Tang Quan, Dou Jun, Zhao Feng-shu, Chu Li-li, Pan Meng, Wang Yong-fang
Department of Pathogenic Biology and Immunology, School of Basic Medical Science, Southeast University, Nanjing 210009, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Jul;23(7):591-4.
To investigate immune effect in mice on the basis of BCG priming and DNA vaccine boosting, and to provide a new strategy for development of new type of anti-tuberculosis DNA vaccine further.
After mice were inoculated with BCG of 1x10(6) clone formation unit each three weeks, 100 microg of DNA vaccine was injected intramuscularly in mice two times at 3 week intervals. The proliferative responses of murine cytotoxic T lymphocytes (CTL), natural killer (NK) and spleen cell to antigen 85A(Ag85A) were measured by MTT method respectively. The antibody titers and IFN-gamma level from the immunized mice were detected by ELISA.
The proliferative responses of CTL and spleen cell to Ag85A, as well as IFN-gamma level in mice immunized with prime-boost strategy were significantly increased respectively compared with the control mice immunized with blank plasmid or BCG only. Although NK activity was a little higher in mice immunized with prime-boost strategy than that of immunized with blank plasmid mice, it was still lower than that of mice immunized with BCG alone. The titer of the specific antibody against Ag85A in mice immunized with prime-boost strategy was also higher than that of mice immunized with DNA vaccine alone.
The immune strategy of BCG-prime and Ag85A/GM-CSF DNA vaccine boost improve immune effect, especially the Th1 cellular immune response increase obviously. This study provides the possibility of further research for investigating protective function in immunized mice challenged by Mycobacterium tuberculosis.
探讨在卡介苗(BCG)初免基础上DNA疫苗加强免疫对小鼠的免疫效果,为新型抗结核DNA疫苗的研发提供新策略。
每3周给小鼠接种1×10⁶克隆形成单位的BCG,3周后间隔3周给小鼠肌肉注射100μg DNA疫苗2次。分别采用MTT法检测小鼠细胞毒性T淋巴细胞(CTL)、自然杀伤细胞(NK)及脾细胞对抗原85A(Ag85A)的增殖反应。采用ELISA法检测免疫小鼠的抗体效价及干扰素-γ水平。
与仅接种空白质粒或仅接种BCG的对照小鼠相比,采用初免-加强策略免疫的小鼠CTL和脾细胞对Ag85A的增殖反应以及干扰素-γ水平均显著升高。虽然采用初免-加强策略免疫的小鼠NK活性略高于接种空白质粒的小鼠,但仍低于单独接种BCG的小鼠。采用初免-加强策略免疫的小鼠抗Ag85A特异性抗体效价也高于单独接种DNA疫苗的小鼠。
BCG初免和Ag85A/GM-CSF DNA疫苗加强免疫的免疫策略可提高免疫效果,尤其是Th1细胞免疫反应明显增强。本研究为进一步研究免疫小鼠在受到结核分枝杆菌攻击时的保护作用提供了可能性。