Advanced immunophenotyping: A powerful tool for immune profiling, drug screening, and a personalized treatment approach

Front Immunol. 2023 Mar 24:14:1096096. doi: 10.3389/fimmu.2023.1096096. eCollection 2023.

Abstract

Various autoimmune diseases are characterized by distinct cell subset distributions and activation profiles of peripheral blood mononuclear cells (PBMCs). PBMCs can therefore serve as an ideal biomarker material, which is easily accessible and allows for screening of multiple cell types. A detailed understanding of the immune landscape is critical for the diagnosis of patients with autoimmune diseases, as well as for a personalized treatment approach. In our study, we investigate the potential of multi-parameter spectral flow cytometry for the identification of patients suffering from autoimmune diseases and its power as an evaluation tool for in vitro drug screening approaches (advanced immunophenotyping). We designed a combination of two 22-color immunophenotyping panels for profiling cell subset distribution and cell activation. Downstream bioinformatics analyses included percentages of individual cell populations and median fluorescent intensity of defined markers which were then visualized as heatmaps and in dimensionality reduction approaches. In vitro testing of epigenetic immunomodulatory drugs revealed an altered activation status upon treatment, which supports the use of spectral flow cytometry as a high-throughput drug screening tool. Advanced immunophenotyping might support the exploration of novel therapeutic drugs and contribute to future personalized treatment approaches in autoimmune diseases and beyond.

Keywords: PBMC (peripheral blood mononucleated cells); T cells; drug screening; flow cytometry; immunophenotyping.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Autoimmune Diseases* / diagnosis
  • Autoimmune Diseases* / drug therapy
  • Drug Evaluation, Preclinical
  • Humans
  • Immunophenotyping
  • Leukocytes, Mononuclear*
  • Precision Medicine

Grants and funding

This work was supported by the FWF Special Research Program F70 (F07003). The authors declare that this study received funding from GlaxoSmithKline and Lilly. MBo received funding from GlaxoSmithKline. LG received a young scientist grant sponsored by Lilly. The funders was not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication.