Annals of Thoracic Medicine Official publication of the Saudi Thoracic Society, affiliated to King Saud University
 
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ORIGINAL ARTICLE
Year : 2015  |  Volume : 10  |  Issue : 4  |  Page : 279-283

Rapid detection of circulating fibrocytes by flowcytometry in idiopathic pulmonary fibrosis


1 Department of Medicine, King Khalid University Hospital, College of Medicine, King Saud University, Riyadh, Canada
2 Department of Pathology, King Khalid University Hospital, College of Medicine, King Saud University, Riyadh, Canada
3 Firestone Institute for Respiratory Health, Department of Medicine, McMaster University, Hamilton, Ontario, Canada

Correspondence Address:
Esam H Alhamad
Pulmonary Division, Department of Medicine (38), College of Medicine, King Saud University, P. O. Box-2925, Riyadh-11461
Canada
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DOI: 10.4103/1817-1737.157294

PMID: 26664567

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Background: 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. Objective: 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. Methods: 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. Results: 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). Conclusions: 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.


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