乙型肝炎核酸疫苗的安全性研究

发布时间：2018-03-27 22:39

本文选题：核酸疫苗　切入点：乙型肝炎　出处：《第二军医大学》2005年博士论文

【摘要】：核酸疫苗是90年代兴起的新型疫苗,已在多种疾病的防治中显示出巨大的潜能。然而核酸疫苗是一个新生事物,目前还存在许多尚未解决的问题,其中安全性的问题最为重要,直接关系到核酸疫苗的应用。全面评价乙肝DNA疫苗的安全性,我们进行了以下的毒理研究:1.急性毒性试验本研究分别考察了乙肝DNA疫苗肌肉注射或皮下注射免疫小鼠或豚鼠的急性毒性,结果表明乙肝DNA疫苗在小鼠或豚鼠上未见任何明显急性毒性反应。半数致死量是推荐临床剂量的几百倍。 2.重复剂量的长期毒性试验分别用三个剂量组的DNA疫苗(4.5m乙肝g/kg、1.5mg/kg、0.5mg/kg)肌注免疫小鼠56天(每1周给药2次,之后每周1次),从实验开始至停药后30天内进行毒性观察。结果表明小鼠体重、食量及粪便与对照组之间未见明显改变。血液学及临床生化指标均属正常范围内波动,未发现与受试药相关的毒性反应。尸检、器官重量检查未发现有与受试药物有关的毒性改变。观察了在上述给药条件下小鼠的多项疫苗免疫指标,发现乙肝DNA疫苗未发现有产生免疫耐受的迹象;不能导致自身免疫;不会诱导抗核抗体;不会引起慢性炎症及非特异性的免疫激活等免疫病理状况。以PCR法检测质粒DNA的染色体整合情况,结果表明1μg基因组DNA中整合质粒低于10拷贝(即PCR的最大灵敏度);说明质粒DNA整合到宿主细胞基因组上并使细胞发生恶性转化的可能性微乎其微。 3.致突变及生殖毒性实验致突变试验的结果表明乙肝DNA疫苗在-S9和+S9系统对CHL细胞染色体畸变均小于5%。乙肝DNA疫苗对染色体无畸变作用。乙肝DNA疫苗不引起母体毒性和胚胎毒性的无毒剂量(NOAEL)大于5.4 mg/kg。
[Abstract]:Nucleic acid vaccine (Nic acid vaccine) is a new vaccine that emerged in 1990s and has shown great potential in the prevention and treatment of many diseases. However, nucleic acid vaccine is a new thing, and there are still many unsolved problems, among which safety is the most important. Directly related to the application of nucleic acid vaccine. Comprehensive evaluation of the safety of hepatitis B DNA vaccine, we conducted the following toxicological study: 1. Acute toxicity test. In this study, the acute toxicity of mice or guinea pigs immunized with hepatitis B DNA vaccine was investigated by intramuscular or subcutaneous injection, respectively. The results showed that there was no obvious acute toxicity of hepatitis B DNA vaccine in mice or guinea pigs, and the half lethal dose was several hundred times of the recommended clinical dose. 2. Long-term toxicity test of repeated doses. Mice were immunized with DNA vaccine of 4.5 mg 路kg ~ (-1) mg 路kg ~ (-1) 路kg ~ (-1) and 0.5 mg 路kg ~ (-1) 路kg ~ (-1) respectively for 56 days (twice every week, then once a week). The toxicity was observed from the beginning of the experiment to 30 days after the withdrawal of the drug. There was no significant change in food intake and feces between the control group and the control group. The hematological and clinical biochemical indexes fluctuated within the normal range, and no toxic reactions related to the tested drug were found. No toxic changes related to the tested drugs were found in organ weight test. The immune indexes of mice were observed under the above administration conditions. It was found that the hepatitis B DNA vaccine had no signs of producing immune tolerance and could not lead to autoimmunity. ANA is not induced, chronic inflammation and non-specific immune activation are not induced. The chromosome integration of plasmid DNA was detected by PCR method. The results showed that the integration plasmid in 1 渭 g genomic DNA was less than 10 copies (i.e., the maximum sensitivity of PCR), indicating that the possibility of integration of plasmid DNA into host cell genome and malignant transformation of the cell was very little. 3. Mutagenicity and reproductive toxicity. The results of mutagenicity test showed that the chromosomal aberration of CHL cells by HBV DNA vaccine was less than 5% in the -S9 and S9 systems. There was no chromosomal aberration of HBV DNA vaccine. The nontoxic dose of HBV vaccine did not cause maternal toxicity and embryotoxicity, and the dose of NOAELs was more than 5.4 mg / kg.
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392.1

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