Antigen-specific CD8+ T cell analysis

Antigen-specific CD8⁺ T cell analysis provides information on the repertoire of antigen-specific responses. Furthermore, the effect of compounds or drug candidates on antigen-specific CD8⁺ T cell responses can be assayed.

What can Sanquin do for your T cell projects?

We can help determine the amount of antigen/tetramer positive CD8^₊ T cells by performing heterotetramer combinatorial coding (HTCC). Furthermore, we can assess antigen-specific CD8 T cell proliferation, cytokine production and cellular phenotype, either directly ex vivo or after in vitro priming or in vitro culture.

CD8⁺ T cell repertoire determination: HTCC

Sanquin offers a high-throughput screening of your patient samples to detect antigen-specific T cells in, e.g., vaccination studies or cell therapy. Thus far, the number of antigen-specific T cell populations that could be detected in a single sample was limited. This precludes a comprehensive analysis of disease-associated or therapy-induced T cell responses. In particular for clinical samples, this can be a considerable disadvantage. Heterotetramer Combinatorial Coding (HTCC) entails staining of CD8⁺ T cells with fluorescently-labelled multimers. This allows for the detection of antigen-specific CD8⁺ T cells that recognize an HLA-binding peptide of interest. By making use of Sanquin’s combinatorial coding method, HTCC allows for the detection and characterization of multiple antigen-specific CD8⁺ T cells simultaneously within the same sample. This allows you to make the most out of precious (patient) material.

Advantage of combinatorial coding

Combinatorial coding is a new high-tech method to circumvent the limitation of sample size. With this technique, monitoring of HLA-peptide complex recognition by T cells can be performed quickly in small sample volumes.

41 different HLA/peptide combinations can be analyzed simultaneously in one blood sample of only 4-8 mL blood  to broadly screen for immunogenic peptides. This is in sharp contrast to conventional methods using more than 200 mL of whole blood for the same analysis.

Combinatorial coding can be used for e.g.:

• combining phenotypical and functional markers with antigen specificity.
• detecting antigen-specific T cells during clinical interventions, such as vaccination studies or cell therapy.
• screening of the immunogenicity of your peptides of interest.

Assessing antigen-specific CD8⁺ T cell responses

By obtaining information on the functionality of CD8⁺ T cells in cellular assays, we can identify the magnitude and type of immune response that has been, or will be, elicited by your treatment. Furthermore, we can identify the effect of compounds on antigen-specific CD8⁺ T cell responses, for instance on cytokine production or cell proliferation. We can also assist in the characterization of antigen-specific CD8⁺ T cells during in vitro culture with compounds, or validate biomarkers.