IL-23、IL-17与移植免疫的相关性研究
发布时间：2018-08-25 09:47
【摘要】： 目的:探讨白介素23(IL-23)、白介素17(IL-17)在小鼠皮肤移植后不同免疫状态下外周循环血浆中及移植物组织病理的变化;了解IL-23、IL-17与调节T细胞(Treg cell)在不同免疫状态下的相互作用;探讨IL-23/IL-17通路在小鼠皮肤移植后的免疫调节作用。为临床移植后排斥反应的预防、诊断和治疗转归及免疫耐受提供理论指导。 方法:小鼠皮肤移植动物模型的建立(n=10):第1组、同种同基因移植组(BALB/c→BALB/c)、第2组、同种异基因移植组(C57BL/6→BALB/c),第3组、供体脾细胞输注组(C57BL/6→BALB/c)。观察三组不同时间移植皮瓣外观变化及镜下三组移植皮肤HE切片的病理改变;采用ELISA方法检测小鼠血浆IL-17、IL-23的浓度;RT-PCR反应检测移植皮肤及脾脏的IL-23mRNA、IL-17mRNA的表达;流式细胞仪检测脾脏Treg细胞百分比。 结果:三组小鼠移植后大体外观及HE染色镜下所见:同基因移植组小鼠移植皮术后皮片生长完好,无排斥反应发生;异基因小鼠移植皮出现典型的急性排斥反应;术前输注脾细胞异基因小鼠移植组,排斥反应明显减轻。 三组小鼠皮肤移植后,IL-17、IL-23及IL-23mRNA、IL-17mRNA术后第1天内均表达升高,差异无统计学意义(P＞0.05);随着移植时间的延长,2、3组IL-17、IL-23及IL-23mRNA、IL-17mRNA的表达明显升高,与第1组相比差异有统计学意义(P＜0.01);2与3组术后3、5、7天的IL-23、IL-23mRNA的表达与1组相比,差异有统计学意义(P＜0.05);2、3组相比无统计学意义(P＞0.05);三组术后第3、5、7天的IL-17、IL-17mRNA表达结果显示:1、3组与第2组之间相比差异均有统计学意义(P＜0.01)。1、3组之间无统计学差异(P＞0.05)。 流式细胞术检测显示:三组CD4~+FoxP3~+Treg细胞术后第1天占CD4~+T细胞的比例分别为(0.9±0.6)%、(0.7±0.5)%、(0.8±0.4)%,差异无统计学意义(P＞0.05)。在皮肤移植术后第7天,第3组CD4~+FoxP~(3+)Treg占CD~(4+)T细胞的比例高于2组,其分别为(5.1±2.8)%和(4.54±2.5)%,差别有统计学意义(P=0.00):Th17/CD4~+FoxP3~+Treg比值在第2组中的表达高于第3组差别有统计学意义(P=0.00)。 结论: (1)、移植后早期发生急性排斥反应时,IL-23,IL-17及IL-23mRNA、IL-17mRNA处于高表达水平;无排斥反应条件下IL-23、IL-17及IL-23mRNA、IL-17mRNA早期升高,随着炎症减轻其数值降低,IL-23,IL-17可作为预测和纠正排斥反应发生的观察指标。 (2)、在急性排斥反应模型中IL-23与IL-17呈正相关性表达,IL-23作为IL-17的上游因子,对Th17细胞的成熟及其IL-17分泌有调控作用。抑制IL-23的分泌可降低IL-17的产生,可能降低排斥反应的发生。 (3)、受体输注供体脾细胞,可明显延长移植物的存活,其机制可能与IL-17的分泌降低CD4~+FoxP3~+Treg增多有关。 (4)、受体输注供体脾细胞可刺激Treg细胞生成,Thl7/CD4~+FoxP3~+Treg比值可能影响移植后的免疫状态。
[Abstract]:Objective: to investigate the pathological changes of interleukin-23 (IL-23) and interleukin-17 (IL-17) in peripheral circulation plasma and grafts after skin transplantation in mice, and to understand the interaction between IL-23,IL-17 and T cell (Treg cell) in different immune states. To investigate the immunomodulatory effect of IL-23/IL-17 pathway after skin transplantation in mice. To provide theoretical guidance for the prevention, diagnosis and treatment of rejection and immune tolerance after transplantation. Methods: the animal model of skin transplantation in mice was established: group 1, BALB/c BALB/c, group 2, C57BL/6 BALB/c, group 3, C57BL/6 BALB/c. The changes of skin flap appearance at different times and the pathological changes of HE sections of transplanted skin in three groups were observed, and the expression of IL-23mRNA,IL-17mRNA in the transplanted skin and spleen was detected by ELISA method, the concentration of IL-17,IL-23 in plasma and the expression of IL-23mRNA,IL-17mRNA in spleen and skin were detected by RT-PCR. The percentage of splenic Treg cells was measured by flow cytometry. Results: the gross appearance of the allogeneic mice after transplantation and the results of HE staining were as follows: the skin grafts in the same gene transplantation group grew well and no rejection occurred, and the allogeneic mice skin grafts showed typical acute rejection. The rejection of splenocyte allogeneic mice was significantly alleviated before operation. The expression of IL-17 IL-23 and IL-23 and IL-23mRNA,IL-17mRNA were increased in all the three groups after skin transplantation (P > 0. 05), and the expression of IL-17,IL-23 and IL-23mRNA,IL-17mRNA in the 3 groups increased significantly with the prolongation of transplantation time, and the expression of IL-17, IL-23 and IL-23mRNA,IL-17mRNA in the 3 groups was significantly higher than that in the control group (P > 0. 