Protocol to assess the suppression of T-cell proliferation by human MDSC


Bruger A. M. , Vanhaver C., Bruderek K., Amodio G., Tavukcuoglu E., ESENDAĞLI G., ...More

TUMOR IMMUNOLOGY AND IMMUNOTHERAPY - CELLULAR METHODS, PT B, vol.632, pp.155-192, 2020 (SCI-Expanded) identifier identifier

Abstract

Inhibition of T-cell proliferation is the most common approach to assess human myeloid-derived suppressor cell (MDSC) functions. However, diverse methodologies hinder the comparison of results obtained in different laboratories. In this chapter, we present a T-cell proliferation assay procedure based on allogeneic MDSC and T-cells that is potentially suitable to multi-center studies. The T-cells are isolated from non-cancerous donors and frozen for later use in different research groups. We observed that pure thawed T-cells showed poor proliferative capacities. To retain proliferation, T-cell-autologous mature dendritic cells are supplemented after thawing. MDSC are isolated from clinical samples and represent the sole variant between assays. Flow cytometry is used to assess T-cell proliferation by the dilution of a tracking dye.