本文選題：急性移植物抗宿主病　切入點：異基因造血干細(xì)胞移植　出處：《第三軍醫(yī)大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:異基因造血干細(xì)胞移植(allogeneic hematopoietic stem cell transplant,allo-HSCT)是治療血液惡性疾病的有效手段。急性移植物抗宿主病(acute graft-versus-host disease,a GVHD)是allo-HSCT常見的并發(fā)癥,嚴(yán)重影響了移植患者的存活率及生存質(zhì)量,同時也是allo-HSCT患者非復(fù)發(fā)死亡的主要原因。現(xiàn)階段,機(jī)體受損靶器官的臨床表現(xiàn)和病理檢查是a GVHD診斷的主要依據(jù),但由于其臨床癥狀多變且缺乏特異性,在臨床診療中有創(chuàng)的病理檢查常常因為各種原因而無法進(jìn)行,導(dǎo)致其診斷的及時性受到影響。因此,找到快捷、無創(chuàng)、實用的實驗室方法,讓a GVHD患者在疾病早期就可以得到診斷并盡早治療,從而更有效地控制a GVHD是臨床上迫切需要解決的問題。蛋白質(zhì)芯片技術(shù)是一門微機(jī)械、微電子并與生命科學(xué)相結(jié)合的新興技術(shù),具有高特異性、高靈敏度、自動化、微型化等特點。本研究采用夾心法膜蛋白質(zhì)芯片技術(shù)檢測異基因造血干細(xì)胞移植術(shù)后發(fā)生a GVHD和無a GVHD患者的22種血清生物標(biāo)志物水平變化,初步篩選出和a GVHD發(fā)生發(fā)展及轉(zhuǎn)歸相關(guān)的分子標(biāo)志物,為后續(xù)a GVHD的臨床早期干預(yù)和預(yù)后評估奠定基礎(chǔ)。方法:連續(xù)定點收集于2014年11月至2015年11月在第三軍醫(yī)大學(xué)新橋醫(yī)院血液科進(jìn)行allo-HSCT患者移植-7d、+7d、+14d、+28d、+56d、+100d血清標(biāo)本。選取10名發(fā)生a GVHD的患者作為實驗組,在未發(fā)生a GVHD患者中隨機(jī)抽取10名作為對照組。采用蛋白質(zhì)芯片技術(shù)檢測兩組患者在移植前后不同時間點22種血清生物標(biāo)志物(IL-1β、IL-2、IL-4、IL-6、IL-7、IL-8、IL-10、IL-12p40、IL-17A、IL-21、IL-22、IL-23、MIF、TNF-α、TGF-β、IFN-γ、Elafin、ST2、TLR4、TNFR1、v WF、TF)的含量。采用AAH-CUST的數(shù)據(jù)分析軟件對芯片檢測結(jié)果進(jìn)行數(shù)據(jù)預(yù)分析,并進(jìn)行t檢驗,數(shù)據(jù)以“均數(shù)±標(biāo)準(zhǔn)差(?)”表示。結(jié)果:1.蛋白芯片檢測發(fā)現(xiàn),a GVHD組和無a GVHD組相比較,移植-7d,a GVHD組和無a GVHD組各血清生物標(biāo)志物之間無顯著差異(P0.05);移植+7d,a GVHD組IL-2、IL-17A、ST2、TLR4水平顯著高于無a GVHD組(P0.05);移植+14d,a GVHD組IL-2、IL-4、IL-17A、IL-21、IL-23、TNFR1水平顯著高于無a GVHD組(P0.05),而IFN-γ水平顯著低于無a GVHD組(P0.05);移植+28d,IL-2、IL-7、IL-17A、TNFR1,a GVHD組水平顯著高于無a GVHD組(P0.05),IFN-γ水平顯著低于無a GVHD組(P0.05);移植+56d,IL-1β、IL-2、IL-4、IL-7、IL-17A、IL-21、IL-23、ST2、TF,a GVHD組水平顯著高于無a GVHD組(P0.05);移植+100d,a GVHD組IL-7、IL-23、TF水平顯著高于無a GVHD組(P0.05)。2.Ⅲ-Ⅳ級a GVHD與II級a GVHD比較:移植-7d,IL-8血清水平前者較后者顯著增高(P0.05);移植+7d,IL-1β、IL-22血清水平前者較后者顯著增高(P0.05);移植+14d,兩組間各血清生物標(biāo)志物水平無顯著差異(P0.05);移植+28d,TLR4血清水平前者較后者顯著增高(P0.05);移植+56d,TNFR1血清水平前者較后者顯著增高(P0.05);移植+100d,IL-21血清水平前者較后者顯著下降(P0.05)。3.a GVHD組患者在治療前后22個血清生物標(biāo)志物比較,發(fā)現(xiàn)a GVHD患者治療有效控制后TLR4水平較高峰時出現(xiàn)顯著下降(P0.05)。4.激素療效差和療效好的a GVHD患者相比較:移植-7d,IL-7血清水平前者較后者顯著降低(P0.05);移植+7d,IL-17A血清水平前者較后者顯著降低(P0.05);移植+14d,兩組間各血清生物標(biāo)志物水平無顯著差異(P0.05);移植+28d,IL-23血清水平前者較后者顯著降低(P0.05);移植+56d,IL-6、IL-12p40血清水平前者較后者顯著增高(P0.05);移植+100d,IL-8、IL-22、ST2、TNFR1血清水平前者較后者顯著增高(P0.05)。結(jié)論:1.蛋白質(zhì)芯片檢測發(fā)現(xiàn)a GVHD組較無a GVHD組共有12個血清生物標(biāo)志物(IL-1β、IL-2、IL-4、IL-7、IL-17A、IL-21、IL-23、IFN-γ、ST2、TLR4、TNFR1、TF)在移植前后各不同時間點表達(dá)有顯著差異,其和a GVHD的發(fā)生發(fā)展緊密相關(guān),可將其作為a GVHD早期診斷及干預(yù)的輔助指標(biāo)。