论文部分内容阅读
目的比较不同活化试剂、不同载体蛋白质及不同免疫程序对W135群脑膜炎球菌结合物在小鼠体内的免疫原性的影响。方法分别采用溴化氰(CNBr)和1-氰基-4-二甲基氨基吡啶-四氟硼酸盐(CDAP)为活化试剂,以己二酸二酰肼(ADH)为连接臂制备W135群脑膜炎球菌多糖(group W135 meningococcal polysaccharide,PSW135)衍生物;以碳二亚胺(EDAC)作为缩合剂,将多糖衍生物分别与破伤风类毒素(tetanus toxoid,TT)、白喉类毒素(diphtheria toxoid,DT)和重组铜绿假单胞菌外毒素A(recombinant Pseudomonas aeruginosa exotoxin A,r EPA)3种蛋白质共价结合,制备6种PSW135蛋白质结合物;比较4针免疫程序(0、2、4、8周)和3针免疫程序(0、4、8周)对结合物在小鼠体内免疫原性的影响。结果两种活化试剂均能对多糖进行有效活化,且不会破坏多糖的抗原性;制备的6种结合物在小鼠体内均能产生良好的免疫原性,且具有显著的剂次加强效应;采用间隔2周的3针免疫程序和2.5μg/只的免疫剂量,不同类型的活化试剂和载体蛋白质对结合物的免疫原性无显著影响,但以DT为载体蛋白质的结合物的免疫加强效应要弱于以TT和r EPA为载体的结合物;延长免疫间隔时间可显著提高结合物在小鼠体内的免疫原性。结论初步建立了适用于W135群脑膜炎球菌结合疫苗的结合工艺,并为结合物在小鼠体内的免疫原性研究提供了一种新的候选免疫程序。
Objective To compare the immunogenicity of different W135 meningococcal conjugates in mice with different activating reagents, different carrier proteins and different immune programs. Methods W135 was prepared by using cyanogen bromide (CNBr) and 1 - cyano - 4 - dimethylaminopyridine - tetrafluoroborate (CDAP) as activating reagent and adipic acid dihydrazide (ADH) Group polysaccharide derivative of group W135 meningococcal polysaccharide (PSW135); and polysaccharide derivative of tetrasus toxoid (TT), diphtheria toxoid (DT) and recombinant Pseudomonas aeruginosa exotoxin A (r EPA) were used to prepare six kinds of PSW135 protein conjugates. The four-needle immunization protocol (0,2,4 , 8 weeks) and 3-needle immunization schedule (0, 4, and 8 weeks) on the immunogenicity of the conjugate in mice. Results Both of the activation reagents could effectively activate polysaccharides without damaging the antigenicity of polysaccharides. The six kinds of conjugates produced good immunogenicity in mice and had significant dose-enhancing effect. Using a 3-dose immunization program at 2-week intervals and an immunizing dose of 2.5 [mu] g per vaccine, different types of activating reagent and carrier protein had no significant effect on the immunogenicity of the conjugate, but the immunopotentiating effect of the conjugates with DT as carrier protein Is weaker than the conjugate with TT and r EPA; prolonging the immunization interval can significantly improve the immunogenicity of the conjugate in mice. Conclusion The binding process of meningococcal conjugate vaccine of group W135 was preliminarily established and a new candidate immunization protocol was provided for the study of the immunogenicity of the conjugate in mice.