趋化因子I-TAC在移植排斥中免疫学效应的体外研究
发布时间：2019-05-16 00:19
【摘要】： 目的研究CXC家族趋化因子I-TAC在内皮细胞上的表达及其多种免疫学功能,同时用免疫抑制剂霉酚酸(MPA)对I-TAC的表达和移植免疫学功能进行干预,从而探讨I-TAC在器官移植排斥反应中新的作用以及MPA新的作用靶点。 方法采用RT-PCR半定量分析检测人脐静脉内皮细胞株ECV304的I-TAC mRNA的表达。ELISA检测同种异体外周血单个核细胞(PBMC)混合培养上清中IFN-γ和TNF-α浓度。通过体外趋化实验观察I-TAC对PBMC的迁移作用。采用黏附实验检测I-TAC促PBMC黏附功能。应用MTT法测定I-TAC对PBMC、ECV304细胞增殖的影响。应用流式细胞术检测CXCR3在PBMC表面的表达。 结果ECV304细胞在IFN-γ作用8h后表达I-TAC mRNA,TNF-α对IFN-γ的这种作用具有协同效应,且呈剂量依赖性;同种异体PBMC的混合细胞培养上清中测定到高浓度的IFN-γ和TNF-α,且能诱导ECV304细胞I-TAC mRNA的表达。I-TAC对PBMC具有趋化作用,且在I-TAC浓度为200ng/ml时趋化功能最强;同时I-TAC具有促PBMC增殖和黏附作用,且呈剂量依赖性,对内皮细胞I-TAC具有抑制增殖作用。I-TAC的受体CXCR3在ECV304细胞上也有表达。混合培养后PBMC上的CXCR3表达与单独培养相比明显提高。 MPA对IFN-γ诱导ECV304细胞I-TAC mRNA表达以及I-TAC对PBMC趋化功能具有一定抑制作用,同时还发现MPA能够降低CXCR3在PBMC上的表达。结论IFN-γ能够诱导ECV304细胞表达I-TAC mRNA,TNF-α对IFN-γ的诱导作用具有协同效应。I-TAC具有趋化、促增殖、促黏附等多重功能,它不仅是T细胞穿过内皮细胞进入移植物的关键因素,而且对淋巴细胞和内皮细胞的功能具有调节作用。 免疫抑制剂MPA不仅对I-TAC的表达及其趋化功能具有抑制作用,而且对其受体CXCR3表达也有一定的影响。MPA的这些综合活性可能是MMF发挥抗移植排斥作用的机制之一。
[Abstract]:Objective to study the expression of CXC family chemokine I-TAC in endothelial cells and its immunological functions, and to intervene the expression of I-TAC and the immunological function of transplantation with mycophenolate mofetil (MPA), an immunosuppressant. In order to explore the new role of I-TAC in organ transplantation rejection and the new target of MPA. Methods the expression of I-TAC mRNA in human umbilical vein endothelial cell line ECV304 was detected by RT-PCR semi-quantitative analysis, and the concentrations of IFN- 纬 and TNF- 伪 in (PBMC) mixed culture medium of allogenic peripheral blood mononuclear cells were detected by Elisa. The migration effect of I-TAC on PBMC was observed by chemotactic assay in vitro. The adhesion function of PBMC promoted by I-TAC was detected by adhesion test. The effect of I-TAC on the proliferation of PBMC,ECV304 cells was measured by MTT assay. The expression of CXCR3 on PBMC surface was detected by flow cytometry. Results the expression of I-TAC mRNA,TNF- 伪 in ECV304 cells for 8 h had synergistic effect on IFN- 纬 in a dose-dependent manner. High concentrations of IFN- 纬 and TNF- 伪 were detected in the mixed cell culture medium of allogenic PBMC, and I-TAC could induce the expression of I-TAC mRNA in ECV304 cells. I-TAC had chemotactic effect on PBMC, and the chemotactic function was the strongest when the concentration of I-TAC was 200ng/ml. At the same time, I-TAC can promote the proliferation and adhesion of PBMC in a dose-dependent manner, and inhibit the proliferation of endothelial cells I-TAC. I-TAC receptor CXCR3 is also expressed in ECV304 cells. The expression of CXCR3 on PBMC in mixed culture was significantly higher than that in culture alone. MPA could inhibit the expression of I-TAC mRNA in ECV304 cells induced by IFN- 纬 and the chemotactic function of PBMC induced by I-TAC. At the same time, it was also found that MPA could decrease the expression of CXCR3 on PBMC. Conclusion the expression of I-TAC mRNA,TNF- 伪 in ECV304 cells induced by IFN- 纬 has synergistic effect on the induction of IFN- 纬. I-TAC has many functions, such as chemotaxis, proliferation, adhesion and so on. It is not only a key factor for T cells to pass through endothelial cells into the graft, but also plays a regulatory role in the function of lymphocytes and endothelial cells. Immunosuppressive agent MPA has inhibitory effect not only on the expression of I-TAC and its chemotactic function, but also on the expression of its receptor CXCR3. These comprehensive activities of MPA may be one of the mechanisms of MMF against transplantation rejection.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392.4
