本文選題：巨噬細(xì)胞 + 嵌合體　； 參考：《蘇州大學(xué)》2007年碩士論文

【摘要】： 目的:在豚鼠到大鼠實(shí)驗(yàn)動物模型中,利用脂質(zhì)體包裹c(diǎn)lodronate清除受體巨噬細(xì)胞后,采用非清髓性的預(yù)處理方案誘導(dǎo)建立混合嵌合體,研究巨噬細(xì)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(chǔ)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(jī)制。 方法:將SD大鼠(受體)30只隨機(jī)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(yù)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c(diǎn)lodronate空脂質(zhì)體。BMC移植后24小時(shí)用免疫組化法檢查受體大鼠肝臟、脾臟中巨噬細(xì)胞清除情況;用流式細(xì)胞儀法測定BMC移植后24小時(shí)內(nèi)不同時(shí)間點(diǎn)受體外周血中豚鼠細(xì)胞比例、24小時(shí)時(shí)受體脾、骨髓中豚鼠細(xì)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(xì)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(chǔ)上,行豚鼠到大鼠的心臟移植。實(shí)驗(yàn)隨機(jī)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c(diǎn)lodronate脂質(zhì)體組,n=5)。流式細(xì)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(shí)間、檢測移植物病理組織學(xué)。 結(jié)果:輸注包裹c(diǎn)lodronate脂質(zhì)體后24小時(shí)受體大鼠肝脾(紅髓及邊緣帶)中巨噬細(xì)胞消失,21天后與正常對照相比無差異。BMC移植后24小時(shí)內(nèi)各時(shí)間點(diǎn),C組大鼠外周血中豚鼠細(xì)胞比例均高于A、B組; 24小時(shí)時(shí)大鼠脾、骨髓中豚鼠細(xì)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(shí)CD3+、CD4+和CD8+T細(xì)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(yīng)答反應(yīng)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(yīng),II、III、IV、V、VI各組移植心存活時(shí)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(yīng)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c(diǎn)lodronate的方法可建立清除巨噬細(xì)胞動物模型。2、巨噬細(xì)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(xì)胞可提高嵌合率。3、建立混合嵌合體雖然不能影響T細(xì)胞亞群的分布,但能使受者T細(xì)胞對供者抗原反應(yīng)性降低,也可延緩與T細(xì)胞活化相關(guān)的IgG抗體升高。4、運(yùn)用CVF克服超急性排斥反應(yīng)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(shí)間延長,并與嵌合水平成正相關(guān)。
[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