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

作者：Barabas Sascha^*; Gary Regina; Bauer Tanja; Lindner Juha; Lindner Petra; Weinberger Birgit; Jilg Wolfgang; Wolf Hans; Deml Ludwig

来源：PLoS Pathogens, 2008, 4(11): e1000198.

DOI：10.1371/journal.ppat.1000198

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.

• 出版日期2008-11
• 单位河北医科大学