MHC multimer staining for detection of antigen-specific T cells

We believe the most powerful strategy for mapping antigen-specific T cells is staining with MHC multimers.

Common MHC multimer staining

Our specialty within detection of antigen-specific T cell responses and the most exact method for enumerating and profiling these is the MHC multimer staining for flow cytometry. In addition to staining with the MHC multimers we co-stain for T cell lineage markers and often further characterize the antigen-specific T cells in regard to e.g. their maturation phenotype and further markers. For an example see our slide on Deep Profiling of Antigen-specific T cells.

Upon treatment with immunotherapies in human or murine cohorts, the enumeration and activation of certain antigen-specific T cells is important to evaluate the treatment-induced immune response by comparing pre- and post-treatment samples.
Multiple different antigen-specific T cells populations can be detected using MHC multimer staining and flow cytometry in samples originating from the preclinical or clinical setting.

MHC multimer staining has a high sensitivity and specificity for detecting antigen-specific T cells and has been heavily used for investigating epitope spreading and tracking immunotherapy-induced immune responses in both the preclinical and clinical setting in the past. We have also used MHC multimer staining for quality control of T cell receptor-transduced cellular products and in investigational selection of cellular product candidates. Furthermore, MHC multimer staining can be used for testing the effect of repeated freeze/thaw cycles for such a cellular product.

MHC multimer staining utilizes the pMHC-TCR recognition

Common MHC Multimer Staining_pMHC interaction

Antigen-specific T cells recognize specific peptide-MHC (pMHC) complexes on the cell surface and bind to their corresponding T cell receptor (TCR). MHC multimers, which are recombinantly produced pMHC complexes coupled with fluorescent labels are used for detecting antigen-specific T cell responses in fluid samples, e.g. from blood, tumor-infiltrating lymphocytes or cultured cells by flow cytometry. MHC multimer staining are used to detect the presence of antigen-specific T cells and can be combined with our immunophenotyping assays, e.g. to detect the maturation profile of antigen-specific T cells. As IFN-γ-releasing assays such as ELISPOT or intracellular cytokine staining detect only the functional antigen-specific T cells, we highly recommend using MHC multimer staining for achieving validation and a more detailed insight in the antigen-specific T cell population. This is our most exact assay for detection of antigen-specific T cells and requires knowledge about the tissue type of the individual or animal in question.

We ensure the procurement of high-quality MHC monomers and multimers from our distributor Immudex.

For specialized purposes we offer the highly multiplexed dCODE™ Dextramer platform, enabling screening of more than 1000 different antigen-specific T cell responses in parallel. For more information see our dedicated site on this innovative method.

Applications of MHC multimer staining by flow cytometry

  • Detection, enumeration and isolation of antigen-specific T cells in fluid cell samples (PBMCs, tumor-infiltrating lymphocytes or from cell cultures)
  • Antigen-specific immune monitoring of patients in clinical trials for immunotherapy and vaccine development
  • Quality control/quality assurance and selection of cellular products expressing TCRs with certain T cell specificities
  • In vitro experiments in discovery and preclinical development phases for hit-lead optimization
  • Validation of neo-antigen-specific T cell responses in samples from animal models or human patients
  • Investigation of epitope mapping for selection of antigens and epitope spreading for detection of treatment-induced antigen-specific T cell responses.

 

We offer MHC multimer staining analyses for human and murine samples.

Learn more about
MULTIPLEX MHC MULTIMER STAINING

Learn how the MHC multimer dCODE dextramer platform can be used within both immune-oncology and the field of autoimmune diseases, and how ImmuMap offer analyses using DNA barcoded MHC complexes for human samples.