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

【摘要】：免疫系統(tǒng)主要由固有免疫系統(tǒng)和適應(yīng)性免疫系統(tǒng)兩大部分組成,其中,固有免疫系統(tǒng)與適應(yīng)性免疫系統(tǒng)又是通過化學(xué)介質(zhì)、趨化因子與細(xì)胞因子的相互作用而有機(jī)的結(jié)合在一起的。對(duì)電離輻射對(duì)免疫細(xì)胞表面分子的表達(dá)以及細(xì)胞因子的分泌的影響進(jìn)行研究,對(duì)于深入的了解輻射免疫效應(yīng)的本質(zhì)有著十分重要的意義。 本文在以往關(guān)于輻射免疫學(xué)的基礎(chǔ)之上,檢測(cè)電離輻射對(duì)體外培養(yǎng)的小鼠巨噬細(xì)胞和T淋巴細(xì)胞表達(dá)CD40蛋白分子的影響,從而對(duì)電離輻射對(duì)機(jī)體的免疫系統(tǒng)的影響做進(jìn)一步的研究。本文主要采用流式細(xì)胞技術(shù)檢測(cè)人巨噬細(xì)胞和T淋巴細(xì)胞在高、低不高劑量的電離輻射下,CD40蛋白分子的表達(dá)情況,同時(shí)還采用RT-PCR技術(shù),對(duì)CD40蛋白分子的mRNA的表達(dá)情況進(jìn)行檢測(cè)。結(jié)果顯示,人巨噬細(xì)胞的CD40蛋白分子在低劑量輻射條件下,其表達(dá)量明顯升高,而在高劑量輻射條件下,CD40的表達(dá)則受到抑制；同時(shí),在高、低劑量輻射條件下,CD40蛋白分子的：mRNA的表達(dá)量都有升高的趨勢(shì)。而人T淋巴細(xì)胞的CD40蛋白分子的表達(dá)量,在低劑量輻射條件下,其表達(dá)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(dá)。說明低劑量電離輻射對(duì)機(jī)體的免疫系統(tǒng)有興奮效應(yīng)而高劑量電離輻射則對(duì)機(jī)體的免疫系統(tǒng)有抑制作用。 本文采用構(gòu)建逆轉(zhuǎn)錄病毒載體,并將其轉(zhuǎn)染至原代細(xì)胞中,以解決原代細(xì)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(zhì)粒phTERTGPlox中獲取端粒酶逆轉(zhuǎn)錄酶催化亞基(hTERT),并將其克隆至逆轉(zhuǎn)錄病毒載體pbabe-GFP中,再將其轉(zhuǎn)染至宿主細(xì)胞B16細(xì)胞中,通過熒光微光顯微鏡觀察其熒光蛋白的表達(dá)情況,進(jìn)而確認(rèn)hTERT的表達(dá)狀況。擬將構(gòu)建的pbabe-gfp-hTERT逆轉(zhuǎn)錄病毒載體轉(zhuǎn)進(jìn)原代宿主免疫細(xì)胞中,使其變成永生化細(xì)胞,然后檢測(cè)其輻射免疫應(yīng)答反應(yīng)。
[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(xué)位授予單位】：長春工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 宋述強(qiáng),程天民,林遠(yuǎn);電離輻射對(duì)傷口液中細(xì)胞因子變化的影響及苯妥英鈉的作用[J];第三軍醫(yī)大學(xué)學(xué)報(bào);1998年03期

2 羅燦,蘇旭,付士波,劉建香,鞠桂芝;不同劑量X射線全身照射對(duì)小鼠脾細(xì)胞轉(zhuǎn)鐵蛋白受體表達(dá)的影響[J];輻射防護(hù);1999年04期

3 蘇旭,羅燦,付士波,劉建香,鞠桂芝,劉樹錚;電離輻射對(duì)小鼠脾細(xì)胞IL-2Rα表達(dá)的影響[J];輻射研究與輻射工藝學(xué)報(bào);1999年01期

4 孫義敏,劉樹錚;小鼠全身照射后輔佐細(xì)胞與脾淋巴細(xì)胞間的相互作用[J];輻射研究與輻射工藝學(xué)報(bào);2000年02期

5 金順子,何淑杰,劉樹錚;不同劑量X射線照射對(duì)小鼠巨噬細(xì)胞表達(dá)CD80和CD86的影響[J];輻射研究與輻射工藝學(xué)報(bào);2001年02期

6 黃建安,顧冬梅,莊羽美,康蘇婭,張學(xué)光;CD40分子在肺癌中的表達(dá)及其臨床意義[J];江蘇醫(yī)藥;2003年09期

7 侯殿俊,喬健維,劉偉,李潔清,商希梅,張憲黨;低劑量電離輻射對(duì)人體體液免疫刺激作用的探討[J];中國預(yù)防醫(yī)學(xué)雜志;2003年02期

8 張學(xué)光,朱劍昆;細(xì)胞因子與急性輻射損傷的防護(hù)和治療[J];中華放射醫(yī)學(xué)與防護(hù)雜志;2001年06期

9 劉樹錚;低劑量輻射興奮效應(yīng)發(fā)生機(jī)理若干問題的探討[J];中華放射醫(yī)學(xué)與防護(hù)雜志;2003年06期

10 蘇旭，張迎春，，劉樹錚;不同劑量照射對(duì)胸腺細(xì)胞四個(gè)亞組的影響[J];中華放射醫(yī)學(xué)與防護(hù)雜志;1995年02期

本文編號(hào)：1451368