Plasma membrane proteomes of differentially matured dendritic cells identified by LC–MS/MS combined with iTRAQ labelling

• Published in: Journal of proteomics 2012-01, Vol.75 (3), p.938-948
• Main Authors: Ferret-Bernard, Stéphanie, Castro-Borges, William, Dowle, Adam A., Sanin, David E., Cook, Peter C., Turner, Joseph D., MacDonald, Andrew S., Thomas, Jerry R., Mountford, Adrian P.
• Format: Article
• Language: English
• Subjects: adenosinetriphosphatase; antigens; Biological and medical sciences; clathrin; Dendritic cell; dendritic cells; Diverse techniques; flow cytometry; Fundamental and applied biological sciences. Psychology; Immunology; iTRAQ; Life Sciences; malate dehydrogenase; metabolism; Molecular and cellular biology; Parasitic helminth; plasma membrane; Plasma membrane proteomics; protein folding; proteomics; pyruvate kinase; transporters
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2011.10.010

Dendritic cells (DCs) play a pivotal role in polarising Th lymphocyte subsets but it is unclear what molecular events occur when DCs generate Th2-type responses. Here, we analysed plasma membrane-enriched fractions from immature, pro-Th1 and pro-Th2 DCs and used a combination of iTRAQ labelling and LC–MS/MS to quantify changes in the proteomes. Analysis was performed on triplicate biological samples and changes verified by flow cytometry. MHC class II molecules and CD29 were up-regulated in pro-Th1 DCs whilst CD18 and CD44 were up-regulated in pro-Th2 DCs. One of the most down-regulated molecules in pro-Th1 DCs was YM-1 whilst the greatest decrease in pro-Th2 DCs was NAP-22. Other molecules up-regulated in pro-Th2 DC compared to pro-Th1 DCs included some potentially involved in protein folding during antigen processing (clathrin and Rab-7), whilst other non-membrane proteins such as enzymes/transporters related to cell metabolism (malate dehydrogenase, pyruvate kinase, and ATPase Na+/K+) were also recorded. This suggests that pro-Th2 DCs are more metabolically active while pro-Th1 DCs have a mature ‘end state’. Overall, although several molecules were preferentially expressed on pro-Th2 DCs, our proteomics data support the view of a ‘limited maturation’ of pro-Th2 DCs compared to pro-Th1 DCs. [Display omitted] ► Analysis of plasma membrane-enriched proteomes of differentially matured DCs. ► Relative expression determined by iTRAQ labelling combined with LC–MS/MS. ► Differential expression of immune associated proteins by pro-Th2 versus pro-Th1 DCs. ► Data support ‘limited maturation’ but not ‘unique’ phenotype of pro-Th2 DCs.