本文選題：干燥綜合征　切入點：Ro60　出處：《廈門大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：干燥綜合征是一種自身免疫病,它影響外分泌腺而導(dǎo)致眼干和口干等癥狀。這個疾病的致病自身抗原還不清楚,所以目前還缺乏一個好的通過主動免疫抗原而建立的動物模型。干燥綜合征病人的一個主要特點是血清中存在抗SSA/Ro60自身抗體,且該自身抗體是診斷干燥綜合征標(biāo)準(zhǔn)之一。本研究旨在通過主動免疫Ro60抗原表位多肽,建立類似人類干燥綜合征的小鼠模型并探索Ro60抗原在干燥綜合征發(fā)病機制中的作用。 方法：利用Titermax和CFA當(dāng)佐劑,分別與Ro60_316-335,Ro60_480-494抗原表位短肽混合后腳掌或皮下免疫BALB/c小鼠,對照組用PBS與各佐劑混合后免疫。利用ELISA的方法,我們檢測血清抗Ro60多肽IgG抗體以及它的亞型還有血清中的細(xì)胞因子的表達(dá)情況。在免疫前,免疫后45天和免疫后90/120天,分別檢測各組小鼠唾液和淚液分泌量以及體重變化情況。實驗在免疫后90/120天結(jié)束,從小鼠體內(nèi)收獲的淚腺,唾液腺和其他組織,用來做組織化學(xué)和免疫學(xué)的評估。用免疫熒光的方法來鑒定組織中的炎癥細(xì)胞浸潤情況。同時,通過免疫熒光的方法來檢測組織中自身抗體的沉積情況。最后,.通過轉(zhuǎn)移免疫小鼠的IgG到正常小鼠來檢驗被免疫小鼠體內(nèi)抗體的致病性。 結(jié)果：Ro60_316-335短肽與Titermax佐劑免疫的BALB/c小鼠,在免疫后90天出現(xiàn)了類似干燥綜合征的癥狀。小鼠的淚液分泌量減少,淚腺組織上出現(xiàn)明顯的黑色斑點,病理學(xué)顯示淚腺中腺泡形態(tài)改變,腺泡萎縮并出現(xiàn)多處空腔。血清中產(chǎn)生了抗Ro60_316-335短肽的自身抗體,淚腺組織中也可以觀察到有自身抗體IgG沉淀。同時,細(xì)胞因子水平發(fā)生明顯變化,其中促炎癥因子IFN-γ和IL-17的水平有明顯的提高。同時,在淚腺組織中可以觀察到T細(xì)胞,B細(xì)胞,巨噬細(xì)胞和中心粒細(xì)胞浸潤。與免疫Ro60_316-335短肽的小鼠不同的是：免疫Ro60_480-494多肽免疫小鼠和對照組小鼠并沒出現(xiàn)以上的癥狀。最后,如果把佐劑改變?yōu)镃FA佐劑后,Ro60_316-335多肽免疫的小鼠僅產(chǎn)生抗Ro60_316-335多肽的自身抗體,但并無其它病理和功能改變。而轉(zhuǎn)移免疫小鼠的IgG到正常小鼠并不能導(dǎo)致小鼠外分泌功能降低。 結(jié)論：通過Ro60_316-335短肽與Titermax佐劑免疫,我們成功地建立了一個新的干燥綜合征的小鼠模型。該模型為進(jìn)一步研究干燥綜合征的發(fā)病機制及診斷治療方法奠定了基礎(chǔ)。同時,這個模型的成功建立也證明了針對Ro60的自身免疫反應(yīng)在干燥綜合征的發(fā)病過程中起到了一定的作用。而這個作用不僅和Ro60蛋白上特定的T細(xì)胞多肽表位相關(guān),而且和輔助免疫的佐劑相關(guān)。
[Abstract]:Objective: Sjogren's syndrome is an autoimmune disease that affects exocrine glands and causes symptoms such as dry eyes and dry mouth. So there is still a lack of a good animal model established by active immune antigens. One of the main characteristics of Sjogren's syndrome patients is the presence of autoantibodies to SSA/Ro60 in the serum. The aim of this study was to establish a mouse model similar to human Sjogren's syndrome and to explore the role of Ro60 antigen in the pathogenesis of Sjogren's syndrome. Methods: Titermax and CFA were used as adjuvants to immunize BALB/c mice on foot or subcutaneously respectively after mixing with Ro60316-335RoS60 480-494 epitope short peptides, while the control group was immunized with PBS and adjuvants. ELISA method was used. We detected the expression of serum anti Ro60 polypeptide IgG antibody, its subtype and cytokines in the serum. Before immunization, 45 days after immunization and 90/120 days after immunization, The changes of saliva and tear secretion and body weight of each group were measured respectively. After 90/120 days of immunization, lacrimal glands, salivary glands and other tissues were harvested from the mice. Used for histochemical and immunological assessments. Immunofluorescence was used to identify the infiltration of inflammatory cells in the tissue. At the same time, Autoantibodies in tissues were detected by immunofluorescence method. Finally, the pathogenicity of antibodies in immunized mice was examined by transferring IgG from immunized mice to normal mice. Results in BALB/c mice immunized with Titermax adjuvant, the symptoms of Sjogren's syndrome appeared 90 days after immunization. The amount of tear secretion in mice decreased, and there were obvious black spots in lacrimal gland tissue. Pathological changes in the lacrimal gland showed acinar morphology, acinar atrophy and multiple cavities. Autoantibodies against Ro60_316-335 short peptides were produced in serum, and autoantibodies IgG precipitated in lacrimal gland tissues. The levels of cytokines, IFN- 緯 and IL-17, were significantly increased. At the same time, T cell B cells were observed in lacrimal gland. Macrophages and central granulocytes infiltrate. Unlike mice immunized with Ro60_316-335 peptides, the mice immunized with Ro60_480-494 peptides did not show these symptoms. If the adjuvant was changed to CFA adjuvant, the mice immunized with Ro60316-335 polypeptide only produced autoantibodies against Ro60_316-335 polypeptide, but there were no other pathological and functional changes. However, the mice transferred to normal mice could not reduce the exocrine function of mice. Conclusion: we successfully established a new Sjogren syndrome mouse model by immunizing with Ro60_316-335 short peptide and Titermax adjuvant, which laid a foundation for further study on pathogenesis and diagnosis and treatment of Sjogren syndrome. The successful establishment of this model also demonstrated that the autoimmune response to Ro60 may play a role in the pathogenesis of Sjogren's syndrome, which is related not only to the specific T cell polypeptide epitopes on the Ro60 protein. And associated with adjuvants of adjuvant immunity.
【學(xué)位授予單位】：廈門大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R-332;R593.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉莉,張奉春;SSA/Ro抗原的臨床及生物學(xué)特性[J];中華風(fēng)濕病學(xué)雜志;2001年06期

