Flow-Cytometric Immune Function Methodology

Flow-Cytometric Immune Function 41

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Flow-Cytometric Immune Function Methodologyfor Human Peripheral Blood Dendritic Cells

Kerstin Willmann

1. Introduction

The following describes the use of ow cytometry to detect the immune response of circulating CD11c+ and CD11c– (CD123 high, pre-DC2) dendritic cells on a single-cell level (1–4). Flow cytometry is an analytical and prepara-tive tool for immunology (5,6). Its use has rapidly advanced the accurate quanti- cation of peripheral blood cell populations. Today, it facilitates the assessment of patient diagnosis and treatment (7,8). Recent studies have demonstrated the importance of investigating not only the presence but also the functionality of speci c leukocyte subsets (9–12). Several dendritic cell (DC) populations are currently being investigated in various disease states, and the need to understand their functional roles is well recognized (13–22). The eld of DC biology has developed slowly over the past three decades. This is the result, in part, of their low frequency, the lack of speci c markers, and their fast turnover rate upon isolation (17,19,22,23). In spite of these limitations, the importance of DCs as the professional antigen-presenting cells of the immune system is well understood (17,Cs present antigen to naive and memory (4,17–19,22,26–28). DCs affect T-cell responses not only by direct contact during antigen presentation but also through cytokine production (21,29–31).The DC immune function assay is derived with modi cations from methods that have been described earlier to measure cytokine secretion in T cells and in monocytes (32–37). It presents a well-optimized tool to detect the immune

From: Methods in Molecular Biology, vol. 215:

Cytokines and Colony Stimulating Factors: Methods and ProtocolsEdited by: D. Körholz and W. Kiess © Humana Press Inc., Totowa, NJ

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