ECMS-油佐剂对兔波氏杆菌灭活苗免疫增强作用的研究

Immune enhancement of ECMS-oil adjuvant on vaccination of inactivated Bordetella in rabbits

为了研究ECMS-矿物油混合佐剂对兔支气管败血波氏杆菌灭活疫苗的免疫增强作用,在灭活的波氏杆菌中加入不同的佐剂,制备波氏杆菌灭活苗。试验A：选取个体质量1.0 kg左右的健康1月龄新西兰肉兔60只,随机分为以下五组,其中第1～2组为不同剂量ECMS-油佐剂组,第3组为矿物油佐剂疫苗组,第4组为铝胶疫苗组,第5组为无佐剂对照组。各组分别于免疫前及免疫后不同时间进行耳静脉采血,分离血清,用间接ELISA法以及微量凝集法（MAT）检测各组兔血清抗体水平,并对试验兔进行攻毒感染,检测疫苗保护率。试验B：选取个体质量1.0 kg左右的健康1月龄新西兰肉兔32只,平均分为以下四组,分别为ECMS800 g-油佐剂组、矿物油佐剂疫苗组、铝胶疫苗组、无佐剂对照组。于免疫后37 d,无菌心脏采集抗凝血进行淋巴细胞转化试验（MTT）,检测各组淋巴细胞增殖情况。结果显示,免疫后13 d开始产生抗体,ECMS-油组和油佐剂组抗体水平均在41 d达到最高,并持续到184 d,三组间差异不显著（P〈0.05）,但与铝胶和无佐剂组差异显著（P〈0.05）,且ECMS800 g-油组均略高于其他两组;MTT结果表明Con A和LPS刺激下,ECMS800 g-油佐剂组的刺激指数显著（P〈0.05）高于其它各组;ECMS800 g-油佐剂组的攻毒保护率高达100%。结果表明,ECMS和油佐剂两者混合使用可能存在协同作用,可有效提高兔支气管败血波氏杆菌灭活疫苗的免疫效果。

In order to investigate the immune enhancement effect of ECMS mixed with mineral oil adjuvant on inactivated Bordetella bronchiseptica（ Bb） vaccine in rabbits,different doses of ECMS-mineral oil,mineral oil and aluminum adjuvants were mixed with inactivated Bb vaccine. Experiment A： Sixty rabbits were divided into 5 groups（ n = 12）. Group 1-2 were immunized with Bb vaccine containing 750 l mineral oil added with 400 g or 800 g of ECMS; Group 3 were immunized with Bb vaccine containing 750 l mineral oil; Group 4 were immunized with Bb vaccine mixed with 100 l alumina gel; Group 5 were immunized with Bb vaccine alone. Blood samples were collected at different time points post-immunization. Humoral immune responses against whole bacteria protein were evaluated by indirect ELISA and micro-agglutination assay. All the experimental rabbits were challenged with Bb in the end. Experiment B： Thirty-two rabbits were divided into 4groups（ n = 8）,including ECMS 800 g- mineral oil＋Bb vaccine,mineral oil＋Bb vaccine,alumina gel＋Bb vaccine and Bb vaccine groups.Blood samples was collected,and cell mediated immune response was evaluated by lymphocyte transformation test 37 d post-immunization.The results showed that： ECMS- mineral oil and mineral oil groups could significantly improve the humoral immune levels of rabbits than other groups. Each group began to produce antibody on 13 d post-immunization. Antibody levels reached the highest on 41 d post-immunization and the high level of antibody continued to maintain until 184 d. Antibody levels against whole bacteria protein in ECMS- mineral oil and mineral oil groups were significantly higher than those in other groups between 13 and 184 d post-immunization. No significant difference was found between ECMS- mineral oil and mineral oil groups,but ECMS800 g- mineral oil group was slightly higher than ECMS400 g- mineral oil or mineral oil group. MTT showed that,stimulation index（ SI） in ECMS800 g- mineral oil group with PHA and LPS stimulation was significant higher than that in other groups. Protection rate in ECMS800 g- mineral oil group was 100%,much higher than the other groups in challenge experiment. It suggests ECMS mixed mineral oil adjuvant can effectively improve the immune effect of inactivated Bb vaccine in rabbits.