TY - JOUR
T1 - Plasma membrane proteomes of differentially matured dendritic cells identified by LC-MS/MS combined with iTRAQ labelling
AU - Ferret-Bernard, Stéphanie
AU - Castro-Borges, William
AU - Dowle, Adam A.
AU - Sanin, David E.
AU - Cook, Peter
AU - Turner, Joseph D.
AU - MacDonald, Andrew S.
AU - Thomas, Jerry R.
AU - Mountford, Adrian P.
N1 - 071762, Wellcome Trust, United Kingdom072255, Wellcome Trust, United KingdomBB/C516328, Biotechnology and Biological Sciences Research Council, United KingdomBBS/B/08531, Biotechnology and Biological Sciences Research Council, United Kingdom
PY - 2012/1/4
Y1 - 2012/1/4
N2 - 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. © 2011.
AB - 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. © 2011.
KW - Dendritic cell
KW - ITRAQ
KW - Parasitic helminth
KW - Plasma membrane proteomics
U2 - 10.1016/j.jprot.2011.10.010
DO - 10.1016/j.jprot.2011.10.010
M3 - Article
C2 - 22040742
SN - 1874-3919
VL - 75
SP - 938
EP - 948
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 3
ER -