Th17细胞在人早孕期母—胎界面功能性调节作用
发布时间：2018-08-18 19:24
【摘要】：生理妊娠类似于同种移植,作为同种移植物的胚胎在母体存活直至分娩,实际上反映母体对胚胎的免疫耐受；母体对胚胎的免疫排斥则导致妊娠失败。揭示母-胎免疫耐受的确切机制,将对人类自然流产等妊娠疾患的防治具有重要意义；并对移植免疫学和肿瘤免疫学的研究将产生推动作用。 母-胎界面主要包括滋养细胞、蜕膜基质细胞、蜕膜腺上皮细胞以及免疫细胞。多年来人们从不同角度研究母-胎界面发生的生物学事件,以阐述母-胎免疫调节机制,其中包括母-胎界面Th2型免疫优势的形成,及调节性T细胞(Treg)扩增及功能机制。Th17细胞作为新近研究发现的一群T辅助细胞亚群,参与多种自身免疫性疾病及移植物抗宿主病的发生,我们研究发现人早孕期间蜕膜Th17细胞数量显著增加。我们以此为切入点,以阐明母-胎界面Th17细胞的来源及其在母-胎免疫调节中的作用。 第一部分人早孕期母-胎界面及外周Th17细胞亚群增加 目的了解人妊娠期间外周血及母-胎界面Th17细胞比例的变化。 方法收集正常早、中、晚孕妇女以及正常非孕育龄期妇女外周血,正常早孕妇女蜕膜组织以及非孕妇女子宫内膜组织,采用流式细胞术分析外周血及蜕膜免疫细胞中Th17细胞的比例。 结果妊娠期妇女外周血、母-胎界面以及正常非孕妇女外周血、子宫内膜中均存在Thl7细胞；而且早孕妇女外周血Th17细胞数量显著增加,随着妊娠进展,其比例逐渐下降,至晚孕期达非孕期水平。与正常非孕妇女子宫内膜组织相比,早孕妇女蜕膜组织Th17细胞亚群显著增加,且显著高于外周血。 结论Th17细胞可能参与正常妊娠的维持 第二部分人母-胎界面Th17细胞与Treg细胞的分化发育 目的解析母-胎界面局部微环境对Th17细胞分化发育的影响 方法流式细胞术分析母-胎界面T细胞的表型。滋养细胞、蜕膜基质细胞与蜕膜naive CD4+T细胞间接接触共培养或者滋养细胞、蜕膜基质细胞或滋养细胞与蜕膜基质细胞共培养体系培养上清存在下,采用anti-CD3抗体和anti-CD28抗体活化磁珠分选的母-胎界面naive CD4+T细胞。妊娠相关激素处理蜕膜naiveCD4+T细胞,流式细胞术分析Th17细胞的比例。 结果母-胎界面存在naive CD4+T细胞。滋养细胞、DSC分别,或两者共培养均显著抑制蜕膜naive CD4+T细胞分化发育为Th17细胞。用母-胎界面主要功能细胞条件培养液处理蜕膜naive CD4+T细胞显示,母-胎界面主要功能细胞条件培养液可显著抑制Thl7细胞的分化,促进Treg细胞的优势分化。妊娠相关激素不影响Th17细胞的分化。 结论母-胎界面微环境有利于naive CD4+T细胞分化发育为Treg细胞,而不利于分化发育为Th17细胞,呈现Treg偏移。 第三部分人蜕膜基质细胞募集通过分泌CCL2和CCL20外周Th17细胞 目的探讨母-胎界面Th17细胞的来源 方法磁珠分选外周血CD4+T细胞,采用滋养细胞、蜕膜基质细胞或两者共培养上清液对Th17细胞进行趋化试验,流式细胞术分析趋化至下室的Th17细胞的数量。 结果DSC条件培养液募集至下室的Th17细胞数约为外周的2.7倍；而滋养细胞上清及滋养细胞与DSC直接接触共培养上清募集至下室的Th17细胞数分别是外周的1.1倍及1.8倍。而在DSC培养上清液中加入anti-CCL2中和性抗体后,可显著抑制DSC培养上清对Th17细胞的趋化作用。 结论蜕膜基质细胞通过分泌CCL2募集外周血Th17细胞到达蜕膜局部第四部分人母-胎界面Th17细胞促进滋养细胞增殖及侵袭 目的探讨Th17细胞对滋养细胞生物学行为的调控作用 方法免疫组织化学及免疫细胞化学检测母-胎界面主要功能细胞IL-17R的表达。磁珠分选naive CD4+T细胞,体外诱导Th17细胞分化,制备Th17细胞培养上清,处理滋养细胞,检测滋养细胞增殖及侵袭能力,流式细胞术分析滋养细胞的凋亡 结果滋养细胞、蜕膜基质细胞均表达IL-17R。rhIL-17可以剂量依赖性的方式促进滋养细胞的增殖及侵袭；Th17细胞培养上清亦可促进滋养细胞增殖及侵袭；加入IL-17中和抗体后,可显著抑制滋养细胞增殖及侵袭能力。Th17细胞培养上清处理滋养细胞后,滋养细胞凋亡率下降,从2.23%±0.15%下降至1.88%±0.42%；加入IL-17中和抗体后,导致滋养的凋亡率恢复升高至2.0%±0.11%。 结论Th17细胞通过分泌细胞因子IL-17调控滋养细胞的生物学行为,参与早孕胎盘形成。
[Abstract]:Physiological pregnancy is similar to allogeneic transplantation. Embryos as allografts survive in the mother until childbirth, which in fact reflects the maternal immune tolerance to the embryo; maternal immune rejection of the embryo leads to pregnancy failure. It will also promote the study of transplantation immunology and tumor immunology.
