Division of Pediatric Hematology/Oncology/Blood and Marrow Transplant, Department of Pediatrics, BC Children's Hospital, University of British Columbia, Vancouver, Canada; Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, Vancouver, Canada.
Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, Vancouver, Canada.
Biol Blood Marrow Transplant. 2019 Mar;25(3):451-458. doi: 10.1016/j.bbmt.2018.11.007. Epub 2018 Nov 14.
Previous studies have reported single B cell-related chronic graft-versus-host disease diagnostic (cGVHD) biomarkers, such as B cell-activating factor (BAFF), CD21, and immature B cells, but research on the performance of biomarker combinations and the covariate effect of steroids is lacking. The primary objective of this study was to determine the most accurate combination of B cell populations using cell surface staining flow cytometry in an independent cohort of patients with cGVHD. Secondary objectives included assessing the effect of corticosteroid use at sample collection on the makeup and accuracy of the diagnostic panel and identifying the mechanism underlying low surface expression of BAFF receptor (BAFF-R) on B cells in cGVHD. Flow cytometry analysis was performed in an adult cohort of post-HCT patients with cGVHD onset (n = 44) and time-matched recipients without cGVHD (n = 63). We confirmed that the onset of cGVHD was associated with higher soluble BAFF (sBAFF) levels, elevated CD27CD10CD21 CD19 B cell and classical switched memory B cell counts, and reduced transitional and naïve B cell counts. The highest single B cell population area under the receiver operating characteristic (ROC) curve (AUC) was .72 for transitional type 1 CD21 B cells. We also showed a significant inverse relationship between sBAFF and surface BAFF-R expression caused by sBAFF modulation of BAFF-R. Steroid use at sample collection influenced the significance of the sBAFF:B cell ratio, naïve and marginal zone-like B cells. The optimal combination of B cell subsets most significantly associated with cGVHD onset with or without concurrent corticosteroid use resulted in ROC AUCs of .87 and .84, respectively. Transitional and CD21 B cells were the only populations present in both panels; however, analyzing only these populations resulted in ROC AUCs of .79 and .78, respectively. This suggests that the inclusion of other populations and use of different panels depending on steroid use is necessary to achieve better accuracy. sBAFF was not a component of either panel. These novel B cell profiles could be tested prospectively in patients post-HSCT and could lead to focused mechanistic studies.
先前的研究报告了单一 B 细胞相关的慢性移植物抗宿主病诊断（cGVHD）生物标志物，如 B 细胞激活因子（BAFF）、CD21 和未成熟 B 细胞，但关于生物标志物组合的性能和类固醇的协变量效应的研究尚缺乏。本研究的主要目的是在另一组 cGVHD 患者中使用细胞表面染色流式细胞术确定最准确的 B 细胞群体组合。次要目标包括评估在样本采集时使用皮质类固醇对诊断面板的组成和准确性的影响，以及确定 cGVHD 中 BAFF 受体（BAFF-R）在 B 细胞表面低表达的机制。在一组发生 cGVHD 的成人移植后患者队列（n=44）和时间匹配的无 cGVHD 接受者（n=63）中进行了流式细胞术分析。我们证实 cGVHD 的发生与可溶性 BAFF（sBAFF）水平升高、CD27CD10CD21 CD19 B 细胞和经典转换记忆 B 细胞计数升高以及过渡性和幼稚 B 细胞计数降低有关。单一 B 细胞群体受试者工作特征（ROC）曲线下面积（AUC）最高的是 1 型转换性 CD21 B 细胞，为 0.72。我们还表明，由于 sBAFF 对 BAFF-R 的调节，sBAFF 与表面 BAFF-R 表达之间存在显著的负相关关系。在样本采集时使用类固醇会影响 sBAFF：B 细胞比值、幼稚和边缘区样 B 细胞的意义。在伴有或不伴有皮质类固醇使用的情况下，与 cGVHD 发生最显著相关的 B 细胞亚群的最佳组合导致 ROC AUC 分别为 0.87 和 0.84。过渡性和 CD21 B 细胞是两个面板中都存在的唯一群体；然而，仅分析这些群体导致 ROC AUC 分别为 0.79 和 0.78。这表明需要根据类固醇的使用情况包含其他群体和使用不同的面板，以达到更好的准确性。sBAFF 不是两个面板中的一个组成部分。这些新的 B 细胞谱可以在移植后患者中进行前瞻性测试，并可能导致有针对性的机制研究。
