使用他克莫司的肝移植受者T淋巴细胞亚群及CD28家族共刺激分子的分析

目的 观察使用他克莫司(Tac)的肝移植受者外周血中T淋巴细胞亚群及CD28家族共刺激分子表达的变化及其意义.方法 以接受同种异体肝移植的受者20例为Tac组,术后采用Tac、霉酚酸酯(或硫唑嘌呤)和糖皮质激素预防排斥反应;以等待肝移植的终末期肝脏疾病患者20例为肝病对照组;以健康志愿者20名为正常对照组.测定正常对照组志愿者和肝病对照组患者的外周血中T淋巴细胞亚群及其CD28、可诱导共刺激分子(ICOS)、CD152和程序性死亡因子1(PD-1)的表达;分别测定Tac组使用Tac后2周、1个月、2个月和3个月时外周血中T淋巴细胞亚群及其CD28、ICOS、CD152和PD-1的表达.结果 肝病对照组、正常对照组和Tac组各时点间的CD3+ T淋巴细胞的差异无统计学意义(P>0.05).随着使用Tac时间的延长,Tac组CD4+ T淋巴细胞持续下降,至使用Tac 3个月时,明显低于正常对照组(P<0.05);CD8+ T淋巴细胞逐渐增加至正常水平.Tac组T淋巴细胞亚群表面ICOS分子表达持续低于正常对照组(P<0.05),CD4+ T淋巴细胞表面CD28分子表达恢复至正常水平(P>0.05),而CD8+ T淋巴细胞表面CD28分子持续低表达(P<0.05).同时,Tac组T淋巴细胞亚群表面CD152分子和PD-1分子的表达均持续高于正常对照组(P<0.05).结论 肝移植患者接受以Tac为主的免疫抑制治疗,其T淋巴细胞亚群平衡紊乱得到纠正,T淋巴细胞亚群表面CD28分子和ICOS分子的表达下调,CD152分子和PD-1分子的表达上调,通过共刺激信号的调节来达到抑制排斥反应的目的.     

联合应用CTLA-4Ig及抗ICOS单克隆抗体延长小鼠移植胰岛存活时间

目的探讨共刺激信号阻断剂细胞毒T淋巴细胞相关抗原4免疫球蛋白（CTLA-4Ig）及抗共同刺激分子ICOS单克隆抗体（ICOSmAb）对移植胰岛功能的影响。方法以BALB／c小鼠为供者，C57BL／6糖尿病小鼠为受者，进行同种胰岛细胞移植。将移植后的小鼠随机分成4组，每组10只。ICOS组：移植后1、3、5d腹腔内注射ICOSmAb 100μg／kg；CTLA4组：移植后0、2、4d腹腔内注射CTLA-4Ig50μg／kg；联合阻断组：移植后腹腔注射CTLA-4Ig和ICOSmAb，用法同CTLA4组和ICOS组；对照组：单纯胰岛移植，不注射CTLA-4Ig和ICOSmAb。观察术后移植物存活时间和移植胰岛的病理改变；逆转录聚合酶链法（RT-PCR）检测移植胰岛组织中白细胞介素2（IL-2）、白细胞介素10（IL-10）mRNA的表达情况；应用流式细胞仪检测CD4^＋、CD8^＋T淋巴细胞表达情况。结果联合阻断组的小鼠移植胰岛存活时间较其它3组明显延长，移植胰岛的细胞形态经光镜检查接近正常。联合阻断组与其它3组比较，IL-2mRNA表达减少，差异有统计学意义（P〈0．05），IL-10mRNA的表达差异无统计学意义（P〉0．05）；移植术后21d，CD4^＋、CD8^＋T淋巴细胞表达上调不明显。结论应用CTLA-4Ig和ICOSmAb联合阻断CD28和共同刺激分子ICOS，可以明显的抑制排斥反应，延长移植胰岛的存活时间及存活率。     

