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

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 C

AU - Turner, Joseph D

AU - MacDonald, Andrew S

AU - Thomas, Jerry R

AU - Mountford, Adrian P

N1 - Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.

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.

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.

UR - http://www.scopus.com/inward/record.url?scp=84355166546&partnerID=8YFLogxK

U2 - 10.1016/j.jprot.2011.10.010

DO - 10.1016/j.jprot.2011.10.010

M3 - Article

C2 - 22040742

VL - 75

SP - 938

EP - 948

JO - JOURNAL OF PROTEOMICS

JF - JOURNAL OF PROTEOMICS

IS - 3

ER -