肾移植受者CD4~+CD25~+CD127~-T reg细胞变化与急性排斥反应的关系以及免疫抑制治疗对其影响
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摘要
目的根据检测CD4~+CD25~+CD1271~(lo/-)调节性T细胞(简称T reg细胞)水平在肾移植受者近期外周血中的动态变化,探讨T reg细胞水平和急性排斥反应、免疫抑制剂治疗之间的关系。
方法应用抗CD4、抗CD25和抗CD127三联单克隆抗体通过流式细胞仪(FCM)分析,动态监测52例肾移植受者(其中男24例,女28例;亲属活体供肾6例,尸体供肾46例)外周血中T reg细胞。分别在术前1-2天、术后3天、术后1周、术后2周、术后3周、术后4周取外周静脉血样本。所有的受者根据术后4周内是否发生急性排斥反应分为急性排斥组和移植肾功能稳定组;根据术后免疫抑制方案的不同(steroids+MMF+CsA或steroids+MMF+FK506)分为CsA组和FK506组。对比术前和术后近期T reg细胞水平的变化,移植肾功能稳定组和急性排斥组T reg细胞水平的变化,CsA组和FK506组T reg细胞水平的变化。
结果移植术前急性排斥组和肾功能稳定组的T reg细胞水平无明显差异(P＞0.05)。术后3天肾功能稳定组的T reg细胞水平迅速升高到达峰值,急性排斥组也升高,但不如稳定组明显,因此术后3天急性排斥组的T reg细胞水平低于稳定组(P＜0.05)。术后3天至术后1周两组T reg细胞水平均急剧下降,这段时间刚好是我们研究的群体开始发生急性排斥反应的高峰时期,因此急性排斥反应可能与T reg细胞水平急剧下降有关。术后1周稳定组和排斥组的T reg细胞水平最接近(P＞0.05),1周后稳定组保持平稳,排斥组继续下降。术后4周稳定组的T reg细胞水平开始回升,但排斥组仍保持在较低水平。除了术后第1次检测的T reg细胞高于术前外,术后其他时间测得的T reg细胞水平均低于术前。术后CsA组和FK506组的T reg细胞水平变化无明显的差异(P＞0.05)。
结论肾移植术后近期外周血中T reg细胞水平出现变化。检测肾移植受者近期外周血中T reg细胞水平有助于急性排斥反应的早期诊断,并有判断预后的指示。
Objective To investigate the relationship of T reg cell levels between acute rejection and immune-suppressive therapy on the basis of the changes of T reg cells in peripheral blood of renal transplant recipients in the near future.
Methods The levels of peripheral blood T reg cells were sequentially measured by flow cytometry using anti-CD4,anti-CD25 and anti-CD127 monoclone antibodies in 52 renal allograft recipients(24 male,28 female;6 living-related donor,46 cadaveric donor).At days 1-2 before transplantation samples were obtained and afterwards at days +3 and at weeks +1,+2,+3,+4.All of the recipients were divided into two groups:acute rejection group and stable kidney function group according to whether an acute rejection occurred within the first 4 weeks posttransplant,and divided into two groups:CsA group and FK506 group base on immunosuppressant: steroids+MMF+CsA or steroids+MMF+FK506.We compared the changes of T reg cell levels pretransplant and short courses posttransplant,the changes of T reg cell levels between the acute rejectors and the stables,and the changes of T reg cell levels between CsA group and FK506 group.
Results Before transplantation,the levels of T reg cells had no differrence in acute rejection group and normal group(P＞0.05).In the first 3 days after renal transplantation,the levels of T reg cells in stable allograft group soared to the peak,T reg cell levels in acute rejection group also increased but not so much as that high of stable allograft group,so the levels of T reg cells in stable allograft group was significantly higher than that of acute rejection group at the third day after operation(P＜0.05).From the third day to the end of the first week after transplantation,a sharp decrease of the T reg fraction was observed in both groups when majority acute rejection began in the cohort we studied.At the end of the first week after operation, the levels of T reg cells was closest between acute rejection group and stable allograft group,then levelled off in the stable allograft group but remained downward in acute rejection group.There was a relative increase of T reg cell levels in a nonrejector at the end of the fourth week posttransplant,but still kept in low levels in a rejector.Except the level of T reg cells at the first time after transplantation higher than that of before transplantation,all of others are lower than that of before transplantation.There was no obvious statistic difference between the CsA group and FK506 group(P＞0.05).
Conclusion The levels of T reg cells in peripheral blood of renal transplant recipients varied at different time in the near future.Detecting the levels of T reg cells was helpful to diagnose acute rejection at early time and to judge prognosis.
