文章基本信息

标题：Sortase-mediated modification of αDEC205 affords optimization of antigen presentation and immunization against a set of viral epitopes
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作者：Lee Kim Swee ; Carla P. Guimaraes ; Sharvan Sehrawat 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2013
卷号：110
期号：4
页码：1428-1433
DOI：10.1073/pnas.1214994110
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：A monoclonal antibody against the C-type lectin DEC205 (DEC205) is an effective vehicle for delivery of antigens to dendritic cells through creation of covalent DEC205-antigen adducts. These adducts can induce antigen-specific T-cell immune responses or tolerance. We exploit the transpeptidase activity of sortase to install modified peptides and protein-sized antigens onto the heavy chain of DEC205, including linkers that contain nonnatural amino acids. We demonstrate stoichiometric site-specific labeling on a scale not easily achievable by genetic fusions (49 distinct fusions in this report). We conjugated a biotinylated version of a class I MHC-restricted epitope to unlabeled DEC205 and monitored epitope generation upon binding of the adduct to dendritic cells. Our results show transfer of DEC205 heavy chain to the cytoplasm, followed by proteasomal degradation. Introduction of a labile dipeptide linker at the N terminus of a T-cell epitope improves proteasome-dependent class I MHC-restricted peptide cross-presentation when delivered by DEC205 in vitro and in vivo. We also conjugated DEC205 with a linker-optimized peptide library of known CD8 T-cell epitopes from the mouse {gamma}-herpes virus 68. Animals immunized with such conjugates displayed a 10-fold reduction in viral load.
关键词：antibody conjugates ; vaccines ; vaccine development ; protein engineering