Objective::The maternal-fetal interface undergoes dynamic changes to allow the fetus to grow and develop in the uterus.The interaction between decidualγδT cells and trophoblasts plays a pivotal role during successfu...Objective::The maternal-fetal interface undergoes dynamic changes to allow the fetus to grow and develop in the uterus.The interaction between decidualγδT cells and trophoblasts plays a pivotal role during successful pregnancy;however,their physiological functions in early-term human pregnancy are still not completely illustrated.This study was undertaken to illustrate the functional roles of CXCL16/CXCR6 to prevent pregnancy loss via the crosstalk between decidualγδT cells and HTR8/SVneo trophoblast cells.Methods::The percentile of CXCR6+γδT cells in the peripheral blood from normal female and recurrent spontaneous abortion(RSA)patients was analyzed by flow cytometry.The expression of CXCR6 was detected in decidual immune cells via flow cytometry,and the expression of CXCL16 was analyzed in HTR8/SVneo trophoblast cells and lentivirus(LV)-HTR8/SVneo trophoblast cells via enzyme-linked immunosorbent assay.Reverse transcriptase-polymerase chain reaction was used to verify the expression of the CXCL16 gene in LV-HTR8/SVneo trophoblast cells.Expression of granzyme B and cytokines and proliferation of decidualγδT cocultured with HTR8/SVneo trophoblast cells were analyzed by flow cytometry.Invasion of HTR8/SVneo trophoblast cells was assessed via Matrigel transwell assay.Adoptive transfer was induced in vivo further to illustrate that the normal expression of CXCL16/CXCR6 could prevent pregnancy loss.Results::The percentile of CXCR6+γδT cells in the peripheral blood from RSA patients was lower than normal pregnancies.The expression of CXCR6 was highest in the decidualγδT cells among decidual immune cells,and the expression of CXCL16 increased as the amount of HTR8/SVneo trophoblast cells increased.Expression of granzyme B in the decidualγδT cells was downregulated by cocultured with HTR8/SVneo cells dependent of CXCL16,and HTR8/SVneo trophoblast cells induced the Th2 cytokines production in the decidualγδT cells.Both the expression of CXCR6 in the decidualγδT cells and proliferation of the decidualγδT cells were promoted by HTR8/SVneo trophoblast cells.On the other hand,decidualγδT cells enhanced the invasion of HTR8/SVneo trophoblast cells and thus promoted embryo implantation.In vivo study was taken further and shown that low expression of CXCL16/CXCR6 results in pregnancy loss because of dialog disorder between decidualγδT cells and trophoblasts.Conclusions::Low expression of CXCL16/CXCR6 results in pregnancy loss because of the dialog disorder between decidualγδT cells and trophoblasts,and it showed a light on the effective strategy of adoptive transfer of CXCR6+γδT cells on the treatment of RSA.This observation provides a scientific basis on which a potential strategy can be applied to the early-detect and treatment of RSA.展开更多
目的探讨s CXCL16对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)细胞体外生物学特征的影响,并分析其初步机制。方法采用CCK8法观察DLBCL细胞株增殖能力;应用Transwell法检测细胞的迁徙能力,全基因组表达谱芯片筛选不同干...目的探讨s CXCL16对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)细胞体外生物学特征的影响,并分析其初步机制。方法采用CCK8法观察DLBCL细胞株增殖能力;应用Transwell法检测细胞的迁徙能力,全基因组表达谱芯片筛选不同干预组差异表达的基因;运用Western blot检测差异基因的蛋白表达。结果外源性s CXCL16重组蛋白对DLBCL细胞增殖影响差异无显著性,但可促进DLBCL细胞自身的迁移并具有剂量依赖性,使用CXCR6抗体可阻断该作用;外源性添加重组蛋白s CXCL16可影响DLBCL细胞多种基因表达,其中趋化因子配体/受体途径相关的13种基因出现上调; Western blot结果证实外源性添加s CXCL16重组蛋白可上调TNFRSF12A蛋白表达,而使用CXCR6抗体则降低其表达。结论 s CXCL16可能通过其自身受体CXCR6,以剂量依赖性模式促进DLBCL细胞的迁移,并调控多种趋化因子配体/受体的表达。展开更多
基金This study was supported by the National Natural Science Foundation of China(NSFC)(No.81300552,92057119,31970798)the Innovation-oriented Science and Technology Grant from NPFPC Key Laboratory of Reproduction Regulation(CX2017-2)+1 种基金the Program for Zhuoxue of Fudan University(JIF157602)the Support Project for Original Personalized Research of Fudan University.
