【摘要】： 誘騙受體3(DcR3,又稱TR6)是新近發(fā)現(xiàn)的一種可溶性腫瘤壞死因子受體(TNFR)超家族成員,可與Fas、HVEM、DR3競爭結(jié)合其相應(yīng)配體FasL、LIGHT及TL1A,但并不介導(dǎo)細胞凋亡,在細胞的生長、分化、死亡以及免疫調(diào)節(jié)等方面發(fā)揮重要作用,與惡性腫瘤、自身免疫疾病及移植排斥等密切相關(guān)。 本文將構(gòu)建的原核表達質(zhì)粒pET28a(+)/DcR3轉(zhuǎn)化入大腸桿菌(E.coli) BL21中,高效誘導(dǎo)表達并純化DcR3融合蛋白,純化并成功復(fù)性的蛋白分別免疫家兔和BALB/c小鼠,獲得了效價高、特異性強的抗人DcR3多克隆抗體(pAb)和5株單克隆抗體(mAb)。以pAb為包被抗體,以生物素化的mAb為檢測抗體,引入生物素-親合素系統(tǒng)(BAS),建立雙抗體夾心BAS-ELISA方法。利用該方法檢測到特發(fā)性肺纖維化(IPF)、小細胞肺癌及慢性阻塞性肺疾病(COPD)急性加重期患者血清DcR3水平明顯高于健康對照。為進一步研究DcR3的功能、應(yīng)用及探討其在臨床相關(guān)疾病發(fā)病機制中的作用奠定基礎(chǔ)。
[Abstract]:Decoy receptor 3 (DcR3, also known as TR6) is a newly discovered soluble tumor necrosis factor receptor (TNFR) superfamily member. It can compete with Fas, HVEM, DR3 to bind its corresponding ligands FasL, LIGHT and TL1A, but does not mediate cell apoptosis, plays an important role in cell growth, differentiation, death and immune regulation, and plays an important role in malignant tumor, itself. Immune diseases and transplant rejection are closely related.
The prokaryotic expression plasmid pET28a (+) / DcR3 was transformed into E. coli BL21. The fusion protein of DcR3 was highly induced and purified. The purified and renatured proteins were immunized to rabbits and BALB / c mice respectively. The polyclonal antibodies against human DcR3 (pAb) and five monoclonal antibodies (mAb) with high titer and specificity were obtained. The serum levels of DcR3 in patients with idiopathic pulmonary fibrosis (IPF), small cell lung cancer (SCLC) and chronic obstructive pulmonary disease (COPD) at acute exacerbation stage were significantly higher than those in healthy controls. To study the function of DcR3 and lay a foundation for its application in the pathogenesis of clinical related diseases.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2008
【分類號】：R392

【參考文獻】

相關(guān)期刊論文 前8條

1 李平,黃昌明,張祥福,盧輝山,楊發(fā)端,張魯榕;DcR3和Ki-67在胃癌的表達及臨床意義[J];福建醫(yī)科大學學報;2004年01期

2 鐘智強;蘭小鵬;楊湘越;李衛(wèi)濱;張魯榕;朱忠勇;;DcR3基因在原發(fā)性肺癌的擴增和表達及臨床意義[J];福州總醫(yī)院學報;2007年03期

3 陳罡;羅殿中;;原發(fā)性肝細胞癌中誘捕受體3的過度表達及臨床意義[J];廣東醫(yī)學;2006年10期

4 李連賀;胡俊蘭;趙瑞力;岳卓立;;喉癌組織中DcR3蛋白的表達及臨床意義[J];臨床耳鼻咽喉頭頸外科雜志;2007年12期

5 黃志衛(wèi),馬忠森,王秀麗,吳艷峰,赫國志;外周血單核細胞DcR3基因過度表達在間質(zhì)性肺疾病發(fā)病中的意義[J];中華結(jié)核和呼吸雜志;2003年05期

6 吳艷峰,楊俊玲,馬忠森,董春玲,秦彥國;DcR_3基因與特發(fā)性肺纖維化肺功能的相關(guān)性研究[J];中國老年學雜志;2004年10期

7 王秀麗,尹金植,黃志衛(wèi),馬忠森;特發(fā)性肺間質(zhì)纖維化病人外周血單核細胞中DcR_3基因表達的意義[J];中國老年學雜志;2004年12期

8 ;Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis[J];World Journal of Gastroenterology;2005年38期

，

本文編號：2182962