeys produced high IgG titer (100) 7 days after the first immunization, and the high level antibody titer (鈮，
本文编号：2162180
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