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目的:研究编码结核分枝杆菌分泌蛋白Ag85B的基因疫苗pTB30m和pTB30s对免疫动物的保护作用。方法:以基因疫苗pTB30m和pTB30s肌注免疫BALB/c小鼠。免疫完成6 wk后,用5×105 CFU的MTB H37Rv毒株经小鼠尾静脉攻击感染。同时用尼龙毛柱分离基因免疫BALB/c小鼠的T细胞,并以5×106 T细胞/只小鼠过继免疫正常BALB/c小鼠,立即用105 CFU的MTB毒株经小鼠尾静脉攻击感染。4 wk后分别计数脾脏中的细菌负荷。结果:与生理盐水对照组相比较,pTB30m及pTB30s质粒免疫组BALB/c小鼠脾脏中的细菌负荷均减少,分别为0.645(log10 CFU,P<0.01)和0.839(log10CFU,P<0.001);而空质粒对照组小鼠脾脏中的细菌负荷减少较少。经质粒pTB30m和pTB30s免疫的BALB/c小鼠的T细胞,过继免疫的正常BALB/c小鼠,对攻击感染的MTB H37Rv毒株在脾脏中的增殖具有部分抑制作用。结论:pTB30s免疫的BALB/c小鼠,对MTB H37 Rv毒株攻击的保护作用优于pTB30m质粒免疫,有望进一步用于结核病的防治研究。
Objective: To study the protective effect of gene vaccines pTB30m and pTB30s encoding Mycobacterium tuberculosis secreting protein Ag85B on the immunized animals. Methods: BALB / c mice were immunized with gene vaccines pTB30m and pTB30s. After 6 weeks of immunization, 5 × 10 5 CFU of MTB H37Rv strain was challenged via the tail vein of mice. BALB / c mice were immunized with nylon-wool column and T cells of BALB / c mice were immunized and normal BALB / c mice were adoptively immunized with 5 × 10 6 T cells / mouse. Immediately, 105 CFU of MTB strain was transplanted via tail vein of mice Attack the infection. After 4 weeks, the bacterial load in the spleen was separately counted. Results: Compared with the saline control group, the bacterial load in the BALB / c mice spleens of pTB30m and pTB30s immunized groups decreased 0.645 (log10 CFU, P <0.01) and 0.839 (log10 CFU, P <0.001), respectively. The empty plasmid control mice spleen bacterial load reduction less. T cells from adoptively immunized BALB / c mice immunized with plasmids pTB30m and pTB30s, and BALB / c mice adoptively immunized, partially inhibited the proliferation of challenged MTB H37Rv strains in the spleen. CONCLUSION: The protective effect of pTB30s immunized BALB / c mice against MTB H37 Rv strain is superior to that of pTB30m plasmid, which is expected to be further used in the prevention and treatment of tuberculosis.