2 趙巖,賈寧,魏麗,王治國,戚務(wù)芳,郭曉萍,高巖,曾小峰,張奉春,唐福林,董怡;原發(fā)性干燥綜合征2002年國際分類(診斷)標(biāo)準(zhǔn)的臨床驗證[J];中華風(fēng)濕病學(xué)雜志;2003年09期

3 湯艷春 ,王吉波 ,孫雪輝 ,王少坤 ,袁威玲;原發(fā)性干燥綜合征唇腺組織中細(xì)胞凋亡及相關(guān)基因表達(dá)的研究[J];中華風(fēng)濕病學(xué)雜志;2004年09期

4 張奉春,ＪｏｈｎＢ．Ｈａｒｌｅｙ,Ｒ．ＨａｌＳｃｏｆｉｅｌｄ,董怡;60 kD人SSA抗原表位免疫反應(yīng)的特異性[J];中華風(fēng)濕病學(xué)雜志;1998年01期

5 王睿;張麗;邢恩鴻;孫宇揚;楊澤;;Th1、Th2樣細(xì)胞因子在干燥綜合征模型小鼠頜下腺組織中mRNA表達(dá)的研究[J];中國老年保健醫(yī)學(xué);2006年03期

6 李麗杰;楊奎真;李明義;李延團(tuán);郝峰強;;角鯊烯作為新型佐劑的免疫效果研究[J];中國海洋藥物;2013年01期

7 鄒新樂;李想;徐凱彪;陳巧林;曾令文;;毒蕈堿受體3抗原表位多肽誘導(dǎo)干燥綜合征動物模型的研究[J];現(xiàn)代免疫學(xué);2009年01期

8 羅豐年,張汗承;干燥綜合征患者免疫功能的研究[J];眼科研究;1996年03期

9 王巖艷;劉怡欣;劉鋼;;干燥綜合征研究進(jìn)展[J];現(xiàn)代臨床醫(yī)學(xué);2007年S1期

10 王松靈，，朱宣智，鄒兆菊;舍格倫綜合征的診斷標(biāo)準(zhǔn)及分型的評價[J];中華口腔醫(yī)學(xué)雜志;1994年06期

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