The maternal-fetal interface mainly includes trophoblasts, decidual stromal cells, decidual gland epithelial cells and immune cells. Over the years, biological events at the maternal-fetal interface have been studied from different perspectives to elucidate the mechanisms of maternal-fetal immune regulation, including the formation of Th2 immunodominance at the maternal-fetal interface, and the expansion and function of regulatory T cells (Treg). Mechanisms. Th17 cells, as a subset of T helper cells found recently, are involved in many autoimmune diseases and graft versus host disease. We found that the number of Th17 cells in human decidua increases significantly during early pregnancy. The role of.
The first part is the increase of maternal fetal interface and peripheral Th17 cell subsets in early pregnancy.
Objective to investigate the ratio of Th17 cells in peripheral blood and maternal fetal interface during pregnancy.
Methods Peripheral blood, decidual tissue and endometrial tissue were collected from normal early, middle and late pregnant women and normal non-pregnant women. The proportion of Th17 cells in peripheral blood and decidual immune cells was analyzed by flow cytometry.
Results Thl7 cells were found in peripheral blood, maternal-fetal interface and normal non-pregnant women's endometrium during pregnancy, and the number of Thl7 cells in peripheral blood of early pregnant women increased significantly. With the progress of pregnancy, the proportion of Thl7 cells decreased gradually and reached the level of non-pregnant women's endometrium at the late pregnancy. The Th17 cell subsets of female Decidua Tissue increased significantly, and was significantly higher than that of peripheral blood.
Conclusion Th17 cells may be involved in the maintenance of normal pregnancy.
The second part is the differentiation and development of Th17 cells and Treg cells at the maternal fetal interface.
Objective to analyze the effect of maternal fetal interface microenvironment on differentiation and development of Th17 cells.
Methods The phenotype of T cells at maternal-fetal interface was analyzed by flow cytometry. The magnetic beads were activated by anti-CD3 antibody and anti-CD28 antibody in the presence of the supernatant of the co-culture system of decidual stromal cells, decidual stromal cells and decidual naive CD4 + T cells, decidual stromal cells and decidual stromal cells. Nave CD4+T cells were selected from the maternal-fetal interface. Pregnancy-related hormones were used to treat naive CD4+T cells in decidua. The proportion of Th17 cells was analyzed by flow cytometry.
