原发性肾小球疾病CD19+、CD5+、CD3+、CD25+的表达

目的:了解原发性肾小球疾病患者外周血淋巴细胞免疫表型的表达,并探讨其意义.方法:采用多种单克隆抗体及流式细胞仪,对14例原发性肾小球疾病患者淋巴细胞免疫表型CD19 、CD5 、CD3 、CD25 进行检测.结果:原发性肾小球疾病患者外周血淋巴细胞CD19 、CD5 、CD3 、CD25 阳性细胞数与健康人群相比明显偏低(P＜0.05),临床表现肾病综合征者CD19 、CD3 比非肾病综合征者低(P＜0.05).结论:CD19 、CD5 、CD3 、CD25 可以评价肾小球疾病患者免疫状态及疾病的严重程度.     

长期低剂量免疫抑制对肾移植受者淋巴细胞表型的影响

目的探讨肾移植受者长期低剂量免疫抑制维持用药的免疫学基础.方法采用多种单克隆抗体,以流式细胞仪为工具,对28例长期低剂量免疫抑制维持用药的肾移植受者外周血淋巴细胞表型进行检测,分析细胞表型与临床用药和移植效果的关系,并与常规剂量用药组和慢性移植肾失功能组比较.结果低剂量组CD 95、NK及HLA-DR阳性细胞数目与正常人接近,而在常规剂量组和移植肾失功能组中,CD 95明显升高,NK细胞明显减少.结论CD 95、NK及HLA-DR水平可用于评价长期肾移植受者的免疫状态,并有可能作为调整免疫抑制剂用量的参考指标.     

长期低剂量免疫抑制对肾移植受者淋巴细胞表型的影响

目的：探讨肾移植受者长期低剂量免疫抑制维持用经的免疫学基础。方法：采用多种单克隆抗体，以流式细胞仪为工具，对28例长期低剂量免疫抑制维持用药的肾移植受者外周血淋巴细胞表型进行检测，分析细胞表型与临床用药和移植效果的关系，并与常规剂量药组和慢性移植肾失功能组比较。结果：低剂量组CD95、NK及HLA－DR阳性细胞数目与正常人接的，而在常规剂量组和移植肾失功能组中，CD95明显升高，NK细胞明显减少。结论：CD95、NK及HLA－DR水平可用于评价长期肾移植受的免疫状态，并有可能作为调整免疫抑制剂用量的参考指标。     

肾移植早期尿淋巴细胞表型分析的应用价值

目的：探讨尿淋巴细胞表型分析在肾移植早期免疫监测中的临床应用价值。方法：以流式细胞技术对37例肾脏移植患者的105份尿样本进行尿淋巴细胞表型分析,测定CD3、CD4、CD8、CD19、CD25、HLA－DR阳性细胞百分率,其结果在急性排斥反应组和肾功能稳定组之间进行比较。结果：与肾功能稳定组比较,急性排斥反应组尿淋巴细胞表达HLA－DR数增多非常显著（P＜0．01）,CD8和CD25细胞增多显著（P＜0．05）,而CD3、CD4、CD19细胞数变化不显著（P＞0．05）。在诊断移植肾急性排斥反应上,HLA－DR阳性样本的诊断敏感性和特异性分别达86．96％和90．24％,CD8和CD25阳性标本的阴性预测值分别达86．79％和85．71％。结论：尿淋巴细胞表型分析可间接反映出移植肾内的免疫状态,是诊断和鉴别诊断移植肾急性排斥反应的有效方法,适用于肾移植早期的免疫监测。     

