Phenotypic assessment of dendritic cell maturation by cost-effective custom ELISA assays.

Abstract

Dendritic cells (DCs) are professional antigen-presenting cells that are pivotal in operating tumor immunosurveillance and orchestrating anticancer immune responses. Endowed with phagocytic and migratory capacities, DCs can capture and process tumor antigens, travel to lymphoid organs and prime naïve T cells, altogether leading to the clonal expansion of cytotoxic T lymphocytes (CTLs) that can specifically target and lyse cancer cells. Additionally, DCs contribute to the formation of immunological memory, ensuring durable therapeutic effects and long-term surveillance against tumor recurrence. Upon antigen engagement, DCs undergo a maturation process characterized by the production of specific cytokines as well as the increased expression of costimulatory molecules and chemokine receptors on their surface. Here we propose a panel of custom sandwich enzyme linked immunosorbent assays (ELISAs) for assessing DC maturation via the precise quantification of cytokines. This economic approach achieves high precision and reproducibility and can be readily applied in labs equipped with basic molecular cell biology facilities. With appropriate automatization, this protocol can be employed for high-throughput screening campaigns for the discovery of DC maturation modulators.