title: A quantitative proteomic approach for detecting protein profiles of activated human myeloid dendritic cells.
abstract: Dendritic cells (DC) direct the magnitude, polarity and effector function of the adaptive immune response. DC express toll-like receptors (TLR), antigen capturing and processing machinery, and costimulatory molecules, which facilitate innate sensing and T cell activation. Once activated, DC can efficiently migrate to lymphoid tissue and prime T cell responses. Therefore, DC play an integral role as mediators of the immune response to multiple pathogens. Elucidating the molecular mechanisms involved in DC activation is therefore central in gaining an understanding of host response to infection. Unfortunately, technical constraints have limited system-wide 'omic' analysis of human DC subsets collected ex vivo. Here we have applied novel proteomic approaches to human myeloid dendritic cells (mDCs) purified from 100 mL of peripheral blood to characterize specific molecular networks of cell activation at the individual patient level, and have successfully quantified over 700 proteins from individual samples containing as little as 200,000 mDCs. The proteomic and network readouts after ex vivo stimulation of mDCs with TLR3 agonists are measured and verified using flow cytometry.
authors.affiliation: Center for Proteomics and Bioinformatics, Case Western Reserve University, Cleveland, OH 44106, United States. dms73@case.edu
authors.forename: Daniela M
authors.initials: DM
authors.lastname: Schlatzer
authors.forename: Julia
authors.initials: J
authors.lastname: Sugalski
authors.forename: Jean-Eudes
authors.initials: JE
authors.lastname: Dazard
authors.forename: Mark R
authors.initials: MR
authors.lastname: Chance
authors.forename: Donald D
authors.initials: DD
authors.lastname: Anthony
mesh_terms: D000328:Adult; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D022423:Myeloid Cells / Q000276:immunology* / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D051196:Toll-Like Receptor 3 / Q000276:immunology / Q000378:metabolism
chemical_list: C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3