Legris Tristan, Picard Christophe, Todorova Dilyana, Lyonnet Luc, Laporte Cathy, Dumoulin Chloé, Nicolino-Brunet Corinne, Daniel Laurent, Loundou Anderson, Morange Sophie, Bataille Stanislas, Vacher-Coponat Henri, Moal Valérie, Berland Yvon, Dignat-George Francoise, Burtey Stéphane, Paul Pascale
Nephrology Dialysis Renal Transplantation Center, Assistance Publique des Hôpitaux de Marseille, Hospital de la Conception , Marseille , France.
Établissement Français du Sang Alpes Méditerranée, Marseille, France; ADES UMR 7268, CNRS, EFS, Aix-Marseille Université, Marseille, France.
Front Immunol. 2016 Aug 11;7:288. doi: 10.3389/fimmu.2016.00288. eCollection 2016.
Although kidney transplantation remains the best treatment for end-stage renal failure, it is limited by chronic humoral aggression of the graft vasculature by donor-specific antibodies (DSAs). The complement-independent mechanisms that lead to the antibody-mediated rejection (ABMR) of kidney allografts remain poorly understood. Increasing lines of evidence have revealed the relevance of natural killer (NK) cells as innate immune effectors of antibody-dependent cellular cytotoxicity (ADCC), but few studies have investigated their alloreactive potential in the context of solid organ transplantation. Our study aimed to investigate the potential contribution of the antibody-dependent alloreactive function of NK cells to kidney graft dysfunction. We first conducted an observational study to investigate whether the cytotoxic function of NK cells is associated with chronic allograft dysfunction. The NK-Cellular Humoral Activation Test (NK-CHAT) was designed to evaluate the recipient and antibody-dependent reactivity of NK cells against allogeneic target cells. The release of CD107a/Lamp1(+) cytotoxic granules, resulting from the recognition of rituximab-coated B cells by NK cells, was analyzed in 148 kidney transplant recipients (KTRs, mean graft duration: 6.2 years). Enhanced ADCC responsiveness was associated with reduced graft function and identified as an independent risk factor predicting a decline in the estimated glomerular filtration rate over a 1-year period (hazard ratio: 2.83). In a second approach, we used the NK-CHAT to reveal the cytotoxic potential of circulating alloantibodies in vitro. The level of CD16 engagement resulting from the in vitro recognition of serum-coated allogeneic B cells or splenic cells was further identified as a specific marker of DSA-induced ADCC. The NK-CHAT scoring of sera obtained from 40 patients at the time of transplant biopsy was associated with ABMR diagnosis. Our findings indicate that despite the administration of immunosuppressive treatments, robust ADCC responsiveness can be maintained in some KTRs. Because it evaluates both the Fab recognition of alloantigens and Fc-driven NK cell activation, the NK-CHAT represents a potentially valuable tool for the non-invasive and individualized evaluation of humoral risk during transplantation.
尽管肾移植仍然是终末期肾衰竭的最佳治疗方法，但它受到供体特异性抗体（DSA）对移植血管的慢性体液攻击的限制。导致肾移植抗体介导排斥反应（ABMR）的补体非依赖性机制仍知之甚少。越来越多的证据表明自然杀伤（NK）细胞作为抗体依赖性细胞毒性（ADCC）的先天免疫效应器具有相关性，但很少有研究在实体器官移植的背景下研究它们的同种异体反应潜力。我们的研究旨在调查NK细胞的抗体依赖性同种异体反应功能对肾移植功能障碍的潜在贡献。我们首先进行了一项观察性研究，以调查NK细胞的细胞毒性功能是否与慢性移植功能障碍相关。NK细胞-细胞体液激活试验（NK-CHAT）旨在评估NK细胞对同种异体靶细胞的受体和抗体依赖性反应性。在148名肾移植受者（KTRs，平均移植时间：6.2年）中分析了NK细胞识别利妥昔单抗包被的B细胞后导致的CD107a/Lamp1(+)细胞毒性颗粒的释放。增强的ADCC反应性与移植功能降低相关，并被确定为预测1年内估计肾小球滤过率下降的独立危险因素（风险比：2.83）。在第二种方法中，我们使用NK-CHAT在体外揭示循环同种异体抗体的细胞毒性潜力。体外识别血清包被的同种异体B细胞或脾细胞导致的CD16结合水平进一步被确定为DSA诱导的ADCC的特异性标志物。在移植活检时从40名患者获得的血清的NK-CHAT评分与ABMR诊断相关。我们的研究结果表明，尽管给予了免疫抑制治疗，但在一些KTRs中仍可维持强大的ADCC反应性。由于它评估了同种异体抗原的Fab识别和Fc驱动的NK细胞激活，NK-CHAT代表了一种潜在有价值的工具，用于移植期间体液风险的非侵入性和个体化评估。
