摘要
试验旨在探讨免疫刺激复合物基质(ISCOM-基质)作为猪支原体肺炎168株活疫苗佐剂使用的可行性,并考察其配合疫苗肌肉注射时的免疫效力和攻毒保护效力。首先以Quil A、胆固醇和磷脂为原料,采用透析法制备ISCOM-基质,然后将制备好的ISCOM-基质与活疫苗混合孵育,检验其对活疫苗的毒性。于体外用ISCOM-matrix直接刺激或与刀豆蛋白共同刺激淋巴细胞,观察细胞的增殖应答情况。而后将添加ISCOM-matrix佐剂的猪支原体肺炎活疫苗肌肉注射免疫动物,检测血清抗体水平及淋巴细胞增殖应答水平,于免疫后8周攻毒,评价攻毒保护效力。结果表明,所制备的ISCOM-基质不影响活疫苗的活力,并且在体外可直接或协同刀豆蛋白刺激淋巴细胞产生明显的增殖反应,最低起效浓度为0.01 ng/ml;当作为佐剂与活疫苗配合肌肉注射免疫时,可以显著诱导动物对疫苗抗原的细胞免疫应答,体液免疫应答亦有一定程度增强。用肺炎支原体强毒攻毒后25 d剖检,免疫组病变明显较攻毒对照组减轻,显示出较好的攻毒保护效果。
This experiment aims to study the possibility of ISCOM-matrix（immune stimulating complex-matrix） to be used as adjuvant of the live attenuated vaccine strain 168 against mycoplasmal pneumonia of pig,and to evaluate the immune response and protection after intramuscular inoculation.ISCOM-matrix was prepared with Quil A,cholesterol and phosphatidylcholine through dialysis.The prepared ISCOM-matrix was incubated with live vaccine to test its toxicity to Mycoplasma hyopneumoniae.The response of lymphocytes to the in vitro stimulation of ISCOM-matrix alone or ISCOM-matrix combined with Con A was observed.The live vaccine adjuvanted with ISCOM-matrix was used to inoculate pigs intramuscularly.Immune response was evaluated by detecting lymphocyte proliferation and serum antibody level.Challenge was performed at 8 weeks after the first immunization to evaluate protection efficacy of the vaccine.The results were as follows.The prepared ISCOM-matrix showed no influence on CCU（colour change unit） of the live vaccine,and it significantly induced proliferation of lymphocyte in vitro both stimulated by itself and co-stimulated with Con A.Intramuscular inoculation with the live vaccine mixed with ISCOM-matrix induced obviously specific cellular immune response and humoral immune response.After challenge,pigs of the inoculated group showed less pathological changes than the ones with no immunization.
出处
《江苏农业学报》
CSCD
北大核心
2011年第6期1310-1315,共6页
Jiangsu Journal of Agricultural Sciences
基金
江苏省博士后科研资助计划项目(0902063C)
中国博士后科学基金面上资助项目(20090461075)
江苏省农业科技自主创新基金项目[CX(10)215]
国家农业科技成果转化基金项目(2009GB2C100129)
关键词
免疫刺激复合物基质
猪支原体肺炎
活疫苗
肌肉注射
ISCOM-matrix
mycoplasmal pneumonia of swine
live vaccine
intra muscular immunization
