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研究携带RA5 3 8的重组腺病毒Ad RA5 3 8潜在的副作用 ,评价其安全性 ,为其进一步进入临床试验提供依据。方法 体外病毒扩增实验 :预先用Ad RA5 3 8感染HeLa细胞 ,制备细胞提取液后反复两次感染HeLa细胞 ,观察Ad RA5 3 8的繁殖、扩增以及对HeLa细胞的作用 ,逆转录 聚合酶链反应 (RT PCR)检测其DNA的转录 ;确定Ad RA5 3 8对作为正常细胞的人胚肺二倍体细胞 2BS的感染能力 ,绘制细胞生长曲线 ,观察Ad RA5 3 8对正常细胞的毒性 ;给BalB/C小鼠腹腔注射重组腺病毒后 ,每日观察一般状况 ,化验肝、肾功能 ,PCR检测重组腺病毒在重要脏器中的分布 ,显微镜观察重要脏器的病理学变化。结果 Ad RA5 3 8是一复制缺陷型腺病毒 ,能高效感染 2BS细胞 ,但并不影响其生长 ,小鼠体内应用后 ,并无急性毒性症状发生 ,肝、肾功能没有变化 ,在肝、肾、胃、脾内可检测到腺病毒的分布 ,但仅在注射 10 9pfu第 6天、第 12天的小鼠肝组织内可观察到轻微炎症。结论 Ad RA5 3 8应用是安全的 ,可以进入临床试验。
To study the potential side effects of the recombinant adenovirus Ad RA5 38 carrying RA53 8 and evaluate its safety, so as to provide basis for further clinical trials. Methods In vitro virus amplification experiments: HeLa cells were pre-infected with Ad RA 3 3 8 and HeLa cells were infected twice after preparing cell extracts to observe the proliferation and proliferation of Ad RA 3 38 and HeLa cells. Reverse transcription polymerase Chain reaction (RT PCR) was used to detect the DNA transcription; Ad RA53 8 was used to determine the infectivity of human embryonic lung diploid cells 2BS as normal cells, and cell growth curves were drawn to observe the toxicity of Ad RA5 38 to normal cells. After intraperitoneal injection of recombinant adenovirus into BalB/C mice, the general condition was observed daily to test liver and kidney function. The distribution of recombinant adenovirus in important organs was detected by PCR, and the pathological changes of important organs were observed with a microscope. Results Ad RA58-8 is a replication-defective adenovirus, which can effectively infect 2BS cells, but does not affect its growth. After the application in mice, there is no acute toxicity symptoms. There is no change in liver and kidney function in the liver and kidney. Adenovirus distribution was detected in the stomach and spleen, but mild inflammation was observed only in the liver tissue of mice injected with 10 9 pfu on the 6th and 12th days. Conclusion The application of Ad RA5 3 8 is safe and can be entered into clinical trials.