Necker-Enfants Malades Institute, Institut National de la Santé et de la Recherche Médicale (INSERM) U1151, University of Paris, Paris, France.
Department of Nephrology and Kidney Transplantation, Necker Hospital, Assistance Publique-Hôpitaux de Paris (AP-HP), Paris, France.
J Am Soc Nephrol. 2021 Dec;32(12):3231-3251. doi: 10.1681/ASN.2021050689.
After kidney transplantation, donor-specific antibodies against human leukocyte antigen donor-specific antibodies (HLA-DSAs) drive antibody-mediated rejection (ABMR) and are associated with poor transplant outcomes. However, ABMR histology (ABMRh) is increasingly reported in kidney transplant recipients (KTRs) without HLA-DSAs, highlighting the emerging role of non-HLA antibodies (Abs).
W e designed a non-HLA Ab detection immunoassay (NHADIA) using HLA class I and II-deficient glomerular endothelial cells (CiGEnCHLA) that had been previously generated through CRISPR/Cas9-induced and gene disruption. Flow cytometry assessed the reactivity to non-HLA antigens of pretransplantation serum samples from 389 consecutive KTRs. The intensity of the signal observed with the NHADIA was associated with post-transplant graft histology assessed in 951 adequate biopsy specimens.
W e sequentially applied CRISPR/Cas9 to delete the and genes to obtain a CiGEnCHLA clone. CiGEnCHLA cells remained indistinguishable from the parental cell line, CiGEnC, in terms of morphology and phenotype. Previous transplantation was the main determinant of the pretransplantation NHADIA result (<0.001). Stratification of 3-month allograft biopsy specimens (=298) according to pretransplantation NHADIA tertiles demonstrated that higher levels of non-HLA Abs positively correlated with increased glomerulitis (=0.002), microvascular inflammation (=0.003), and ABMRh (=0.03). A pretransplantation NHADIA threshold of 1.87 strongly discriminated the KTRs with the highest risk of ABMRh (=0.005, log-rank test). A multivariate Cox model confirmed that NHADIA status and HLA-DSAs were independent, yet synergistic, predictors of ABMRh.
The NHADIA identifies non-HLA Abs and strongly predicts graft endothelial injury independent of HLA-DSAs.
肾移植后，针对人类白细胞抗原供体特异性抗体（HLA-DSAs）的供体特异性抗体会引发抗体介导的排斥反应（ABMR），并与较差的移植结果相关。然而，越来越多的肾移植受者（KTRs）在没有 HLA-DSAs 的情况下报告出现 ABMR 组织学（ABMRh），这凸显了非 HLA 抗体（Abs）的新兴作用。
我们使用通过 CRISPR/Cas9 诱导的 和 基因敲除先前生成的 HLA I 类和 II 类缺陷肾小球内皮细胞（CiGEnCHLA）设计了一种非 HLA Ab 检测免疫测定法（NHADIA）。流式细胞术评估了 389 例连续 KTR 移植前血清样本对非 HLA 抗原的反应性。在 951 例充分活检标本中评估 NHADIA 观察到的信号强度与移植后移植物组织学相关。
我们依次应用 CRISPR/Cas9 敲除 和 基因以获得 CiGEnCHLA 克隆。CiGEnCHLA 细胞在形态和表型方面与亲本细胞系 CiGEnC 仍然无法区分。先前的移植是移植前 NHADIA 结果的主要决定因素（<0.001）。根据移植前 NHADIA 三分位数对 3 个月移植物活检标本进行分层（=298）表明，较高水平的非 HLA Abs 与肾小球肾炎增加（=0.002）、微血管炎症（=0.003）和 ABMRh（=0.03）呈正相关。移植前 NHADIA 阈值为 1.87 可强烈区分具有最高 ABMRh 风险的 KTR（=0.005，对数秩检验）。多变量 Cox 模型证实 NHADIA 状态和 HLA-DSAs 是 ABMRh 的独立但协同的预测因子。
NHADIA 可识别非 HLA Abs，并独立于 HLA-DSAs 强烈预测移植物内皮损伤。
