Zakliczyński Michał, Zakliczyńska Helena, Klimczak Aleksandra, Nozyński Jerzy, Kozłowska Katarzyna, Trzcińska Iwona, Przybylski Roman, Wojarski Jacek, Durmała Jacek, Lange Andrzej, Zembala Marian
Dept. of Cardiac Surgery & Transplantation, Silesian Center for Heart Disease, Zabrze, Poland.
Ann Transplant. 2003;8(1):13-8.
Aim of this study was to find features characteristic for steroid resistant cellular rejection (SRR) of the transplanted heart, using phenotypic identification of cells creating infiltrates in endomyocardial biopsies (EMBs) obtained before and after high dose steroids treatment.
146 heart transplant recipients, treated with cyclosporine-A, azathioprine and prednisone, were taken under consideration. EMB results > or = 3A (ISHLT) were considered significant rejection, requiring treatment with 1 g i.v. methylprednizolone for 3 days followed by oral prednisone. SRR was diagnosed in case of increased grade of rejection in control EMB, lack of improvement in 2 consecutive EMBs or increasing hemodynamic compromise. SRR was found in 15 pts. (study group). Control group consisted of remaining 131 pts. Paraffin-embedded blocks containing EMB samples from 9 pts. from study group and randomly chosen 14 pts. from control group were used (2 EMBs per pt.). Significant rejection was present in the first EMB, the second EMB was performed 7 days after beginning of the treatment. In the study group, first 2 EMBs creating a sequence of SRR were analysed. Following antigens were identified: CD45RO (T-cells), CD8 (cytotoxic T-cells), CD20 (B-cells), and CD95 (marker of apoptosis). DR expression and macrophages presence were also quantified.
CD45RO was predominant phenotype before and after treatment in both groups. Higher quantity of CD20 cells were observed in study group, however its number increased after treatment in control group. CDB-cells and macrophages were present in low amounts, that did not react to treatment. CD95 positive cells were present only in 3 EMBs. None of above differences was statistically significant. DR expression staining showed no difference either in biopsies taken before steroid treatment or after completing of high dose steroid therapy.
Phenotype identification of cells infiltrating myocardium of the transplanted heart was not sufficient to predict or characterise steroid resistant rejection.
本研究的目的是通过对高剂量类固醇治疗前后获取的心肌内膜活检（EMB）中形成浸润的细胞进行表型鉴定，找出移植心脏类固醇抵抗性细胞排斥反应（SRR）的特征。
研究对象为146例接受环孢素A、硫唑嘌呤和泼尼松治疗的心脏移植受者。EMB结果≥3A（国际心脏和肺移植学会标准）被视为显著排斥反应，需要静脉注射1g甲泼尼龙治疗3天，随后口服泼尼松。如果对照EMB中排斥反应等级增加、连续两次EMB无改善或血流动力学损害加重，则诊断为SRR。15例患者（研究组）被发现存在SRR。对照组由其余131例患者组成。使用了来自研究组9例患者和对照组随机选择的14例患者的包含EMB样本的石蜡包埋块（每位患者2次EMB）。首次EMB存在显著排斥反应，治疗开始7天后进行第二次EMB。在研究组中，分析了形成SRR序列的前两次EMB。鉴定了以下抗原：CD45RO（T细胞）、CD8（细胞毒性T细胞）、CD20（B细胞）和CD95（凋亡标志物）。还对DR表达和巨噬细胞的存在进行了定量。
两组治疗前后CD45RO均为主要表型。研究组中观察到CD20细胞数量较多，但对照组中其数量在治疗后增加。CD8细胞和巨噬细胞数量较少，对治疗无反应。仅在3次EMB中存在CD95阳性细胞。上述差异均无统计学意义。DR表达染色在类固醇治疗前或高剂量类固醇治疗完成后的活检中均未显示差异。
对移植心脏心肌浸润细胞进行表型鉴定不足以预测或表征类固醇抵抗性排斥反应。
