Peptide-MHC tetramer staining is a common practice with both R&D and clinical applications in infectious disease investigations, vaccine development, immunotherapy and autoimmunity research, and transplantation. Experimentally, this method can be used to measure detailed T cell responses and functions. Clinically, peptide-MHC tetramer staining is used to count, track, and phenotype T cells during infections and cancer, and to monitor the effectiveness of therapeutics.

While it is possible to apply tetramer staining techniques to analyze antigen-specific T cells found in circulating peripheral blood, these analyses may not reflect the true immune response. Because the percentage of T cells specific for a given peptide and HLA molecule is very low in circulation, the detection of antigen-specific T cells in PBMC may not be very convincing. Less than 1% of cells in PBMC are specific for a given antigen, so any changes or differences in frequency will appear insignificant.

 

Applying Antigen-Specific T Cells for More Accurate Analysis

To increase confidence in staining antigen-specific T cells in peripheral blood, run peptide-MHC tetramer staining experiments using antigen-specific T cells either as a positive control or as a spike to determine the limit of detection.

Our antigen-specific T cell lines are exciting tools for understanding T cell function, antigen presentation, and interaction with tumors and microbes. They can also be used as controls to understand the sensitivity of various methods used to measure the immune response, including tetramer staining.

The peptide-MHC tetramer reagents can detect T cells specific for a given peptide epitope by flow cytometric analysis. They consist of HLA molecules and peptides that are bound together in groups of 4 (tetramers), 5 (pentamers), or even 10 (dextramers). Putting several T cell receptor ligands together improves the binding of T cell antigen receptors, and adding a fluorescent label allows for detection and analysis using a flow cytometer.

Since the percentage of tetramer-positive cells is very high in our antigen-specific T cell lines, they can be added to a population of interest at defined cell numbers to determine the lower limit of detection.

On the left are PBMC from donor 153 before in vitro stimulation. On the right is the same staining of our antigen-specific T cells from donor 153. CD8+, CMV pp65 tetramer positive are found in the upper right quadrant.

 

 

Get More Specific

Antigen-specific T cells can make your peptide-MHC tetramer staining experiments more significant and impactful. We have T cell lines specific to various autoimmune diseases and tumor types, including SARS-CoV-2, influenza, CMV, HPV, HER2, MAGE A10, MART-1, NY-ESO-1, and WT-1.

You can always request a custom-developed line for your exact research needs.

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