本文選題：巨噬細胞 + 嵌合體��； 參考：《蘇州大學》2007年碩士論文

【摘要】： 目的:在豚鼠到大鼠實驗動物模型中,利用脂質(zhì)體包裹clodronate清除受體巨噬細胞后,采用非清髓性的預處理方案誘導建立混合嵌合體,研究巨噬細胞在建立非協(xié)調(diào)性異種混合嵌合體中的作用。并在此基礎上,行豚鼠到大鼠的腹部心臟移植,研究混合嵌合體的形成對受體的免疫功能影響,探討異種免疫耐受的機制。 方法:將SD大鼠(受體)30只隨機分成3組:所有大鼠受體接受60Coγ射線全身照射,經(jīng)環(huán)磷酰胺預處理,照射后4h內(nèi)經(jīng)尾靜脈輸注BMC 2x108/ml×0.8ml。A組僅輸注豚鼠BMC;B組輸注豚鼠BMC前輸注不包含clodronate空脂質(zhì)體;C組輸注豚鼠BMC前輸注包裹clodronate空脂質(zhì)體。BMC移植后24小時用免疫組化法檢查受體大鼠肝臟、脾臟中巨噬細胞清除情況;用流式細胞儀法測定BMC移植后24小時內(nèi)不同時間點受體外周血中豚鼠細胞比例、24小時時受體脾、骨髓中豚鼠細胞比例,以及移植后21天、35天嵌合率;于BMC移植后21天,行MLR測定,并用流式細胞儀法測定T淋巴亞群的變化。 在成功建立混合嵌合體的基礎上,行豚鼠到大鼠的心臟移植。實驗隨機分為六組:I組(空白組,n=5)、II組(CVF組,n=5)III組(CVF+clodronate脂質(zhì)體組,n=5)、IV組(CVF+豚鼠BMC移植組,n=5)、V組(CVF+豚鼠BMC移植+空脂質(zhì)體組,n=5)、VI組(CVF+豚鼠BMC移植+包裹clodronate脂質(zhì)體組,n=5)。流式細胞儀法檢測移植前后大鼠抗豚鼠IgG水平;記錄移植物平均存活時間、檢測移植物病理組織學。 結(jié)果:輸注包裹clodronate脂質(zhì)體后24小時受體大鼠肝脾(紅髓及邊緣帶)中巨噬細胞消失,21天后與正常對照相比無差異。BMC移植后24小時內(nèi)各時間點,C組大鼠外周血中豚鼠細胞比例均高于A、B組; 24小時時大鼠脾、骨髓中豚鼠細胞比例,C組亦明顯高于A、B兩組;比較21天及35天嵌合水平,C組明顯高于A、B兩組。21天時CD3+、CD4+和CD8+T細胞比例均下降,各組CD4/CD8比值與對照組相比無差異。MLR顯示耐受大鼠的免疫應答反應性降低,C組較其他組比最低。 心臟移植后,I組移植心表現(xiàn)為超急性排斥反應,II、III、IV、V、VI各組移植心存活時間分別為:306±45分鐘、314±51分鐘、366±35分鐘、372±25分鐘、424±32分鐘,病理均表現(xiàn)為急性血管排斥反應改變。移植后抗豚鼠IgG水平均有輕微升高,VI組上升幅度最小。 結(jié)論:1、利用脂質(zhì)體包裹clodronate的方法可建立清除巨噬細胞動物模型。2、巨噬細胞在BMC移植后迅速減少中起重要作用,在BMC移植前去除巨噬細胞可提高嵌合率。3、建立混合嵌合體雖然不能影響T細胞亞群的分布,但能使受者T細胞對供者抗原反應性降低,也可延緩與T細胞活化相關的IgG抗體升高。4、運用CVF克服超急性排斥反應的情況下,混合嵌合體能使豚鼠到大鼠心臟移植供心存活時間延長,并與嵌合水平成正相關。
[Abstract]:Objective: in guinea pig to rat model, the mixed chimerism was induced by non-myeloablative pretreatment after clodronate was encapsulated with liposome to clear the macrophages. To study the role of macrophages in the establishment of discordant heterologous chimerism. On this basis, the abdominal heart transplantation from guinea pigs to rats was performed to study the effect of mixed chimerism on the immune function of the receptors, and to explore the mechanism of xenogeneic tolerance. Methods: thirty SD rats were randomly divided into three groups: all the receptors were irradiated by 60Co 緯 -ray and pretreated with cyclophosphamide. Within 4 hours after irradiation, BMC 2x108/ml 脳 0.8ml.A group was injected via caudal vein. The liver of recipient rats was examined by immunohistochemistry at 24 hours after transplantation of clodronate empty liposome, clodronate empty liposome, and clodronate empty liposome, without clodronate empty liposome in group C, without clodronate empty liposome. The clearance of macrophages in spleen, the percentage of guinea pig cells in peripheral blood of recipients at 24 hours after BMC transplantation and the percentage of guinea pig cells in bone marrow at 24 hours after BMC transplantation were determined by flow cytometry. The changes of T lymphoid subsets were measured by flow cytometry and MLR were measured 21 days after BMC transplantation. On the basis of successful establishment of mixed chimerism, cardiac transplantation from guinea pig to rat was performed. The experiment was randomly divided into six groups: group 鈪，

本文編號：1784137