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目的研究结核分枝杆菌Ag85A/ESAT-6嵌合基因DNA疫苗及其与抗痨药物联合治疗小鼠结核病的效果。方法用结核分枝杆菌临床分离的利福平和异烟肼敏感株HB240-1尾静脉注射17～19g的6～8周龄雌性BALB/c小鼠后,将小鼠随机均匀地分为5组,感染后第3d开始,分别用pVAX1空载体(A组)、利福平和异烟肼(B组)、利福平、异烟肼和Ag85A/ESAT-6嵌合基因DNA疫苗(C组)、Ag85A/ESAT-6嵌合基因DNA疫苗(D组)、HSP65基因DNA疫苗(E组)治疗40d,每组10只小鼠。治疗结束后2w,分别取肺、肝和脾观察病理改变、称取重量、做菌落计数。结果小鼠感染5w后,肺内菌量可达到1.8×106CFU,脾内菌量达到1.7×106CFU。治疗结束后2w,B组和C组肺、脾脏器重量指数显著低于D组和E组,D组和E组脾脏器重量指数低于A组,但差别不显著。A组肺脏病变程度最重,病变范围80%～100%,干酪样坏死灶为弥漫性;D组和E组肺脏病变范围50%,干酪样坏死灶为片状;B组和C组肺脏病变程度较轻,病变范围较小,干酪样坏死灶为点状。A组脾脏9只重度肿大;B组和C组脾脏未见明显病变;D组和E组脾脏各有4只重度肿大。与A组相比,E组、D组、B组和C组肺脏菌落数分别减少了45%、50%、98.9%、99%;脾脏菌落数依次减少了50%、55%、99.2%、98.4%。与pVAX1空载体治疗组相比,各治疗组均有不同程度的疗效,Ag85A/ESAT-6嵌合基因疫苗组与HSP65基因疫苗组疗效相当。结论Ag85A/ESAT-6嵌合基因DNA疫苗组联合化疗的疗效明显强于Ag85A/ESAT6嵌合基因DNA疫苗单独应用。
Objective To study the DNA vaccine of Mycobacterium tuberculosis Ag85A / ESAT-6 chimeric gene and its efficacy in combination with anti-tuberculosis drugs in the treatment of tuberculosis in mice. Methods Seventeen to 19g female BALB / c mice aged 6-8 weeks were injected into tail vein of Mycobacterium tuberculosis clinically isolated rifampicin and isoniazid sensitive strain HB240-1. The mice were randomly divided into 5 groups , The third day after infection, DNA vaccine (group C) was obtained by using pVAX1 empty vector (group A), rifampicin and isoniazid (group B), rifampicin, isoniazid and Ag85A / ESAT- , Ag85A / ESAT-6 chimeric DNA vaccine (group D) and HSP65 DNA vaccine (group E) for 40 days. After treatment 2w, respectively, take the lung, liver and spleen pathological changes were observed, weighed, do colony count. Results After 5 weeks of infection in mice, the amount of bacteria in the lungs reached 1.8 × 106CFU, and the intra-spleen bacteria reached 1.7 × 106CFU. At 2 weeks after treatment, the weight index of lung and spleen in group B and C were significantly lower than those in group D and E, and those in group D and E were lower than those in group A, but the difference was not significant. A group of lung lesions the most severe lesions ranging from 80% to 100%, caseous necrosis was diffuse; D group and E group of lung lesions range 50%, caseous necrosis lesions for the sheet; B group and C group of lung lesions To a lesser extent, the lesser extent of the lesion, the caseous necrosis was punctate. A group of 9 spleen severe enlargement; B group and C group spleen no obvious lesions; D group and E group spleen each have 4 severe enlarged. Compared with group A, the number of lung colonies in group E, group D, group B and group C decreased by 45%, 50%, 98.9% and 99%, respectively; the number of colonies in spleens decreased by 50%, 55% and 99.2% 98.4%. Compared with the pVAX1 empty vector treatment group, each treatment group has different degrees of efficacy, Ag85A / ESAT-6 chimeric gene vaccine group and HSP65 gene vaccine group efficacy. Conclusion The curative effect of combination chemotherapy with Ag85A / ESAT-6 chimeric DNA vaccine group is obviously stronger than DNA vaccine with Ag85A / ESAT6 chimeric gene alone.