可诱导性共刺激分子单抗联合细胞毒T淋巴细胞4Ig在大鼠胰腺移植中的作用

目的 探讨可诱导性共刺激分子（ICOS）单抗、细胞毒T淋巴细胞4ig（CTLA41g）阻断共刺激通路对于异基因大鼠胰腺移植的作用及其机制。方法 建立胰腺移植模型，分A组（对照组），B组（CTLA4Ig组），C组（抗ICOS单抗组），D组（联合CTLA4Ig和抗ICOS单抗组）。术后1、4,7d监测血糖，第7天取胰腺做苏木素-伊红染色，脾脏做流式细胞检测CD3^＋CD8^＋T细胞、CD4^＋T细胞、CD4^＋CD25^＋T细胞。结果 与A组比较：B、C、D组排斥反应较A组明显减弱，显著延长了移植物的存活时间。CD3^＋CD8^＋T细胞计数B、C、D与A组比较均有减少，差异有统计学意义（P〈0．01）。CD4^＋T细胞B、D与A组的差异有统计学意义（P〈0．05）。CD4^＋CD25^＋T细胞各组间逐渐升高，差异有统计学意义（P〈0．01）。结论 抗ICOS单抗和CTLA4Ig能有效地抑制大鼠胰腺移植排斥反应，延长移植物存活时间，且联合应用比单一应用更有效。其可能机制为诱导了CD4^＋CD25^＋T细胞的产生。     

胃癌外周血淋巴细胞亚群表达水平对患者生存率的影响

目的探讨胃癌患者外周血淋巴细胞亚群表达与生存率的关系。方法用流式细胞仪检测833例胃癌首诊患者外周血的淋巴细胞亚群CD3^＋、CD4^＋、CD8^＋、CD4^＋／CD8^＋、CD19^＋、CD25^＋、CD44^＋及NK^＋细胞．并根据96名健康对照者的平均检测值分为高表达组和低表达组．比较各亚群高表达组与低表达组患者的生存率。结果与健康对照者相比．胃癌患者CD3^＋、CD8^＋低表达。而CD4^＋,CDl9^＋,CD25^＋、CD4^＋／CD8^＋、CD44^＋,NK^＋高表达（P〈0．05）。CD19^＋高表达与低表达者分别为444例和389例,3年生存率为36．4％和18．5％,差异有统计学意义（P〈0．05）：而其他7种淋巴细胞亚群表达水平则与患者生存率无关（均P〉0．05）。结论与健康人群相比,胃癌患者外周血淋巴细胞亚群发生显著变化．其中CDl9^＋高表达患者具有明显的生存优势。     

肾病综合征患儿外周血单个核细胞共刺激分子的表达及意义

目的：探讨共刺激分子的表达与特发性肾病综合征（INS）的关系。方法：采用流式细胞仪直接计数法测定INS患儿和健康对照者外周血CD28、CD152、CD80和CD86的表达。结果：初发肾病组外周血单个核细胞表面CO80、CD86、CD28和CD152的表达均较正常对照组明显增高，初发肾病组CD28^＋／CD152^＋和CD^80^＋／CD86^＋均较正常对照组明显降低。与初发肾病组相比，激素完全效应组和激素部分效应组CD80、CD86．CD28和CD152的表达均明显降低；CD28^＋／CD152^＋和CD80^＋／CD86^＋则与初发肾病组无统计学差异。激素完全效应组CD80、CD86、CD28和CD152的表达，CD28^＋／CD152^＋和CD80^＋／CD86^＋均和激素部分效应组无统计学差异。结论：抗原递呈细胞表面抑制细胞活性的配体CD80的表达增高不足以抑制促进细胞活性的配体CD86异常的增高表达可能是促进INS的发生因素之一，糖皮质激素可影响INS外周血单个核细胞共刺激分子的表达，但与其激素治疗效应无关。     

