Testing for HLA/peptide tetramer-binding to the T cell receptor complex on human T lymphocytes

• Published in: Results in immunology Vol. 2; pp. 88 - 96
• Main Authors: Weder, Pauline, Schumacher, Ton N.M., Spits, Hergen, Luiten, Rosalie M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 2012; Elsevier
• Subjects: Antigen-specificity; HLA/peptide tetramer; Immune response; Patients; T lymphocytes; T lymphocytes; MFI; HLA/peptide tetramer; Immune response; APC; PE; GAM-Ig; Patients; mAb; Antigen-specificity; APC, allophycocyanin; PE, phycoerythrin; GAM-Ig, goat anti-mouse immunoglobulin; mAb, monoclonal antibody; MFI, mean fluorescence intensity
• ISSN: 2211-2839; 2211-2839
• DOI: 10.1016/j.rinim.2012.04.001

HLA/peptide tetramers are frequently used for ex vivo monitoring of disease- or vaccine-induced T cell immune responses and for T cell epitope identification. However, when low-levels HLA/peptide tetramer-positive T cell populations are encountered, it is difficult to ascertain whether this represents a true T cell receptor (TCR)-mediated interaction or background signal. To address this issue, we have developed a method for both HLA class I and class II tetramer assays to confirm tetramer-binding to the TCR/CD3 complex. Preincubation of T cells with anti-CD3 mAb SPV-T3b and subsequent crosslinking interferes with the binding of HLA/peptide tetramers to the TCR/CD3 complex and thereby indicates to what extent HLA/peptide tetramer binds through interaction with TCR/CD3 complex. SPV-T3b pretreatment results in a 2- to 10-fold decrease in tetramer-binding intensity to antigen-specific CD8+ or CD4+ T cells, whereas background reactivity of HLA/peptide tetramers containing HIV-derived peptide in HIV-negative donors remained unchanged. SPV-T3b pretreatment forms a valuable tool to verify tetramer-based detection of antigen-specific T cells during the monitoring of immune responses in clinical studies.