Hospices Civils de Lyon, Hôpital Edouard Herriot, Service de Transplantation, Néphrologie et Immunologie Clinique, Lyon, France; INSERM U1111, Centre International de Recherche en Infectiologie (CIRI), Ecole Normale Supérieure de Lyon, Lyon, France; Université Lyon 1, Faculté de medecine Lyon-Est, Lyon, France.
Unité d'Analyse d'Images Biologiques, Institut Pasteur Paris, France.
Kidney Int. 2017 Jul;92(1):214-226. doi: 10.1016/j.kint.2017.01.011. Epub 2017 Mar 18.
Antibody-mediated rejection is associated with heterogeneous kidney allograft outcomes. Accurate evaluation of risk for graft loss at time of diagnosis is necessary to offer personalized treatment. In contrast with serological and molecular assessment, morpho-histological evaluation of antibody-mediated rejection lesions has not significantly evolved. This relies on Banff classifications designed to be of diagnostic discriminatory power rather than prognostic and face quantitative and qualitative limitations. Here we developed a method of Computer-assisted Analysis of Graft Inflammation (CAGI) to improve the classification of allograft inflammation. Digitization of immunostained biopsy sections, image processing and algorithm-driven analysis allowed quantification of macrophages, T cells, B cells, and granulocytes per unit surface of interstitium, capillaries or glomeruli. CAGI was performed on biopsy specimens of 52 patients with extensively phenotyped antibody-mediated rejection. Macrophage numbers in capillaries and interstitium, but not Banff scores or the amount of other immune cell subsets, correlated with donor-specific antibody (DSA) mean fluorescence intensity and DSA-C3d status. The quantity of macrophages in the interstitium and DSA-C3d status were the only independent predictors for significant allograft loss at the time of antibody-mediated rejection diagnosis (hazard ratio 3.71 and 2.34, respectively). A significant strategy integrating the DSA-C3d assay and the quantification of interstitial macrophages allowed identification of three groups with distinct renal prognosis: DSA-C3d, DSA-C3d/macrophages-low and DSAC3d/macrophages-high. Thus, CAGI brings a missing piece to the antibody-mediated rejection puzzle by identifying morpho-histological processes that bridge in vitro parameters of DSA pathogenicity and graft loss. Hence, this approach could be useful in future integrated strategies of risk evaluation.
抗体介导的排斥反应与异体肾移植物的异质性结局相关。为了提供个性化的治疗，在诊断时准确评估移植物丢失的风险是必要的。与血清学和分子评估相比，抗体介导的排斥反应病变的形态学评估并没有显著发展。这依赖于设计用于诊断区分能力而非预后的 Banff 分类，并且面临定量和定性的限制。在这里，我们开发了一种计算机辅助分析移植物炎症（CAGI）的方法，以改善异体移植物炎症的分类。免疫染色活检切片的数字化、图像处理和算法驱动的分析允许定量分析间质、毛细血管或肾小球单位表面积的巨噬细胞、T 细胞、B 细胞和粒细胞。对 52 例具有广泛表型抗体介导排斥反应的患者的活检标本进行了 CAGI 检测。毛细血管和间质中的巨噬细胞数量，但不是 Banff 评分或其他免疫细胞亚群的数量，与供体特异性抗体（DSA）平均荧光强度和 DSA-C3d 状态相关。间质中巨噬细胞的数量和 DSA-C3d 状态是抗体介导排斥反应诊断时显著移植物丢失的唯一独立预测因素（危险比分别为 3.71 和 2.34）。一种整合 DSA-C3d 检测和间质巨噬细胞定量的重要策略，可以识别出具有不同肾脏预后的三个组：DSA-C3d、DSA-C3d/巨噬细胞低和 DSA-C3d/巨噬细胞高。因此，CAGI 通过识别连接 DSA 致病性和移植物丢失的体外参数的形态学过程，为抗体介导的排斥反应难题提供了一个缺失的部分。因此，这种方法在未来的风险评估综合策略中可能有用。
