FK506处理的供者树突状细胞在诱导大鼠同种移植免疫耐受中的作用

发布时间：2018-03-07 12:44

本文选题：FK506　切入点：未成熟树突状细胞　出处：《青岛大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的：研究FK506处理的供者未成熟树突状细胞(immature dendritic cell，imDC)对大鼠同种异体器官移植免疫耐受的影响，探讨imDC诱导免疫耐受的机制。方法：联合应用低剂量重组大鼠GM-CSF(1.5ng／ml)和IL-4(4 ng／ml)诱导Wistar大鼠骨髓细胞，培养出大量未成熟树突状细胞(imDC)，并分别用LPS和FK506加以处理，利用形态学及FCM鉴定其成熟度。将SD大鼠分为3组：第1组为对照组，术前7天尾静脉注射生理盐水1.0ml；第2组为未经FK506处理组，术前7天尾静脉注射未经FK506处理的imDC 2×10~6(1.0ml)；第3组为FK506处理组，术前7天注射FK506处理的imDC 2×10~6(1.0ml)。以Wistar大鼠为供体，SD大鼠为受体，行颈部心脏移植，观察移植心脏存活时间、移植后大鼠与Wistar大鼠及第三品系Lewis大鼠单向MLR、心肌病理及血清IL-2、IL-4、IL-10、IFN-r的水平变化。结果：1、流式细胞仪分析结果显示所培养的细胞表面低表达CD80、CD86，LPS的刺激使这些分子表达明显上调，因此确定所培养的细胞是imDC。FK506能够抑制DC表面CD80和CD86分子的上调。 2、45例大鼠颈部心脏移植模型，手术成功率80.0％。H检验显示，3组移植心脏存活时间的差异有高度统计学意义(P＜0.01)；q检验显示，3组间两两比较，，1、2组，1、3组间差异有高度统计学意义(P＜0.01)，2、3组间差异有统计学意义(P＜0.05)。 3、供体大鼠脾细胞作为刺激细胞时，未经FK506处理组大鼠MLR的SI值(3.48±0.84)，FK506处理组SI值(3.59±0.68)，均明显低于对照组(6.83±0.47)，差异有高度统计学意义(P＜0.01)，2、3组间SI值无明显差异。第三品系Lewis大鼠脾细胞作为刺激细胞时，未经FK506处理组SI值(6.57±0.51)，FK506处理组SI值(6.67±0.66)，与对照组(6.58±1.03)之间无明显差异(P＞0.05)。 4、方差分析示各组大鼠血清IL-2、IFN-r、IL-4、IL-10浓度间差异有高度统计学意义(P＜0.01)。S-N-K检验显示，2、3组IL-2水平于第7天时无明显差异，IFN-r水平于第14天时无明显差异(P＞0.05)；其余各组间差异均有高度统计学意义(P＜0.01)。第2、3组代表Th1的IL-2、IFN-r的水平明显降低，而代表Th2的IL-4、IL-10水平明显增高。 5、对照组的移植心脏病理显示心肌明显充血、水肿，大量炎性细胞浸润，心肌细胞明显变性坏死；未经FK506处理组的心肌淋巴细胞漫润明显减少，心肌细胞无明显变性、坏死；FK506处理组几乎见不到炎性细胞浸润以及心肌细胞变性和坏死。结论：1、FK506对DC的功能成熟有明显的抑制作用。 2、imDC能够诱导同种异体大鼠心脏移植免疫耐受；FK506处理的imDC能够加强这种免疫耐受。 3、imDC所诱导的免疫耐受是供者特异性的。 4、imDC可能主要通过调节T细胞免疫应答类型(Th1至Th2的免疫偏移)、诱导调节性T细胞(regulatory T cells，T-reg)和诱导T细胞失能等途径来参与免疫耐受的形成。
[Abstract]:Aim: to study the effect of immature dendritic dendritic cells treated with FK506 on the tolerance of allogeneic organ transplantation in rats, and to explore the mechanism of imDC induced immune tolerance. Methods: bone marrow cells of Wistar rats were induced by low dose recombinant rat GM-CSF 1.5 ng / ml and IL-4(4 ng / ml. A large number of immature dendritic cells were cultured and treated with LPS and FK506, respectively. The SD rats were divided into three groups: the control group was treated with 1.0 ml saline at the end of 7 days before operation, the second group was the group without FK506 treatment, and the control group was divided into three groups: the control group was injected intravenously with physiological saline 7 days before operation, and the second group was treated with FK506. On the 7th day before operation, imDC 2 脳 10 ~ (6) 0 ml without FK506 was injected intravenously 7 days before operation, while in the third group, imDC 2 脳 10 ~ (6) 1 ml of FK506 was injected 7 days before operation. Wistar rats were used as the recipient, and the survival time of the transplanted heart was observed. After transplantation, the changes of unidirectional MLR, cardiomyopathology and the level of IL-10 and IFN-r in serum of Wistar rats and third strain Lewis rats were observed. Results: the results of flow cytometry analysis showed that the low expression of CD80, CD86 and LPS on the surface of the cultured cells significantly up-regulated the expression of these molecules. It was concluded that the cultured cells could inhibit the upregulation of CD80 and CD86 molecules on DC surface by imDC.FK506. (2) the cervical heart transplantation model of 45 rats, The success rate of operation was 80.0%. H test showed that the survival time of cardiac transplantation was significantly different among the three groups (P < 0.01). The results showed that there was a significant difference between the two groups (P < 0.01). There was a significant difference among the three groups (P < 0.01). There was a significant difference among the three groups (P < 0.05). 3, when donor rat spleen cells were used as stimulators, The SI value of MLR in the untreated group was 3.48 卤0.84FK506 (3.59 卤0.68), which was significantly lower than that in the control group (6.83 卤0.47). There was no significant difference in SI between the three groups (P < 0.01). There was no significant difference between the SI value of the untreated group and the control group (6.57 卤0.51) and the control group (6.67 卤0.66) (P > 0.05). (4) the analysis of variance showed that there was a significant difference in the concentration of IL-10 between the two groups (P < 0.01). S-N-K test showed that there was no significant difference in the level of IL-2 between the three groups on the 7th day. There was no significant difference in the level of IFN-r between the three groups on the 14th day (P > 0.05), and there was a high difference among the other groups. The level of IL-2tIFN-r, which represents Th1, was significantly decreased in group 2 (P < 0.01), and the level of IFN-r in group 2 was significantly lower than that in group 2 (P < 0.01). The level of IL-10, which represents Th2, was significantly increased. 5. The heart grafts in the control group showed obvious congestive heart, edema, large number of inflammatory cells infiltration, obvious degeneration and necrosis of myocardial cells, but no obvious degeneration of myocardial cells in the group without FK506 treatment. In FK506 treated group, inflammatory cell infiltration, degeneration and necrosis of cardiomyocytes were almost absent. Conclusion FK506 has a significant inhibitory effect on the functional maturation of DC. (2) imDC treated with FK506 can enhance the immune tolerance of allogeneic rat heart transplantation induced by MDC. The immune tolerance induced by MDC is donor specific. 4imDC may be involved in the formation of immune tolerance by regulating T cell immune response type from Th1 to Th2, inducing regulatory T cell regulatory T cell T-regn and inducing T cell disability.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392.4

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