A Six-Color Flow Cytometric Assay for the Analysis of Peripheral Blood Dendritic Cells

被引：20

作者：

Giannelli, Stefania ^{[1
]}

Taddeo, Adriano ^{[1
]}

Presicce, Pietro ^{[1
]}

Villa, Maria Luisa ^{[1
]}

Della Bella, Silvia ^{[1
]}

机构：

[1] Univ Milan, Immunol Lab, Dipartimento Sci & Tecnol Biomed, LITA, I-20090 Segrate, MI, Italy

来源：

CYTOMETRY PART B-CLINICAL CYTOMETRY | 2008年 / 74B卷 / 06期

关键词：

peripheral blood dendritic cells; myeloid dendritic cells; plasmacytoid dendritic cells; costimulatory molecules; multiparametric flow cytometry;

D O I：

10.1002/cyto.b.20434

中图分类号：

R446 [实验室诊断]; R-33 [实验医学、医学实验];

学科分类号：

1001 ;

摘要：

Background: Flow cytometric analysis of peripheral blood dendritic cells (PBDCs) and their myeloid (mDCs) and plasmacytoid (pDCs) subsets is a less invasive procedure that is acquiring growing clinical relevance. Because dendritic cells (DCs) lack unique lineage markers, current methods that are based on 3- or 4-color assays do not allow multiparametric analysis of DC subsets. In this study a dedicated 6-color assay was developed. Methods: mDCs and pDCs were counted and characterized for the expression of activation/maturation markers by using a single-platform 6-color assay. Whole-blood samples from 20 healthy controls were directly stained with either C080-FITC/CD40-PUlineage-PerCP-Cy5.5/CD123-PE-Cy7/CD11c-APC/HLA-DR-AFC-Cy7 or CD86-FITC/CD83-PE/lineage-PerCP-Cy5.5/CD123-PE-Cy7/CDllc-APC/HLA-DR-APC-Cy7 combination, in the presence of commercial fluorospheres. A dual-platform 3-color assay currently in use was run in parallel for comparison. Results: The 6-color assay provided mDCs and pDCs counts similar to counts obtained by the 3-color assay. Only the 6-color assay could show differential expression of activation markers by mDCs and pDCs, with pDCs expressing lower levels of costimulatory molecules and HLA-DR, but higher levels of CD83. Conclusions: The 6-color assay described here may be a sensitive tool for assessing possible variations in the number and features of mDCs and pDCs whose reciprocal balance is critical in understanding the more detailed orchestration of immune responses. 0 2008 Clinical Cytometry Society

引用

页码：349 / 355

页数：7

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