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Utilizing pMHC Multimers to Study Antigen-Specific T Cells and T Cell Exhaustion in Chronic Viral Infections
K Keetz1, M Knapp1, J Schulze Zur Wiesch1
1Infectious Diseases Unit, I. Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
This chapter presents a detailed protocol for peptide-major histocompatibility complex (pMHC) multimer staining and magnetic activated cell separation, offering a step-by-step guide to isolating and studying low-frequency antigen-specific T cells. This methodological approach provides a means to evaluate the phenotype and function of antigen-specific T cells, potentially facilitating the assessment of novel therapeutic interventions and aiding in understanding T cell exhaustion in chronic viral diseases. Despite challenges such as the necessity of knowing the epitope and HLA restrictions as well as the intricate nature of the pMHC multimer staining process, this protocol serves as a fundamental tool for both basic research and translational applications in chronic viral infections.
This chapter presents a detailed protocol for peptide-major histocompatibility complex (pMHC) multimer staining and magnetic activated cell separation, offering a step-by-step guide to isolating and studying low-frequency antigen-specific T cells. This methodological approach provides a means to evaluate the phenotype and function of antigen-specific T cells, potentially facilitating the assessment of novel therapeutic interventions and aiding in understanding T cell exhaustion in chronic viral diseases. Despite challenges such as the necessity of knowing the epitope and HLA restrictions as well as the intricate nature of the pMHC multimer staining process, this protocol serves as a fundamental tool for both basic research and translational applications in chronic viral infections.
