重组粘着素用于百日咳疫苗开发的评价研究

发布时间：2018-03-31 17:05

本文选题：百日咳　切入点：重组粘着素　出处：《昆明理工大学》2009年硕士论文

【摘要】：PRN(百日咳粘着素,Pertactin,PRN)是无细胞百日咳组分疫苗的重要组成部分。天然PRN纯化工艺复杂,制备成本高,用重组PRN代替天然PRN是开发新型百日咳疫苗的发展趋势。本论文对重组PRN的免疫原性、抗原性和保护力进行评价,并研究其激发机体免疫应答的类型。重组PRN和天然PRN交叉检测重组PRN组和天然PRN组两次免疫的PRN抗体,两组的抗体滴度无显著性差异(p0.05)。此结果表明重组PRN具有较好的免疫原性和抗原性。百日咳的攻击实验表明,重组PRN免疫小鼠后可以产生一定的保护力,抵抗百日咳攻击菌18323的感染。在三个免疫剂量中,中剂量的保护率最高。将重组PRN和天然PRN分别与百日咳疫苗原液、FIM2和FIM3组合模拟无细胞百日咳组分疫苗,进行百日咳攻击实验。结果表明,重组PRN+原液+FIM2+FIM3组高剂量的存活率和天然PRN+原液+FIM2+FIM3组高剂量的相同,均为最高值93.75%,而中剂量和低剂量的存活率高于天然PRN+原液+FIM2+FIM3组的。细胞因子的检测结果表明,重组PRN组分泌较高水平的Th2型细胞因子IL-4和IL-12,而Th1型细胞因子IL-2,IL-10和IFN-γ的分泌量较低,说明重组PRN可以激发体液免疫和细胞免疫,但以体液免疫应答为主。天然PRN组的应答类型与重组PRN组相同。模拟疫苗重组PRN+原液+FIM2+FIM3组和天然PRN+原液+FIM2+FM3组也以体液免疫为主要的应答方式。通过对IgGl和IgG2a的测定表明,重组PRN组、天然PRN组、重组PRN+原液+FIM2+FIM3组和天然PRN+原液+FIM2+FIM3组既产生IgGl又产生IgG2a,且IgGl高于IgG2a,说明以上抗原可以激发机体产生一定程度的体液免疫和细胞免疫,但体液免疫为主要的免疫应答类型。本文的研究标明,重组PRN完全有可能代替天然PRN应用于百日咳组分疫苗。此研究为研发新型的多组分百日咳疫苗提供了有效的实践依据。
[Abstract]:PRN (pertussis adhesives) is an important component of acellular pertussis vaccine. The purification process of natural PRN is complex and the preparation cost is high. It is a trend to develop new pertussis vaccine with recombinant PRN instead of natural PRN. In this paper, the immunogenicity, antigenicity and protective power of recombinant PRN were evaluated, and the types of immune response induced by recombinant PRN were studied. The antibody titers of recombinant PRN and natural PRN were not significantly different from those of PRN group and natural PRN group. The results showed that the recombinant PRN had good immunogenicity and antigenicity. The results of pertussis attack test showed that there was no significant difference in the titer of PRN antibody between the two groups, and the results showed that the recombinant PRN had good immunogenicity and antigenicity. After immunizing mice with recombinant PRN, it can produce a certain protective effect against the infection of 18323 pertussis. The recombinant PRN and natural PRN were combined with pertussis vaccine extract (FIM2) and FIM3 to simulate the acellular pertussis component vaccine. The results showed that the recombinant PRN and natural PRN were used to simulate the acellular pertussis component vaccine. The high dose survival rate of FIM2 FIM3 group was the same as that of FIM2 FIM3 group, which was the highest value of 93.75%, but the survival rate of medium and low dose of FIM2 FIM3 group was higher than that of FIM2 FIM3 group. The results of cytokine detection showed that the recombinant PRN group secreted higher levels of Th2 type cytokines IL-4 and IL-12, while Th1 type cytokines IL-2, IL-10 and IFN- 纬 secreted less, suggesting that recombinant PRN could stimulate humoral and cellular immunity. But humoral immune response was dominant. The type of response in natural PRN group was the same as that in recombinant PRN group. Humoral immunity was also the main response mode in FIM2 FIM3 group and FIM2 FM3 group. The determination of IgGl and IgG2a showed that the recombinant PRN group, the natural PRN group, The recombinant PRN FIM2 FIM3 group and the natural PRN FIM2 FIM3 group produced both IgGl and IgG2a, and the IgGl was higher than IgG2a, indicating that the above antigens could stimulate humoral and cellular immunity to some extent, but humoral immunity was the main type of immune response. The research in this paper indicates that recombinant PRN can replace natural PRN in the application of pertussis component vaccine. This study provides an effective practical basis for the development of a new multicomponent pertussis vaccine.
【学位授予单位】：昆明理工大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392

【参考文献】