Immunology Dpt. and Biostatistic Unit, Hospital Universitario Ramón y Cajal, IRYCIS, Ctra. de Colmenar Viejo km 9.100, 28034 Madrid, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
Immunology and Neurology Dpt., Universitat Pompeu Fabra, Institut Hospital del Mar d'Investigacions Mèdiques (IMIM), Carrer del Dr. Aiguader 88, 08003 Barcelona, Spain; Red Española de Esclerosis Múltiple (REEM), Spain.
Clin Chim Acta. 2019 Jan;488:135-142. doi: 10.1016/j.cca.2018.11.008. Epub 2018 Nov 5.
Percentages of blood CD19+CD5+ B cells and CD8+perforin+ T lymphocytes can predict response to Interferon (IFN)-beta treatment in relapsing-remitting multiple sclerosis (RRMS) patients. We aimed to standardize their detection in a multicenter study, prior to their implementation in clinical practice.
Fourteen hospitals participated in the study. A reference centre was established for comparison studies. Peripheral blood cells of 105 untreated RRMS patients were studied. Every sample was analyzed in duplicate in the participating centre and in the reference one by flow cytometry. When needed, participating centres corrected fluorescence compensations and negative cut-off position following reference centre suggestions. Concordance between results obtained by participating centres and by reference one was evaluated by intraclass correlation coefficients (ICC) and Spearman correlation test. Centre performance was measured by using z-scores values.
After results review and corrective actions implementation, overall ICC was 0.86 (CI: 0.81-0.91) for CD19+CD5+ B cell and 0.89 (CI: 0.85-0.93) for CD8+ perforin+ T cell quantification; Spearman r was 0.92 (0.89-0.95; p <0.0001) and 0.92 (0.88-0.95; p <0.0001) respectively. All centres obtained z-scores≤0.5 for both biomarkers.
Homogenous percentages of CD19+CD5+ B cells and CD8 perforin+ T lymphocytes can be obtained if suitable compensation values and negative cut-off are pre-established.
血液中 CD19+CD5+B 细胞和 CD8+穿孔素+T 淋巴细胞的百分比可预测复发缓解型多发性硬化症（RRMS）患者对干扰素（IFN）-β治疗的反应。我们旨在通过一项多中心研究来标准化这些检测，以便在临床实践中实施。
14 家医院参与了这项研究。建立了一个参考中心用于比较研究。研究了 105 例未经治疗的 RRMS 患者的外周血细胞。每个样本在参与中心和参考中心均由流式细胞术进行了两次分析。如果需要，参与中心根据参考中心的建议，对荧光补偿和阴性截止位置进行修正。采用组内相关系数（ICC）和 Spearman 相关检验评估参与中心和参考中心获得的结果的一致性。采用 z 分数值衡量中心的性能。
经过结果审查和校正措施的实施，CD19+CD5+B 细胞的总 ICC 为 0.86（95%CI：0.81-0.91），CD8+穿孔素+T 细胞的 ICC 为 0.89（95%CI：0.85-0.93）；Spearman r 分别为 0.92（0.89-0.95;p<0.0001）和 0.92（0.88-0.95;p<0.0001）。两个生物标志物的所有中心均获得 z 分数≤0.5。
如果预先确定合适的补偿值和阴性截止值，则可获得同质的 CD19+CD5+B 细胞和 CD8+穿孔素+T 淋巴细胞的百分比。
