Top

Published in:

01-01-2013 | Original article

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

Authors: Gabriela A. Pizzurro, Florencia P. Madorsky Rowdo, Luciana M. Pujol-Lereis, Luis A. Quesada-Allué, Andrea M. Copati, María P. Roberti, Jean-Luc Teillaud, Estrella M. Levy, María M. Barrio, José Mordoh

Published in: Cancer Immunology, Immunotherapy | Issue 1/2013

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.

Available only for authorised users

Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392(6673):245–252. doi:10.1038/32588 PubMedCrossRef

Steinman RM, Turley S, Mellman I, Inaba K (2000) The induction of tolerance by dendritic cells that have captured apoptotic cells. J Exp Med 191(3):411–416PubMedCrossRef

Delamarre L, Mellman I (2011) Harnessing dendritic cells for immunotherapy. Semin Immunol 23(1):2–11. doi:10.1016/j.smim.2011.02.001 PubMedCrossRef

Palucka K, Ueno H, Banchereau J (2011) Recent developments in cancer vaccines. J Immunol 186(3):1325–1331. doi:10.4049/jimmunol.0902539 PubMedCrossRef

Green DR, Ferguson T, Zitvogel L, Kroemer G (2009) Immunogenic and tolerogenic cell death. Nat Rev Immunol 9(5):353–363. doi:10.1038/nri2545 PubMedCrossRef

Albert ML, Sauter B, Bhardwaj N (1998) Dendritic cells acquire antigen from apoptotic cells and induce class I-restricted CTLs. Nature 392(6671):86–89. doi:10.1038/32183 PubMedCrossRef

Ferlazzo G, Semino C, Spaggiari GM, Meta M, Mingari MC, Melioli G (2000) Dendritic cells efficiently cross-prime HLA class I-restricted cytolytic T lymphocytes when pulsed with both apoptotic and necrotic cells but not with soluble cell-derived lysates. Int Immunol 12(12):1741–1747PubMedCrossRef

Chin L, Garraway LA, Fisher DE (2006) Malignant melanoma: genetics and therapeutics in the genomic era. Genes Dev 20(16):2149–2182. doi:10.1101/gad.1437206 PubMedCrossRef

Dutton-Regester K, Hayward NK (2012) Reviewing the somatic genetics of melanoma: from current to future analytical approaches. Pigment Cell Melanoma Res. doi:10.1111/j.1755-148X.2012.00975.x PubMed

10.

Castle JC, Kreiter S, Diekmann J, Lower M, van de Roemer N, de Graaf J, Selmi A, Diken M, Boegel S, Paret C, Koslowski M, Kuhn AN, Britten CM, Huber C, Tureci O, Sahin U (2012) Exploiting the mutanome for tumor vaccination. Cancer Res. doi:10.1158/0008-5472.CAN-11-3722 PubMed

11.

Palucka AK, Ueno H, Connolly J, Kerneis-Norvell F, Blanck JP, Johnston DA, Fay J, Banchereau J (2006) Dendritic cells loaded with killed allogeneic melanoma cells can induce objective clinical responses and MART-1 specific CD8 + T-cell immunity. J Immunother 29(5):545–557. doi:10.1097/01.cji.0000211309.90621.8b PubMedCrossRef

12.

Nouri-Shirazi M, Banchereau J, Bell D, Burkeholder S, Kraus ET, Davoust J, Palucka KA (2000) Dendritic cells capture killed tumor cells and present their antigens to elicit tumor-specific immune responses. J Immunol 165(7):3797–3803PubMed

13.

Berard F, Blanco P, Davoust J, Neidhart-Berard EM, Nouri-Shirazi M, Taquet N, Rimoldi D, Cerottini JC, Banchereau J, Palucka AK (2000) Cross-priming of naive CD8 T cells against melanoma antigens using dendritic cells loaded with killed allogeneic melanoma cells. J Exp Med 192(11):1535–1544PubMedCrossRef

14.

Michaud M, Martins I, Sukkurwala AQ, Adjemian S, Ma Y, Pellegatti P, Shen S, Kepp O, Scoazec M, Mignot G, Rello-Varona S, Tailler M, Menger L, Vacchelli E, Galluzzi L, Ghiringhelli F, di Virgilio F, Zitvogel L, Kroemer G (2011) Autophagy-dependent anticancer immune responses induced by chemotherapeutic agents in mice. Science 334(6062):1573–1577. doi:10.1126/science.1208347 PubMedCrossRef

15.

