噬菌体D29的免疫原性及安全性评价

Evaluation on Immunogenicity and Security of Mycobacteriophage D29

目的对噬菌体D29的免疫原性及安全性进行评价。方法将BALB／C小鼠随机分为噬菌体D29滴鼻暴露组（剂量9×10^8PFU）、噬菌体D29皮下注射组（剂量9×10^8PFU），phagebuffer滴鼻组、phagebuffer皮下注射组、生理盐水滴鼻组、生理盐水皮下注射组、空白对照组，每组动物第一周第一天、第三天暴露一次，以后每周第一天暴露一次，共暴露6周。观察暴露期间噬菌体中和抗体的动态变化、暴露6周后血清中IL-2、IL-4、IL-12、IFN-1、TNF-α和NO的含量及CD4／CD8比例变化，小鼠碳粒廓清率变化、在饲养过程中各组动物的进食量以及活动、体重、生存情况，以及暴露结束后解剖取得脏器作病理学比较。结果D29滴鼻暴露组在第7天即产生低效价的中和抗体，第14天抗体效价下降，第28天之后不能检测到中和抗体，D29皮下注射组中和抗体从第7天开始产生，第21天上升，之后趋于稳定，其余组均无中和抗体产生。各组小鼠血清中细胞因子、CIM／CD8比例、碳粒廓清率以及饲养过程中各组动物的进食量以及活动、体重、生存情况均无明显差异。病理学上各实验组脏器除脾脏外均有轻微充血、水肿表现。结论D29具有抗原特性，可刺激机体产生中和抗体，但不会对机体的安全产生威胁。

Objective To evaluate the immunogenicity and security of Mycobacteriophage D29. Methods BALB/c mice were randomly divided into 7 groups： drop D29 group （9 × 10^8 PFU） , subcutaneous injection D29 group（9 × 10^8 PFU） , drop phage buffer group, subcutaneous injection phage buffer group, drop normal saline group, subcutaneous injection nmTnal saline group and blank control group, the mice were dealt with the methods as above at the first and the third day at the first week, then at the first day each week until 6 weeks. To observe the dynamic changes in the serum titer of neutralizing antibodies against bacteriophages, the changes in serum levels of IL-2, IL-4, IL-12,IFN-7,TNF- α, NO, CD4/CD8 ratio, the changes of the carbon particle clearance level and the food-intake, movement, weight, survival during the trial, and to compare the pathology of visceral organs at the sixth week. Results Neutralizing antibodies against the bacteriophages began to appear in the serum of the mice at the 7^th day,decreased at the 14^th day,and could not be detected at the 28^th day in the drop D29 group,though in the subcutaneous injection D29 group it increased at the 21^st day and began to be steady,there were not any neutralizing antibodies detected in the other groups. The various cell factors, CD4/CD8 ratio,carhon particle clearance level,the food-intake, movement,weight, survival had no significant differences in all the groups. Except the spleen, other organs displayed slightly hyperemia and edema in all experiment groups. Conclusion D29 had the immunogenicity of its own and could stimulate organism producing neutralizing antibodies, but it would not threaten the life security.