摘要
目的比较C群脑膜炎球菌多糖蛋白质结合物(简称结合物)的相对分子质量大小和免疫剂量对其在小鼠体内免疫原性的影响,为结合物的分子大小的质控和结合疫苗成品免疫剂量的选择提供实验依据。方法利用CDAP活化法制备C群脑膜炎球菌结合物,通过硫酸铵盐析进行纯化,然后利用Sepharose CL-4B凝胶过滤层析分析,并根据化学检测结果将结合物分为KD0-0.2(组分1)、KD0.2-0.4(组分2)、KD0.4-0.7(组分3)等3个不同相对分子质量组分,每份分别采用1.0μg、0.2μg的免疫剂量免疫小鼠,并对血清进行ELISA分析。结果采用1.0μg免疫剂量时,3种不同相对分子质量的C群脑膜炎球菌结合物均能产生较高水平的抗体,具有典型的加强效应,第2剂免疫即可产生高水平的抗体,而第2、3剂免疫后的抗体水平差异无统计学意义;采用0.2μg免疫剂量时,三种不同相对分子质量的C群脑膜炎球菌结合物只有在第3剂免疫后才能产生较高水平抗体,第1、2剂免疫后的抗体水平差异无统计学意义,组分1和组分2在第3剂免疫后产生的抗体水平优于组分3;对于相同相对分子质量结合物,用1.0μg免疫小鼠产生的抗体水平优于0.2μg免疫组,而且有显著的统计学意义。结论高剂量时,多糖相对分子质量大小对C群脑膜炎球菌结合物的免疫原性没有显著性影响;低剂量时,小相对分子质量结合物对其免疫原性有影响。同等相对分子质量时,1.0μg比0.2μg的免疫剂量可以激发更高的抗体水平。
Objective The mice were immunized by group C meningococcal conjugates with different molecular sizes and different doses,then the immunogenicity and related influence were compared and analyzed. Based on the experiment result,the scientific basis was provided in quality control of molecular size for conjugates and optimal selection of dose for group C meningococcal conjugate vaccine. Methods Group C meningococcal conjugates were prepared by CDAP activation and purified by ammonium sulfate precipitation,and divided into KD0. 0- 0. 2（ part Ⅰ）,KD0. 2- 0. 4（ part Ⅱ）,KD0. 4- 0. 7（ part Ⅲ）,then processed and analysed by Sepharose CL-4B filtration. Immunized mice with 1. 0 μg or 0. 2 μg dose of each part,and immunized sera were analyzed by ELISA. Results Three conjugates with different molecular size in1. 0 μg dose stimulated high levels of Ig G antibodies,with a typical enhance effect. The conjugates evoked higher levels of Ig G antibodies after the second dose. There was not a statistical difference in antibody levels generated between the second and third dose. Immunized with 0. 2 μg dose in mice,three conjugates stimulated higher level antibodies only after the third dose. There was not a statistical difference in antibody levels generated by the first and second immunization. The partⅠ and part Ⅱconjugates produced higher levels of antibody than part Ⅲ after the third immunization. As immunized with same molecular size conjugates,1. 0 μg dose groups produced antibody level was higher than 0. 2 μg dose ones,which had a significant statistical differences in antibody levels. Conclusion In high dose immunization groups,it had no apparent influence to the immunogenicity from three conjugates with different molecular size,and immunogenicity could be influenced by conjugate with small molecular size at a low dose. In comparison,the higher immunoresponse could be induced by immunization with 1. 0 μg dose than 0. 2 μg dose.
出处
《微生物学免疫学进展》
2016年第1期7-13,共7页
Progress In Microbiology and Immunology
基金
国家"重大新药创制"专项(2013ZX09402-302-215)
甘肃省科技重大专项(0801NKDA003)
