Flow cytometry evaluation of the immunomodulatory potential of peptides targeting the BTLA-HVEM complex

The spreadsheets show raw data associated with the evaluation of the immunomodulatory potential of five peptides targeting the BTLA-HVEM complex on the activity of T cells. Methods: Magnetically isolated T cells from healthy donor-derived venous blood samples (n=10) were cultured with the examined compounds for 3 and 5 days. There were two types of culture conditions: T cells were either exposed only to the examined peptides or stimulated with ImmunoCult™ Human CD3/CD28 T Cell Activator and exposed to the tested peptides. Subsequently, T cells were subjected to cytometric phenotypic analysis and the evaluation of proliferation and apoptosis. Moreover, the expression of BTLA and HVEM was analyzed on CD4+ T cells and CD8+ T cells. T cell proliferation rate was based on VPD450 staining, while annexin V and 7-AAd were used to evaluate apoptosis of these cells. The activation status of T cells was assessed based on the expression of activation markers CD25, CD69, and CD62L. T-cell memory compartment analysis was based on the expression of CCR7 (CD197) and CD45RA, and the following subpopulations were identified: naive T cells CD45RA+CCR7+, central memory T cells CD45RA-CCR7+, effector memory T cells CD45RA-CCR7- and effector memory re-expressing CD45RA T cells CD45RA+CCR7-(TEMRA). Data acquisition was performed with BD FACSVerse™ Flow Cytometer (Becton Dickinson, USA). The results were analyzed with FlowJo software version 10.8.1 (Becton Dickinson, USA). Spreadsheet part 1 - shows data regarding the activation status of T cells and the expression of BTLA and HVEM on T cells. Spreadsheet part 2 - shows data regarding the proliferation and apoptosis of T cells and the distribution of T cell memory compartment subpopulations. More detailed information can be found in the original paper: https://doi.org/10.1016/j.ejps.2023.106677