阿来佐单抗行肾移植免疫诱导治疗的有效性和安全性

目的 评价阿来佐单抗行肾移植免疫诱导治疗的有效性和安全性.方法 将89例肾移植受者随机分为试验组(43例)和对照组(46例).试验组于肾移植术前和术后24 h内分别静脉滴注阿来佐单抗15 mg,对照组不接受免疫诱导治疗.受者术后常规应用环孢素A(或他克莫司)+吗替麦考酚酯+泼尼松预防排斥反应.统计两组术后12月内的移植肾功能、急性排斥反应发生率、感染发生率、移植肾功能延迟恢复发生率、移植肾存活率及淋巴细胞计数,并用ImmuKnowTM免疫细胞功能测定法检测受者CD4+T淋巴细胞的三磷酸腺苷(ATP)值.结果 术后12个月内试验组7.0%(3/43)的受者发生病理证实的急性排斥反应,明显低于对照组的23.9%(11/46,P＜0.05).试验组和对照组总体的感染发生率为别为39.5%(17/43)和30.4%(14/46,P＞0.05),两组机会性感染的发生率分另为23.2%(10/43)和17.4%(8/46,P＞0.05).术后3个月内,试验组淋巴细胞计数低于对照组;术后6个月内,试验组CD4+T淋巴细胞ATP值低于对照组.结论 阿来佐单抗行肾移植免疫诱导治疗可维持受者的免疫抑制状态,未见严重不良反应.     

肾移植受者血他克莫司浓度对外周血自然杀伤细胞及其受体的影响

目的 研究肾移植受者血他克莫司(Tac)浓度对外周血自然杀伤(NK)细胞及其受体的影响.方法 将2007年12月至2009年7月间的60例受者纳入研究,术后受者均采用以Tac为基础的免疫抑制方案.根据术后6个月监测到的血Tac浓度将受者分为低浓度组和高浓度组[各为30例,术后6个月时血Tac浓度分别为(6.84±1.72)和(11.88±2.59)μg/L],另以20名健康志愿者作为对照组.术前和术后6个月,采用流式细胞术检测NK细胞及其抑制性受体(CD85j和CD158d)和活化性受体( CD94、NKG2D)的表达情况,采用酶联免疫吸附试验法检测免疫耐受分子分泌型HLA-G5( sH LA-G5)的表达水平.结果 术前低浓度组和高浓度组受者外周血NK细胞绝对值均较对照组显著降低(P＜0.05),术后6个月时低浓度组和高浓度组NK细胞比例及绝对值较对照组均显著降低(P＜0.05),低浓度组NK细胞绝对值显著高于高浓度组(P＜0.05).术前两组间CD85j、CD158d、CD94、NKG2D表达的差异均无统计学意义(P＞0.05);术后6个月时低浓度组和高浓度组CD85j和CD158d的表达较术前升高,CD94和NKG2D的表达下降,而低浓度组CD85j和CD158d的表达显著高于高浓度组(P＜0.05).经Spearman系数统计,CD85j和CD158d与sHLA-G5呈正相关(P＜0.01),NKG2D与sHLA-G5呈负相关(P＜0.01).结论 肾移植受者血Tac浓度与外周血NK细胞数量及其受体的表达具有相关性,低血Tac浓度受者的NK细胞数量及其抑制性受体的表达升高,仍然能有效保护移植肾功能.     

测定CD3 8+ CD8+ T淋巴细胞在监测肾移植后巨细胞病毒感染中的作用

目的观察CD38+CD8+T淋巴细胞水平在肾移植后巨细胞病毒感染患者体内的变化,探讨其监测肾移植后巨细胞病毒活动性感染的可能性.方法分别应用流式细胞术和免疫组织化学方法测定56例肾移植受者手术前后的CD38+CD8+T淋巴细胞水平和巨细胞病毒白细胞抗原,并将两者结果进行比较.结果肾移植术前所有患者巨细胞病毒白细胞抗原均为阴性,其(CD38+CD8+)/CD8+的平均比值为0.11±0.05;肾移植后检测到有14例患者巨细胞病毒白细胞抗原阳性,出现阳性的时间为术后(32.7±16.6)d,(CD38+CD8+)/CD8+的比值在术后(29.6±8.4)d出现了有显著意义的升高,平均数值为0.43±0.21.这些患者接受静脉滴注丙氧鸟苷治疗后巨细胞病毒白细胞抗原转阴,(CD38+CD8+)/CD8+平均比值下降为0.16±0.09.治疗前后CD38+CD8+T淋巴细胞水平比较差异有显著性意义(P＜0.05).结论CD38+CD8+T淋巴细胞水平的检测结合巨细胞病毒白细胞抗原检查有助临床监测肾移植后CMV活动性感染.     

