Dendritic Cell subsets: Functional analysis and TLR Phenotyping (98.13)

Abstract

Abstract In-depth analysis of subtypes of Dendritic Cells (DCs), both myeloid (mDC) and plasmacytoid (pDC) subsets, has been developed through Toll-like Receptor (TLR) Phenotyping by Flow Cytometry. Sets of TLR specific mAbs are combined with cell lineage and DC markers like CD11c and CD123 for mDC and pDC, in a simple assay for consistent DC analysis. DC subsets in peripheral blood differ in origin, phenotype, and function as analyzed by cytokines, chemokines or pro-inflammatory factors produced upon interaction with pathogens. DC response to invading pathogens depends upon expression of a spectrum of receptors, such as TLRs, which are expressed on the cell surface and in intracellular endosomes. TLR signaling in DC subtypes results in production of cytokines, chemokines and pro-inflammatory factors that elicit and control adaptive immune responses. TLR Phenotyping analysis includes development of 4 color Flow Cytometry assay cocktails for TLR expression of DCs to characterize mDC or pDC in human peripheral blood. In combination with activation and functional markers such as cytokines or chemokines the assay system provides a new research tool to identify changes in TLR expression in steady state or pathogen infected conditions and during differentiation of monocytes, DCs, B cells/B cell lines (EBV transformation), or T cells. The approach also provides additional information on TLR signaling and downstream pro-inflammatory mediator production.