Abstract
Immune surveillance of infected or tumor cells by CD8+ T cells requires that MHC class I molecules present a diverse repertoire of peptides on the cell surface. Even a few copies of individual peptides among this mixture are sufficient for recognition by the antigen receptors of appropriate CD8+ T cells. Here we describe a method for biochemical analysis of the naturally processed peptides in living cells. The peptide mixture in cell extracts is fractionated using reverse phase high performance liquid chromatography and detected by the activation of CD8+ T cell hybridomas. The results provide information on the structure and amount of the peptides and yield insights into the mechanisms that generate the naturally processed peptides.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Shastri N, Cardinaud S, Schwab SR, Serwold T, Kunisawa J (2005) All the peptides that fit: the beginning, the middle, and the end of the MHC class I antigen-processing pathway. Immunol Rev 207:31–41
Falk K, Rotzschke O, Stevanovic S, Jung G, Rammensee H-G (1991) Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules. Nature 351:290–296
Van Bleek GM, Nathenson SG (1990) Isolation of an endogenously processed immunodominant viral peptide from the class I H-2Kb molecule. Nature 348:213–216
Rötzschke O, Falk K, Wallny HJ, Faath S, Rammensee HG (1990) Characterization of naturally occurring minor histocompatibility peptides including H-4 and H-Y. Science 249(4966):283–287
Rammensee HG, Bachmann J, Stevanovic S (1997) MHC ligands and peptide motifs. Landes Bioscience, Austin, TX
Kunisawa J, Shastri N (2003) The group II chaperonin TRiC protects proteolytic intermediates from degradation in the MHC class I antigen processing pathway. Mol Cell 12(3):565–576
Kunisawa J, Shastri N (2006) Hsp90alpha chaperones large proteolytic intermediates in the MHC class I antigen processing pathway. Immunity 24:523–534
Falk K, Rötzschke O, Rammensee HG (1990) Cellular peptide composition governed by major histocompatibility complex class I molecules. Nature 348(6298):248–251
Malarkannan S, Goth S, Buchholz DR, Shastri N (1995) The role of MHC class I molecules in the generation of endogenous peptide/MHC complexes. J Immunol 154(2):585–598
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Kanaseki, T., Shastri, N. (2013). Biochemical Analysis of Naturally Processed Antigenic Peptides Presented by MHC Class I Molecules. In: van Endert, P. (eds) Antigen Processing. Methods in Molecular Biology™, vol 960. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-218-6_15
Download citation
DOI: https://doi.org/10.1007/978-1-62703-218-6_15
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-217-9
Online ISBN: 978-1-62703-218-6
eBook Packages: Springer Protocols