Background Chimerism analysis is an important tool for the surveillance of post-transplant engraftment. It offers the possibility of identifying impending graft rejection and recurrence of underlying malignant or non-...Background Chimerism analysis is an important tool for the surveillance of post-transplant engraftment. It offers the possibility of identifying impending graft rejection and recurrence of underlying malignant or non-malignant disease. Here we investigated the quantitative chimerism kinetics of 21 relapsed leukemia patients after allogeneic hematopoietic stem cell transplantation （HSCT）. Methods A panel of 29 selected sequence polymorphism （SP） markers was screened by real-time polymerase chain reaction （RT-PCR） to obtain the informative marker for every leukemia patient. Quantitative chimerism analysis of bone marrow （BM） samples of 21 relapsed patients and 20 patients in stable remission was performed longitudinally. The chimerisms of BM and peripheral blood （PB） samples of 14 patients at relapse were compared. Results Twenty-one patients experienced leukemia relapse at a median of 135 days （range, 30-720 days） after transplantation. High recipient chimerism in BM was found in all patients at relapse, and increased recipient chimerism in BM samples was observed in 90% （19/21） of patients before relapse. With 0.5% recipient DNA as the cut-off, median time between the detection of increased recipient chimerism and relapse was 45 days （range, 0-120 days）, with 76% of patients showing increased recipient chimerism at least 1 month prior to relapse. Median percentage of recipient DNA in 20 stable remission patients was 0.28%, 0.04%, 0.05%, 0.05%, 0.08%, and 0.05% at 1, 2, 3, 6, 9, and 12 months, respectively, after transplantation. This was concordant with other specific fusion transcripts and fluorescent in situ hybridization examination. The recipient chimerisms in BM were significantly higher than those in PB at relapse （P=-0.001）. Conclusions This SP-based RT-PCR assay is a reliable method for chimerism analysis. Chimerism kinetics in BM can be used as a marker of impending leukemia relapse, especially when no other specific marker is available. Based on our findings, we recom展开更多
目的探讨肌腱蛋白-C(TNC)与肝移植术后早期急性排斥反应(AR)的关系。方法将6只Brown Norway(BN)大鼠和16只Lewis大鼠分为AR组(Lewis→BN,供、受体各6只)和对照组(Lewis→Lewis,供、受体各5只)。对两组大鼠移植肝组织进行病理学检查,采用...目的探讨肌腱蛋白-C(TNC)与肝移植术后早期急性排斥反应(AR)的关系。方法将6只Brown Norway(BN)大鼠和16只Lewis大鼠分为AR组(Lewis→BN,供、受体各6只)和对照组(Lewis→Lewis,供、受体各5只)。对两组大鼠移植肝组织进行病理学检查,采用Banff分级评估排斥活动指数(RAI);采用免疫组织化学(免疫组化)染色、蛋白质印迹法检测两组大鼠移植肝TNC蛋白表达情况;采用酶联免疫吸附试验(ELISA)检测血清TNC表达水平,并分析其与RAI评分的相关性。结果大鼠肝移植术后7 d移植肝病理学结果显示AR组RAI评分高于对照组;免疫组化结果显示术后7 d AR组移植肝TNC阳性细胞分布多于对照组;蛋白质印迹法结果显示术后7 d AR组移植肝TNC蛋白相对表达量高于对照组(t=5.112,P=0.007);ELISA结果显示术后7 d AR组血清TNC表达水平高于对照组(t=3.152,P=0.012),且血清TNC表达水平与RAI评分呈正相关(r=0.7909,P=0.004)。结论肝移植术后TNC表达水平与AR相关,TNC有可能成为临床肝移植术后早期AR诊断和治疗的新靶点。展开更多
目的建立外周血单核细胞检测心脏移植免疫排斥相关基因的研究方法，探讨基因在心脏移植术后免疫排斥监测中的临床价值。方法应用实时荧光定量RT-PCR技术，以SYBR Green Ⅰ作荧光染料，GAPDH作内对照，观察心脏移植术后不同时期病人外周...目的建立外周血单核细胞检测心脏移植免疫排斥相关基因的研究方法，探讨基因在心脏移植术后免疫排斥监测中的临床价值。方法应用实时荧光定量RT-PCR技术，以SYBR Green Ⅰ作荧光染料，GAPDH作内对照，观察心脏移植术后不同时期病人外周血单个核细胞中与免疫排斥相关基因的mRNA表达水平，并与病人术前基因表达水平对照。结果使用Primer Premier 5．0和Primer Express 3．0软件成功进行引物设计，并建立稳定检测的RT-PCR方法。心脏移植术前外周血单个核细胞16个与免疫排斥相关候选基因的mRNA的相对表达量与正常对照组比较差异无统计学意义（P〉0．05）。1TGA4、FKB、IL1R2、PF4、ITGAM、TGFB1、RHOU7种基因可能与机体免疫排斥状态有更大相关性。结论成功建立了荧光定量RT-PCR方法，检测外周血单个核细胞基因mRNA表达水平简便快速、特异、重复性好、可信度高具有良好的检测心脏移植早期排斥反应的应用前景。展开更多