05). The expression of IL-23,IL-23mRNA in group 2 and group 3 was significantly higher than that in group 1 (P < 0. 01). There was no significant difference in IL-17,IL-17mRNA expression between the 3 groups (P < 0. 05), and there was no significant difference between the 3 groups (P < 0. 01). There was no significant difference in the expression of IL-17,IL-17mRNA between the 3 groups (P < 0. 01) and the second group (P > 0. 05), and there was no significant difference between the 3 groups (P < 0. 01). The results of IL-17,IL-17mRNA expression on the 5th day after operation showed that there was no significant difference between the 3 groups (P < 0. 01). Flow cytometry showed that the ratio of CD4~ FoxP3~ Treg cells to CD4~ T cells was (0.9 卤0.6), (0.7 卤0.5) and (0.8 卤0.4) in the first day after operation, respectively. There was no significant difference between the three groups (P > 0. 05). On the 7th day after skin transplantation, the ratio of CD4~ FoxP~ (3) Treg to CD~ (4) T cells in group 3 was higher than that in group 2 (5.1 卤2.8%) and (4.54 卤2.5), respectively. The difference was statistically significant (P < 0.001). The ratio of CD4~ FoxP~ (3) to CD~ (4) T cells in group 2 was significantly higher than that in group 3 (P < 0. 00). Conclusion: (1) the expression of IL-23 IL-17 and IL-23mRNA,IL-17mRNA was high in the early stage of acute rejection after transplantation, and the level of IL-23,IL-17 and IL-23mRNA,IL-17mRNA increased in the early stage without rejection. IL-23 IL-17 can be used as an index to predict and correct rejection with the decrease of inflammation. (2) in acute rejection model, IL-23 was positively correlated with IL-17 and IL-23 was expressed as upstream factor of IL-17. It can regulate the maturation and IL-17 secretion of Th17 cells. Inhibiting the secretion of IL-23 can decrease the production of IL-17 and decrease the occurrence of rejection. (3) Transfusion of donor spleen cells can prolong the survival of grafts. The mechanism may be related to the decrease of the secretion of IL-17 and the increase of CD4~ FoxP3~ Treg. (4) the ratio of Thl7 / CD4- FoxP3~ Treg to Thl7 / CD4 ~ FoxP3~ Treg may affect the immune state after transplantation.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
[Abstract]:Objective: to investigate the pathological changes of interleukin-23 (IL-23) and interleukin-17 (IL-17) in peripheral circulation plasma and grafts after skin transplantation in mice, and to understand the interaction between IL-23,IL-17 and T cell (Treg cell) in different immune states. To investigate the immunomodulatory effect of IL-23/IL-17 pathway after skin transplantation in mice. To provide theoretical guidance for the prevention, diagnosis and treatment of rejection and immune tolerance after transplantation. Methods: the animal model of skin transplantation in mice was established: group 1, BALB/c BALB/c, group 2, C57BL/6 BALB/c, group 3, C57BL/6 BALB/c. The changes of skin flap appearance at different times and the pathological changes of HE sections of transplanted skin in three groups were observed, and the expression of IL-23mRNA,IL-17mRNA in the transplanted skin and spleen was detected by ELISA method, the concentration of IL-17,IL-23 in plasma and the expression of IL-23mRNA,IL-17mRNA