本文選題：異基因骨髓移植　切入點：小鼠　出處：《華中科技大學》2010年博士論文　論文類型：學位論文

【摘要】： 目的：建立一個穩(wěn)定可靠的異基因骨髓移植(allogeneic bone marrow transplantation, allo-BMT)小鼠急性移植物抗宿主病(acut graft-versus-host disease, aGVHD)動物模型,為后期實研究驗allo-BMT后aGVHD和移植耐受提供理想的實驗平臺。 方法：將供者C57BL/6J (H-2b)雄性小鼠骨髓細胞和脾細胞按不同比例(1:1和1：2)混合,輸入接受致死性全身照射(8.5Gy)的受鼠雌性BALB/C (H-2d)小鼠以引起不同程度的aGVHD,檢測嵌合體情況,根據(jù)受鼠的一般情況、臨床表現(xiàn)、生存期和組織病理學等判斷aGVHD程度。 結(jié)果：混合不同比例的供鼠骨髓細胞和脾細胞allo-BMT的小鼠均發(fā)生了aGVHD,但出現(xiàn)aGVHD的時間和程度均有差異。其中骨髓與脾細胞1：2組小鼠可在相對集中的時間內(nèi)觀察到典型的aGVHDI臨床和病理改變,所有小鼠均在14-24d死亡。骨髓與脾細胞1:1組aGVHD程度較輕,移植后4周小鼠均全部存活。 結(jié)論：骨髓細胞和脾細胞比例為1:2組比1:1組出現(xiàn)更典型的aGVHD,其發(fā)病和死亡時間較集中。選定TBI8.5Gy及細胞移植的混合比例(1:2)構建小鼠aGVHD模型,為后續(xù)實驗提供理想模型。 目的：探討不同方法誘導的受者來源樹突狀細胞(dendritic cells, DCs)輸注對異基因骨髓移植(allogeneic bone marrow transplantation, allo-BMT)小鼠急性移植物抗宿主病(acut graft-versus-host disease,aGVHD)和造血重建的影響。 方法：受鼠為雌性BALB/C (H-2d)小鼠,供鼠為雄性C57BL/6J (H-2b)小鼠。無菌分離受鼠骨髓細胞(bone marrow cell, BMC),與GM-CSF共培養(yǎng)得到未成熟樹突狀細胞(immature dendritic cells, imDCs);受鼠BMC與GM-CSF、IL-10共培養(yǎng)得到IL-10-DCs;用配對性免疫球蛋白樣受體B (Paired immunoglobin-like receptor B, PIR-B)慢病毒載體轉(zhuǎn)染imDCs得到PIR-B-DCs。受鼠隨機分為四組,在移植骨髓后分別予尾靜脈注射imDCs、IL-10-DCs、PIR-B-DCs和RPMI1640培養(yǎng)液。以移植后aGVHD臨床表現(xiàn),肝臟、小腸、皮膚病理組織改變,生存期為觀察指標并進行組間比較。 結(jié)果：PIR-B-DCs移植組、IL-10-DCs移植組、imDCs移植組和單純移植組受鼠平均生存時間為(46.0±13.6)天、(36.4±13.0)天、(21.6±2.8)天和(17.4±3.6)天,p＜0.01；4組小鼠在移植后15天GVHD臨床評分為(5.28±0.27)、(5.26±0.31)、(2.46±0.18)和(0.86±0.21),p0.05。單純移植組病理顯示肝臟匯管區(qū)大量淋巴細胞浸潤,小膽管塌陷破壞,皮膚基底層連續(xù)性中斷,有大量淋巴細胞,腸粘膜下層淋巴細胞浸潤,腸絨毛萎縮變性,PIR-B-DCs移植組病理組織檢查僅有輕微GVHD表現(xiàn)。 結(jié)論：受者來源PIR-B-DCs聯(lián)合骨髓移植能夠明顯延長受鼠生存時間,減輕移植后aGVHD反應,促進造血重建。
[Abstract]:Objective: to establish a stable and reliable animal model of acute graft-versus-host disease graft-versus-host disease (aGVHD) in allogeneic bone marrow transfer (allo-BMTT) mice, and to provide an ideal experimental platform for the later study of aGVHD and transplantation tolerance after allo-BMT. Methods: bone marrow cells and spleen cells of donor C57BL / 6J / H-2b) mice were mixed at different proportions of 1: 1 and 1: 2) and injected into murine female BALB/C / H-2d mice receiving lethal whole-body irradiation for different degrees of aGV HDD, and chimerism was detected. The degree of aGVHD was judged according to the general condition, clinical manifestation, survival time and histopathology of recipient mice. Results: the aGV HDD was found in mice mixed with different proportions of donor bone marrow cells and splenocytes allo-BMT, but the time and degree of aGVHD appeared were different, among which the bone marrow and spleen cells 1: 2 group could be observed in a relatively concentrated time. Typical clinical and pathological changes of aGVHDI, All the mice died from 14 to 24 days. The aGVHD degree of bone marrow and spleen cells 1: 1 group was