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目的采用微量溶血分光光度法测定气管炎疫苗的免疫活性。方法将BALB/c小鼠随机分为5组:空白对照组、阴性对照组及试验1、2、3组,阴性对照组免疫生理盐水,试验1、2、3组分别免疫3批气管炎疫苗,免疫途径为小鼠颈背部皮下,免疫剂量为0.5 ml/只,免疫时间为第1、7、14天,除空白对照组外,所有小鼠第22天经腹腔注射20%的绵羊红细胞(sheep red blood cell,SRBC)悬液,0.2 ml/只。4 d后处死小鼠,取脾脏,制备脾细胞悬液,与SRBC结合,于37℃,在补体的参与下,使抗体形成细胞周围的SRBC溶解,通过酶标仪测定抗体形成细胞介导的溶血反应中,SRBC溶解后释放血红蛋白的量。另设7组小鼠:空白对照组、阴性对照组及试验4、5、6、7、8组,阴性对照组免疫生理盐水,试验4、5、6、7、8组分别免疫不同浓度(6×108、3×108、1.5×108、0.75×108、0.375×108/ml)的气管炎疫苗,按上述方法检测不同浓度气管炎疫苗的免疫活性。结果在抗体形成细胞介导的溶血反应中,3个试验组SRBC溶解后释放的血红蛋白的量均明显高于阴性对照组(P<0.05)。随着气管炎疫苗浓度的降低,SRBC溶解后释放的血红蛋白的量也呈下降趋势,气管炎疫苗浓度在1.5×108/ml以上时,其免疫活性明显高于阴性对照组(P<0.05);低于0.75×108/ml时,与阴性对照组比较,差异无统计学意义(P>0.05)。结论微量溶血分光光度法可用于检测气管炎疫苗的免疫活性。气管炎疫苗浓度在1.5×108/ml以上时,即可增强小鼠免疫功能。
Objective To measure the immunological activity of bronchitis vaccine by using microhemolytic spectrophotometry. Methods BALB / c mice were randomly divided into 5 groups: blank control group, negative control group and experimental groups 1, 2 and 3, and negative control group immunized with physiological saline. Groups 1, 2 and 3 were immunized with 3 groups of bronchitis vaccine The immunization route was subcutaneous in the back of the neck of the mice. The immunization dose was 0.5 ml per mouse and the immunization time was on the 1st, 7th and 14th days. All mice except the blank control group were injected intraperitoneally with 20% sheep erythrocytes sheep red blood cell, SRBC) suspension, 0.2 ml / mouse. After 4 days, the mice were sacrificed and the spleens were harvested. Spleen cell suspensions were prepared and combined with SRBC to lyse the SRBC surrounding the antibody-forming cells at 37 ° C with antibody supplementation. Antibody-forming cell-mediated In hemolytic reaction, the amount of hemoglobin released after SRBC is dissolved. Another 7 mice were set up: blank control group, negative control group and test group 4, 5, 6, 7 and 8, and negative control group immunized with normal saline. Groups 4, 5, 6, 7 and 8 were immunized with different concentrations of 6 × 108,3 × 108,1.5 × 108,0.75 × 108,0.375 × 108 / ml) bronchitis vaccine, according to the above method to detect different concentrations of tracheitis vaccine immunocompetence. Results In antibody -mediated cell-mediated hemolysis, the amount of hemoglobin released from the SRBCs in the three experimental groups was significantly higher than that in the negative control group (P <0.05). With the decrease of the concentration of bronchitis vaccine, the amount of hemoglobin released by SRBC also showed a downward trend. When the concentration of bronchitis vaccine was above 1.5 × 108 / ml, the immunological activity was significantly higher than that of the negative control group (P <0.05). When it was less than 0.75 × 108 / ml, there was no significant difference between the two groups (P> 0.05). Conclusion Michemolytic spectrophotometry can be used to detect the immunological activity of bronchitis vaccine. Bronchitis vaccine concentration of 1.5 × 108 / ml or more, you can enhance immune function in mice.