effect of differential n-linked and o-linked mannosylation on recognition of fungal antigens by dendritic cells微分n-linked和o-linked mannosylation真菌抗原的树突细胞的识别.pdfVIP

Effect of Differential N-linked and O-linked Mannosylation on Recognition of Fungal Antigens by Dendritic Cells 1,2¤a 3 1,2,3¤b Jennifer S. Lam , Haibin Huang , Stuart M. Levitz * 1 Department of Medicine and the Immunology Training Program, Boston University School of Medicine, Boston, Massachusetts, United States of America, 2 Department of Microbiology and the Immunology Training Program, Boston University School of Medicine, Boston, Massachusetts, United States of America, 3 Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, United States of America Background. An experimental approach for improving vaccine efficacy involves targeting antigens to mannose receptors (MRs) on dendritic cells (DCs) and other professional antigen presenting cells. Previously, we demonstrated that mannosylated Pichia pastoris-derived recombinant proteins exhibited increased immunogenicity compared to proteins lacking mannosyla- tion. In order to gain insight into the mechanisms responsible for this observation, the present study examined the cellular uptake of the mannosylated and deglycosylated recombinant proteins. Methodology/Principal Findings. Utilizing transfected cell lines, roles for the macrophage mannose receptor (MMR, CD206) and DC-SIGN (CD209) in the recognition of the mannosylated, but not deglycosylated, antigens were demonstrated. The uptake of mannosylated antigens into murine bone marrow-derived DCs (BMDCs) was inhibited by yeast mannans (YMs), suggesting a mannose-specific C-type lectin receptor-dependent process, while the uptake of deglycosylated antigens remained unaffected. In particular, antigens with both N-linked and extensive O-linked mannosylation showed the highest binding and uptake by BMDCs. Finally, confocal microscopy studies revealed t