方法应用抗CD4、抗CD25和抗CD127三联单克隆抗体通过流式细胞仪(FCM)分析,动态监测52例肾移植受者(其中男24例,女28例;亲属活体供肾6例,尸体供肾46例)外周血中T reg细胞。分别在术前1-2天、术后3天、术后1周、术后2周、术后3周、术后4周取外周静脉血样本。所有的受者根据术后4周内是否发生急性排斥反应分为急性排斥组和移植肾功能稳定组;根据术后免疫抑制方案的不同(steroids+MMF+CsA或steroids+MMF+FK506)分为CsA组和FK506组。对比术前和术后近期T reg细胞水平的变化,移植肾功能稳定组和急性排斥组T reg细胞水平的变化,CsA组和FK506组T reg细胞水平的变化。
结果移植术前急性排斥组和肾功能稳定组的T reg细胞水平无明显差异(P＞0.05)。术后3天肾功能稳定组的T reg细胞水平迅速升高到达峰值,急性排斥组也升高,但不如稳定组明显,因此术后3天急性排斥组的T reg细胞水平低于稳定组(P＜0.05)。术后3天至术后1周两组T reg细胞水平均急剧下降,这段时间刚好是我们研究的群体开始发生急性排斥反应的高峰时期,因此急性排斥反应可能与T reg细胞水平急剧下降有关。术后1周稳定组和排斥组的T reg细胞水平最接近(P＞0.05),1周后稳定组保持平稳,排斥组继续下降。术后4周稳定组的T reg细胞水平开始回升,但排斥组仍保持在较低水平。除了术后第1次检测的T reg细胞高于术前外,术后其他时间测得的T reg细胞水平均低于术前。术后CsA组和FK506组的T reg细胞水平变化无明显的差异(P＞0.05)。
结论肾移植术后近期外周血中T reg细胞水平出现变化。检测肾移植受者近期外周血中T reg细胞水平有助于急性排斥反应的早期诊断,并有判断预后的指示。
Objective To investigate the relationship of T reg cell levels between acute rejection and immune-suppressive therapy on the basis of the changes of T reg cells in peripheral blood of renal transplant recipients in the near future.
Methods The levels of peripheral blood T reg cells were sequentially measured by flow cytometry using anti-CD4,anti-CD25 and anti-CD127 monoclone antibodies in 52 renal allograft recipients(24 male,28 female;6 living-related donor,46 cadaveric donor).At days 1-2 before transplantation samples were obtained and afterwards at days +3 and at weeks +1,+2,+3,+4.All of the recipients were divided into two groups:acute rejection group and stable kidney function group according to whether an acute rejection occurred within the first 4 weeks posttransplant,and divided into two groups:CsA group and FK506 group base on immunosuppressant: steroids+MMF+CsA or steroids+MMF+FK506.We compared the changes of T reg cell levels pretransplant and short courses posttransplant,the changes of T reg cell levels between the acute rejectors and the stables,and the changes of T reg cell levels between CsA group and FK506 group.
Results Before transplantation,the levels of T reg cells had no differrence in acute rejection group and normal group(P＞0.05).In the first 3 days after renal transplantation,the levels of T reg cells in stable allograft group soared to the peak,T reg cell levels in acute rejection group also increased but not so much as that high of stable allograft group,so the levels of T reg cells in stable allograft group was significantly higher than that of acute rejection group at the third day after operation(P＜0.05).From the third day to the end of the first week after transplantation,a sharp decrease of the T reg fraction was observed in both groups when majority acute rejection began in the cohort we studied.At the end of the first week after operation, the levels of T reg cells was closest between acute rejection group and stable allograft group,then levelled off in the stable allograft group but remained downward in acute rejection group.There was a relative increase of T reg cell levels in a nonrejector at the end of the fourth week posttransplant,but still kept in low levels in a rejector.Except the level of T reg cells at the first time after transplantation higher than that of before transplantation,all of others are lower than that of before transplantation.There was no obvious statistic difference between the CsA group and FK506 group(P＞0.05).
Conclusion The levels of T reg cells in peripheral blood of renal transplant recipients varied at different time in the near future.Detecting the levels of T reg cells was helpful to diagnose acute rejection at early time and to judge prognosis.
引文
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[2] Curotto de Lafaille, M.A.J.J. Lafaille. CD4(+) regulatory T cells in autoimmunity and allergy[J]. Curr. Opin. Immunol. 2002,14:771-778.
[3]Tang Q, K.J. Henriksen,M .Bi, et al. In vitro-expanded antigen-specific regulatory T cells suppress autoimmune diabetes[J]. Exp.Med,2004,199:1455-1465.
[4]von Boehmer, H. Mechanisms of suppression by suppressor T cells[J]. Nat. Immunol, 2005,6:3 38-344.
[5] Thornton, A.M., C.A. Piccirillo, et al.. Activation requirements for the induction of CD4~+CD25~+ T cell suppressor function[J]. Eur.J. Immunol, 2004,34:366-376.
[6] Belghith M., J.A. Bluestone, S. Barriot, et al. TGF-β-dependent mechanisms mediate restoration of self-tolerance induced by antibodies to CD3 in overt autoimmune diabetes[J]. Nat. Med, 2003, 9:1202-1208.
[7]Apostolou. I., H. von Boehmer. In vivo instruction of sup pressor commitment in naive Tcells[ J], Exp. Med. 2004, 199:1401-1408.
[8] Bluestone, J.A.,A.K. Abbas. Natural versus adaptive regulatory T cells[J]. Nat. Rev.Immunol.2003,3:253-257.
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