in spleen and skin were detected by RT-PCR. The percentage of splenic Treg cells was measured by flow cytometry. Results: the gross appearance of the allogeneic mice after transplantation and the results of HE staining were as follows: the skin grafts in the same gene transplantation group grew well and no rejection occurred, and the allogeneic mice skin grafts showed typical acute rejection. The rejection of splenocyte allogeneic mice was significantly alleviated before operation. The expression of IL-17 IL-23 and IL-23 and IL-23mRNA,IL-17mRNA were increased in all the three groups after skin transplantation (P > 0. 05), and the expression of IL-17,IL-23 and IL-23mRNA,IL-17mRNA in the 3 groups increased significantly with the prolongation of transplantation time, and the expression of IL-17, IL-23 and IL-23mRNA,IL-17mRNA in the 3 groups was significantly higher than that in the control group (P > 0. 05). The expression of IL-23,IL-23mRNA in group 2 and group 3 was significantly higher than that in group 1 (P < 0. 01). There was no significant difference in IL-17,IL-17mRNA expression between the 3 groups (P < 0. 05), and there was no significant difference between the 3 groups (P < 0. 01). There was no significant difference in the expression of IL-17,IL-17mRNA between the 3 groups (P < 0. 01) and the second group (P > 0. 05), and there was no significant difference between the 3 groups (P < 0. 01). The results of IL-17,IL-17mRNA expression on the 5th day after operation showed that there was no significant difference between the 3 groups (P < 0. 01). Flow cytometry showed that the ratio of CD4~ FoxP3~ Treg cells to CD4~ T cells was (0.9 卤0.6), (0.7 卤0.5) and (0.8 卤0.4) in the first day after operation, respectively. There was no significant difference between the three groups (P > 0. 05). On the 7th day after skin transplantation, the ratio of CD4~ FoxP~ (3) Treg to CD~ (4) T cells in group 3 was higher than that in group 2 (5.1 卤2.8%) and (4.54 卤2.5), respectively. The difference was statistically significant (P < 0.001). The ratio of CD4~ FoxP~ (3) to CD~ (4) T cells in group 2 was significantly higher than that in group 3 (P < 0. 00). Conclusion: (1) the expression of IL-23 IL-17 and IL-23mRNA,IL-17mRNA was high in the early stage of acute rejection after transplantation, and the level of IL-23,IL-17 and IL-23mRNA,IL-17mRNA increased in the early stage without rejection. IL-23 IL-17 can be used as an index to predict and correct rejection with the decrease of inflammation. (2) in acute rejection model, IL-23 was positively correlated with IL-17 and IL-23 was expressed as upstream factor of IL-17. It can regulate the maturation and IL-17 secretion of Th17 cells. Inhibiting the secretion of IL-23 can decrease the production of IL-17 and decrease the occurrence of rejection. (3) Transfusion of donor spleen cells can prolong the survival of grafts. The mechanism may be related to the decrease of the secretion of IL-17 and the increase of CD4~ FoxP3~ Treg. (4) the ratio of Thl7 / CD4- FoxP3~ Treg to Thl7 / CD4 ~ FoxP3~ Treg may affect the immune state after transplantation.
【学位授予单位】：中国人民解放军军医进修学院
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
【共引文献】
相关期刊论文 前4条
1 孙创斌;胡晋红;朱全刚;刘继勇;刘艳霞;沈奎亚;;苦参碱对普萘洛尔所致豚鼠银屑病样皮损的实验研究[J];解放军药学学报;2009年06期
2 陈昭U，
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