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 B2M and CIITA 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 B2M and CIITA 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 (P<0.001). Stratification of 3-month allograft biopsy specimens (n=298) according to pretransplantation NHADIA tertiles demonstrated that higher levels of non-HLA Abs positively correlated with increased glomerulitis (P=0.002), microvascular inflammation (P=0.003), and ABMRh (P=0.03). A pretransplantation NHADIA threshold of 1.87 strongly discriminated the KTRs with the highest risk of ABMRh (P=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)，并与移植结果不佳相关。然而，ABMR 组织学 (ABMRh) 在没有 HLA-DSA 的肾移植受者 (KTR) 中的报道越来越多，突出了非 HLA 抗体 (Abs) 的新兴作用。

我们使用 HLA I 类和 II 类缺陷型肾小球内皮细胞 (CiGEnC) 设计了一种非 HLA 抗体检测免疫测定 (NHADIA)HLA)，之前是通过 CRISPR/Cas9 诱导的B2M和CIITA基因破坏产生的。流式细胞术评估了来自 389 个连续 KTR 的移植前血清样本对非 HLA 抗原的反应性。用 NHADIA 观察到的信号强度与在 951 个足够的活检标本中评估的移植后移植物组织学相关。

我们依次应用 CRISPR/Cas9 删除B2M和CIITA基因以获得 CiGEnCHLA克隆。CiGEnCHLA 细胞在形态和表型方面与亲代细胞系 CiGEnC 无法区分。既往移植是移植前 NHADIA 结果的主要决定因素 ( P <0.001)。根据移植前 NHADIA 三分位数对 3 个月同种异体移植活检标本 ( n = 298) 进行分层表明，较高水平的非 HLA 抗体与肾小球炎 ( P = 0.002)、微血管炎症 ( P = 0.003) 和 ABMRh ( P =0.03)。1.87 的移植前 NHADIA 阈值强烈区分了具有最高 ABMRh 风险的 KTR（P=0.005，对数秩检验）。多变量 Cox 模型证实 NHADIA 状态和 HLA-DSA 是 ABMRh 的独立但协同的预测因子。

NHADIA 识别非 HLA 抗体，并强烈预测独立于 HLA-DSA 的移植物内皮损伤。