Results There were naive CD4+T cells in the maternal-fetal interface. Trophoblasts, DSCs, or co-cultures significantly inhibited the differentiation and development of decidual naive CD4+T cells into Th17 cells. Cell differentiation promotes the differentiation of Treg cells. Pregnancy related hormones do not affect the differentiation of Th17 cells.
Conclusion Maternal-fetal microenvironment is beneficial to the differentiation and development of naive CD4+T cells into Treg cells, but not to Th17 cells, showing Treg migration.
In the third part, human decidual stromal cells are recruited by secreting CCL2 and CCL20 peripheral Th17 cells.
Objective to investigate the origin of Th17 cells in maternal fetal interface.
Methods Peripheral blood CD4+T cells were sorted by magnetic beads. Th17 cells were chemotaxis by trophoblast, decidual stromal cells or co-culture supernatant. The number of Th17 cells chemotaxis to the lower chamber was analyzed by flow cytometry.
Results The number of Th17 cells in DSC conditioned medium was about 2.7 times that of peripheral cells, while the number of Th17 cells in supernatant and co-culture supernatant was 1.1 times and 1.8 times that of peripheral cells, respectively. The chemotaxis effect of supernatant on Th17 cells.
Conclusion Decidual stromal cells can promote the proliferation and invasion of trophoblasts by secreting CCL2 to recruit Th17 cells from peripheral blood to the fourth part of human maternal-fetal interface.
Objective to investigate the regulation of Th17 cells on the biological behavior of trophoblast cells.
Methods The expression of IL-17R was detected by immunohistochemistry and immunocytochemistry. Nave CD4+T cells were sorted by magnetic beads, and Th17 cells were induced to differentiate in vitro. The culture supernatant of Th17 cells was prepared. Trophoblasts were treated. The proliferation and invasiveness of trophoblasts were detected. The apoptosis of trophoblasts was analyzed by flow cytometry.
Results Both trophoblasts and decidual stromal cells expressed IL-17R.rhIL-17 in a dose-dependent manner, and the supernatant of Th17 cells also promoted the proliferation and invasion of trophoblasts. The apoptotic rate of trophoblasts decreased from 2.23%+0.15% to 1.88%+0.42% and the apoptotic rate of trophoblasts recovered to 2.0%+0.11% after adding IL-17 neutralizing antibody.
Conclusion Th17 cells regulate the biological behavior of trophoblasts by secreting cytokine IL-17 and participate in placenta formation in early pregnancy.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
本文编号：2190443
[Abstract]:Physiological pregnancy is similar to allogeneic transplantation. Embryos as allografts survive in the mother until childbirth, which in fact reflects the maternal immune tolerance to the embryo; maternal immune rejection of the embryo leads to pregnancy failure. It will also promote the study of transplantation immunology and tumor immunology.
The maternal-fetal interface mainly includes trophoblasts, decidual stromal cells, decidual gland epithelial cells and immune cells. Over the years, biological events at the maternal-fetal interface have been studied from different perspectives to elucidate the mechanisms of maternal-fetal immune regulation, including the formation of Th2 immunodominance at the maternal-fetal interface, and the expansion and function of regulatory T cells (Treg). Mechanisms. Th17 cells, as a subset of T helper cells found recently, are involved in many autoimmune diseases and graft versus host disease. We found that the number of Th17 cells in human decidua increases significantly during early pregnancy. The role of.
The first part is the increase of maternal fetal interface and peripheral Th17 cell subsets in early pregnancy.
Objective to investigate the ratio of Th17 cells in peripheral blood and maternal fetal interface during pregnancy.
Methods Peripheral blood, decidual tissue and endometrial tissue were collected from normal early, middle and late pregnant women and normal non-pregnant women. The proportion of Th17 cells in peripheral blood and decidual immune cells was analyzed by flow cytometry.
