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A polychromatic antibody cocktail to investigate phenotypic and functional markers on human myeloid and plasmacytoid dendritic cells (65.3)
Journal article   Peer reviewed

A polychromatic antibody cocktail to investigate phenotypic and functional markers on human myeloid and plasmacytoid dendritic cells (65.3)

Pooja Jain, Michael Betts, Sharron Manuel and George Makedonas
The Journal of immunology (1950), v 186(1_Supplement), pp 65-65.3
01 Apr 2011
DOI: https://doi.org/10.4049/jimmunol.186.Supp.65.3

Abstract

Abstract Antigen presentation is a critical feature of adaptive immunity and essential in self versus nonself discrimination as well as in stimulating immune responses against pathogens. Dendritic cells (DCs) are considered to be the most potent antigen-presenting cells and have long been recognized as key regulators of the immune system. Because DCs are such crucial cells of the immune system, an extensive assessment of their quality in infected individuals is critical. Therefore, we have standardized a unique DC cocktail containing 12 different functional DC markers using an LSR II polychromatic flow cytometer. To this point, we have tested our newly standardized cocktail on the fresh, frozen, and cultured peripheral blood mononuclear cell (PBMCs) as well as on monocyte-derived DCs (MDDCs), a widely used surrogate system for primary blood DCs. This study emphasizes the practicality of utilizing frozen versus fresh cells, especially in rare disease states, wherein obtaining fresh material is virtually impossible. Furthermore, we focus on the feasibility of PBMCs compared to whole blood as starting material. To our knowledge this is the first comprehensive DC phenotyping protocol that uses an extensive list of functional markers (>8). Collectively, these investigations possess great potential to enable investigators to perform immune cell monitoring and develop diagnostic and therapeutic strategies.

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