不同免疫抑制方案对肾移植术受者CD4~+ Foxp3~+ 调节性T细胞的影响

目的 探讨不同免疫抑制剂方案对肾移植术受者外周血CD4~+ Foxp3~+调节性T细胞(regulatory T cells,Treg)表达水平的影响.方法 定群研究了2006年1月至2008年1月在本移植中心接受初次移植50例随访满1年肾移植受者,分为钙调神经蛋白抑制组(钙调神经蛋白抑制剂+吗替麦考酚酯+强的松)19例,其中环孢素组10例,他克莫司组9例;雷帕霉素组(雷帕霉素+吗替麦考酚酯+强的松)31例.另取20例行规律血液透析终末期肾病患者为对照组.采用流式细胞仪的方法检测3组外周血CD4~+ Foxp3~+ Treg占CD4~+ T细胞的比例,比较各组间表达水平与不同免疫抑制方案的关系.结果 钙调神经蛋白抑制剂组、雷帕霉素组和终末期肾病组3组年龄、性别比无统计学差异(P>0.05).钙调神经蛋白抑制剂组、雷帕霉素组2组冷缺血时间、HLA错配率、群体反应性抗体(PRA)和急性排斥反应发生率无统计学差异(P>0.05).雷帕霉素组和终末期肾病组CD4~+ Foxp3~+ T细胞占CD4~+ T细胞的比例均明显高于钙调神经蛋白抑制组,差异有统计学意义(P<0.01).使用环孢素患者和他克莫司患者外周血中CD4~+ Foxp3~+ T细胞占CD4~+ T细胞的比例之间无显著性差异(P>0.05).结论 肾移植术后服用雷帕霉素组患者外周血CD4~+ Foxp3~+ Treg占CD4~+ T细胞的比例显著高于服用钙调神经蛋白抑制组患者,提示雷帕霉素有助于诱导宿主对移植肾免疫耐受.     

复发性流产患者主动免疫治疗后淋巴细胞免疫表型的变化

目的探讨原因不明复发性流产(URSA)患者行淋巴细胞主动免疫治疗(LIT)后淋巴细胞免疫表型的变化对治疗效果的评估价值。方法采用流式细胞术分析URSA患者LIT前后外周血T淋巴细胞、B淋巴细胞、NK细胞和调节性T细胞免疫表型的变化(P0.05)。结果 25例URSA患者经LIT后成功妊娠16例,治疗后所有URSA患者(n=25)和妊娠成功组(n=16)外周血CD3+T细胞、CD4+HLA-DR+T细胞比例较治疗前均明显增加,CD4+T和CD3-CD56+NK细胞比例明显降低(P0.05);而治疗前后B细胞和Treg细胞、CD56bright CD16-NK、CD56dimCD16+NK、CD3+CD56+NKT及CD69+NK细胞比例则无明显变化(P0.05)。结论 LIT后外周血T细胞、NK细胞的比例发生了明显变化,CD4+T细胞、CD3-CD56+NK细胞比例降低和CD3+T细胞、活化CD4+T细胞增加也许有利于维持妊娠,T细胞和NK细胞的免疫表型有望作为LIT疗效评估的一个重要指标。     

胃癌患者及其手术前后红细胞免疫功能与淋巴细胞功能之间的相关性研究

目的探讨胃癌患者及其手术前后红细胞免疫功能的变化及与淋巴细胞免疫功能之间的相关性.方法72例胃癌患者,20例正常人为对照,检测血红细胞C3b受体花环(RBC-C3bRR)和免疫复合物花环(RBC-ICR),T淋巴细胞亚群、B细胞含量;NK细胞活性、血清循环免疫复合物(CIC)、血清免疫球蛋白和补体.结果(1)胃癌患者RBC-C3bRR显著降低,RBC-ICR显著升高(P＜0.01).(2)术后组RBC-C3bRR显著升高,RBC-ICR显著降低(P＜0.01).(3)胃癌患者CD3+、CD4+、CD4+/CD8+,NK细胞活性,B细胞,IgG、IgA均显著降低,CIC显著升高(P＜0.05或P＜0.01);手术后CD3+、CD4+、CD4+/CD8+、NK细胞活性,免疫球蛋白IgA均有显著提高(P＜0.01);胃癌患者术前组及手术前后RBC-C3bRR变化组与CD3+、CD4+、NK细胞组均呈显著正相关(P＜0.01).结论(1)胃癌患者红细胞免疫粘附功能、淋巴细胞免疫功能低下.(2)手术可使胃癌患者红细胞免疫、淋巴细胞免疫功能明显改善.(3)红细胞免疫粘附功能与T淋巴细胞、NK细胞活性之间关系密切.     

