本文關鍵詞： 巨噬細胞 T淋巴細胞 CD4O hTERT 電離輻射　出處：《長春工業(yè)大學》2011年碩士論文　論文類型：學位論文

【摘要】：免疫系統(tǒng)主要由固有免疫系統(tǒng)和適應性免疫系統(tǒng)兩大部分組成,其中,固有免疫系統(tǒng)與適應性免疫系統(tǒng)又是通過化學介質、趨化因子與細胞因子的相互作用而有機的結合在一起的。對電離輻射對免疫細胞表面分子的表達以及細胞因子的分泌的影響進行研究,對于深入的了解輻射免疫效應的本質有著十分重要的意義。 本文在以往關于輻射免疫學的基礎之上,檢測電離輻射對體外培養(yǎng)的小鼠巨噬細胞和T淋巴細胞表達CD40蛋白分子的影響,從而對電離輻射對機體的免疫系統(tǒng)的影響做進一步的研究。本文主要采用流式細胞技術檢測人巨噬細胞和T淋巴細胞在高、低不高劑量的電離輻射下,CD40蛋白分子的表達情況,同時還采用RT-PCR技術,對CD40蛋白分子的mRNA的表達情況進行檢測。結果顯示,人巨噬細胞的CD40蛋白分子在低劑量輻射條件下,其表達量明顯升高,而在高劑量輻射條件下,CD40的表達則受到抑制；同時,在高、低劑量輻射條件下,CD40蛋白分子的：mRNA的表達量都有升高的趨勢。而人T淋巴細胞的CD40蛋白分子的表達量,在低劑量輻射條件下,其表達量受到抑制,而在高劑量輻射條件下,CD40蛋白分子得到高表達。說明低劑量電離輻射對機體的免疫系統(tǒng)有興奮效應而高劑量電離輻射則對機體的免疫系統(tǒng)有抑制作用。 本文采用構建逆轉錄病毒載體,并將其轉染至原代細胞中,以解決原代細胞在體外培養(yǎng)期間增殖速度慢,生長周期長的問題。本研究首先采用酶切法從質粒phTERTGPlox中獲取端粒酶逆轉錄酶催化亞基(hTERT),并將其克隆至逆轉錄病毒載體pbabe-GFP中,再將其轉染至宿主細胞B16細胞中,通過熒光微光顯微鏡觀察其熒光蛋白的表達情況,進而確認hTERT的表達狀況。擬將構建的pbabe-gfp-hTERT逆轉錄病毒載體轉進原代宿主免疫細胞中,使其變成永生化細胞,然后檢測其輻射免疫應答反應。
[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.
【學位授予單位】：長春工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R392

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