Fas/FasL在强直性脊柱炎患者外周血T细胞亚群上的表达

目的检测强直性脊柱炎（As）患者外周血T细胞亚群上的Fas／FasL的表达水平，探讨Fas／FasL诱导的细胞凋亡在As免疫学发病机制中的作用。方法以临床确诊的60例As患者作为研究对象，同时选择30例正常对照，运用流式细胞仪（FCM）检测其外周血CD3^＋CD4^＋、CD3^＋CD8^＋T细胞亚群上的Fas／FasL表达水平。结果早、晚期AS患者外周血CD3^＋CD4^＋T细胞上的FasL的表达率分别为（0．59％、0．93％），CD3^＋CD8^＋T细胞上的FasL的表达率分别为（2．93％、4．32％），与健康对照组（0．48％、1．14％）比较，其表达率差异具有统计学意义（P〈0．05）；与健康对照组外周血CD3^＋CD4’、CD3^＋CD8’T细胞上的Fas表达率（58．25％、59．91％）比较，早期AS患者外周血CD3^＋CD4^＋T细胞上的Fas的表达率（64．75％）明显升高（P〈0．05），而CD3^＋CD8^＋T细胞上的Fas的表达率（48．64％）明显降低（P〈0．05），Fas在晚期AS患者外周血CD3^＋CD4^＋、CD3^＋CD8^＋T细胞上的表达率（57．63％、56．32％）无明显变化。结论Fas、FasL在外周血T细胞亚群上的表达水平与AS的病情发展阶段相关；Fas、FasL的异常表达所导致T细胞凋亡功能紊乱可能是AS发病的重要机制之一。     

联合阻断CD28和可诱导共刺激分子对小鼠同种异体胰岛移植物急性排斥反应的影响

目的 在小鼠同种异体胰岛移植模型上，通过联合阻断CD28和可诱导共刺激分子（ICOS），观察CTLA-4Ig及ICOS单克隆抗体（ICOSmAb）对胰岛移植物功能的影响。方法将小鼠随机分成4组，每组10只。联合阻断组：分别于移植后0．2、4d和1、3、5d腹腔内注射CTLA-4Ig50μg／kg和ICOSmAb100μg／kg体重组。ICOS组：移植后1、3．5d腹腔内注射ICOSmAb100μg/kg体重组。CTLA4组：移植后0、2、4d腹腔内注射CTLA-4Ig50μg/kg体重组。对照组：单纯胰岛移植，不做CT-LA-4Ig和ICOSmAb处理组。观察术后移植物存活时间和病理改变，RT-PCR检测移植组织中IL-2、IL-10mRNA表达，流式细胞仪检测CD4^＋、CD8^＋T淋巴细胞表达。结果 联合阻断组小鼠移植物存活时间平均为（31．00±6．57）d，较其他3组延长，差异有统计学意义（P〈0．05）；IL-2和IL-10mRNA表达水平分别为（44．23±6．24）和（65．23±11．02），与其他3组比较，IL-2mRNA表达差异有统计学意义（P〈0．05），而IL-10mRNA表达差异无统计学意义（P〉0．05）；CD4^＋、CD8^＋T淋巴细胞荧光阳性百分率分别为（13．73±0．49）％和（14．56±0．31）％，表达上调均不明显；移植胰岛光镜检查接近正常。结论 应用CTLA-4Ig和ICOSmAb联合阻断CD28和ICOS，可以明显抑制胰岛移植物排斥反应。     

慢性特发性荨麻疹患者外周血T及Th淋巴细胞亚群的表达

目的：探讨慢性特发性荨麻疹患者外周血T及辅助性T淋巴细胞（Th）亚群的表达及其在慢性特发性荨麻疹发病机制中的作用。方法：采用流式细胞术检测经四色荧光抗体染色的慢性特发性荨麻疹患者及正常对照外周血CD3^＋、CD4^＋、CD8^＋T淋巴细胞数及CD4^＋／IFN-γ’（Th1）、CD4^＋／IL-4^＋（Th2）细胞含量。结果：慢性特发性荨麻疹组外周血CD3^＋T淋巴细胞数无明显变化、CD4^＋T淋巴细胞数、CD8^＋T淋巴细胞数均降低;CD4^＋／CD8^＋比值增高,差异有统计学意义（P〈0．01）。慢性特发性荨麻疹患者外周血Th1细胞含量、Th1／Th2比值均明显低于正常对照组（P〈0．01,P〈0．05）,Th2细胞含量高于正常对照组（P〈0．01）。结论：慢性特发性荨麻疹患者外周血存在着T及Th淋巴细胞亚群分化失衡,这可能为慢性特发性荨麻疹发病的机制之一。     