Influence of age, sex, and hepatitis C virus infection on peripheral blood lymphocyte subsets in stable kidney transplantation

BACKGROUND: Young age and hepatitis C virus infection (HCVI) are believed to be risk factors in kidney transplantation recipients. The first group is treated empirically with an intensive immunosuppressive regimen, because it is considered to have high immune alloreactivity. The other cohort usually receives a less intensive regimen to avoid excessive immunosuppressive effects. Our aim was to investigate the influence of age, sex, and HCVI on immune status in stable kidney transplant recipients through measurement of peripheral blood lymphocyte subsets. METHODS: Absolute CD3+, CD3+, CD4+, CD3+, CD8+, CD19+, CD16+ CD3- lymphocyte counts and CD4/CD8 ratios were assessed at five time points in 65 stable kidney allograft patients over 12 months. The subsets were compared according to age, sex, and HCVI of the recipients. RESULTS: An inverse association was observed between recipient age and absolute CD19+ and CD3+ CD4+ lymphocyte counts, which was significant at all time points with respect to CD19+ counts, and at three time points with respect to CD3+ CD4+ counts. A significant positive association was observed between recipient age and absolute CD3- CD16+ lymphocyte counts at three time points. Female recipients showed significantly lower CD3+ CD8+ counts and significantly higher CD4/CD8 ratios than male recipients at four time points. HCVI recipients showed significantly lower CD16+ CD3- counts at four time points. CONCLUSIONS: We observed links between immune status and age, sex and HCVI in stable kidney transplant recipients that could offer new insights into recommendations for maintenance immunosuppression.     

Determination of acquired immunodeficiency syndrome (AIDS) after renal transplantation

Diagnosing the acquired immunodeficiency syndrome (AIDS) in transplant recipients can be difficult due to the patient's medication-induced immunosuppressed state. We report two renal allograft recipients who acquired HIV infection at the time of transplantation and later went on to develop multiple opportunistic infections. Careful documentation of HIV antibody status of the donor and recipient, when available, the nature of immunosuppressive therapy used, the type of infections and their timing after transplantation, as well as the patient's absolute T4 lymphocyte count, T cell ratio, and B cell humoral response to infection were used as factors to distinguish between infection related to immunosuppressive therapy and that seen in HIV-induced immunodeficiency. Reduction in immunosuppressive therapy because of the HIV-related immunodeficiency state did not result in allograft rejection. Both patients died of their multiple infections. The determination of AIDS in the transplant recipient has both therapeutic and prognostic significance. This diagnosis should be considered when transplant patients develop unusual infections in relationship to their posttransplant course.     

Quantitation of Epstein-Barr virus DNA in the blood of adult liver transplant recipients

BACKGROUND: Posttransplant lymphoproliferation is most often observed in pediatric transplant recipients who experience primary Epstein-Barr virus (EBV) infection at the time of or after transplantation. Lymphoproliferation is believed to be caused by impaired control of EBV-infected cells, which may be of recipient or donor origin. Most studies of EBV infection and lymphoproliferation have focused on the pediatric age group. METHODS: We have undertaken a prospective study of EBV infection in adult liver transplant recipients. Sequentially collected peripheral blood lymphocytes were examined with a recently developed quantitative polymerase chain reaction assay. The assay quantitates EBV DNA genomic titre over a 5 log10 range. RESULTS: Compared with healthy EBV seropositive people not undergoing immunosuppressive therapy, blood EBV DNA titre is elevated in patients with liver disease before transplantation. Overall, highest titres were observed during the first posttransplant month, and in the context of antilymphocyte therapy. In one patient, lymphoproliferation was associated with high titres which fell during reduction of immunosuppressive therapy. In another patient with lymphoproliferation of donor lymphocyte origin, blood EBV DNA titre was not as high. CONCLUSIONS: EBV proliferation is seen in the context of advanced liver disease and after liver transplantation. EBV DNA quantitation is a useful tool to examine the effects of immunosuppression on EBV-associated lymphoproliferation, and may be an essential technique for programs exploring the merits of EBV adoptive immunotherapy.     

