Rapid detection of circulating fibrocytes by flowcytometry in idiopathic pulmonary fibrosis.
Rapid detection of circulating fibrocytes by flowcytometry in idiopathic pulmonary fibrosis.
复制标题
特发性肺纤维化中流量术快速检测循环纤维细胞。
DOI：
10.4103/1817-1737.157294
复制
发表时间：
2015-10
影响因子：
2.3
通讯作者：
Kolb M
中科院分区：
文献类型：
--
作者：
Alhamad EH;Shakoor Z;Al-Kassimi FA;Almogren A;Gad ElRab MO;Maharaj S;Kolb M
目前用于检测各种肺部和非肺部疾病中描述的外周血中的循环纤维细胞（CF）的方案涉及复杂且耗时的非标准化技术。在特发性肺纤维化（IPF）患者和健康对照者中测试使用基于全血裂解流式细胞术的测定快速检测和定量CF的方法。从33名IPF患者和35名健康对照受试者中采集1毫升乙二胺四乙酸（EDTA）静脉血样。采用全血裂解法，用细胞表面单克隆抗体（CD 34和CD 45）和细胞内标记物（胶原-1）标记外周血白细胞，用于流式细胞术分析。CF定义为共表达I型胶原和CD 34分子的CD 45+细胞。在29例（87.8%）IPF患者和10例（28.5%）对照受试者中，在由CD 45+细胞的侧向散射特性生成的点图中检测到定义明确的高度颗粒状CD 45+细胞群。这些CD 45+细胞被确定为CF的基础上，共表达的胶原蛋白-I和CD 34，没有其他类型的细胞在外周血中标记这些单克隆抗体。IPF患者的CF百分比显著高于健康对照（中位数（范围）：分别为1.37%（0.52-5.65）和1.04%（0.1-1.84）; P = 0.03）。全血裂解法结合荧光激活细胞分选术（FACS）可以检测到明确定义的均匀CF群体。该方法简单，重现性好，可准确快速地估计CF。
Current protocols for detection of circulating fibrocytes (CFs) in peripheral blood described in various pulmonary and nonpulmonary disorders involve complex and time consuming, non standardized techniques. Testing a method to rapidly detect and quantify CFs using whole blood lysis flow cytometry-based assay in patients with idiopathic pulmonary fibrosis (IPF) and healthy controls. One milliliter of venous blood sample in ethylenediaminetetraacetic acid (EDTA) from 33 IPF patients and 35 healthy control subjects was collected. Using whole blood lysis method peripheral blood leukocytes were labeled with monoclonal antibodies for cell surface (CD34 and CD45) and intracellular markers (collagen-1) for flow cytometric analysis. CFs were defined as CD45+ cells coexpressing collagen-I and CD34 molecules. In 29 (87.8%) IPF patients and 10 (28.5%) control subjects, a well-defined highly granular CD45+ cell population was detected in dot plots generated by side scatter properties of CD45+ cells. These CD45+ cells were identified as CFs on the basis of coexpression of collagen-I and CD34; none of the other cell types in the peripheral blood were labeled with these monoclonal antibodies. In IPF patients the percentage of CFs was significantly higher compared to healthy controls (median (range): 1.37% (0.52-5.65) and 1.04% (0.1-1.84), respectively; P = 0.03). Whole blood lysis method combined with fluorescence-activated cell sorting (FACS) allows detecting a well-defined homogeneous population of CFs. This method is simple, reproducible, and provides an accurate and rapid estimation of CFs.