免疫细胞辐射诱导免疫反应的研究

发布时间：2018-01-21 11:36

本文关键词： 巨噬细胞 T淋巴细胞 CD4O hTERT 电离辐射　出处：《长春工业大学》2011年硕士论文　论文类型：学位论文

【摘要】：免疫系统主要由固有免疫系统和适应性免疫系统两大部分组成,其中,固有免疫系统与适应性免疫系统又是通过化学介质、趋化因子与细胞因子的相互作用而有机的结合在一起的。对电离辐射对免疫细胞表面分子的表达以及细胞因子的分泌的影响进行研究,对于深入的了解辐射免疫效应的本质有着十分重要的意义。本文在以往关于辐射免疫学的基础之上,检测电离辐射对体外培养的小鼠巨噬细胞和T淋巴细胞表达CD40蛋白分子的影响,从而对电离辐射对机体的免疫系统的影响做进一步的研究。本文主要采用流式细胞技术检测人巨噬细胞和T淋巴细胞在高、低不高剂量的电离辐射下,CD40蛋白分子的表达情况,同时还采用RT-PCR技术,对CD40蛋白分子的mRNA的表达情况进行检测。结果显示,人巨噬细胞的CD40蛋白分子在低剂量辐射条件下,其表达量明显升高,而在高剂量辐射条件下,CD40的表达则受到抑制；同时,在高、低剂量辐射条件下,CD40蛋白分子的：mRNA的表达量都有升高的趋势。而人T淋巴细胞的CD40蛋白分子的表达量,在低剂量辐射条件下,其表达量受到抑制,而在高剂量辐射条件下,CD40蛋白分子得到高表达。说明低剂量电离辐射对机体的免疫系统有兴奋效应而高剂量电离辐射则对机体的免疫系统有抑制作用。本文采用构建逆转录病毒载体,并将其转染至原代细胞中,以解决原代细胞在体外培养期间增殖速度慢,生长周期长的问题。本研究首先采用酶切法从质粒phTERTGPlox中获取端粒酶逆转录酶催化亚基(hTERT),并将其克隆至逆转录病毒载体pbabe-GFP中,再将其转染至宿主细胞B16细胞中,通过荧光微光显微镜观察其荧光蛋白的表达情况,进而确认hTERT的表达状况。拟将构建的pbabe-gfp-hTERT逆转录病毒载体转进原代宿主免疫细胞中,使其变成永生化细胞,然后检测其辐射免疫应答反应。
[Abstract]:The immune system is mainly composed of the innate immune system and the adaptive immune system. Among them, the inherent immune system and the adaptive immune system are also through the chemical medium. The effects of ionizing radiation on the expression of molecules on the surface of immune cells and the secretion of cytokines were studied. It is of great significance to understand the essence of radiation immune effect. The effects of ionizing radiation on the expression of CD40 protein molecules in mouse macrophages and T lymphocytes in vitro were studied on the basis of previous studies on radiation immunology. In this paper, flow cytometry was used to detect human macrophages and T lymphocytes at high and low doses of ionizing radiation. The expression of CD40 protein and the expression of mRNA of CD40 protein were detected by RT-PCR technique. The expression of CD40 protein in human macrophages increased significantly under low dose radiation, but the expression of CD40 was inhibited under high dose radiation. At the same time, under high and low doses of radiation, the expression of CD40 protein of CD40 protein was increased, while the expression of CD40 protein in human T lymphocytes was higher than that in human T lymphocytes. Its expression was inhibited under low dose radiation and high dose radiation. The high expression of CD40 protein indicated that low dose ionizing radiation had excitatory effect on immune system and high dose ionizing radiation had inhibitory effect on body immune system. The retrovirus vector was constructed and transfected into primary cells to solve the problem of slow proliferation of primary cells during in vitro culture. The first step of this study was to obtain telomerase reverse transcriptase catalytic subunit (hTERT) from plasmid phTERTGPlox by enzyme digestion. It was cloned into the retroviral vector pbabe-GFP and then transfected into the host cell B16 cells. The expression of fluorescent protein was observed by fluorescence microscopes. In order to confirm the expression of hTERT, we intend to transfer the constructed pbabe-gfp-hTERT retrovirus vector into the primary host immune cells to make them immortalized cells. Then the immune response to radiation was detected.
【学位授予单位】：长春工业大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392

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