Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells.
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 immu...
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doaj-77c62d1fcacc4a8abc89a8ed4aff8c582020-11-24T22:00:30ZengPublic Library of Science (PLoS)PLoS ONE1932-62032007-10-01210e100910.1371/journal.pone.0001009Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells.Jennifer S LamHaibin HuangStuart M LevitzAn 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 mannosylation. 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.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 that both mannosylated and deglycosylated P. pastoris-derived recombinant proteins localized in MHC class II+ compartments within BMDCs.Taken together with our previous results, these data suggest that increased uptake by mannose-specific C-type lectin receptors is the major mechanism responsible for the enhanced antigenicity seen with mannosylated proteins. These findings have important implications for vaccine design and contribute to our understanding of how glycosylation affects the immune response to eukaryotic pathogens.http://europepmc.org/articles/PMC1995759?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jennifer S Lam Haibin Huang Stuart M Levitz |
spellingShingle |
Jennifer S Lam Haibin Huang Stuart M Levitz Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. PLoS ONE |
author_facet |
Jennifer S Lam Haibin Huang Stuart M Levitz |
author_sort |
Jennifer S Lam |
title |
Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. |
title_short |
Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. |
title_full |
Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. |
title_fullStr |
Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. |
title_full_unstemmed |
Effect of differential N-linked and O-linked mannosylation on recognition of fungal antigens by dendritic cells. |
title_sort |
effect of differential n-linked and o-linked mannosylation on recognition of fungal antigens by dendritic cells. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2007-10-01 |
description |
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 mannosylation. 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.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 that both mannosylated and deglycosylated P. pastoris-derived recombinant proteins localized in MHC class II+ compartments within BMDCs.Taken together with our previous results, these data suggest that increased uptake by mannose-specific C-type lectin receptors is the major mechanism responsible for the enhanced antigenicity seen with mannosylated proteins. These findings have important implications for vaccine design and contribute to our understanding of how glycosylation affects the immune response to eukaryotic pathogens. |
url |
http://europepmc.org/articles/PMC1995759?pdf=render |
work_keys_str_mv |
AT jenniferslam effectofdifferentialnlinkedandolinkedmannosylationonrecognitionoffungalantigensbydendriticcells AT haibinhuang effectofdifferentialnlinkedandolinkedmannosylationonrecognitionoffungalantigensbydendriticcells AT stuartmlevitz effectofdifferentialnlinkedandolinkedmannosylationonrecognitionoffungalantigensbydendriticcells |
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