Microchimerism in transfused trauma patients is associated with diminished donor-specific lymphocyte response

BACKGROUND: Blood transfusion can result in long-term survival of donor leukocyte subpopulations, or microchimerism, in the peripheral blood of injured patients. Neither injury severity nor the number of transfusions is associated with its occurrence. We sought to determine whether changes in general or antigen-specific lymphocyte activation may be associated with the subsequent development of microchimerism. METHODS: We evaluated 63 transfused trauma patients, which we compared with 10 non-transfused trauma patients and 10 healthy control subjects. Of the 63 transfused patients, 31 were known to have evidence of microchimerism at hospital discharge with real-time quantitative PCR for non-recipient HLA DR alleles. We assessed lymphocyte response to phytohemagglutinin (PHA) using blood sampled upon arrival to the hospital (before transfusion) and at discharge. We performed one-way mixed leukocyte cultures (MLC) with pre-transfusion recipient specimens to assess recipient lymphocyte response to mitomycin-C treated donor cells and vice versa. RESULTS: Lymphocyte response to PHA in microchimeric transfusion recipients was lower at admission (before transfusion) and discharge than in non-microchimeric recipients. Lymphocytes from microchimeric patients had less response to donor cells than did lymphocytes from non-microchimeric patients. Microchimeric patients also more frequently had diminished lymphocyte response to a single blood donor on MLC. CONCLUSIONS: Transfusion-associated microchimerism is correlated with diminished response to mitogen challenge as well as to specific alloantigenic challenges. This microchimerism is predated by diminished lymphocyte response to a specific blood donor in many instances. The blood donor associated with this diminished alloantigenic lymphocyte response may be the source of microchimeric cells present in the recipient.     

Tolerance to vascularized kidney grafts in canine mixed hematopoietic chimeras

BACKGROUND: Recent progress in allogeneic hematopoietic stem cell transplantation provides new methods for reliable engraftment with nonlethal conditioning regimens. These techniques have been successfully applied in the treatment of both malignant and nonmalignant diseases, but have not been fully exploited for their potential to tolerize recipients for organ transplantation. These studies were undertaken to test whether the tolerance of host immune cells toward donor hematopoietic cells in mixed hematopoietic chimeras extends to include a vascularized organ, the kidney. METHODS: Using nonmyeloablative doses of total body irradiation, a short course of immunosuppression, and hematopoietic stem cells from marrow or peripheral blood sources, five dog lymphocyte antigen-identical canines were made to become stable mixed hematopoietic chimeras with no development of graft-versus-host disease or posttransplant lymphoproliferative disorder. Subsequently, renal transplantations were performed between stem cell donor and recipient littermates, and no additional immunosuppressive therapy was given after stem cell transplantation. RESULTS: All mixed chimeric dogs demonstrate different, but stable, levels of donor peripheral blood lymphocyte and granulocyte chimerism. With follow-up of longer than 1 year, all of the mixed chimeric dogs (five/five) have excellent renal function with normal serum creatinines (<1.5 mg/dl) and no pathological evidence of rejection on biopsies. CONCLUSIONS: In a major histocompatibility-matched model, minor antigen differences between donor and recipient are not sufficient to induce a host immune response to a vascularized kidney transplant in mixed hematopoietic chimeras.     

Evidence that indefinite survival of small bowel allografts achieved by a brief course of cyclosporine or FK506 is not due to systemic hyporesponsiveness.

