本文關(guān)鍵詞：CC趨化因子受體5在疾病中的作用研究暨胸腺素α原cDNA多態(tài)性研究　出處：《第二軍醫(yī)大學(xué)》2007年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： CCR5 實(shí)驗(yàn)性自身免疫性心肌炎 T細(xì)胞 單克隆抗體 濾泡性甲狀腺癌 胸腺素α原 cDNA 序列多態(tài)性

【摘要】： CC趨化因子受體5 (cc chemokine receptor 5,CCR5)為細(xì)胞膜蛋白,是趨化因子受體的一種亞型。CCR5主要表達(dá)在單核細(xì)胞、T細(xì)胞等白細(xì)胞上,最近的研究表明其為活化Thl細(xì)胞的表面標(biāo)志物,提示CCR5與自身免疫性疾病的發(fā)生發(fā)展有關(guān)。我們通過研究CCR5在實(shí)驗(yàn)性自身免疫性心肌炎(experimental autoimmune myocarditis, EAM)小鼠模型以及濾泡性甲狀腺癌的表達(dá)情況試圖發(fā)現(xiàn)CCR5在疾病發(fā)生發(fā)展過程中所發(fā)揮的作用。我們采用了RT-PCR、免疫組化、流式細(xì)胞檢測(cè)等經(jīng)典的方法,并成功構(gòu)建了抗體封閉和細(xì)胞過繼等實(shí)驗(yàn)動(dòng)物模型,從分子、細(xì)胞以及動(dòng)物模型等不同層次證明了CCR5在EAM小鼠以及濾泡狀甲狀腺癌的重要性。我們發(fā)現(xiàn)CCR5在EAM小鼠模型的外周血中表達(dá)含量明顯上升,并且在炎性心肌組織中浸潤的炎性細(xì)胞多數(shù)呈CCR5陽性表達(dá);CCR5陽性細(xì)胞過繼小鼠其心肌炎發(fā)病程度明顯高于T細(xì)胞過繼組,而CCR5陰性過繼小鼠發(fā)病程度則明顯降低;以CCR5單克隆抗體封閉EAM小鼠其發(fā)病率明顯下降。 近來的研究表明趨化因子受體參與腫瘤的生長和分化,且與腫瘤的轉(zhuǎn)移有著密切的聯(lián)系。為研究CCR5在濾泡狀甲狀腺癌的發(fā)生或發(fā)展中所起的作用,我們通過對(duì)濾泡性甲狀腺癌中CCR5的表達(dá)情況的分析,發(fā)現(xiàn)在濾泡狀甲狀腺癌組織中,CCR5表達(dá)陽性率明顯高于甲狀腺腺瘤和正常甲狀腺組織(PO.OO1),且濾泡狀甲狀腺癌患者血清血中CCL3, CCL5的濃度顯著高于對(duì)照組(P0.O1)。提示CCR5在濾泡狀甲狀腺癌發(fā)病進(jìn)程中發(fā)揮著重要的作用。 此外通過對(duì)人胸腺素α原cDNA測(cè)序,分析了胸腺素α原的序列多態(tài)性。我們利用RT-PCR技術(shù)從外周血及臍帶血中擴(kuò)增胸腺素原cDNA,純化后與克隆載體pMD18-T連接,經(jīng)克隆測(cè)序,通過與標(biāo)準(zhǔn)序列比對(duì),分析其多態(tài)性。序列分析結(jié)果表明,克隆的ProTα基因的核苷酸序列并不一致。與已報(bào)道的胸腺素α原基因進(jìn)行比較,發(fā)現(xiàn)存在兩種變異。(1)107位單核苷酸變異,以及110-121位和191-205位的核苷酸片段缺失;(2)306位單核苷酸缺失,多見于60-80年齡組。故我們認(rèn)為人胸腺素α原cDNA序列存在多態(tài)性,我們發(fā)現(xiàn)其存在兩種變異序列,但此兩種基因變異并未影響到N-端前28個(gè)肽,并未影響到胸腺素α原的功能。該結(jié)果為胸腺素a原的結(jié)構(gòu)、功能和演化研究提供了信息。
[Abstract]:CC chemokine receptor 5 chemokine receptor 5 (CCR5) is a membrane protein. CCR5 is a subtype of chemokine receptor. CCR5 is mainly expressed on monocyte T cells and other leukocytes. Recent studies have shown that CCR5 is a surface marker for activating Thl cells. It is suggested that CCR5 may be associated with the development of autoimmune diseases. We studied the role of CCR5 in experimental autoimmune myocarditis. Experimental autoimmune myocarditis. EAM mouse model and the expression of follicular thyroid carcinoma try to find out the role of CCR5 in the development of the disease. We used RT-PCR, immunohistochemistry. Flow cytometry and other classical methods, and successfully constructed antibody blocking and cell adoptions of experimental animal models, from the molecules. The importance of CCR5 in EAM mice and follicular thyroid carcinoma was demonstrated at different levels such as cells and animal models. We found that the expression of CCR5 in peripheral blood of EAM mice was significantly increased. And most of the infiltrating inflammatory cells in the inflammatory myocardium showed CCR5 positive expression. The degree of myocarditis in CCR5 positive adoptive mice was significantly higher than that in T cell adoptive mice, but the degree of CCR5 negative adoptive mice was significantly lower. The incidence of CCR5 monoclonal antibody in EAM mice decreased significantly. Recent studies have shown that chemokine receptors are involved in tumor growth and differentiation, and are closely related to tumor metastasis. To study the role of CCR5 in the occurrence or development of follicular thyroid carcinoma. By analyzing the expression of CCR5 in follicular thyroid carcinoma, we found that it was found in follicular thyroid carcinoma. The positive rate of CCR5 expression was significantly higher than that of thyroid adenoma and normal thyroid tissue, and CCL3 in serum of follicular thyroid carcinoma patients. The concentration of CCL5 was significantly higher than that of the control group (P0. 01), which suggested that CCR5 plays an important role in the pathogenesis of follicular thyroid carcinoma. In addition, the sequence polymorphism of human thymosin 偽 proto cDNA was analyzed by sequencing. We amplified thymogen cDNA from peripheral blood and umbilical cord blood by RT-PCR technique. After purification, it was ligated with the clone vector pMD18-T, then cloned and sequenced. The polymorphism was analyzed by alignment with the standard sequence. The nucleotide sequence of the cloned ProT 偽 gene was not consistent with that of the reported thymosin 偽 gene. And deletion of nucleotide fragments at 110-121 and 191-205; The deletion of single nucleotide was found in the 60-80 age group, so we believe that there is polymorphism in the human thymosin 偽 proto cDNA sequence, and we found that there are two variant sequences. However, these two mutations did not affect the function of the first 28 peptides and the function of prothymosin 偽, which provided information for the study of the structure, function and evolution of prothymosin a.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2007
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 汪朝暉,廖玉華,董繼華,李淑莉,王金平,涂源淑;Myosin-Induced Autoimmune Myocarditis in BALB/C Mice[J];Journal of Tongji Medical University;1999年02期

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本文編號(hào)：1402479