Guangming Li1,2, Liang Cheng3, Lishan Su1,2. 1. Division of Virology, Pathogenesis and Cancer, Institute of Human Virology, Baltimore, Maryland. 2. Department of Pharmacology, University of Maryland School of Medicine, Baltimore, Maryland. 3. Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Wuhan University, Wuhan, China.
Abstract
Plasmacytoid dendritic cells (pDCs) are a distinct lineage of bone-marrow-derived cells that reside mainly in blood and lymphoid organs in the steady state but are also present in sites of infection, inflammation, and cancer. The protocols in this article describes (1) detection and quantification of human pDCs in peripheral blood; (2) isolation of human pDCs by magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS); (3) evaluation of human pDC function by stimulation with TLR7 or TLR9 agonists; (4) detection of human pDCs in lymphoid tissues of humanized mice (hu-mice) by flow cytometry; (5) functional study of human pDC in hu-mice in vivo; and (6) specific depletion of human pDCs in vivo in hu-mice using monoclonal antibody targeting human pDCs. These assays thus provide comprehensive methods for phenotypic and functional studies in vitro and for the investigation of human plasmacytoid dendritic cells in hu-mice in vivo.
Plasmacytoid dendritic cells (pDCs) are a distinct lineage of bone-marrow-derived cells that reside mainly in blood and lymphoid organs in the steady state but are also present in sites of infection, inflammation, and cancer. The protocols in this article describes (1) detection and quantification of human pDCs in peripheral blood; (2) isolation of human pDCs by magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS); (3) evaluation of human pDC function by stimulation with TLR7 or TLR9 agonists; (4) detection of human pDCs in lymphoid tissues of humanized mice (hu-mice) by flow cytometry; (5) functional study of human pDC in hu-mice in vivo; and (6) specific depletion of human pDCs in vivo in hu-mice using monoclonal antibody targeting human pDCs. These assays thus provide comprehensive methods for phenotypic and functional studies in vitro and for the investigation of human plasmacytoid dendritic cells in hu-mice in vivo.
Authors: F P Siegal; N Kadowaki; M Shodell; P A Fitzgerald-Bocarsly; K Shah; S Ho; S Antonenko; Y J Liu Journal: Science Date: 1999-06-11 Impact factor: 47.728
Authors: Nicolin Bloch; Meagan O'Brien; Thomas D Norton; Sylvie B Polsky; Nina Bhardwaj; Nathaniel R Landau Journal: AIDS Res Hum Retroviruses Date: 2013-09-04 Impact factor: 2.205
Authors: Curdin Conrad; Josh Gregorio; Yi-Hong Wang; Tomoki Ito; Stephan Meller; Shino Hanabuchi; Sonya Anderson; Neely Atkinson; Pedro T Ramirez; Yong-Jun Liu; Ralph Freedman; Michel Gilliet Journal: Cancer Res Date: 2012-07-31 Impact factor: 12.701
Authors: Calvin C Lo; Jordan A Schwartz; Dylan J Johnson; Monica Yu; Nasra Aidarus; Shariq Mujib; Erika Benko; Martin Hyrcza; Colin Kovacs; Mario A Ostrowski Journal: PLoS One Date: 2012-05-31 Impact factor: 3.240