Abstract:The objective of the present study was to investigate immune effect by using ESAT6, CFP10 DNA vaccine combined with BCG to immunize mice. DNA vaccine pEGFP-N1-ESAT6 and pEGFP-N1-CFP10 was primarily constructed, and 80 mice were randomly divided into eight groups including saline group (SLYS), BCG group (KJM), pEGFP-N1 group (P-N1), pEGFP-N1-ESAT6 group (P-N1-E), pEGFP-N1-CFP10 group (P-N1-C), BCG plus pEGFP-N1 group (K+P-N1), BCG plus pEGFP-N1-ESAT6 group (K+P-N1-E), and BCG plus pEGFP-N1-CFP10 group (K+P-N1-C). Each mouse was intradermally injected with 100 μL BCG, while each mouse was intramuscularly injected with plasmid pEGFP-N1-ESAT6, pEGFP-N1-CFP10 (50 μg). Each group was injected for three times. Purified ESAT6, CFP10 proteins were used as antigen. DOT-ELISA method was used to detect antibodies in the serum of mice. ELISA method was used to detect the change of the IFN-γ in the serum of mice. Antibodies against CFP10 proteins were detected in mouse serum after three immunizations. Antibody against ESAT6 proteins was not detected. However, IFN-γ levels in serum increased significantly. The average content of the measured IFN-γ was significantly higher than that before immunization; the first immunization and the second immunization (P<0.01) was (107.591±7.3281) pg/mL and (95.7503±9.0184) pg/mL after three times immunization with K+P-N1-C and K+P-N1-E. It concluded that Mycobacterium tuberculosis ESAT6 and CFP10 DNA vaccine combined with BCG combination could induce an effective cellular immune response, which lay a foundation for the study of DNA vaccines.
王慧勤, 李跃峰, 李志强, 温书香, 陈鹏博, 赵庆亮, 纪太旺, 曹旭东, 陈创夫. 结核分枝杆菌ESAT6、CFP10基因DNA疫苗对小鼠免疫原性的初步研究[J]. 中国人兽共患病学报, 2014, 30(5): 458-463.
WANG Hui-qin, LI Yue-feng, LI Zhi-qiang, WEN Shu-xiang, CHEN Peng-bo, ZHAO Qing-liang, JI Tai-wang, CAO Xun-dong, CHEN Chuang-fu. Preliminary study of Mycobacterium tuberculosis ESAT6 and CFP10 DNA vaccine immunogenicity in mice. Chinese Journal of Zoonoses, 2014, 30(5): 458-463.
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