Effects of Interleukin-10 on the Protein Expression of Human Mature Dendritic Cells

Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.In view of their exceptional ability to present antigen and to interact with T cells,DC play distinct roles in linking innate and adaptive immune responses and thus become logical targets for cancer immunotherapy.Evidences show that tumor-derived cytokines could impair DCs'biomechanics properties,which lead to inefficacy of DCs-based immune therapies.Our previous studies found that IL-10,as one of the widespread suppressive cytokines from tumor microenvironment(TME),could deteriorate DCs'motility and biomechanics properties while the underlying mechanism is unknown.In this study,CD14+monocytes were induced to differentiate into mature dendritic cells after isolation in vitro using recombinant cytokines IL-4,GM-CSF,LPS,and IFN-γ.And we also have compared the proteomic changes of mDCs treated by IL-10 and control group via two-dimensional electrophoresis combine with MALDI-TOF/TOF MS.Then we analyzed the function of differentially expressed proteins through bioinformatics methods include GO analysis that clarified the biological functions of differential proteins and KEGG analysis which enriched signal pathways of differential proteins to explore the molecular mechanism of IL-10 which has inhibitory effect on mDCs.The results showed that IL-10 significantly affected the morphology of mDCs,especially reducing the number and length of filopodia.Different expressed proteins were analyzed by two-dimensional electrophoresis combined with bioinformatics analysis to enrich for glycolytic signaling pathway,HIF-1 signaling pathway and cytoskeletal binding protein expression changes.The results of two-dimensional electrophoresis were verified by Western blot,and the results showed that the data were reliable.In addition,the intracellular ROS levels were significantly higher in mDCs treated with IL-10,validating the previously enriched HIF-1 signaling pathway.In summary,it indicated that IL-10 may interfered with the oxidative metabolic process,glycolytic metabolism,and expression of cytoskeleton-related proteins in mDCs,and disturbances in these physiological processes resulted in reduced biomechanics properties and motility of mDCs and subsequently impaired their immune functions,making DC-based tumor vaccines less effective than which we desired.Our study reveals alterations in the physiological metabolism of mDCs under IL-10 treatment from the proteome,which lays the foundation for further exploration of the altered state of mDCs in the tumor microenvironment.