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Title: Dynamic quantification of MHC class I-peptide presentation to CD8+ T cells via intracellular cytokine staining.
Austin Authors: Pang, Ken C;Wei, Joe Q Z;Chen, Weisan
Affiliation: T cell Laboratory, Ludwig Institute for Cancer Research, Melbourne Branch, Austin Health, Heidelberg, VIC, 3084, Australia
Issue Date: 20-Feb-2006
Publication information: Journal of Immunological Methods 2006; 311(1-2): 12-8
Abstract: In order to further our basic understanding of antigen processing and presentation as well as to translate that knowledge into clinically effective vaccines and immunotherapies, having appropriate tools to study MHC class I-peptide presentation is highly desirable. Current methods are based upon HPLC fractionation of extracted peptides, monoclonal Ab, multivalent T cell receptors (TCR), T cell hybridomas, TCR transgenic cells, and T cell lines. However, each of these is associated with problems that make them either difficult to apply generally or too insensitive to adequately quantitate antigen presentation. We have developed a method based upon intracellular cytokine staining (ICS) that dynamically and relatively quantitates MHC class I-peptide presentation to CD8+ T cells in a manner that is both widely applicable and highly sensitive. It is well-suited to assess antigen presentation in its early stages, does not require fixation nor labeling of antigen presenting cells (APC), can be used to examine cross-presentation, and is able to directly employ ex vivo T cells which obviates the need for the development and maintenance of T cell lines and hybridomas. Our method represents a simple yet powerful tool that others interested in studying antigen processing and presentation should find of great practical value.
Gov't Doc #: 16516224
DOI: 10.1016/j.jim.2006.01.008
Journal: Journal of immunological methods
Type: Journal Article
Subjects: Animals
Antigen Presentation.immunology
Brefeldin A.chemistry
CD8-Positive T-Lymphocytes.immunology
Coloring Agents.chemistry
Cytokines.immunology.isolation & purification
Cytotoxicity Tests, Immunologic
Dendritic Cells.immunology
Flow Cytometry
Histocompatibility Antigens Class I.immunology
Influenza A virus.immunology
Mice, Inbred BALB C
Mice, Inbred C57BL
Vaccinia virus.immunology
Appears in Collections:Journal articles

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