Extensive flow cytometric immunophenotyping of human PBMC incorporating detection of chemokine receptors, cytokines and tetramers

Characterization of immune cells is essential to advance our understanding of immunology and flow cytometry is an important tool in this context. Addressing both cellular phenotype and antigen-specific functional responses of the same cells is valuable to achieve a more integrated understanding of immune cell behavior and maximizes information obtained from precious samples. Until recently, panel size was limiting, resulting in panels generally focused on either deep immunophenotyping or functional readouts. Ongoing developments in the field of (spectral) flow cytometry have made panels of 30(+) markers more accessible, opening up possibilities for advanced integrated analyses. Here, we optimized immune phenotyping by co-detection of markers covering chemokine receptors, cytokines and specific T cell/peptide tetramer interaction using a 32-color panel. Such panels enable integrated analysis of cellular phenotypes and markers assessing the quality of immune responses and will contribute to our understanding of the immune system.

Automated summary

Characterization of immune cells is crucial for advancing our understanding of immunology and flow cytometry plays a vital role in this. By simultaneously analyzing cellular phenotype and antigen-specific functional responses, we can gain a more comprehensive understanding of immune cell behavior. Recent developments in (spectral) flow cytometry have made it possible to use panels with 30(+) markers, allowing for advanced integrated analyses. In this study, we optimized immune phenotyping using a 32-color panel, enabling the analysis of cellular phenotypes and markers associated with immune response quality and contributing to our understanding of the immune system.