背景:循环纤维细胞数量增加与哮喘控制不充分相关,对现有疗法反应不佳,并增加不良后果的风险。缺乏可靠且临床适用的检测方法,因此无法对作为哮喘预后生物标志物的血细胞计数进行适当评估。该报告涉及使用多参数流式细胞术测定法对全血中的纤维细胞进行计数。
方法:同意的纤维细胞供体是19例哮喘患者,通过目前的治疗得到良好控制,16例难治性哮喘患者,9例短暂不受控制的哮喘患者和14例年龄匹配的正常人。对短暂未控制的哮喘患者进行一次血液采样,两次,间隔一周,在其他科目。该测定在100μl全血中进行,并涉及顺序门控策略和使用单个仪器的绝对纤维细胞计数(单平台测定)。
结果:通过该测定的循环纤维细胞的定量在分析和临床上是有效的。在临床状况稳定的个体中,一周以上的血细胞计数重复性良好.组内相关系数为总变异性的0.939和96.88%,反映了受试对象之间的平均差异。稳定的血液样品可以在处理之前在4°C下储存长达96小时。
结论:用于全血中纤维细胞计数的新测定法是可靠的且临床适用的。
结论:本报告证明了在多中心临床试验中循环纤维细胞计数的第一个优化试验的有效性和可靠性。
BACKGROUND: Elevated numbers of circulating fibrocytes are associated with inadequately controlled asthma, poor response to available therapies, and increased risk of adverse outcomes. The lack of reliable and clinically-applicable assays precludes a proper evaluation of blood fibrocyte count as a prognostic biomarker in asthma. This report concerns the use of a multiparameter flow cytometry assay for the enumeration of fibrocytes in the whole blood.
METHODS: Consenting fibrocyte donors were 19 patients with asthma well controlled by current treatment, 16 patients with treatment-resistant asthma, 9 patients with transiently uncontrolled asthma and 14 age-matched normal individuals. Blood sampling was performed once in patients with transiently uncontrolled asthma and twice, at an interval of one week, in the other subjects. The assay was performed in 100 μl of whole blood and involved a sequential gating strategy and absolute fibrocyte counting with a single instrument (single-platform assay).
RESULTS: The quantification of circulating fibrocytes by this assay was analytically and clinically valid. In individuals with stable clinical conditions, the repeatability of blood fibrocyte counts over one week was good. The intraclass correlation coefficient was 0.939 and 96.88% of the total variability reflected on-average differences among the tested subjects. Stabilized blood samples could be stored at 4 °C for up to 96 h before processing.
CONCLUSIONS: The novel assay for the enumeration of fibrocytes in the whole blood is reliable and clinically applicable.
CONCLUSIONS: This report demonstrates the validity and reliability of the first optimized assay for the enumeration of circulating fibrocytes in multicenter clinical trials.