Background:The use of clinical immunotherapies targeting T-cells in an ever increasing number of patients, in particular cancer patients, begs the question how the success of such therapies can be monitored in future in a reliable way and at reasonable cost. Typically, antigen-specific T-cells are detected by ELISPOT or flow-cytometry (intracellular cytokines), however these methods are relatively difficult to standardize and costly. PepSupTMis a simple, highly standardized, peptide-based cell stimulation assay, which was designed to produce short-term culture supernatants from whole blood.
Method:Sample processing only required a programmable heat block and a simple bench-top centrifuge. Following 16 hours of overnight stimulation of whole blood with a range of different PepMixTMpeptide pools dissolved in optimized PepSupTMmedia, samples were centrifuged and supernatants collected. Samples were analyzed with the MSD V-Plex platform or the BD Cytometric Bead Array. Cytokines originating from T-cells but also other cells were measured.
Results: PepSupTMprovided very similar results as incubation of samples in a standard incubator using CO2-dependent media, however, offered significant advantages in regards to hands-on time and sample handling.
Conclusions:PepSupTMis the ideal basis for monitoring antigen-specific, stimulation-induced mediator secretion in a range of situations, for example in the study of infection, transplantation, or tumour-specific immunity. Because of its high degree of standardization and hence reproducibility, it is particularly well-suited to longitudinal patient monitoring.