2.與II級a GVHD相比,Ⅲ-Ⅳ級a GVHD有5個血清標(biāo)志物(IL-1β、IL-8、IL-22、TLR4、TNFR1)高表達(dá),1個(IL-21)低表達(dá),表明這6個血清標(biāo)志物表達(dá)水平可能和a GVHD嚴(yán)重程度相關(guān),可作為Ⅲ-Ⅳ級a GVHD診斷依據(jù),為臨床針對性治療提供參考。3.TLR4血清水平治療后顯著下降,預(yù)示a GVHD患者的治療療效較好,為a GVHD的免疫靶向治療提供新的預(yù)判手段。4.3個血清標(biāo)志物(IL-7、IL-17A、IL-23)低表達(dá),6個血清標(biāo)志物(IL-6、IL-12p40、IL-8、IL-22、ST2、TNFR1)高表達(dá),可能和a GVHD的激素療效相關(guān),是預(yù)測患者是否對激素耐藥的參考指標(biāo)。
[Abstract]:Objective: allogeneic hematopoietic stem cell transplantation (allogeneic hematopoietic stem cell transplant, allo-HSCT) is an effective therapy for malignant hematological diseases. Acute graft-versus-host disease (acute graft-versus-host disease, a GVHD allo-HSCT) is a common complication of serious impact on transplant patient survival and quality of life, but also the non allo-HSCT patients the main causes of recurrence of death. At present, the clinical manifestations and pathological injury of target organ is the main basis for the diagnosis of GVHD a, but because of its clinical symptoms and complex and lack of specificity in clinical diagnosis and treatment of pathological examination and often for a variety of reasons can not lead to the timely diagnosis, so affected. Find, fast, non-invasive, practical laboratory method, let a GVHD patients early in the disease can be diagnosed and treated as soon as possible, so as to more effectively control A GVHD is an urgent need to solve clinical problem. Protein chip technology is a new technology of micro mechanical, microelectronics and life science and combination, with high specificity, high sensitivity, automation, miniaturization and other characteristics. This research adopts the sandwich method to detect membrane protein chip technology for allogeneic hematopoietic stem cell transplantation after a GVHD and a GVHD with 22 serum biomarker levels, initial screening and a GVHD development and prognosis related molecular markers, which lays the foundation for the clinical early intervention and prognosis of subsequent a GVHD. Methods: collected continuously from November 2014 to November 2015 with allo-HSCT transplantation -7d in the Department of Hematology, Xinqiao Hospital of Third Military Medical University of +7d, +14d, +28d, +56d, +100d. A total of 10 serum samples from patients with a GVHD as the experimental group, the random a did not occur in patients with GVHD 鍙，

本文編號：1605472