bcr-abl融合基因片段和鼠IL-7双表达基因疫苗的免疫保护机制研究
发布时间：2018-10-08 11:58
【摘要】：慢性粒细胞白血病(Chronic myeloid leukemia,CML)具有特征性的bcr-abl融合基因。本实验室成功构建表达bcr-abl基因融合区内大约472bp的融合基因片段的重组表达载体pcDNAbcr-abl,用其免疫小鼠,能在小鼠体内成功诱生针对bcr-abl融合蛋白的特异性抗体。白细胞介素-7(Interleukin-7,IL-7)能增强特异性CTL细胞活性。本课题试图分三部分研究小鼠IL-7基因对bcr-abl融合基因疫苗诱导机体免疫应答的影响。 一、bcr-abl融合基因片段和mlL-7基因双表达载体的构建与鉴定 为了给小鼠IL-7增强bcr-abl融合基因片段基因疫苗诱导特异性免疫应答的研究创造条件,我们将小鼠IL-7基因和bcr-abl融合基因片段分别插入具有两个多克隆位点的真核表达载体pVITR02-mcs,成功构建pVbcr-abl/mIL7双表达质粒。在pVbcr-abl/mIL7质粒中,IL-7基因、bcr-abl融合基因片段分别受不同的启动子调控。我们用pVbcr-abl/mIL7双表达载体转染CHO细胞,分别从蛋白表达水平和mRNA水平证实,bcr-abl融合基因片段和小鼠IL-7在真核细胞CHO中得到高效表达。bcr-abl基因片段和mIL-7基因双表达真核细胞载体的构建为慢性粒细胞白血病bcr-abl融合基因疫苗的研究提供了新的实验研究工具。 二、两种bcr-abl基因疫苗对SP2/0/bcr-abl移植瘤小鼠的影响 在构建bcr-abl基因片段和mIL-7基因双表达载体、表达bcr-abl融合基因片段的SP2/0/bcr-abl细胞的基础上,我们首先用pVbcr-abl、pVbcr-abl/mIL7两种质粒分别免疫BALB/c小鼠,然后用SP2/0/bcr-abl细胞攻击,观察疫苗对SP2/0/bcr-abl
[Abstract]:Chronic myeloid leukemia (Chronic myeloid leukemia,CML) has a characteristic bcr-abl fusion gene. In our laboratory, the recombinant expression vector pcDNAbcr-abl, expressing the fusion gene fragment of approximately 472bp in the fusion region of bcr-abl gene was successfully constructed to immunize mice, and the specific antibody against bcr-abl fusion protein was successfully induced in mice. Interleukin-7 (Interleukin-7,IL-7) can enhance the activity of specific CTL cells. This paper attempts to study the effect of mouse IL-7 gene on immune response induced by bcr-abl fusion gene vaccine in three parts. Construction and identification of a bcr-abl fusion gene fragment and a double expression vector of mlL-7 gene in order to induce specific immunity to mouse IL-7 enhanced bcr-abl fusion gene fragment gene vaccine The study of epidemic response creates conditions, The mouse IL-7 gene and the bcr-abl fusion gene fragment were inserted into the eukaryotic expression vector pVITR02-mcs, with two polyclonal sites respectively to construct the pVbcr-abl/mIL7 double expression plasmid. In pVbcr-abl/mIL7 plasmids, the fragment of bcr-abl fusion gene of IL-7 gene was regulated by different promoters. We transfected CHO cells with pVbcr-abl/mIL7 double expression vector. The expression of bcr-abl fusion gene and mouse IL-7 in eukaryotic cell CHO was confirmed by protein expression level and mRNA level, respectively. The eukaryotic expression vector of bcr-abl gene and mIL-7 gene was constructed as bcr-abl in chronic myeloid leukemia. The research of fusion gene vaccine provides a new experimental research tool. Secondly, the effect of two bcr-abl gene vaccines on SP2/0/bcr-abl transplanted tumor mice was based on the construction of bcr-abl gene fragment and mIL-7 gene expression vector, and the expression of bcr-abl fusion gene fragment in SP2/0/bcr-abl cells. We first immunized BALB/c mice with two plasmids of pVbcr-abl,pVbcr-abl/mIL7, then attacked with SP2/0/bcr-abl cells to observe the effect of vaccine on SP2/0/bcr-abl.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1
本文编号：2256659
[Abstract]:Chronic myeloid leukemia (Chronic myeloid leukemia,CML) has a characteristic bcr-abl fusion gene. In our laboratory, the recombinant expression vector pcDNAbcr-abl, expressing the fusion gene fragment of approximately 472bp in the fusion region of bcr-abl gene was successfully constructed to immunize mice, and the specific antibody against bcr-abl fusion protein was successfully induced in mice. Interleukin-7 (Interleukin-7,IL-7) can enhance the activity of specific CTL cells. This paper attempts to study the effect of mouse IL-7 gene on immune response induced by bcr-abl fusion gene vaccine in three parts. Construction and identification of a bcr-abl fusion gene fragment and a double expression vector of mlL-7 gene in order to induce specific immunity to mouse IL-7 enhanced bcr-abl fusion gene fragment gene vaccine The study of epidemic response creates conditions, The mouse IL-7 gene and the bcr-abl fusion gene fragment were inserted into the eukaryotic expression vector pVITR02-mcs, with two polyclonal sites respectively to construct the pVbcr-abl/mIL7 double expression plasmid. In pVbcr-abl/mIL7 plasmids, the fragment of bcr-abl fusion gene of IL-7 gene was regulated by different promoters. We transfected CHO cells with pVbcr-abl/mIL7 double expression vector. The expression of bcr-abl fusion gene and mouse IL-7 in eukaryotic cell CHO was confirmed by protein expression level and mRNA level, respectively. The eukaryotic expression vector of bcr-abl gene and mIL-7 gene was constructed as bcr-abl in chronic myeloid leukemia. The research of fusion gene vaccine provides a new experimental research tool. Secondly, the effect of two bcr-abl gene vaccines on SP2/0/bcr-abl transplanted tumor mice was based on the construction of bcr-abl gene fragment and mIL-7 gene expression vector, and the expression of bcr-abl fusion gene fragment in SP2/0/bcr-abl cells. We first immunized BALB/c mice with two plasmids of pVbcr-abl,pVbcr-abl/mIL7, then attacked with SP2/0/bcr-abl cells to observe the effect of vaccine on SP2/0/bcr-abl.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1
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