Frank Renee, Dean Stephanie A, Molina Maria R, Kamoun Malek, Lal Priti
Pathology and Laboratory Medicine, Hospital of the University of Pennsylvania, Philadelphia, PA, United States.
Pathology and Laboratory Medicine, Hospital of the University of Pennsylvania, Philadelphia, PA, United States.
Cardiovasc Pathol. 2015 May-Jun;24(3):168-72. doi: 10.1016/j.carpath.2014.11.001. Epub 2014 Nov 7.
Acute antibody-mediated rejection (AMR) is a major complication after heart transplantation, posing a significant risk for allograft failure, cardiac allograft vasculopathy, and poor survival. While the inflammatory milieu of cellular rejection and Quilty lesions is well known, the immunologic components of AMR are not well understood. Our aim was to better define the immunophenotype of infiltrating lymphocytes in biopsies with AMR, specifically in relation to donor-specific antibodies to human leukocyte antigen (HLA) class I, II, or both.
We performed a retrospective analysis of cardiac transplant patients with concurrent endomyocardial biopsies (EMB), donor-specific antibody (DSA) measurements, and immunofluorescence for C4d at our institution (2005-2011). DSA was evaluated against HLA class I and class II specificities pre- and posttransplant using flow cytometry and/or Luminex bead assays. Acute cellular rejection (ACR) and pathologic AMR (pAMR) were based on the International Society for Heart and Lung Transplantation 2005/2013 reports. Immunohistochemical analysis for CD3, CD4, CD8, and CD79a was performed using standard immunohistochemical protocols on one formalin-fixed, paraffin-embedded EMB from each patient. The number of lymphocytes expressing each protein was enumerated microscopically at 400×. Ratios of T:B cells and CD4:CD8 T cells were then calculated for each EMB.
Seventy-nine cardiac transplant patients who had pre- and posttransplant DSA measurements were analyzed. Of these 79 patients, 37 had DSA against HLA class I, HLA class II, or both. Of patients with DSA, the average CD4:CD8 ratio in the EMB was 0.80, while those with only ACR had a CD4:CD8 ratio of 1.49. Interestingly, the T:B cell ratio in patients with and without DSA was 5.7 and 5.5, respectively.
Cardiac transplant patients with DSA against HLA have more CD8 cytotoxic T cells than CD4 helper T cells in the EMB lymphocytic infiltrate compared with patients without DSA against HLA. The inflammatory infiltrate T:B cell ratio was similar in patients both with and without DSA. The relative increase of cytotoxic T cells in EMB while the patient has DSA suggests a possible pathogenic role of these cells and may aid in the diagnosis and treatment of AMR.
急性抗体介导的排斥反应（AMR）是心脏移植后的主要并发症，对移植物功能衰竭、心脏移植物血管病变及生存率低下构成重大风险。虽然细胞排斥反应和奎尔蒂病变的炎症环境已为人熟知，但AMR的免疫成分尚未得到充分了解。我们的目的是更好地确定伴有AMR的活检组织中浸润淋巴细胞的免疫表型，特别是与针对人类白细胞抗原（HLA）I类、II类或两者的供体特异性抗体的关系。
我们对本机构（2005 - 2011年）同时进行心内膜心肌活检（EMB）、供体特异性抗体（DSA）检测及C4d免疫荧光检测的心脏移植患者进行了回顾性分析。使用流式细胞术和/或Luminex微珠分析法评估移植前后针对HLA I类和II类特异性的DSA。急性细胞排斥反应（ACR）和病理性AMR（pAMR）依据国际心肺移植学会2005/2013年报告判定。对每位患者的一份福尔马林固定、石蜡包埋的EMB标本，采用标准免疫组织化学方案进行CD3、CD4、CD8和CD79a的免疫组织化学分析。在400倍显微镜下计数表达每种蛋白的淋巴细胞数量。然后计算每份EMB的T:B细胞比值和CD4:CD8 T细胞比值。
分析了79例进行了移植前后DSA检测的心脏移植患者。在这79例患者中，37例存在针对HLA I类、HLA II类或两者的DSA。有DSA的患者，EMB中的平均CD4:CD8比值为0.80，而仅有ACR的患者CD4:CD8比值为1.49。有趣的是，有和没有DSA的患者T:B细胞比值分别为5.7和5.5。
与没有针对HLA的DSA的患者相比，有针对HLA的DSA的心脏移植患者，其EMB淋巴细胞浸润中CD8细胞毒性T细胞比CD4辅助性T细胞更多。有和没有DSA的患者炎症浸润的T:B细胞比值相似。患者存在DSA时EMB中细胞毒性T细胞的相对增加提示这些细胞可能具有致病作用，并且可能有助于AMR的诊断和治疗。
