發(fā)布時間：2019-03-08 07:53

【摘要】：目的 異種移植被認(rèn)為是有潛力的解決器官供源不足的方法,但異種移植面臨的免疫排斥比同種異體移植更復(fù)雜,移植物抗宿主病(GVHD)更為嚴(yán)重。我們應(yīng)用大鼠-小鼠造血嵌和體模型,進(jìn)一步研究反向骨髓移植對aGVHD的預(yù)防機(jī)制。 方法 1、異種骨髓嵌合體模型的建立:術(shù)前5天SD大鼠(8～10周齡,雌性,SPF級)開始飲含紅霉素(250mg/L)和慶大霉素(320mg/L)抗生素溶液凈化腸道。無菌條件下取BALB/C小鼠(8～10周齡,雌性,SPF級)的骨髓細(xì)胞,制成單細(xì)胞懸液,SD大鼠接受7.5Gy(照射率0.5Gy/min)全身照射(TBI)后4小時內(nèi)經(jīng)尾靜脈輸入上述BALB/c小鼠骨髓細(xì)胞2×10~8/只,48小時后腹腔注射環(huán)磷酰胺50mg/kg(此預(yù)處理方案為非清髓性),將該嵌合了BALB/c小鼠骨髓細(xì)胞的供體大鼠飼養(yǎng)30天后,檢測外周血嵌合率,并做皮片移植,結(jié)果證實小鼠源性骨髓已在大鼠體內(nèi)植活,并誘導(dǎo)出對BALB/c小鼠的特異性耐受。再將該嵌合體SD大鼠的骨髓細(xì)胞移植到目標(biāo)受體(BALB/C小鼠)。 2、實驗分組:A,B,C組以BALB/C(8～10周齡,雌性,SPF級)小鼠為受鼠,D組以無關(guān)第3者B6小鼠為受鼠,腸道凈化后接受9Gy~(60)Coγ射線致死性TBI:A組經(jīng)尾靜脈輸注正常SD大鼠的骨髓細(xì)胞4×10~7/只;B組經(jīng)尾靜脈輸注嵌合體SD大鼠的骨髓細(xì)胞4×10~7/只;C組經(jīng)尾靜脈輸注生理鹽水;D組經(jīng)尾靜脈輸注嵌合體SD大鼠的骨髓細(xì)胞4×10~7/只。觀察各組小鼠aGVHD的臨床表現(xiàn)及生存時間,瀕死小鼠做病理分析。移植后15天測定TNF-α、INF-γ與IL-4的產(chǎn)量,移植后
[Abstract]:Aim xenotransplantation is considered to be a potential method to solve the shortage of organ supply and source, but the immune rejection of xenotransplantation is more complicated than that of allotransplantation, and (GVHD) is more serious. We used the rat-mouse hemopoietic chimerism model to further study the preventive mechanism of reverse bone marrow transplantation (BMT) on aGVHD. Methods 1. Establishment of xenogeneic bone marrow chimera model: SD rats (8 weeks, 10 weeks old, female, 5 days before operation); SPF grade) began to drink erythromycin (250mg/L) and gentamicin (320mg/L) antibiotic solution to purify the intestinal tract. Bone marrow cells of BALB/C mice (8-10-week-old, female, SPF grade) were taken under sterile conditions to make single-cell suspension. The bone marrow cells of the above-mentioned BALB/c mice were injected into the tail vein of SD rats within 4 hours after the whole body irradiation with 7.5Gy (irradiation rate 0.5Gy/min) 2 脳 10 ~ 8 / mouse. 48 hours later, cyclophosphamide 50mg/kg (a non-myeloablative regimen) was injected intraperitoneally. After 30 days of feeding the donor rat chimeric with BALB/c mouse bone marrow cells, the peripheral blood chimerism rate was detected and skin graft was performed. The results showed that mouse-derived bone marrow had been implanted in rats and induced specific tolerance to BALB/c mice. The bone marrow cells of the chimeric SD rats were transplanted to the target receptor (BALB/C mice). 2. The experimental group: a, B, C: BALB/C (8-week-old, female, SPF grade) mice as recipient mice, group D: B-6 mice with no third group as recipient mice, group C (8-10 weeks old, female, SPF grade) mice as recipient mice. After intestinal purification, 4 脳 10 ~ 7 bone marrow cells of normal SD rats were infused through tail vein in 9 Gy ~ (60) Co 緯-ray lethal TBI:A group. Bone marrow cells of chimeric SD rats were infused into group B (4 脳 10 ~ 7), group C (normal saline), and group D (4 脳 10 ~ 7) of chimeric SD rats. The clinical manifestation and survival time of aGVHD in each group were observed, and pathological analysis was made in dying mice. The production of TNF- 偽, INF- 緯 and IL-4 was measured 15 days after transplantation.
【學(xué)位授予單位】：第一軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 唐湘鳳,李春富,裴夫瑜;小鼠→大鼠異種移植耐受模型的建立[J];免疫學(xué)雜志;2003年06期

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本文編號：2436591