2种益生菌菌体OMP对新城疫病毒免疫增强作用  

Antigenicity of the outer membrane protein of two kinds of probiotics to Newcastle disease virus

作　　者：王小娥[1] 朱瑞良[1] 杨春晓[1] 崔金生[1] 肖海军[1] 王允超 岳寿松 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]青岛康地恩生物科技有限公司,山东青岛266061

出　　处：《中国兽医学报》2009年第3期266-271,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(3067114、30740077)

摘　　要：为研究乳酸杆菌、地衣芽孢杆菌菌体OMP的免疫增强作用,本试验利用超声波细胞粉碎器破碎、TritonX-100处理技术提取了乳酸杆菌与地衣芽孢杆菌OMP,通过考马斯亮蓝G-250法测得两者菌体OMP含量,进行了SDS-PAGE电泳检测,然后将提取的菌体OMP以不同的方式、含量作为免疫增强剂与新城疫病毒尿囊液混合后再与油佐剂以1∶3比例制备成免疫原,免疫7日龄雏鸡,通过持续10周的抗体检测、外周淋巴细胞比例(A)及白细胞吞噬指数(B)检测,判断菌体OMP对新城疫病毒具有免疫增强作用。结果,乳酸杆菌与地衣芽孢杆菌菌体OMP含量分别为2 323.5、867.6 mg/L;SDS-PAGE电泳试验得出分子量分别为6.4×104、8.6×104的乳酸杆菌、地衣芽孢杆菌菌体OMP蛋白含量多,带的颜色较深,为起主要作用的蛋白;各试验组HI抗体效价免疫保护期远远高于单独使用新城疫LaSota株灭活油苗的对照组及空白组,差异显著(P<0.01或P<0.05);各试验组外周淋巴细胞比例(A)及白细胞吞噬指数(B)均高于单独使用新城疫LaSota株灭活油苗的对照组及空白组,差异显著(P<0.01或P<0.05);同时还可以得出菌体OMP含量越高则HI抗体效价、淋巴细胞比例(A)、白细胞吞噬指数(B)均增高;不同菌体OMP混合免疫增强作用高于单独使用一种菌体OMP。由此表明乳酸杆菌、地衣芽孢杆菌菌体OMP能显著增强ND疫苗的免疫效果,为今后研制免疫效果更好地ND疫提供了技术支持。To explore extraction and identification of outer membrane protein of Bacillus acidi ldctici and Bacillus licheniformis. We could determine contents of Bacillus licheniformis and Bacillus acidi lactici＇ outer membrane protein with Coomassie brilliant blue G-250, do SDS-PAGE electrophoresis of Bacillus licheniformis and Bacillus acidi lactici ＇ outer membrane protein and prepare outer membrane protein immunity to immunity seven-day-chikens with different does. The different way and content of outer membrane protein with Newcastle Disease LaSota Virus was admixed to add immunolgic adjuvant use 1 ： 3 to immnue seven-day-chickens. Through 10 weeks identification of antibody titer （log2） of different days about different groups and Lymphocyte proportion（A） and phagoteophic percentage of Leucocyte（B）. The result showed that：The countent of outer membrane protein was 2 323.5,867.6 mg/ L separately. The result of SDS-PAGE showed that the important molecular weight of Bacillus acidi lactici and Bacillus licheniformis＇ outer membrane protein is 6.4 × 10^4 , 8.6 × 10^4. The result of percentage of periph. Lymphocyte （A） and exponent of phagoteophic Leucocyte（B） experiment group 6r HI was highest. All experiment groups had higher HI and lymphocyte proportion（A）, phagoteophic percentage of Leucocyte（B） than only using Newcastle Disease La Sota Vaccine,the diversity was marked （P〈0.01 or P〈0.05）. The higher contents of outer membrane protein,the higher antibody titer and ercentage of periph. Lymphocyte（A） and exponent of phagoteophic Leucocyte（B）. So the use of two kind of breaking bacterial cellular wall could have wide market.

关 键 词：乳酸杆菌 地衣芽孢杆菌 外膜蛋白 提取 新城疫病毒 免疫增强作用 

分 类 号：S852.65[农业科学—基础兽医学]