本文编号：2477874
[Abstract]:Objective to study the expression of CXC family chemokine I-TAC in endothelial cells and its immunological functions, and to intervene the expression of I-TAC and the immunological function of transplantation with mycophenolate mofetil (MPA), an immunosuppressant. In order to explore the new role of I-TAC in organ transplantation rejection and the new target of MPA. Methods the expression of I-TAC mRNA in human umbilical vein endothelial cell line ECV304 was detected by RT-PCR semi-quantitative analysis, and the concentrations of IFN- 纬 and TNF- 伪 in (PBMC) mixed culture medium of allogenic peripheral blood mononuclear cells were detected by Elisa. The migration effect of I-TAC on PBMC was observed by chemotactic assay in vitro. The adhesion function of PBMC promoted by I-TAC was detected by adhesion test. The effect of I-TAC on the proliferation of PBMC,ECV304 cells was measured by MTT assay. The expression of CXCR3 on PBMC surface was detected by flow cytometry. Results the expression of I-TAC mRNA,TNF- 伪 in ECV304 cells for 8 h had synergistic effect on IFN- 纬 in a dose-dependent manner. High concentrations of IFN- 纬 and TNF- 伪 were detected in the mixed cell culture medium of allogenic PBMC, and I-TAC could induce the expression of I-TAC mRNA in ECV304 cells. I-TAC had chemotactic effect on PBMC, and the chemotactic function was the strongest when the concentration of I-TAC was 200ng/ml. At the same time, I-TAC can promote the proliferation and adhesion of PBMC in a dose-dependent manner, and inhibit the proliferation of endothelial cells I-TAC. I-TAC receptor CXCR3 is also expressed in ECV304 cells. The expression of CXCR3 on PBMC in mixed culture was significantly higher than that in culture alone. MPA could inhibit the expression of I-TAC mRNA in ECV304 cells induced by IFN- 纬 and the chemotactic function of PBMC induced by I-TAC. At the same time, it was also found that MPA could decrease the expression of CXCR3 on PBMC. Conclusion the expression of I-TAC mRNA,TNF- 伪 in ECV304 cells induced by IFN- 纬 has synergistic effect on the induction of IFN- 纬. I-TAC has many functions, such as chemotaxis, proliferation, adhesion and so on. It is not only a key factor for T cells to pass through endothelial cells into the graft, but also plays a regulatory role in the function of lymphocytes and endothelial cells. Immunosuppressive agent MPA has inhibitory effect not only on the expression of I-TAC and its chemotactic function, but also on the expression of its receptor CXCR3. These comprehensive activities of MPA may be one of the mechanisms of MMF against transplantation rejection.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392.4
【参考文献】
相关期刊论文 前2条
1 郭克泰;趋化因子及其受体的研究新进展[J];国外医学(免疫学分册);2002年01期
2 张锋;一种新的趋化因子—Fractalkine[J];国外医学(免疫学分册);2002年03期
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