Multiplexed quantitative proteomics of CD4 T cell-mediated postsynaptic dendritic cells (DC)

We aimed to characterize the proteome remodeling in DCs induced upon antigen presentation. To do so, we used an in vitro antigen presentation model to generate postsynaptic (psDC) or nonsynaptic (nsDC) DCs similarly to previously reported (Alcaraz-Serna et al., 2021). Bone marrow-derived DCs (BMDCs), which had been activated with LPS and pulsed (psDC) or not (nsDC) with the ovalbumin peptide OVA323-339, were co-cultured with resting CD4+ T cells from OT-II mice to allow cognate antigen-dependent immune synapse formation (psDC) or not (nsDC). DCs were then purified for proteomic analysis by using mass spectrometry based on multiplexed isotopic labeling peptides approach.