噬菌体作为疫苗载体诱导小鼠脾细胞产生细胞因子类型的研究

发布时间：2018-06-24 17:14

本文选题：系统性白念菌感染 + Hsp90　；参考：《东北师范大学》2007年硕士论文

【摘要】： 白色念珠菌是一种寄生于健康人体的正常菌群,在机体的免疫力下降或者正常菌群失调的情况下,就会转变成致病菌。免疫抑制剂的应用增加了病人对念珠菌的易感性,使医院内患者念珠菌感染的发病率不断增加。热休克蛋白90(Hsp90)是白色念珠菌在侵染机体过程中分泌的一种毒性因子,同时,作为念珠菌的一种免疫优势抗原,可以诱导感染机体产生特异性抗体。Matthews和本实验室的工作证明,抗Hsp90的抗体是一种保护性抗体。我们课题组已经证明,噬菌体展示的表位能够诱导小鼠产生体液免疫应答和细胞免疫应答。在念珠菌感染过程中,CD4+T细胞的分化方向是决定机体对念珠菌感染抗性或易感性的重要方面。为了进一步研究噬菌体作为表位载体,诱导小鼠产生细胞因子的类型,本研究利用展示有C表位的杂合噬菌体、野生噬菌体和与匙孔血蓝蛋白(KLH)偶联的化学合成C表位(KLH-C),免疫C57BL/6J小鼠,再采用定量ELISA方法,检测脾细胞培养上清中分泌细胞因子的水平。同时用RT-PCR方法分析了mRNA水平上细胞因子的转录,确定了检测小鼠体内脾中Th1和Th2型细胞因子的转录。得到的结果如下: 1.噬菌体作为疫苗载体可以增强免疫鼠脾细胞Th1型细胞因子的转录。杂合噬菌体、野生噬菌体和KLH-C免疫小鼠体内脾中Th1型细胞因子,IFN-γ、IL-2和IL-12的转录水平相似,与TE对照组比较,差异显著。IL-10在免疫鼠和对照鼠间的转录水平相似。 2.噬菌体作为疫苗载体可以诱导免疫鼠的脾细胞分泌Th1型细胞因子。杂合噬菌体免疫小鼠的脾细胞培养上清中,Th1型细胞因子,IFN-γ和IL-12的蛋白分泌水平与TE对照组比较,具有显著性差异,与KLH-C免疫小鼠相似。Th2型细胞因子,IL-10的分泌水平在免疫组和对照组之间没有差异。因此,噬菌体作为疫苗载体,能够通过增强Th1型细胞因子的转录诱导Th1型细胞因子的产生,与传统的化学载体KLH相比,具有相似的作用。而且其制备简单、省时省力的优点使噬菌体作为表位载体在疫苗研究中具有广阔的前景。
[Abstract]:Candida albicans is a normal group of bacteria parasitic on healthy people, which can be transformed into pathogenic bacteria when the immunity of the body decreases or the normal flora is out of balance. The use of immunosuppressants increases the susceptibility of patients to Candida and increases the incidence of Candida infection in hospitals. Heat shock protein 90 (Hsp90) is a toxic factor secreted by Candida albicans during its infection. As an immune dominant antigen of Candida albicans, heat shock protein 90 (Hsp90) can induce infection to produce specific antibodies. Antibody against Hsp90 is a protective antibody. Our team has demonstrated that phage-displayed epitopes can induce humoral and cellular immune responses in mice. In the process of Candida infection, the differentiation direction of CD4 T cells is an important factor in determining the resistance or susceptibility to Candida infection. In order to further study the type of cytokine production induced by phage as epitope carrier, a hybrid phage with C epitope was used in this study. Wild phages and chemically synthesized C epitopes (KLH-C) coupled with keyhole hemocyanin (KLH) were used to immunize C57BL / 6J mice. The levels of cytokines secreted in the supernatant of splenocyte culture were detected by quantitative Elisa. At the same time, the transcription of cytokines at mRNA level was analyzed by RT-PCR, and the transcription of Th1 and Th2 type cytokines in mouse spleen was determined. The results are as follows: 1. Phage as a vaccine vector can enhance the transcription of Th1 cytokines in spleen cells of immunized mice. The transcription levels of IL-2 and IL-12 in spleen of hybrid phage, wild phage and KLH-C immunized mice were similar to those of te control group. The transcriptional level of IL-10 in immunized mice and control mice was similar. 2. 2. Bacteriophage as a vaccine vector can induce Th1 cytokine secretion from spleen cells of immunized mice. The secretory levels of Th1 type cytokines IFN- 纬 and IL-12 in the supernatant of spleen cell culture of hybrid phage immunized mice were significantly different from those of te control group. The secretory level of IL-10 was similar to that of KLH-C immunized mice. There was no difference between the immunized group and the control group. Therefore, phage, as a vaccine vector, can induce Th1 cytokine production by enhancing the transcription of Th1 cytokines, which is similar to that of traditional chemical vector KLH. Moreover, the advantages of simple preparation, time saving and labor saving make bacteriophage as epitope carrier has a broad prospect in vaccine research.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

【相似文献】