文摘Objective::The maternal-fetal interface undergoes dynamic changes to allow the fetus to grow and develop in the uterus.The interaction between decidualγδT cells and trophoblasts plays a pivotal role during successful pregnancy;however,their physiological functions in early-term human pregnancy are still not completely illustrated.This study was undertaken to illustrate the functional roles of CXCL16/CXCR6 to prevent pregnancy loss via the crosstalk between decidualγδT cells and HTR8/SVneo trophoblast cells.Methods::The percentile of CXCR6+γδT cells in the peripheral blood from normal female and recurrent spontaneous abortion(RSA)patients was analyzed by flow cytometry.The expression of CXCR6 was detected in decidual immune cells via flow cytometry,and the expression of CXCL16 was analyzed in HTR8/SVneo trophoblast cells and lentivirus(LV)-HTR8/SVneo trophoblast cells via enzyme-linked immunosorbent assay.Reverse transcriptase-polymerase chain reaction was used to verify the expression of the CXCL16 gene in LV-HTR8/SVneo trophoblast cells.Expression of granzyme B and cytokines and proliferation of decidualγδT cocultured with HTR8/SVneo trophoblast cells were analyzed by flow cytometry.Invasion of HTR8/SVneo trophoblast cells was assessed via Matrigel transwell assay.Adoptive transfer was induced in vivo further to illustrate that the normal expression of CXCL16/CXCR6 could prevent pregnancy loss.Results::The percentile of CXCR6+γδT cells in the peripheral blood from RSA patients was lower than normal pregnancies.The expression of CXCR6 was highest in the decidualγδT cells among decidual immune cells,and the expression of CXCL16 increased as the amount of HTR8/SVneo trophoblast cells increased.Expression of granzyme B in the decidualγδT cells was downregulated by cocultured with HTR8/SVneo cells dependent of CXCL16,and HTR8/SVneo trophoblast cells induced the Th2 cytokines production in the decidualγδT cells.Both the expression of CXCR6 in the decidualγδT cells and proliferation of the decidualγδT cells were promoted by HTR8/SVneo trophoblast cells.On the other hand,decidualγδT cells enhanced the invasion of HTR8/SVneo trophoblast cells and thus promoted embryo implantation.In vivo study was taken further and shown that low expression of CXCL16/CXCR6 results in pregnancy loss because of dialog disorder between decidualγδT cells and trophoblasts.Conclusions::Low expression of CXCL16/CXCR6 results in pregnancy loss because of the dialog disorder between decidualγδT cells and trophoblasts,and it showed a light on the effective strategy of adoptive transfer of CXCR6+γδT cells on the treatment of RSA.This observation provides a scientific basis on which a potential strategy can be applied to the early-detect and treatment of RSA.
文摘目的探讨s CXCL16对弥漫大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)细胞体外生物学特征的影响,并分析其初步机制。方法采用CCK8法观察DLBCL细胞株增殖能力;应用Transwell法检测细胞的迁徙能力,全基因组表达谱芯片筛选不同干预组差异表达的基因;运用Western blot检测差异基因的蛋白表达。结果外源性s CXCL16重组蛋白对DLBCL细胞增殖影响差异无显著性,但可促进DLBCL细胞自身的迁移并具有剂量依赖性,使用CXCR6抗体可阻断该作用;外源性添加重组蛋白s CXCL16可影响DLBCL细胞多种基因表达,其中趋化因子配体/受体途径相关的13种基因出现上调; Western blot结果证实外源性添加s CXCL16重组蛋白可上调TNFRSF12A蛋白表达,而使用CXCR6抗体则降低其表达。结论 s CXCL16可能通过其自身受体CXCR6,以剂量依赖性模式促进DLBCL细胞的迁移,并调控多种趋化因子配体/受体的表达。