Nestle FO, Alijagic S, Gilliet M, Sun Y, Grabbe S, Dummer R, Burg G, Schadendorf D (1998) Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4(3):328–332PubMedCrossRef

16.

von Euw EM, Barrio MM, Furman D, Levy EM, Bianchini M, Peguillet I, Lantz O, Vellice A, Kohan A, Chacon M, Yee C, Wainstok R, Mordoh J (2008) A phase I clinical study of vaccination of melanoma patients with dendritic cells loaded with allogeneic apoptotic/necrotic melanoma cells. Analysis of toxicity and immune response to the vaccine and of IL-10 -1082 promoter genotype as predictor of disease progression. J Transl Med 6:6. doi:10.1186/1479-5876-6-6 CrossRef

17.

Neidhardt-Berard EM, Berard F, Banchereau J, Palucka AK (2004) Dendritic cells loaded with killed breast cancer cells induce differentiation of tumor-specific cytotoxic T lymphocytes. Breast Cancer Res 6(4):R322–R328. doi:10.1186/bcr794 PubMedCrossRef

18.

von Euw EM, Barrio MM, Furman D, Bianchini M, Levy EM, Yee C, Li Y, Wainstok R, Mordoh J (2007) Monocyte-derived dendritic cells loaded with a mixture of apoptotic/necrotic melanoma cells efficiently cross-present gp100 and MART-1 antigens to specific CD8(+) T lymphocytes. J Transl Med 5:19. doi:10.1186/1479-5876-5-19 CrossRef

19.

Dranoff G, Jaffee E, Lazenby A, Golumbek P, Levitsky H, Brose K, Jackson V, Hamada H, Pardoll D, Mulligan RC (1993) Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity. Proc Natl Acad Sci USA 90(8):3539–3543PubMedCrossRef

20.

Herber DL, Cao W, Nefedova Y, Novitskiy SV, Nagaraj S, Tyurin VA, Corzo A, Cho HI, Celis E, Lennox B, Knight SC, Padhya T, McCaffrey TV, McCaffrey JC, Antonia S, Fishman M, Ferris RL, Kagan VE, Gabrilovich DI (2010) Lipid accumulation and dendritic cell dysfunction in cancer. Nat Med 16(8):880–886. doi:10.1038/nm.2172 PubMedCrossRef

21.

Kawakami Y, Eliyahu S, Delgado CH, Robbins PF, Rivoltini L, Topalian SL, Miki T, Rosenberg SA (1994) Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci USA 91(9):3515–3519PubMedCrossRef

22.

Barrio MM, de Motta PT, Kaplan J, von Euw EM, Bravo AI, Chacon RD, Mordoh J (2006) A phase I study of an allogeneic cell vaccine (VACCIMEL) with GM-CSF in melanoma patients. J Immunother 29(4):444–454. doi:10.1097/01.cji.0000208258.79005.5f PubMedCrossRef

23.

Levy EM, Sycz G, Arriaga JM, Barrio MM, von Euw EM, Morales SB, Gonzalez M, Mordoh J, Bianchini M (2009) Cetuximab-mediated cellular cytotoxicity is inhibited by HLA-E membrane expression in colon cancer cells. Innate Immun 15(2):91–100. doi:10.1177/1753425908101404 PubMedCrossRef

24.

Resnicoff M, Medrano EE, Podhajcer OL, Bravo AI, Bover L, Mordoh J (1987) Subpopulations of MCF7 cells separated by Percoll gradient centrifugation: a model to analyze the heterogeneity of human breast cancer. Proc Natl Acad Sci USA 84(20):7295–7299PubMedCrossRef

25.

Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226(1):497–509PubMed

26.

Aloisi JD SJ, Fried B (1990) Quantification of lipids by one-dimensional TLC on Preadsorbent High Performance Silica Gel Plates J Liq Chrom Relat Tech 13 (20):3949–3961

27.

Sterin-Speziale N, Kahane VL, Setton CP, Fernandez MC, Speziale EH (1992) Compartmental study of rat renal phospholipid metabolism. Lipids 27(1):10–14PubMedCrossRef

28.

