【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(jìng)爭(zhēng)結(jié)合其相應(yīng)配體FasL、LIGHT及TL1A,但并不介導(dǎo)細(xì)胞凋亡,在細(xì)胞的生長(zhǎng)、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(dá)質(zhì)粒pET28a(+)/DcR3轉(zhuǎn)化入大腸桿菌(E.coli) BL21中,高效誘導(dǎo)表達(dá)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(jià)高、特異性強(qiáng)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(cè)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(cè)到特發(fā)性肺纖維化(IPF)、小細(xì)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(duì)照。為進(jìn)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(jī)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(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

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