MCP Tips for better browsing
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

A more recent version of this article appeared on May 1, 2006.
This Article
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
T500014-MCP200v1
5/5/902    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Glossary
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Meiring, H. D.
Right arrow Articles by van Els, C. A C. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Meiring, H. D.
Right arrow Articles by van Els, C. A C. M.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Submitted on June 29, 2005
Revised on January 18, 2006
Accepted on January 23, 2006

Stable isotope tagging of epitopes-a highly selective strategy for the identification of MHC class I-associated peptides induced upon viral infection

H. D. Meiring, E. C. Soethout, M C. M. Poelen, D. Mooibroek, R. Hoogerbrugge, H. Timmermans, C. J. Boog, A. J. R. Heck, A. P. J. M. de Jong, and C. A C. M. van Els

Vaccine Research, Netherlands Vaccine Institute, Bilthoven 3720 AL

Corresponding Author: Ernst.Soethout{at}nvi-vaccin.nl

Identification of peptides presented in MHC class I molecules after viral infection is of strategic importance for vaccine development. Until recently, mass spectrometric identification of virus-induced peptides was based on comparative analysis of peptide pools isolated from uninfected and virus-infected cells. Here, we report on a powerful strategy aiming at the rapid, unambiguous identification of naturally processed MHC class I-associated peptides, which are induced by viral infection. The methodology, Stable Isotope Tagging of Epitopes (SITE), is based on metabolic labeling of endogenously synthesized proteins during infection. This is accomplished by culturing virus-infected cells with stable isotope labeled amino acids that are expected to be anchor residues (i.e. residues of the peptide that have amino acid side chains that bind into pockets lining the peptide-binding groove of the MHC class I molecule) for the HLA allele of interest. Subsequently, these cells are mixed with an equal number of non-infected cells, which are cultured in normal medium. Finally, peptides are acid-eluted from immunoprecipitated MHC molecules and subjected to 2-dimensional nanoscale LC-MS analysis. Virus-induced peptides are identified through computer-assisted detection of characteristic, binomially distributed ratios of labeled and unlabeled molecules. Using this approach we identified novel measles virus and respiratory syncytial virus epitopes as well as infection-induced self-peptides in several cell types, showing that SITE is a unique and versatile method for unequivocal identification of disease-related MHC class I epitopes.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Immunol.Home page
V. S. Meyer, W. Kastenmuller, G. Gasteiger, M. Franz-Wachtel, T. Lamkemeyer, H.-G. Rammensee, S. Stevanovic, D. Sigurdardottir, and I. Drexler
Long-Term Immunity against Actual Poxviral HLA Ligands as Identified by Differential Stable Isotope Labeling
J. Immunol., November 1, 2008; 181(9): 6371 - 6383.
[Abstract] [Full Text] [PDF]

Home page
 JEMHome page
M.-H. Fortier, E. Caron, M.-P. Hardy, G. Voisin, S. Lemieux, C. Perreault, and P. Thibault
The MHC class I peptide repertoire is molded by the transcriptome
J. Exp. Med., March 17, 2008; 205(3): 595 - 610.
[Abstract] [Full Text] [PDF]

Home page
 J. Immunol.Home page
J. Heidema, M. V. Lukens, W. W. C. van Maren, M. E. A. van Dijk, H. G. Otten, A. J. van Vught, D. B. M. van der Werff, S. J. P. van Gestel, M. G. Semple, R. L. Smyth, et al.
CD8+ T Cell Responses in Bronchoalveolar Lavage Fluid and Peripheral Blood Mononuclear Cells of Infants with Severe Primary Respiratory Syncytial Virus Infections
J. Immunol., December 15, 2007; 179(12): 8410 - 8417.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
M. Marcilla, J. J. Cragnolini, and J. A. Lopez de Castro
Proteasome-independent HLA-B27 Ligands Arise Mainly from Small Basic Proteins
Mol. Cell. Proteomics, May 1, 2007; 6(5): 923 - 938.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
 All ASBMB Journals   Journal of Biological Chemistry 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.