文摘Background Chimerism analysis is an important tool for the surveillance of post-transplant engraftment. It offers the possibility of identifying impending graft rejection and recurrence of underlying malignant or non-malignant disease. Here we investigated the quantitative chimerism kinetics of 21 relapsed leukemia patients after allogeneic hematopoietic stem cell transplantation （HSCT）. Methods A panel of 29 selected sequence polymorphism （SP） markers was screened by real-time polymerase chain reaction （RT-PCR） to obtain the informative marker for every leukemia patient. Quantitative chimerism analysis of bone marrow （BM） samples of 21 relapsed patients and 20 patients in stable remission was performed longitudinally. The chimerisms of BM and peripheral blood （PB） samples of 14 patients at relapse were compared. Results Twenty-one patients experienced leukemia relapse at a median of 135 days （range, 30-720 days） after transplantation. High recipient chimerism in BM was found in all patients at relapse, and increased recipient chimerism in BM samples was observed in 90% （19/21） of patients before relapse. With 0.5% recipient DNA as the cut-off, median time between the detection of increased recipient chimerism and relapse was 45 days （range, 0-120 days）, with 76% of patients showing increased recipient chimerism at least 1 month prior to relapse. Median percentage of recipient DNA in 20 stable remission patients was 0.28%, 0.04%, 0.05%, 0.05%, 0.08%, and 0.05% at 1, 2, 3, 6, 9, and 12 months, respectively, after transplantation. This was concordant with other specific fusion transcripts and fluorescent in situ hybridization examination. The recipient chimerisms in BM were significantly higher than those in PB at relapse （P=-0.001）. Conclusions This SP-based RT-PCR assay is a reliable method for chimerism analysis. Chimerism kinetics in BM can be used as a marker of impending leukemia relapse, especially when no other specific marker is available. Based on our findings, we recom
文摘目的探讨肌腱蛋白-C(TNC)与肝移植术后早期急性排斥反应(AR)的关系。方法将6只Brown Norway(BN)大鼠和16只Lewis大鼠分为AR组(Lewis→BN,供、受体各6只)和对照组(Lewis→Lewis,供、受体各5只)。对两组大鼠移植肝组织进行病理学检查,采用Banff分级评估排斥活动指数(RAI);采用免疫组织化学(免疫组化)染色、蛋白质印迹法检测两组大鼠移植肝TNC蛋白表达情况;采用酶联免疫吸附试验(ELISA)检测血清TNC表达水平,并分析其与RAI评分的相关性。结果大鼠肝移植术后7 d移植肝病理学结果显示AR组RAI评分高于对照组;免疫组化结果显示术后7 d AR组移植肝TNC阳性细胞分布多于对照组;蛋白质印迹法结果显示术后7 d AR组移植肝TNC蛋白相对表达量高于对照组(t=5.112,P=0.007);ELISA结果显示术后7 d AR组血清TNC表达水平高于对照组(t=3.152,P=0.012),且血清TNC表达水平与RAI评分呈正相关(r=0.7909,P=0.004)。结论肝移植术后TNC表达水平与AR相关,TNC有可能成为临床肝移植术后早期AR诊断和治疗的新靶点。
文摘目的建立外周血单核细胞检测心脏移植免疫排斥相关基因的研究方法，探讨基因在心脏移植术后免疫排斥监测中的临床价值。方法应用实时荧光定量RT-PCR技术，以SYBR Green Ⅰ作荧光染料，GAPDH作内对照，观察心脏移植术后不同时期病人外周血单个核细胞中与免疫排斥相关基因的mRNA表达水平，并与病人术前基因表达水平对照。结果使用Primer Premier 5．0和Primer Express 3．0软件成功进行引物设计，并建立稳定检测的RT-PCR方法。心脏移植术前外周血单个核细胞16个与免疫排斥相关候选基因的mRNA的相对表达量与正常对照组比较差异无统计学意义（P〉0．05）。1TGA4、FKB、IL1R2、PF4、ITGAM、TGFB1、RHOU7种基因可能与机体免疫排斥状态有更大相关性。结论成功建立了荧光定量RT-PCR方法，检测外周血单个核细胞基因mRNA表达水平简便快速、特异、重复性好、可信度高具有良好的检测心脏移植早期排斥反应的应用前景。
