【作者】 秦卫兵；

【导师】 谭锦泉； 蒋建伟； 肖昕；

【作者基本信息】 安徽医科大学 ， 免疫学， 2003， 硕士

【摘要】 目的：探讨CpG ODN对免疫抑制小鼠、老年鼠、孕鼠及其新生小鼠的免疫刺激作用；筛选对人脐血淋巴细胞免疫刺激作用较强的一种CpG ODN；观察其对早产儿脐血淋巴细胞的免疫刺激作用及刺激NK细胞对K₅₆₂细胞的杀伤活性，为开发适合人类新型疫苗佐剂奠定基础。方法：选用公认的对小鼠作用最强的CpG 1826与乙肝疫苗经左侧胫前肌同时肌注到免疫抑制小鼠、老年鼠和孕鼠体内，两周后同剂量加强免疫一次，用ELISA方法检测抗HBsAg抗体、IL-12水平；取各模型小鼠的脾脏做病理切片，行H-E染色，在光学显微镜镜下观察脾脏的细胞变化；按CpG ODN的设计原则设计6种非硫代化CpG ODN，用MTT法筛选对人类免疫细胞刺激作用较强的CpG ODN；同时用MTT法观察CpG ODN对早产儿脐血淋巴细胞增殖反应；用LDH法观察经CpG ODN刺激过的NK细胞对K₅₆₂细胞的杀伤情况。结果：CpG ODN能使免疫抑制小鼠、老年鼠产生抗HBsAg抗体、IL-12水平升高。但20μg CpG ODN不能明显提高孕鼠产生抗HBsAg抗体水平，也不能明显增强其新生小鼠的体液免疫力。用MTT法筛选出一条CpG ODN4，发现其对足月儿和早产儿脐血淋巴细胞具有较强的免疫刺激作用，而且对足月儿脐血淋巴细胞增殖反应比早产儿的强，对K₅₆₂细胞具有明显的杀伤作用。结论：CpG ODN能提高免疫抑制小鼠、老年鼠的免疫能力，但20μg CpG ODN尚不能明显增强孕鼠及新生小鼠的体液免疫力。人特异性CpG ODN对人脐血淋巴细胞具有较强的免疫刺激作用。更多还原

【Abstract】 Objective: To investigate immunostimulatory activity of CpG ODN against immunosuppression mice, aged mice, pregnant mice and their newly born mice; To screen a CpG ODN that its immunostimulatory activity against human cord blood lymphocytes is stronger than others; To observe immunostimulatory activity of CpG ODN against premature infants cord blood lymphocytes and to observe its cytolysis against Kse2 cells. Methods: All mice were received an injection into the left tibialis anterior muscle with HBsAg vaccine or both HBsAg vaccine and CpG 1826 that was generally accepted as the optimal CpG ODN in mice, then mice were boosted at 2 weeks with the same formulation as was used at prime. The specific antibody IgG and IL-12 were measured by ELISA. Spleen lymphocytes evaluation was undertaken using hematoxylin and eosin-stained paraffin sections in light microscope. We designed six phosphorothioate CpG ODNs according to the law of CpG ODN design , one CpG ODN that its immunostimulatory activity against human cord blood lymphocytes is stronger than others was screened by MTT. Immunostimulatory activity of CpG ODN against premature infants cord blood lymphocytes was observed by MTT and its cytolysis against K562 cells was observed by LDH. Results: There was an significant increase in anti-HBsAg IgG and IL-12 when CpG ODN was mixed with HBsAg vaccine in immunity suppression model mice; there were also significant increases in anti-HBsAg IgG and IL-12 when 10 g and 20 g CpG ODN were mixed with HBsAg vaccine in aged mice. But 20 g CpG ODN cann’t enhance the immunity of pregnant mice and their newly born mice. We screened out CpG ODN4 that has highly immunostimulatory activity against term infants and premature infants cord blood lymphocytes. We also showed that the immunostimulatory activity of CpG ODN4against term infants cord blood lymphocytes was stronger than those of premature infants. We observed CpG ODN4 had strong cytolytic activity against K562 cells. Conclusions: CpG ODN can improve the immunity of immunosuppression mice, aged mice, but 20 g CpG ODN cann’t enhance the immunity of pregnant mice and its newly born mice. Human specific CpG ODN has highly immunostimulatory activity against human cord blood lymphocytes.更多还原