Urea-Mediated Cross-Presentation of Soluble Epstein-Barr Virus BZLF1 Protein

PLoS Pathogens

Regina Gary¹  Birgit Weinberger²  Juha Lindner³  Tanja Bauer³  Petra Lindner³  Sascha Barabas³  Ludwig Deml³  Wolfgang Jilg³  Hans Wolf³ 

[1] Department of Hematology and Oncology, University of Erlangen-Nuernberg, Germany;Institute for Biomedical Aging Research, Austrian Academy of Sciences, Austria;Institute of Medical Microbiology, University of Regensburg, Germany

关键词: Urea;    T cells;    Antigen-presenting cells;    Cytotoxic T cells;    Enzyme-linked immunoassays;    Dendritic cells;    B cells;    Monocytes;   

DOI  :  10.1371/journal.ppat.1000198

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Soluble extracellular proteins usually do not enter the endogenous human leukocyte antigen (HLA) I–dependent presentation pathway of antigen-presenting cells, strictly impeding their applicability for the re-stimulation of protein-specific CD8+ cytotoxic T lymphocytes (CTL). Here we present for the Epstein-Barr virus (EBV) BZLF1 a novel strategy that facilitates protein translocation into antigen-presenting cells by its solubilisation in high molar urea and subsequent pulsing of cells in presence of low molar urea. Stimulation of PBMC from HLA-matched EBV-seropositive individuals with urea-treated BZLF1 but not untreated BZLF1 induces an efficient reactivation of BZLF1-specific CTL. Urea-treated BZLF1 (uBZLF1) enters antigen-presenting cells in a temperature-dependent manner by clathrin-mediated endocytosis and is processed by the proteasome into peptides that are bound to nascent HLA I molecules. Dendritic cells and monocytes but also B cells can cross-present uBZLF1 in vitro. The strategy described here has potential for use in the development of improved technologies for the monitoring of protein-specific CTL.

【 授权许可】

CC BY