The immunological status of Lewis (LEW) recipients of indefinitely surviving (greater than 400 days) orthotopic Brown-Norway (BN) small bowel allografts was investigated 1 to 1 1/2 years after cessation of immunosuppressive therapy with either cyclosporine or FK506 and compared with recipients of syngeneic grafts. A normal proliferative response (as measured by a mixed lymphocyte culture) of recipient peripheral lymph node lymphocytes in response to the donor-specific (BN) and the third-party (ACI) antigen, was observed in all experimental groups. Cytolytic T cell generation (as measured by a standard 51Cr-release cytotoxicity assay) in response to the donor-specific (BN) and the third-party (ACI) antigen was observed also in all groups. A FACS analysis of allograft-recipient splenocytes showed no evidence for systemic lymphoid chimerism. BN or ACI skin grafts transplanted onto recipients of allogeneic and syngeneic small bowel grafts were rejected completely in 12-17 days, while the intestinal grafts remained functional. Immunohistologic evaluation of the allografts, using anti-BN class I and anti-Lewis class II monoclonal antibodies showed anti-BN staining on the epithelial and endothelial structures, whereas the mononuclear cells in the lamina propria stained positively with the anti-LEW monoclonal antibody. However, lymphoid depletion and scarring of Peyer's patches and mesenteric lymph nodes as well as focal obliterative mesenteric arteriopathy, indicative of an indolent chronic rejection, were observed. These data demonstrate that recipients of indefinitely surviving small bowel allografts remain immune competent and do not retain the intestinal graft on the basis of specific hyporesponsiveness to the donor antigens.     

食蟹猴腹主动脉补丁缝合术后的免疫排斥反应监测

目的  建立腹主动脉补丁缝合术的异种移植模型以及术后免疫排斥监测平台。方法  将野生型巴马猪的颈动脉,修剪成约2.5 cm×1.0 cm大小的梭形补丁,缝合于食蟹猴腹主动脉,未给予免疫抑制剂。观察受体猴一般情况。术后1年采用病理学检查观察移植物动脉补丁的形态学变化。分别在术前和术后7、14、28、49 d采集受体猴血液样本,利用猪的红细胞、外周血单核细胞（PBMC）检测受体猴血清中针对猪抗原的IgM和IgG抗体水平。采用血常规五分类法和流式细胞术检测受体猴血液中淋巴细胞计数。结果  3只移植猴存活状态良好。术后1年动脉补丁所在的血管壁外侧组织呈现深红色,苏木素-伊红（HE）染色显示有大量的红细胞和血小板沉积,其中有淋巴细胞的浸润。以猪的红细胞和PBMC为靶细胞,血清中的抗猪IgM和IgG抗体水平在术后28 d达到峰值,术后49 d时水平有所下降。淋巴细胞、T细胞亚群也是在术后28 d达到峰值,49 d开始回落。结论  动脉补丁缝合术是一种便捷可靠的异种移植模型。没有免疫抑制剂的条件下,受体维持了正常的生理状态。移植物可有效激活受体的免疫系统,诱导抗猪抗体的产生,引发细胞免疫排斥反应,因此该模型可用于监测异种移植过程中的免疫排斥反应。     

Psychomotor performance in lung transplant recipients: simple reaction time.

BACKGROUND: Many transplant recipients report difficulty completing fine motor activities such as eating, writing and manipulating buttons. These impairments are thought to stem from the immunosuppressive medications being taken by these patients. The purpose of this study was to examine central and peripheral processes and the force regulation involved in producing appropriate and quality movement in lung transplant recipients. METHODS: Fifty-one right-handed subjects were recruited from 3 study groups (17 in each group): Group 1, lung transplant recipients (LTR); Group 2, subjects with advanced emphysema; and Group 3, healthy adult controls. Each subject completed a fine motor and gross motor simple reaction time task. Central processing was examined by measuring pre-motor time, peripheral processing was measured by motor time, and force regulation was measured using movement time. A 3 x 5 multivariate analysis of co-variance (MANCOVA) was utilized to examine group differences, with the performance on the 6-minute walk test serving as co-variant. Correlation analyses were conducted to examine the relationship between medication and psychomotor performance. RESULTS: The lung transplant recipient group exhibited a longer movement time and a trend toward longer pre-motor times. There was also a significant relationship between medication and movement time. CONCLUSIONS: The results support the hypothesis that lung transplant recipients have deficits in psychomotor performance, which is consistent with the literature showing that immunosuppressive medications and hypoxia have adverse effects on skeletal muscle. This line of research is relevant to the restoration of function and improvement in quality of life of LTR.     

