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IL-23、IL-17與移植免疫的相關(guān)性研究

發(fā)布時間:2018-08-25 09:47
【摘要】: 目的:探討白介素23(IL-23)、白介素17(IL-17)在小鼠皮膚移植后不同免疫狀態(tài)下外周循環(huán)血漿中及移植物組織病理的變化;了解IL-23、IL-17與調(diào)節(jié)T細(xì)胞(Treg cell)在不同免疫狀態(tài)下的相互作用;探討IL-23/IL-17通路在小鼠皮膚移植后的免疫調(diào)節(jié)作用。為臨床移植后排斥反應(yīng)的預(yù)防、診斷和治療轉(zhuǎn)歸及免疫耐受提供理論指導(dǎo)。 方法:小鼠皮膚移植動物模型的建立(n=10):第1組、同種同基因移植組(BALB/c→BALB/c)、第2組、同種異基因移植組(C57BL/6→BALB/c),第3組、供體脾細(xì)胞輸注組(C57BL/6→BALB/c)。觀察三組不同時間移植皮瓣外觀變化及鏡下三組移植皮膚HE切片的病理改變;采用ELISA方法檢測小鼠血漿IL-17、IL-23的濃度;RT-PCR反應(yīng)檢測移植皮膚及脾臟的IL-23mRNA、IL-17mRNA的表達(dá);流式細(xì)胞儀檢測脾臟Treg細(xì)胞百分比。 結(jié)果:三組小鼠移植后大體外觀及HE染色鏡下所見:同基因移植組小鼠移植皮術(shù)后皮片生長完好,無排斥反應(yīng)發(fā)生;異基因小鼠移植皮出現(xiàn)典型的急性排斥反應(yīng);術(shù)前輸注脾細(xì)胞異基因小鼠移植組,排斥反應(yīng)明顯減輕。 三組小鼠皮膚移植后,IL-17、IL-23及IL-23mRNA、IL-17mRNA術(shù)后第1天內(nèi)均表達(dá)升高,差異無統(tǒng)計(jì)學(xué)意義(P>0.05);隨著移植時間的延長,2、3組IL-17、IL-23及IL-23mRNA、IL-17mRNA的表達(dá)明顯升高,與第1組相比差異有統(tǒng)計(jì)學(xué)意義(P<0.01);2與3組術(shù)后3、5、7天的IL-23、IL-23mRNA的表達(dá)與1組相比,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);2、3組相比無統(tǒng)計(jì)學(xué)意義(P>0.05);三組術(shù)后第3、5、7天的IL-17、IL-17mRNA表達(dá)結(jié)果顯示:1、3組與第2組之間相比差異均有統(tǒng)計(jì)學(xué)意義(P<0.01)。1、3組之間無統(tǒng)計(jì)學(xué)差異(P>0.05)。 流式細(xì)胞術(shù)檢測顯示:三組CD4~+FoxP3~+Treg細(xì)胞術(shù)后第1天占CD4~+T細(xì)胞的比例分別為(0.9±0.6)%、(0.7±0.5)%、(0.8±0.4)%,差異無統(tǒng)計(jì)學(xué)意義(P>0.05)。在皮膚移植術(shù)后第7天,第3組CD4~+FoxP~(3+)Treg占CD~(4+)T細(xì)胞的比例高于2組,其分別為(5.1±2.8)%和(4.54±2.5)%,差別有統(tǒng)計(jì)學(xué)意義(P=0.00):Th17/CD4~+FoxP3~+Treg比值在第2組中的表達(dá)高于第3組差別有統(tǒng)計(jì)學(xué)意義(P=0.00)。 結(jié)論: (1)、移植后早期發(fā)生急性排斥反應(yīng)時,IL-23,IL-17及IL-23mRNA、IL-17mRNA處于高表達(dá)水平;無排斥反應(yīng)條件下IL-23、IL-17及IL-23mRNA、IL-17mRNA早期升高,隨著炎癥減輕其數(shù)值降低,IL-23,IL-17可作為預(yù)測和糾正排斥反應(yīng)發(fā)生的觀察指標(biāo)。 (2)、在急性排斥反應(yīng)模型中IL-23與IL-17呈正相關(guān)性表達(dá),IL-23作為IL-17的上游因子,對Th17細(xì)胞的成熟及其IL-17分泌有調(diào)控作用。抑制IL-23的分泌可降低IL-17的產(chǎn)生,可能降低排斥反應(yīng)的發(fā)生。 (3)、受體輸注供體脾細(xì)胞,可明顯延長移植物的存活,其機(jī)制可能與IL-17的分泌降低CD4~+FoxP3~+Treg增多有關(guān)。 (4)、受體輸注供體脾細(xì)胞可刺激Treg細(xì)胞生成,Thl7/CD4~+FoxP3~+Treg比值可能影響移植后的免疫狀態(tài)。
[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 鹵0.6), (0.7 鹵0.5) and (0.8 鹵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 鹵2.8%) and (4.54 鹵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.
【學(xué)位授予單位】:中國人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級別】:博士
【學(xué)位授予年份】:2009
【分類號】:R392

【共引文獻(xiàn)】

相關(guān)期刊論文 前4條

1 孫創(chuàng)斌;胡晉紅;朱全剛;劉繼勇;劉艷霞;沈奎亞;;苦參堿對普萘洛爾所致豚鼠銀屑病樣皮損的實(shí)驗(yàn)研究[J];解放軍藥學(xué)學(xué)報(bào);2009年06期

2 陳昭U,

本文編號:2202506


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