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豚鼠延髓c |
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张燕华 沙朝晖 李 峻 吴俊梅
摘 要 目的和方法：观察钩体外膜疏水蛋白OmpL39的免疫保护作用和对豚鼠中枢c-fos基因表达的影响，分别用OmpL39与钩体死菌苗(WLCV)、生理盐水(NS)对照免疫豚鼠，用强毒017株钩体活菌攻击，处死动物后用新鲜延髓切片观察对比。结果：OmpL39在豚鼠体内能产生高效价的阳性抗体，免疫保护率为100%。OmpL39免疫组豚鼠中枢c-fos基因表达面积比为0.0086%±0.0003%，颗粒计数为463.00±1.34个，较WLCV和NS组显著减少(P＜0.05)。结论：OmpL39有较强的免疫保护作用，对豚鼠中枢c-fos基因的表达有抑制作用。
主题词 钩端螺旋体属；免疫法；基因，FOS
Changes of the expression of the c-fos of the bulb of quinea pigs
in the immune response of OmpL39 from leptospira
ZHANG Yan-Hua, SHA Zhao-Hui, LI Jun, WU Jun-Mei
Department of Pathophysiology of West China University of Medical Sciences, Chengdu(610041)
Abstract AIM and METHODS To study the immunoprotection and the related mechanism of OmpL39 and the effect of idiosyncrasy antigen on the c-fos expression in the bulb of the guinea pigs. We immunized the guinea pigs separately with OmpL39, whole Leptospira cell vaccine(WLCV) and normal saline (NS) and then attacked them with virulent strain 017.RESULTS:OmpL39 could produce high-valence positive antibody (MAT 1:＞3200). The immunoprotective rate was 100%. The c-fos gene expression granule area rate was 0.0086%±0.0003%, and the granule count was 4.63±1.34. The c-fos gene expression of the guinea pigs of OmpL39 group was remarkably less than that of the control group (WLCV and NS) (P＜0.05).CONCLUSION:OmpL39 could specifically inhibit the expressions the c-fos in the bulb of the guinea pigs. It was more effective in immunoprotection than WLCV.
MeSH Leptospira; Immunization; Genes,FOS
我们曾报道，赖型钩体强毒017株外膜疏水蛋白OmpL39是稳定的免疫原［1,2］。参照《中国生物制品规程》钩体疫苗制造及检定规程［3］，用纯化的OmpL39与钩体死菌苗、生理盐水对照免疫豚鼠，研究OmpL39的免疫保护作用，并对OmpL39抗原特异性刺激影响豚鼠延髓c-fos基因的表达进行了观察，以期为OmpL39基因疫苗的研制及免疫保护机制提供实验和理论依据。
材料与方法
一、钩体菌液：
问号状黄疸出血群赖型017株钩体，由卫生部成都生物制品研究所钩体菌苗室提供。
二、钩体OmpL39的抽提、分离和纯化：
参照Haake等［4］的方法用TritonX－114(TX-114)抽提分离017株钩体疏水OMP，经制备性SDS-PAGE电泳分离纯化相对分子量为39×103疏水外膜蛋白(O[1] [2] 下一页
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