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Establishing MHC Class I Peptide Motifs
Major histocompatibility complex (MHC) class I peptide motifs are used on a regular basis to identify and predict MHC class I ligands and CD8+ T-cell epitopes. This approach is above all an invaluable tool for the identification of disease-associated epitopes. As a matter of fact, the vast majority of T-cell epitopes discovered during the past two decades was identified by means of epitope prediction. Here we describe the steps which are necessary to establish MHC class I peptide motifs and to compose a reliable scoring matrix for epitope prediction. As an example, a scoring matrix for the prediction of HLA-B*35-presented ...
Source: Springer protocols feed by Immunology - December 3, 2012 Category: Allergy & Immunology Source Type: news

Biochemical Large-Scale Identification of MHC Class I Ligands
The large-scale identification of MHC class I presented peptides is indispensable for gaining insight into the fundamental rules of immune recognition as well as it is an invaluable tool in identifying potential targets for the immunotherapy of disease. In this chapter we briefly review the existing strategies for the analysis of MHC ligandomes and provide an in-depth protocol for the immunoaffinity purification of MHC class I presented peptides from primary tissues or cells. (Source: Springer protocols feed by Immunology)
Source: Springer protocols feed by Immunology - December 3, 2012 Category: Allergy & Immunology Source Type: news

Determining the Activity of the Transporter Associated with Antigen Processing in the Compartments of the Secretory Pathway
Peptide-receptive MHC class I molecules and the TAP (transporter associated with antigen processing) peptide transporter are known to leave the ER and cycle through the cis side of the Golgi apparatus. The amount, and the extent of the activity, of TAP in post-ER compartments is likely to vary between different cell types. Here we describe a convenient microscopic assay to determine it. (Source: Springer protocols feed by Immunology)
Source: Springer protocols feed by Immunology - December 3, 2012 Category: Allergy & Immunology Source Type: news

Evaluation of Viral Interference with MHC Class I-Restricted Antigen Processing and Presentation Using a Flow Cytometry-Based Approach
The peptide content of MHC class I molecules present at the cell surface is monitored by surveilling CD8+ cytotoxic T cells. In case of a viral infection, a proportion of the MHC class I molecules will carry peptides derived from viral proteins. This allows the CD8+ T cells to recognize and eliminate virus-infected cells. This highly sensitive detection system of the host is counteracted by viruses, which have acquired functions to downregulate cell surface expression of MHC class I molecules. In this chapter, we describe a flow cytometry-based method to identify viral gene product(s) responsible for evasion from MHC class...
Source: Springer protocols feed by Immunology - December 3, 2012 Category: Allergy & Immunology Source Type: news