Deng M C, Eisen H J, Mehra M R, Billingham M, Marboe C C, Berry G, Kobashigawa J, Johnson F L, Starling R C, Murali S, Pauly D F, Baron H, Wohlgemuth J G, Woodward R N, Klingler T M, Walther D, Lal P G, Rosenberg S, Hunt S
Columbia University, New York, NY, USA.
Am J Transplant. 2006 Jan;6(1):150-60. doi: 10.1111/j.1600-6143.2005.01175.x.
Rejection diagnosis by endomyocardial biopsy (EMB) is invasive, expensive and variable. We investigated gene expression profiling of peripheral blood mononuclear cells (PBMC) to discriminate ISHLT grade 0 rejection (quiescence) from moderate/severe rejection (ISHLT > or = 3A). Patients were followed prospectively with blood sampling at post-transplant visits. Biopsies were graded by ISHLT criteria locally and by three independent pathologists blinded to clinical data. Known alloimmune pathways and leukocyte microarrays identified 252 candidate genes for which real-time PCR assays were developed. An 11 gene real-time PCR test was derived from a training set (n = 145 samples, 107 patients) using linear discriminant analysis (LDA), converted into a score (0-40), and validated prospectively in an independent set (n = 63 samples, 63 patients). The test distinguished biopsy-defined moderate/severe rejection from quiescence (p = 0.0018) in the validation set, and had agreement of 84% (95% CI 66% C94%) with grade ISHLT > or = 3A rejection. Patients >1 year post-transplant with scores below 30 (approximately 68% of the study population) are very unlikely to have grade > or = 3A rejection (NPV = 99.6%). Gene expression testing can detect absence of moderate/severe rejection, thus avoiding biopsy in certain clinical settings. Additional clinical experience is needed to establish the role of molecular testing for clinical event prediction and immunosuppression management.
通过心内膜心肌活检（EMB）进行排斥反应诊断具有侵入性、成本高且结果存在差异。我们研究了外周血单个核细胞（PBMC）的基因表达谱，以区分国际心脏和肺移植学会（ISHLT）0级排斥反应（静止期）与中度/重度排斥反应（ISHLT≥3A）。对患者进行前瞻性随访，在移植后随访时采集血样。活检标本由当地按照ISHLT标准进行分级，并由三位对临床数据不知情的独立病理学家进行分级。通过已知的同种免疫途径和白细胞微阵列鉴定出252个候选基因，并针对这些基因开发了实时聚合酶链反应（PCR）检测方法。使用线性判别分析（LDA）从一个训练集（n = 145个样本，107例患者）中得出一个11基因的实时PCR检测方法，将其转换为一个评分（0 - 40），并在一个独立的数据集（n = 63个样本，63例患者）中进行前瞻性验证。在验证集中，该检测方法能够区分活检确定的中度/重度排斥反应与静止期（p = 0.0018），并且与ISHLT≥3A级排斥反应的一致性为84%（95%置信区间66% - 94%）。移植后超过1年且评分低于30分的患者（约占研究人群的68%）极不可能发生≥3A级排斥反应（阴性预测值 = 99.6%）。基因表达检测能够检测出不存在中度/重度排斥反应，从而在某些临床情况下避免进行活检。需要更多的临床经验来确立分子检测在临床事件预测和免疫抑制管理中的作用。