Monitoring systemic donor lymphocyte macrochimerism to aid the diagnosis of graft-versus-host disease after liver transplantation

BACKGROUND: The diagnosis of graft-versus-host disease (GvHD) after liver transplantation can be difficult because early symptoms are often nonspecific. In this study, the presence of donor lymphocyte macrochimerism in recipient peripheral blood was examined as a diagnostic aid for GvHD after cadaveric donor liver transplantation. METHODS: Between 1996 and 2002, 33 liver transplant recipients with a clinical suspicion of GvHD (skin rash, diarrhea, pyrexia, pancytopenia, or anemia, without an obvious alternative cause) were investigated for peripheral blood donor lymphocyte macrochimerism. Donor macrochimerism was determined at the time of first clinical presentation by a low-sensitivity polymerase chain reaction (PCR) to detect donor human leukocyte antigen (HLA) alleles using genomic DNA extracted from recipient peripheral blood. Where donor HLA alleles were detected, the percentage of donor T cells was quantified by two-color flow cytometric analysis using antibodies specific for mismatched donor and recipient HLA alleles. The relationship between the presence or absence of donor lymphocyte macrochimerism and final diagnoses based on clinical and histological criteria was examined. RESULTS: Seven of the 33 patients were PCR positive for donor HLA alleles. All had macrochimerism, with donor T lymphocyte levels ranging from 4% to 50% of circulating lymphocytes. All seven patients had normal liver function tests, skin rash, and diagnosis of GvHD histologically confirmed by skin or gut biopsies. Twenty-six patients were PCR negative, and, in 23, an alternative diagnosis was eventually established. The remaining three patients made a rapid and spontaneous recovery with no further symptoms suggestive of GvHD. CONCLUSIONS: Donor lymphocyte macrochimerism was present in all patients in whom the diagnosis of GvHD was confirmed. In patients with symptoms consistent with GvHD and a negative PCR for donor HLA, an alternative diagnosis was eventually established or the patients recovered spontaneously. Detection of donor HLA alleles in recipient peripheral blood by PCR is a useful diagnostic tool for GvHD after liver transplantation.     

混合培养的供、受者骨髓细胞过继回输延长小鼠移植心脏存活时间

目的 将供、受者骨髓细胞经混合培养后过继回输,以观察其对同种异体移植心脏存活时间和受者免疫功能的影响.方法 取Balb/c小鼠和C57BL/6J小鼠的骨髓细胞,进行混合培养.配制含Balb/c小鼠和C57BL/6J小鼠脾淋巴细胞的混合淋巴细胞反应体系(MLR)以及含Balb/c小鼠和C3H小鼠脾淋巴细胞的MLR,分别加入混合培养的骨髓细胞,观察其对MLR中细胞增殖的影响.以C57BL/6J小鼠为供者,Balb/c小鼠为受者行腹腔异位心脏移植,实验分为4组:(1)移植对照组,受者仅进行心脏移植,不作其他处理;(2)实验对照组,心脏移植后给予西罗莫司灌胃;(3)实验组,移植手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司;(4)第三方对照组,受者接受C3H小鼠的移植心脏,手术结束前注射混合培养的骨髓细胞1×10~7个,术后给予西罗莫司.记录移植心脏存活时间;移植心脏停跳当日,取受者外周血,检测CD4~+ CD25~+ T淋巴细胞的比例及供者来源的H-2K~b细胞的比例.结果 加入混合培养的骨髓细胞后,Balb/c和C57BL/6J的MLR的淋巴细胞增殖率低于Balb/c和C3H的MLR.实验组移植心脏的存活时间长于其他3组(P<0.05).实验组CD4~+CD25~+T淋巴细胞的百分率高于其他3组(P<0.05).实验组外周血中H-2K~b细胞的比例高于其他3组(P<0.05).结论 受者输注混合培养的供、受者骨髓细胞可在一定程度上调节免疫应答,延长小鼠移植心脏的存活时间,该作用具有供者抗原特异性.