Research
Print page Print page
Switch language
The Capital Region of Denmark - a part of Copenhagen University Hospital
Published

More tricks with tetramers: a practical guide to staining T cells with peptide-MHC multimers

Research output: Contribution to journalJournal articleResearchpeer-review

DOI

  1. Frequency and immunophenotype of IL10-producing regulatory B cells in optic neuritis

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Dietary gluten alters the balance of pro-inflammatory and anti-inflammatory cytokines in T cells of BALB/c mice

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides

    Research output: Contribution to journalJournal articleResearchpeer-review

  4. Integrative analysis correlates donor transcripts to recipient autoantibodies in primary graft dysfunction after lung transplantation

    Research output: Contribution to journalJournal articleResearchpeer-review

  1. Genome-wide CRISPR-Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1

    Research output: Contribution to journalJournal articleResearchpeer-review

  2. Tertiary lymphoid structures improve immunotherapy and survival in melanoma

    Research output: Contribution to journalJournal articleResearchpeer-review

  3. The real-world outcome of metastatic melanoma: Unknown primary vs. known cutaneous

    Research output: Contribution to journalJournal articleResearchpeer-review

  4. Long-Term Exposure to Inflammation Induces Differential Cytokine Patterns and Apoptosis in Dendritic Cells

    Research output: Contribution to journalJournal articleResearchpeer-review

View graph of relations

Analysis of antigen-specific T-cell populations by flow cytometry with peptide-MHC (pMHC) multimers is now commonplace. These reagents allow the tracking and phenotyping of T cells during infection, autoimmunity and cancer, and can be particularly revealing when used for monitoring therapeutic interventions. In 2009, we reviewed a number of 'tricks' that could be used to improve this powerful technology. More recent advances have demonstrated the potential benefits of using higher order multimers and of 'boosting' staining by inclusion of an antibody against the pMHC multimer. These developments now allow staining of T cells where the interaction between the pMHC and the T-cell receptor is over 20-fold weaker (K(D) > 1 mm) than could previously be achieved. Such improvements are particularly relevant when using pMHC multimers to stain anti-cancer or autoimmune T-cell populations, which tend to bear lower affinity T-cell receptors. Here, we update our previous work to include discussion of newer tricks that can produce substantially brighter staining even when using log-fold lower concentrations of pMHC multimer. We further provide a practical guide to using pMHC multimers that includes a description of several common pitfalls and how to circumvent them.

Original languageEnglish
JournalImmunology
Volume146
Issue number1
Pages (from-to)11-22
Number of pages12
ISSN0019-2805
DOIs
Publication statusPublished - Sep 2015

    Research areas

  • Antibodies, CD8-Positive T-Lymphocytes, Flow Cytometry, Fluorescent Dyes, Histocompatibility Antigens Class I, Histocompatibility Antigens Class II, Humans, Major Histocompatibility Complex, Peptides, Protein Multimerization, Receptors, Antigen, T-Cell, alpha-beta, Staining and Labeling

ID: 45943970