TY - JOUR

T1 - Development of an In Vitro Assay to Assess Pharmacological Compounds and Reversion of Tumor-Derived Immunosuppression of Dendritic Cells

AU - Andersen, Mikkel Møller

AU - Larsen, Jesper

AU - Hansen, Morten

AU - Pedersen, Anders Elm

AU - Gad, Monika

PY - 2021

Y1 - 2021

N2 - Background: Cancer immunotherapies have achieved much success and have become the forefront treatment of cancers previously associated with poor prognosis. However, a major challenge in cancer immunotherapies remains the heterogeneity of the immunoregulatory capacities of cancers, and not all patients of a given cancer responds to current therapeutic strategies. To address this issue and to facilitate the development of new pharmacological compounds, we here describe an in vitro model of dendritic cell suppression by cancer cells. Methods: We treated monocyte-derived dendritic cells with conditioned medium from cancer cell lines and assessed their maturation using ELISA and flow cytometry. In addition, we assessed their ability to induce T cell activation and differentiation. Results: We found that both the phenotypic and functional maturation of dendritic cells was suppressed by the conditioned medium. The expression of IL-12p70, TNF-α, CD80, CD83, and CD86 was significantly reduced by conditioned medium from the 786-O and HeLa cell lines, and CD4+ T cells had a weaker TH1 phenotype with significantly decreased expression of IFN-γ and T-bet following co-culturing. Furthermore, we use our model to characterize the differential immunoregulatory capacities of primary cancers by using conditioned medium of cultured primary cancer cells. Conclusion: This model can be used to screen pharmacological compounds seeking to alleviate the immunosuppression of the tumor microenvironment and can furthermore be used to investigate the immunoregulatory capacities of primary cancer cells, which could be a helpful prognostic tool following tumor resection.

AB - Background: Cancer immunotherapies have achieved much success and have become the forefront treatment of cancers previously associated with poor prognosis. However, a major challenge in cancer immunotherapies remains the heterogeneity of the immunoregulatory capacities of cancers, and not all patients of a given cancer responds to current therapeutic strategies. To address this issue and to facilitate the development of new pharmacological compounds, we here describe an in vitro model of dendritic cell suppression by cancer cells. Methods: We treated monocyte-derived dendritic cells with conditioned medium from cancer cell lines and assessed their maturation using ELISA and flow cytometry. In addition, we assessed their ability to induce T cell activation and differentiation. Results: We found that both the phenotypic and functional maturation of dendritic cells was suppressed by the conditioned medium. The expression of IL-12p70, TNF-α, CD80, CD83, and CD86 was significantly reduced by conditioned medium from the 786-O and HeLa cell lines, and CD4+ T cells had a weaker TH1 phenotype with significantly decreased expression of IFN-γ and T-bet following co-culturing. Furthermore, we use our model to characterize the differential immunoregulatory capacities of primary cancers by using conditioned medium of cultured primary cancer cells. Conclusion: This model can be used to screen pharmacological compounds seeking to alleviate the immunosuppression of the tumor microenvironment and can furthermore be used to investigate the immunoregulatory capacities of primary cancer cells, which could be a helpful prognostic tool following tumor resection.

KW - dendritic cells

KW - Drug screening

KW - immunosuppression

KW - in vitro assay

KW - tolerance

U2 - 10.1080/08820139.2020.1778024

DO - 10.1080/08820139.2020.1778024

M3 - Journal article

C2 - 32573300

AN - SCOPUS:85087352309

VL - 50

SP - 527

EP - 543

JO - Immunological Investigations

JF - Immunological Investigations

SN - 0882-0139

IS - 5

ER -