alexa Caveats in the design of MHC class I tetramer antigen-specific T lymphocytes dissociation assays.
Microbiology

Microbiology

Journal of Microbial & Biochemical Technology

Author(s): Wang XL, Altman JD

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Abstract Relative avidities of antigen-specific T cells for major histocompatibility complex peptide complexes (MHCp) have recently been measured by MHC tetramer dissociation assays, but there is no consensus on the methodologies used for these experiments. While we do not question the conclusions reached in previous studies, in this paper we discuss the caveats that are present in the design of all MHCp tetramer dissociation protocols, and we propose a set of criteria that should be met in the evaluation of appropriate methodologies. We find that it is necessary to use specific reagents to compete with rebinding of the labeled tetramer, but that when either intact anti-MHC antibodies or cold MHC tetramers are used, the dissociation rates are dependent upon the concentrations of the competitor. In contrast, we demonstrate that apparent dissociation rates are independent of the competitor concentration when blocking anti-MHC Fab fragments are used, suggesting that these are the most appropriate reagents to use for tetramer dissociation experiments.
This article was published in J Immunol Methods and referenced in Journal of Microbial & Biochemical Technology

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