他克莫司联合青藤碱抑制大鼠心脏移植急性排斥反应

目的 探讨青藤碱（SIN）对大鼠心脏移植急性排斥反应抑制作用的机制，并评价青藤碱与他克莫司（FK506）联合作用的效果。方法 以PVG大鼠为供者，DA大鼠为受者，建立同种异体心脏移植模型。将受者随机分为4组，每组20只。对照组：采用生理盐水3ml·kg^-1·d^-1灌胃；SIN组：腹腔注射SIN 30mg·kg^-1·d^-1；FK506组：采用FK506 0．25mg·kg^-1·d^-1灌胃；联合组：腹腔注射SIN 30mg·kg^-1·d^-1并联合应用FK506 0．25mg·kg^-1·d^-1灌胃。各组均在术后24h内用药，用药时间共7d。观察各组移植心存活时间，术后第7d取部分受者的移植心做病理检查，检测外周血中肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、T细胞亚群、一氧化氮合酶（iNOS）等的浓度。结果 对照组移植心平均存活时间为（6．0±1．054）d，FK506组为（10．2±2．2）d，SIN组为（9．5±1．84）d，联合组为（16．2±2．1）d，联合组与其他3组比较，差异有统计学意义（P〈0．05）。SIN组、FK506组及联合组与对照组比较，外周血中TNF-α、IFN-γ、iNOS的表达明显减少（P〈0．05），CD4^＋T细胞亚群增殖率也明显下降（P〈0．05）。其中联合组的效果更优于SIN组和FK506组（P〈0．05）。结论 SIN能明显地抑制大鼠心脏移植急性排斥反应的发生，与FK506联合应用有协同作用。     

尿毒症患者和肾移植受者外周血调节性T细胞表达的意义

目的探讨尿毒症患者和肾移植受者外周血CD4^＋CD127^-调节性T细胞（CD4^＋CD127^- Treg）的表达水平及意义。方法采用流式细胞术测定13例尿毒症患者（尿毒症组）、13例肾移植受者（肾移植组）和20例健康志愿者（对照组）外周血CD4^＋CD127^- Treg和CD4^＋CD25^＋CD127^- Treg占CD4^＋T细胞的比例。结果尿毒症组和肾移植组CD4^＋细胞中CD4^＋CD127^- Treg的比例明显低于对照组（P〈0．05，P〈0．01）；肾移植组CD4^＋CD25^＋CD127^- Treg的比例明显低于对照组（P〈0．05）。结论尿毒症患者外周血CD4^＋CD127^- Treg数量降低，免疫功能紊乱。肾移植受者外周血CD4^＋CD127^- Treg和CD4^＋CD25^＋CD127^-Treg数量降低，免疫反应性增强。     

Baohuoside-1, a novel immunosuppressive molecule, inhibits lymphocyte activation in vitro and in vivo

BACKGROUND: We evaluated the in vitro and in vivo immunosuppressive effects of baohuoside-1 (B1), a novel flavonoid isolated from Epimedium davidii. METHODS: Proliferation assay was used to determine the antiproliferative properties on T-cell and B-cell proliferation. Flow cytometry analysis was applied to detect changes of phenotypes on activated cells. RESULTS: B1 inhibits the lymphocyte proliferation activated by polyclonal mitogens and mixed lymphocyte reaction with a 50% inhibitory concentration of low micromolar concentration. Also, B1 suppressed T-cell activation in T cell receptor/CD3-mediated signaling pathways in a dose- and time-dependent manner. The suppression of B1 was not simply a result of a toxic effect and was recovered by withdrawing the drug. B1 down-regulated the expression of some phenotype molecules. In Ca(2+)-independent or -dependent antigen stimulation, although B1 had different inhibitive patterns on CD69 expression stimulated by phorbol 12-myristate 13-acetate (PMA) or Ca2+ ionophore, it inhibited T-cell proliferation induced by CD3/CD28 or PMA/ionomycin and partially blocked that induced by PMA/CD28. Interestingly, an additive inhibition between B1 and tacrolimus (FK506) was found in the CD69 expression stimulated by PMA/CD28 and PMA/ionomycin. Similarly, this immunosuppression by combination therapy was observed in a heart transplantation model in vivo and might act through an immunosuppressive mechanism different from FK506. CONCLUSIONS: B1, whose mechanism of action is not similar to that of FK506, has selectively immunosuppressive effects on T-cell and B-cell activation in vitro and effectively prevents rat heart allograft rejection in vivo.     

