DNA疫苗治疗小鼠耐多药结核病的研究

中国防痨杂志 ›› 2009, Vol. 31 ›› Issue (11): 649-652.

DNA疫苗治疗小鼠耐多药结核病的研究

梁艳¹；吴雪琼¹；李忠明²；张俊仙¹；李宁¹；阳幼荣¹；余琦¹；白雪娟¹；宋晶莹¹；王兰¹；史迎昌¹；刘洁¹；刘成龙¹；朱琰¹；徐雪玉¹

1.中国人民解放军第309医院全军结核病研究所北京 100091；2.上海海规生物科技有限公司上海 200436

出版日期:2009-11-10 发布日期:2011-11-03
基金资助:
国家重大传染病专项资助（基金编号2008ZX10003013）；中国人民解放军总医院第二附属医院基金资助项目（基金编号2007AY04）。

Therapeutic effects of DNA vaccines in a mouse model of multi-drug resistant Mycobacterium tuberculosis infection

Liang Yan¹，Wu Xueqiong¹， Li Zhongming²,Zhang Junxian¹,Li Ning¹,Yang Yourong¹,Yu Qi¹,Bai Xuejuan¹,Song Jingying¹,Wang Lan¹,Shi Yingchang¹,Liu Jie¹,Liu Chenglong¹,Zhu Yan¹,Xu Xueyu¹

1. Tuberculosis Research Institute，the 309^th Hospital of PLA, Beijing 100091, China；2. Shanghai H&G Biotechnology company, Shanghai 200436, China

Online:2009-11-10 Published:2011-11-03

摘要/Abstract

摘要： 目的研究结核分枝杆菌DNA 疫苗治疗小鼠耐多药结核病的效果。方法用结核分枝杆菌耐利福平和异烟肼临床分离株HB361尾静脉注射60只雌性BALB/c小鼠后，将小鼠随机均匀地分为6组，感染后第3d开始，分别用生理盐水（A组）、pVAX1载体（B组）、利福平（C组）、HSP65 DNA疫苗（D组）、Ag85A DNA 疫苗（E组）、Ag85A/ESAT6嵌合DNA疫苗（F组）治疗60d，DNA疫苗每隔15d肌肉注射1次，共5次。治疗结束后3周，分别取肺和脾观察病理改变、称取重量、做菌落计数。结果治疗结束后3周，与对照组比较，D组、E组和F组肺脏病变有不同程度减轻，病变局限，2/3区域可见正常的肺泡结构，肺泡轮廓相对清晰，细胞分布均匀。与A组相比，D组、E组和F组肺脏菌落数分别减少了0.34、0.50 和0.26 logs；脾脏菌落数依次减少了0.37、0.46 和0.28 logs（P<0.05，P<0.01）。结论Ag85A DNA 疫苗治疗小鼠耐多药结核病效果优于HSP65 DNA和Ag85A/ESAT6嵌合DNA疫苗。

关键词: 结核, 抗多种药物性/治疗, 疫苗, DNA, 分枝杆菌, 结核

Abstract: ObjectiveTo evaluate the therapeutic effects of DNA vaccines in mouse models of M. tuberculosis infection to establish new immunotherapeutic agents to treat MDR-TB. Methods60 female BALB/C mice were infected intravenously via the tail vein with 220000 CFU of clinical isolate M. tuberculosis HB361 which was resistant to RFP and INH, then randomly divided into 6 groups and treated as follow at the third day after infection for 60 days: saline, plasmid vector, rifampin, HSP65 plasmid DNA, Ag85A plasmid DNA vaccines，Ag85A/ESAT-6 chimeric plasmid DNA vaccines. DNA vaccines were injected intramuscularly 5 times at 15 days intervals. The lungs and spleens from the mice were taken and their pathological changes, weight and number of mycobacterial colony were examined at 3 weeks after the end of treatment. ResultsThe histopathological changes of lung showed that the lung lesions were slight and limited, there were relatively clear profile and normal structure of the alveoli in the treatment groups compared with the control groups. Compared with vector group, HSP65, Ag85A, Ag85A/ESAT-6 DNA vaccines groups reduced the pulmonary bacterial loads by 0.30, 0.50, 0.26 logs, and reduced the splenic bacterial loads by 0.37, 0.46, and 0.28 logs (P<0.05 or P<0.01), respectively. ConclusionThe immunotherapeutic effects of Ag85A DNA was significantly stronger than that of chimeric Ag85A/ESAT6 DNA，HSP65 DNA vaccine in the mouse model of multi-drug resistant tuberculosis.

Key words: tuberculosis,multidrug-resistant/therapy, vaccines,DNA, mycobacterium tuberculosis

梁艳,吴雪琼,李忠明,张俊仙,李宁,阳幼荣,余琦,白雪娟,宋晶莹,王兰,史迎昌,刘洁,刘成龙,朱琰,徐雪玉. DNA疫苗治疗小鼠耐多药结核病的研究[J]. 中国防痨杂志, 2009, 31(11): 649-652.

Liang Yan¹，Wu Xueqiong¹， Li Zhongming²,Zhang Junxian¹,Li Ning¹,Yang Yourong¹,Yu Qi¹,Bai Xuejuan¹,Song Jingying¹,Wang Lan¹,Shi Yingchang¹,Liu Jie¹,Liu Chenglong¹,Zhu Yan¹,Xu Xueyu¹. Therapeutic effects of DNA vaccines in a mouse model of multi-drug resistant Mycobacterium tuberculosis infection[J]. Chinese Journal of Antituberculosis, 2009, 31(11): 649-652.