mild, and all the mice survived 4 weeks after transplantation. Conclusion: the ratio of bone marrow cells to spleen cells in group 1: 2 is more typical than that in group 1: 1, and the onset and death time is more concentrated. The aGVHD model of mice was constructed by selecting the mixed ratio of TBI8.5Gy and cell transplantation (1: 2) to provide an ideal model for further experiments. Aim: to investigate the effects of dendritic cells (DCs) induced by different methods on allogeneic bone marrow transfer (allo-BMTM) and hematopoietic reconstitution in mice with acute graft-versus-host disease, acut graft-versus-host disease, aGVHD. Methods: the recipient mice were female BALB/C (H-2d) mice. The donor mice were male C57BL / 6J ~ (H-2b) mice. Bone marrow cells were isolated from bone marrow cells and co-cultured with GM-CSF to obtain immature dendritic cells dendritic cells, imDCsN; recipient BMC co-cultured with GM-CSFFIL-10 to obtain IL-10-DCs; matched immunoglobulin-like receptor B Paired immunoglobin-like receptor. The imDCs vector was transfected with B, PIR-B-DCs.The recipient mice were randomly divided into four groups. After bone marrow transplantation, we injected imDCssil IL-10-DCssil PIR-B-DCs and RPMI1640 culture medium into caudal vein, respectively. The clinical manifestations of aGVHD, pathological changes of liver, small intestine and skin, survival time were observed and compared among the groups. Results the average survival time of the two groups were 46.0 鹵13.6) and 36.4 鹵13.0) days and 21.6 鹵2.8 days and 17.4 鹵3.6 days, respectively. The GVHD clinical scores were 5.28 鹵0.27 + 5.26 鹵0.31 鹵0.18) and 0.86 鹵0.21 p0.05respectively. The clinical scores of GVHD were 5.28 鹵0.27, 2.46 鹵0.18) and 0.86 鹵0.21p0.05in the 15 days after transplantation, respectively. The clinical scores of GVHD were 5.28 鹵0.27 (2.46 鹵0.18) and 0.86 鹵0.21 (p0.05) respectively in the 10 ~ (-10) -DCs transplantation group and in the control group. The clinical scores of GVHD were 5.28 鹵0.27 (2.46 鹵0.18) and 0.86 鹵0.21 (p0.05) respectively. A large number of lymphocytic infiltration in the canal area, In the small bile duct collapse, the continuity of the basal layer of the skin was interrupted, there were a large number of lymphocytes and submucous lymphocytes infiltrated in the intestinal submucosa. The pathological findings of PIR-B-DCs transplantation group showed only slight GVHD findings. Conclusion: recipient PIR-B-DCs combined with bone marrow transplantation can significantly prolong the survival time of recipient mice, alleviate aGVHD response after transplantation, and promote hematopoietic reconstitution.
【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2010
【分類號】：R392

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