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High lipid content of irradiated human melanoma cells does not affect cytokine-matured dendritic cell function

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Abstract

Gamma irradiation is one of the methods used to sterilize melanoma cells prior to coculturing them with monocyte-derived immature dendritic cells in order to develop antitumor vaccines. However, the changes taking place in tumor cells after irradiation and their interaction with dendritic cells have been scarcely analyzed. We demonstrate here for the first time that after irradiation a fraction of tumor cells present large lipid bodies, which mainly contain triglycerides that are several-fold increased as compared to viable cells as determined by staining with Oil Red O and BODIPY 493/503 and by biochemical analysis. Phosphatidyl-choline, phosphatidyl-ethanolamine and sphingomyelin are also increased in the lipid bodies of irradiated cells. Lipid bodies do not contain the melanoma-associated antigen MART-1. After coculturing immature dendritic cells with irradiated melanoma cells, tumor cells tend to form clumps to which dendritic cells adhere. Under such conditions, dendritic cells are unable to act as stimulating cells in a mixed leukocyte reaction. However, when a maturation cocktail composed of TNF-alpha, IL-6, IL-1beta and prostaglandin E2 is added to the coculture, the tumor cells clumps disaggregate, dendritic cells remain free in suspension and their ability to efficiently stimulate allogeneic lymphocytes is restored. These results help to understand the events following melanoma cell irradiation, shed light about interactions between irradiated cells and dendritic cells, and may help to develop optimized dendritic cell vaccines for cancer therapy.

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Acknowledgments

This work has been supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), Fundación Sales, Fundación Cáncer (FUCA), Fundación Pedro F. Mosoteguy, Fundación María Calderón de la Barca, Argentina and by INSERM (Institut National de la Santé et de la Recherche Médicale), by an Evaluation-Orientation de la Coopération Scientifique Sud (ECOS Sud)/Secretaría de Promoción Científica y Tecnológica (SEPCyT) exchange program grant (A03S03) from the Argentine SEPCyT and the French Ministère des Affaires Etrangères and by a grant from an INSERM/CONICET exchange program. JM, MMB, EML and LAQA are members of CONICET. FPMR, LMPL and MPR are fellows of the same Institution. GAP is a fellow of ANPCyT. JLT is a member of INSERM. AMC was supported by FUCA.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Centro de Investigaciones Oncológicas-Fundación Cáncer, Instituto Alexander Fleming, Cramer 1180, 1426, Buenos Aires, Argentina

    Gabriela A. Pizzurro, Andrea M. Copati, María P. Roberti, Estrella M. Levy, María M. Barrio & José Mordoh

  2. Fundación Instituto Leloir-IIBBA, CONICET, Av. Patricias Argentinas 435, 1405, Buenos Aires, Argentina

    Florencia P. Madorsky Rowdo, Luciana M. Pujol-Lereis, Luis A. Quesada-Allué & José Mordoh

  3. INSERM UMR-S 872, Centre de Recherche des Cordeliers, Université Pierre et Marie Curie, Université Paris Descartes, 75006, Paris, France

    Jean-Luc Teillaud

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  1. Gabriela A. Pizzurro
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Correspondence to José Mordoh.

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Pizzurro, G.A., Madorsky Rowdo, F.P., Pujol-Lereis, L.M. et al. High lipid content of irradiated human melanoma cells does not affect cytokine-matured dendritic cell function. Cancer Immunol Immunother 62, 3–15 (2013). https://doi.org/10.1007/s00262-012-1295-4

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