Application of immune cell functional assay in monitoring immune status in renal transplant recipients

Objective To evaluate the value of immune cell functional assay (ImmuKnow CD4+ T cell ATP assay) in monitoring immune status in renal recipients. Methods A total of 131 adult renal transplant recipients who received transplantation for the first time were under investigation. According to the dynamic monitoring ATP concentration before operation, 2 week, 1, 3, 6 months after operation and during infect or rejection, samples were divided into the following groups: health control group (HC), pretransplant (Pre-Tx) group, stable (Tx) group, infect group, acute rejection (AR) group, acute kidney injury (AKI) group. Immune cell functions were detected by ImmuKnow CD4+ T cell ATP assay. Lymphocyte subsets (CD4+/CD8+) were analysed and serum concentrations of FK506 were tested. Mixed lymphocyte reaction(MLR) was analysed. Results The ATP concentration was no significant difference between Pre-Tx and HC group. The ATP concentration of 2 weeks, 1 months after operation were significantly higher than Pre-Tx group (P＜0.01). After 3 months, 6 months follow-up, the ATP concentration stabilized with time. The ATP concentration of AR group was significantly higher than other three groups (Tx, infect and AKI group, all P＜0.05). The correlation coefficient between the ATP concentration and MLR, CD4+/CD8+, FK506 level were R2＝0.0072, R2＝10-6, R2＝0.004 respectively (all P﹥0.05). Conclusions The cell-mediated immunity of recipients is relatively strongger during the first month after transplantation. The ATP concentration is not related to the levels of MLR, CD4+/ CD8+, FK506. ImmuKnow ATP assay is a valuable predictor in acute rejection diagnosis.     

Acceptance of islet allografts in the liver of mice by blockade of an inducible costimulator

BACKGROUND: An inducible costimulator (ICOS) has been found to be a novel costimulator for T-cell activation, although its precise role in transplant immunobiology remains unclear. This study determined whether ICOS plays an essential role in rejection of intrahepatic islet allografts in streptozotocin-induced diabetic mice. METHODS: Mononuclear cells in the liver of mice were isolated and examined by flow cytometry with respect to expression of ICOS in association with rejection, and the effects of in vivo treatment with an anti-ICOS antibody on survival of intrahepatic islet allografts were determined.RESULTSFlow cytometric analysis of mononuclear cells in the liver of normal untreated mice revealed that ICOS is expressed on CD4+CD3int natural killer T cells. The expression of ICOS was up-regulated on CD4+CD3bright T cells and expanded CD8 T cells in the liver in association with rejection. Posttransplant short-term administration of anti-ICOS antibody alone produced a significant prolongation of islet allograft survival. Administration of the antibody in conjunction with a subtherapeutic regimen of FK506 prevented rejection, leading to the acceptance of islet allografts. CONCLUSION: ICOS has an essential role in rejection of intrahepatic islet allografts and the blockade of ICOS interaction might be a novel approach for preventing islet allograft rejection.     