Results Thl7 cells were found in peripheral blood, maternal-fetal interface and normal non-pregnant women's endometrium during pregnancy, and the number of Thl7 cells in peripheral blood of early pregnant women increased significantly. With the progress of pregnancy, the proportion of Thl7 cells decreased gradually and reached the level of non-pregnant women's endometrium at the late pregnancy. The Th17 cell subsets of female Decidua Tissue increased significantly, and was significantly higher than that of peripheral blood.
Conclusion Th17 cells may be involved in the maintenance of normal pregnancy.
The second part is the differentiation and development of Th17 cells and Treg cells at the maternal fetal interface.
Objective to analyze the effect of maternal fetal interface microenvironment on differentiation and development of Th17 cells.
Methods The phenotype of T cells at maternal-fetal interface was analyzed by flow cytometry. The magnetic beads were activated by anti-CD3 antibody and anti-CD28 antibody in the presence of the supernatant of the co-culture system of decidual stromal cells, decidual stromal cells and decidual naive CD4 + T cells, decidual stromal cells and decidual stromal cells. Nave CD4+T cells were selected from the maternal-fetal interface. Pregnancy-related hormones were used to treat naive CD4+T cells in decidua. The proportion of Th17 cells was analyzed by flow cytometry.
Results There were naive CD4+T cells in the maternal-fetal interface. Trophoblasts, DSCs, or co-cultures significantly inhibited the differentiation and development of decidual naive CD4+T cells into Th17 cells. Cell differentiation promotes the differentiation of Treg cells. Pregnancy related hormones do not affect the differentiation of Th17 cells.
Conclusion Maternal-fetal microenvironment is beneficial to the differentiation and development of naive CD4+T cells into Treg cells, but not to Th17 cells, showing Treg migration.
In the third part, human decidual stromal cells are recruited by secreting CCL2 and CCL20 peripheral Th17 cells.
Objective to investigate the origin of Th17 cells in maternal fetal interface.
Methods Peripheral blood CD4+T cells were sorted by magnetic beads. Th17 cells were chemotaxis by trophoblast, decidual stromal cells or co-culture supernatant. The number of Th17 cells chemotaxis to the lower chamber was analyzed by flow cytometry.
Results The number of Th17 cells in DSC conditioned medium was about 2.7 times that of peripheral cells, while the number of Th17 cells in supernatant and co-culture supernatant was 1.1 times and 1.8 times that of peripheral cells, respectively. The chemotaxis effect of supernatant on Th17 cells.
Conclusion Decidual stromal cells can promote the proliferation and invasion of trophoblasts by secreting CCL2 to recruit Th17 cells from peripheral blood to the fourth part of human maternal-fetal interface.
Objective to investigate the regulation of Th17 cells on the biological behavior of trophoblast cells.
Methods The expression of IL-17R was detected by immunohistochemistry and immunocytochemistry. Nave CD4+T cells were sorted by magnetic beads, and Th17 cells were induced to differentiate in vitro. The culture supernatant of Th17 cells was prepared. Trophoblasts were treated. The proliferation and invasiveness of trophoblasts were detected. The apoptosis of trophoblasts was analyzed by flow cytometry.
Results Both trophoblasts and decidual stromal cells expressed IL-17R.rhIL-17 in a dose-dependent manner, and the supernatant of Th17 cells also promoted the proliferation and invasion of trophoblasts. The apoptotic rate of trophoblasts decreased from 2.23%+0.15% to 1.88%+0.42% and the apoptotic rate of trophoblasts recovered to 2.0%+0.11% after adding IL-17 neutralizing antibody.
Conclusion Th17 cells regulate the biological behavior of trophoblasts by secreting cytokine IL-17 and participate in placenta formation in early pregnancy.
【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392
【参考文献】
相关期刊论文 前2条
1 陈巧英,李大金,金莉萍,朱晓勇,贺银燕,王明雁;人早孕期外周及蜕膜CD4~+CD25~+调节性T细胞变化[J];现代免疫学;2005年05期
2 黄煜;李大金;;人早孕蜕膜基质细胞及免疫活性细胞趋化因子受体CXCR6的表达[J];中华微生物学和免疫学杂志;2006年04期
，本文编号：2190443
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2190443.html
最近更新
教材专著
