Zhang Hai, Shi Chang-hong, Xue Ying, Bai Yin-lan, Wang Li-mei, Xu Zhi-kai
Laboratory Animal Research Center, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jul;22(4):443-6.
To study murine humoral and cellular immune response induced by fusion protein ESAT6-CFP10 and to examine its protective efficacy against M. tuberculosis (MTB) in mice.
BALB/c mice were immunized subcutaneously on the back with fusion protein ESAT6-CFP10 that was transferred to nitro cellulose (NC) membrane beforehand. Stimulation index (SI) of the spleen lymphocytes of the immunized mice was measured by MTT colorimetry. The level of IFN-gamma and IL-2 and CTL upon antigen-specific stimulation were detected. The vaccinated BALB/c mice were intravenously infected with MTB H37Rv (10(5) CFU/mouse). Four weeks later the number of CFU in spleens was determined.
The titer of serum specific antibody in BALB/c mice immunized with fusion protein ESAT6-CFP10 was 1:6,400. The SI of fusion protein immunized group (1.90+/-0.15) was significantly higher than that of saline-immunized group (0.9+/-0.15). The level of IFN-gamma and IL-2 induced by the fusion protein was 1.792+/-19 ng/L and 0.211+/-11 ng/L respectively, which was significantly higher than that of saline-immunized group and lower than that of BCG-immunized group. The specific killing activity of splenocytes was 36%. Compared with the saline-immunized mice (bacterial load was 6.51+/-0.13), MTB number (bacterial load was 5.24+/-0.15) was reduced dramatically in the spleens of BALB/c mice immunized with the fusion protein, but the protective efficacy of the mice immunized with BCG was higher than that of ESAT6-CFP10 vaccinated group.
Fusion protein ESAT6-CFP10 can be used as a candidate for novel vaccines.
研究融合蛋白ESAT6-CFP10诱导的小鼠体液免疫和细胞免疫反应,并检测其对小鼠结核分枝杆菌(MTB)的保护效力。
将预先转移至硝酸纤维素(NC)膜上的融合蛋白ESAT6-CFP10皮下注射免疫BALB/c小鼠背部。采用MTT比色法检测免疫小鼠脾淋巴细胞的刺激指数(SI)。检测抗原特异性刺激后IFN-γ、IL-2水平及CTL。对免疫后的BALB/c小鼠静脉注射MTB H37Rv(10⁵CFU/只)。4周后测定脾脏中的CFU数量。
用融合蛋白ESAT6-CFP10免疫的BALB/c小鼠血清特异性抗体效价为1:6400。融合蛋白免疫组的SI(1.90±0.15)显著高于生理盐水免疫组(0.9±0.15)。融合蛋白诱导的IFN-γ和IL-2水平分别为1.792±19 ng/L和0.211±11 ng/L,显著高于生理盐水免疫组且低于卡介苗免疫组。脾细胞的特异性杀伤活性为36%。与生理盐水免疫小鼠(细菌载量为6.51±0.13)相比,用融合蛋白免疫的BALB/c小鼠脾脏中的MTB数量(细菌载量为5.24±