Regulatory effect of FK506 on CD152 and PD-1 in the liver allorecipients

AIM: We dynamically observed the expression of CD152 and PD-1 on T-cell surfaces in peripheral blood of liver allorecipients to explore the regulatory effect of FK506 on negative costimulatory molecules. METHODS: We evaluated 20 liver allorecipients, 19 end-stage liver disease patients, and 20 healthy volunteers for FK506 concentrations measured by an enzyme-multiplied immunoassay technique and flow cytometry to determine T-cell subsets as well as CD152 and PD-1 expression. RESULTS: After liver transplantation, the frequency of CD4+ T cells gradually decreased to significantly lower level than those in the disease controls (P < .05). CD8+ T cells in each treatment group were obviously higher than those in the disease controls (P < .05). The expression of CD152 on CD4+ and CD8+ T cells was greater than those in healthy controls (P < .05); and at 2 and 4 weeks, higher than those in disease controls (P < .05). The expression of PD-1 on CD4+ T cells from the 2 weeks after treatment was significantly greater than that in healthy controls (P < .05), and that on CD8+ T cells increased obviously from the 4 weeks compared with disease controls (P < .05). CONCLUSION: FK506 up-regulated the expression of CD152 and PD-1 on the T-cell surface inhibiting proliferation and activation of effector T cells, favoring the survival of allorecipients.     

骨化三醇在抑制大鼠同种肢体移植排斥反应中的作用

目的 探讨应用骨化三醇(1,25-二羟维生素D;简称:VD5)在抑制肢体移植排斥反应中的作用.方法 以Wistar大鼠为供者,SD大鼠为受者,建立同种肢体移植模型.随机将受者分为4组,每组12只.(1)对照组:术后不用免疫抑制剂,仅以15 ml·kg-1·d-1.生理盐水灌服.(2)他克莫司(FK506)组:将FK506用生理盐水稀释为0.5 mg/ml,术后前2周的用量为1.0 mg·kg-1·d-1,术后第3周起,每周灌服2次.(3)VD3组:将骨化三醇用生理盐水稀释为0.125 μg/ml,术后用量为2.5μg·kg-1·d-1,连续灌服4周.(4)VD3+FK506组:术后联合应用骨化三醇及FK506,用药方法和用量与FK506组和VD3组相同.术后观察各组受者移植肢体排斥反应发生的时间和存活时间;流式细胞仪检测T淋巴细胞亚群CD4+、CD8+细胞的百分率.结果 对照组、FK506组、VD3组以及VD3+FK506组肢体移植后排斥反应发生的时间分别为:(3.50±0.50)d、(13.13±1.50)d、(10.63±0.38)d和(29.25±0.63)d;移植肢体的存活时间分别为:(8.50±0.50)d、(26.25±1.50)d、(17.25±0.38)d和(62.00±0.63)d;与对照组比较,VD3组排斥反应发生的时间和移植肢体的存活时间均明显延长,差异有统计学意义(P＜0.01);VD3+FK506组抗排斥反应效果更佳,与FK506组和VD3组比较,差异有统计学意义(P＜0.01).VD3组T淋巴细胞亚群CD4+/CD8+细胞的比值明显低于对照组,VD3+FK506组又明显低于VD3组和FK506组(P＜0.01).结论 骨化三醇能明显减轻同种肢体移植排斥反应,延长移植肢体的存活时间;骨化三醇与FK506联合应用效果更佳,二者具有协同作用.     

他克莫司与来氟米特对异种胰岛移植术后T细胞亚群的影响

目的 研究他克莫司(FK506)和来氟米特(Lef)对异种胰岛移植后急性排斥反应的抑制作用。方法 将大鼠胰岛移植于糖尿病小鼠肾被膜下。观察移植后不用药对照组、单独应用FK506组、单独应用Lef组以及联合应用FK506和Lef组的胰岛存活情况；并于移植后第5d检测外周血T细胞亚群分布和血清IL-2含量。结果 单独用药组与不用药对照组比较，胰岛存活时间明显延长；联合用药组又明显长于单独用药组。各用药组外周血CD4 T淋巴细胞明显低于未用药组。结论 FK506与Lef通过抑制CD4 T淋巴细胞的增殖而阻止或延缓异种胰岛移植的急性排斥反应。     

Effects of a calcineurin inhibitor, FK506, and a CCR5/CXCR3 antagonist, TAK-779, in a rat small intestinal transplantation model

