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Cell surface levels of MHC II loaded with peptide is controlled by ubiquitination. In this study, we have examined how MHC II ubiquitination impacts immunity using MHC IIKRKI\/KI mice expressing mutant MHC II molecules that are unable to be ubiquitinated. Numbers of conventional DC (cDC) 1, cDC2 and plasmacytoid DCs were significantly reduced in MHC IIKRKI\/KI spleen, with the remaining MHC IIKRKI\/KI DCs expressing an altered surface phenotype. Whereas Ag uptake, endosomal pH, and cathepsin protease activity were unaltered, MHC IIKRKI\/KI cDC1 produced increased inflammatory cytokines and possessed defects in Ag proteolysis. Immunization of MHC IIKRKI\/KI mice identified impairments in MHC II and MHC class I presentation of soluble, cell-associated and\/or DC-targeted OVA via mAb specific for DC surface receptor Clec9A (anti-Clec9A-OVA mAb). Reduced T cell responses and impaired CTL killing was observed in MHC IIKRKI\/KI mice following immunization with cell-associated and anti-Clec9A-OVA. Immunization of MHC IIKRKI\/KI mice failed to elicit follicular Th cell responses and generated barely detectable Ab to anti-Clec9A mAb-targeted Ag. In summary, MHC II ubiquitination in DCs impacts the homeostasis, phenotype, cytokine production, and Ag proteolysis by DCs with consequences for Ag presentation and T cell and Ab-mediated immunity.<\/jats:p>","DOI":"10.4049\/jimmunol.2001426","type":"journal-article","created":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T16:50:36Z","timestamp":1633107036000},"page":"2255-2264","source":"Crossref","is-referenced-by-count":20,"title":["MHC Class II Ubiquitination Regulates Dendritic Cell Function and Immunity"],"prefix":"10.1093","volume":"207","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2023-7964","authenticated-orcid":false,"given":"Kayla R","family":"Wilson","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne , Parkville, Victoria 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