目的:小儿肝移植(pLT)后,儿童接受终身免疫抑制,因为用于评估排斥反应概率的可靠生物标志物很少.在多中心(n=7)前瞻性临床队列研究中,我们旨在表征pLT后第一年可溶性和细胞免疫介质的纵向动态,并确定与结局相关的早期生物标志物.
方法:使用配对的基于Luminex的多重技术和流式细胞术,我们表征了可溶性免疫介质的纵向动力学(SIM,n=50)和244名患者的血液中的免疫细胞在一年以上的8次访问:之前,7/14/21/28天,pLT后3/6/12个月。
结果:基于SIM资料的患者无监督聚类揭示了与临床结果相关的6个独特的SIM特征。从与改善结果相关的3个签名中,其中一项与一年的无排斥反应生存率和稳定的移植物功能相关,其特征是促炎性反应水平较低(CXCL8/9/10/12,CCL7,SCGF-β,sICAM-1),高水平的再生(SCF,TNF-β),和促凋亡(TRAIL)SIM(全部,p<0.001,倍数变化>100)。值得注意的是,此SIM签名在pLT后两周出现,并在全年保持稳定,指出其作为一种新型早期生物标志物的潜力,用于最小化或戒断免疫抑制。在这些病人的血液中,CD56brightNK细胞的频率较高(p<0.01),一个已知的标志也与手术耐受性pLT患者有关,被检测到。基于识别的SIM特征预测排斥反应的模型的一致性为0.715和0.795,结合与生活相关的移植作为协变量,分别。
结论:SIM血液特征可以在pLT后的第一年进行非侵入性和早期评估排斥风险,为改善治疗选择铺平了道路。
OBJECTIVE: After pediatric liver transplantation (pLT), children undergo life-long immunosuppression since reliable biomarkers for the assessment of rejection probability are scarce. In the multicentre (n=7) prospective clinical cohort \"ChilSFree\" study, we aimed to characterize longitudinal dynamics of soluble and cellular immune mediators during the first year after pLT and identify early biomarkers associated with outcome.
METHODS: Using paired Luminex-based multiplex technique and flow cytometry, we characterized longitudinal dynamics of soluble immune mediators (SIM, n=50) and immune cells in the blood of 244 patients at 8 visits over one year: before, 7/14/21/28 days, 3/6/12 months after pLT.
RESULTS: The unsupervised clustering of patients based on SIM profiles revealed 6 unique SIM signatures associated with clinical outcome. From 3 signatures linked to improved outcome, one was associated with one-year-long rejection-free survival and stable graft function and was characterized by low levels of pro-inflammatory (CXCL8/9/10/12, CCL7, SCGF-β, sICAM-1), high levels of regenerative (SCF, TNF-β), and pro-apoptotic (TRAIL) SIM (all, p<0.001, fold change >100). Of note, this SIM signature appeared two weeks after pLT and remained stable over the entire year, pointing towards its potential as a novel early biomarker for minimizing or weaning immunosuppression. In the blood of these patients, a higher frequency of CD56bright NK cells (p<0.01), a known hallmark also associated with operationally tolerant pLT patients, was detected. The concordance of the model for prediction of rejection based on identified SIM signatures was 0.715, and 0.795, in combination with living-related transplantation as co-variate, respectively.
CONCLUSIONS: SIM blood signatures may enable the non-invasive and early assessment of rejection risks in the first year after pLT, paving the way to improved therapeutic options.