The effects of FK506, and TAK-779, antagonists of CCR5 and CXCR3, were investigated using a rat intestinal transplantation model. Small intestines from DA rats were heterotopically transplanted into LEW rats. The recipients were treated with FK506 (1 mg/kg/day, day 0-5) and TAK-779 (10 mg/kg/day, day 0-10). Graft survival and immunological responses to these materials were estimated by mixed lymphocyte reactions and IFN-?? production. The expression of chemokine receptors on lymphocytes was also examined. The average duration of survival was 7.0 ± 0.3, 12.0 ± 1.0, 9.8 ± 0.5 and 18.0 ± 1.5 days in the allogeneic, FK506, TAK-779 and the two-drug combined groups, respectively. Cell proliferative responses and IFN-?? production were suppressed to a significant extent in the FK506 group compared with the TAK-779 group. In addition, the two-drug combination showed a tendency for stronger suppression than FK506 alone, correlated with in vivo and histopathological data. The numbers of both CD4⁺ and CD8⁺ cells were significantly suppressed in the blood of the recipients of both the FK506 and the TAK-779 groups, and in Peyer's patches of the graft of the TAK-779 group, but the FK506 group was not, as evidenced by FACS analysis. In addition, double-staining of graft-infiltrating lymphocytes showed a significant reduction in lymphocyte numbers, expressing CCR5 and CXCR3 in the TAK-779 group, but not evident in the FK506 group, compared to the allogeneic group.While FK506 suppresses cell proliferation and effecter function, it has less effect on the expression of CCR5 and CXCR3 in lymphocytes. Further exploration of the effects of a combined therapy with TAK-779 could represent a novel treatment for intestinal transplantation.     

Detection of alloreactive T cells by flow cytometry: a new test compared with limiting dilution assay

BACKGROUND: Frequencies of alloreactive T cells determined by limiting dilution assays (LDA) may not adequately reflect the donor-reactive immune status in transplant recipients. To reevaluate LDA frequencies, we developed a flow cytometry test for direct determination of alloreactive T-cell frequencies and compared these frequencies with classical LDA estimates of frequencies. METHODS: For determination of frequencies by flow cytometry, peripheral blood lymphocytes (or lymphocytes taken from primary mixed lymphocyte culture) were stimulated with either Epstein-Barr virus-transformed lymphoblastoid cell lines or T cell-depleted spleen cells and stained for intracellular interferon (IFN)-gamma production and CD69. In lung transplant recipients, frequencies of IFN+ alloreactive T cells were compared with LDA frequencies, that is, cytotoxic T lymphocyte precursors and helper T lymphocyte precursors. RESULTS: With flow cytometry, alloreactive T cells were detected after overnight allostimulation as IFN-gamma CD69bright cells (range, 0.1-0.58% and 0.1-0.66% of total CD4 and CD8 cells, respectively). Frequencies increased 25-fold or more when lymphocytes were prestimulated in primary mixed lymphocyte culture before testing. After lung transplantation, mean donor-specific IFN+ CD8 T-cell frequencies did not decrease as mean donor-specific LDA cytotoxic T lymphocyte precursor frequencies, whereas no difference was seen in pretransplantation samples or third-party-specific frequencies at both time points. Mean frequencies of IFN+ CD4 did not differ from helper T lymphocyte precursors at both time points, but frequencies did not correlate. CONCLUSIONS: The flow cytometry test allows a direct measurement of alloreactive T-cell frequencies and demonstrates a discrepancy between donor-specific IFN+ CD8 T-cell frequencies and LDA CLTp after transplantation. This may be a result of the existence of "functional diverse" alloreactive T cells or of activation-induced cell death of donor-reactive T cells during long (LDA) culturing, which is avoided in the flow cytometry test.     

肺移植术后稳定状态受者T淋巴细胞亚群的动态变化分析

目的 分析肺移植术后稳定状态受者1年内T淋巴细胞亚群的动态变化及影响因素.方法 收集行同种异体肺移植手术且术后处于稳定状态的41例受者的临床资料.采用流式细胞术检测受者术前、术后2周及每个月(术后1年内)外周血T淋巴细胞亚群绝对值和比值.分析受者年龄、性别、体质量指数(BMI)、手术方式、原发性移植物功能障碍(PGD)...