Samsa MM, Mondotte JA, Iglesias NG, Assuncao-Miranda I, Barbosa-Lima G, Da Poian AT, Bozza PT, Gamarnik AV (2009) Dengue virus capsid protein usurps lipid droplets for viral particle formation. PLoS Pathog 5(10):e1000632. doi:10.1371/journal.ppat.1000632 PubMedCrossRef

29.

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRef

30.

Kruth HS (1984) Localization of unesterified cholesterol in human atherosclerotic lesions. Demonstration of filipin-positive, oil-red-O-negative particles. Am J Pathol 114(2):201–208PubMed

31.

Kuhajda FP (2000) Fatty-acid synthase and human cancer: new perspectives on its role in tumor biology. Nutrition 16(3):202–208PubMedCrossRef

32.

De Maziere AM, Muehlethaler K, van Donselaar E, Salvi S, Davoust J, Cerottini JC, Levy F, Slot JW, Rimoldi D (2002) The melanocytic protein Melan-A/MART-1 has a subcellular localization distinct from typical melanosomal proteins. Traffic 3(9):678–693PubMedCrossRef

33.

Steinman RM, Gutchinov B, Witmer MD, Nussenzweig MC (1983) Dendritic cells are the principal stimulators of the primary mixed leukocyte reaction in mice. J Exp Med 157(2):613–627PubMedCrossRef

34.

Jonuleit H, Kuhn U, Muller G, Steinbrink K, Paragnik L, Schmitt E, Knop J, Enk AH (1997) Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions. Eur J Immunol 27(12):3135–3142. doi:10.1002/eji.1830271209 PubMedCrossRef

35.

Toh HC, Wang WW, Chia WK, Kvistborg P, Sun L, Teo K, Phoon YP, Soe Y, Tan SH, Hee SW, Foo KF, Ong S, Koo WH, Zocca MB, Claesson MH (2009) Clinical benefit of allogeneic melanoma cell lysate-pulsed autologous dendritic cell vaccine in MAGE-positive colorectal cancer patients. Clin Cancer Res 15(24):7726–7736. doi:10.1158/1078-0432.CCR-09-1537 PubMedCrossRef

36.

Walther TC, Farese RV Jr (2009) The life of lipid droplets. Biochim Biophys Acta 1791(6):459–466. doi:10.1016/j.bbalip.2008.10.009 PubMedCrossRef

37.

Murphy DJ (2001) The biogenesis and functions of lipid bodies in animals, plants and microorganisms. Prog Lipid Res 40(5):325–438PubMedCrossRef

38.

Accioly MT, Pacheco P, Maya-Monteiro CM, Carrossini N, Robbs BK, Oliveira SS, Kaufmann C, Morgado-Diaz JA, Bozza PT, Viola JP (2008) Lipid bodies are reservoirs of cyclooxygenase-2 and sites of prostaglandin-E2 synthesis in colon cancer cells. Cancer Res 68(6):1732–1740. doi:10.1158/0008-5472.CAN-07-1999 PubMedCrossRef

39.

Swinnen JV, Brusselmans K, Verhoeven G (2006) Increased lipogenesis in cancer cells: new players, novel targets. Curr Opin Clin Nutr Metab Care 9(4):358–365. doi:10.1097/01.mco.0000232894.28674.30 PubMedCrossRef

40.

Kuhajda FP (2006) Fatty acid synthase and cancer: new application of an old pathway. Cancer Res 66(12):5977–5980. doi:10.1158/0008-5472.CAN-05-4673 PubMedCrossRef

41.

Bozza PT, Viola JP (2010) Lipid droplets in inflammation and cancer. Prostaglandins Leukot Essent Fatty Acids 82(4–6):243–250. doi:10.1016/j.plefa.2010.02.005 PubMedCrossRef

42.

Bostrom P, Andersson L, Rutberg M, Perman J, Lidberg U, Johansson BR, Fernandez-Rodriguez J, Ericson J, Nilsson T, Boren J, Olofsson SO (2007) SNARE proteins mediate fusion between cytosolic lipid droplets and are implicated in insulin sensitivity. Nat Cell Biol 9(11):1286–1293. doi:10.1038/ncb1648 PubMedCrossRef

43.

Al-Saffar NM, Titley JC, Robertson D, Clarke PA, Jackson LE, Leach MO, Ronen SM (2002) Apoptosis is associated with triacylglycerol accumulation in Jurkat T-cells. Br J Cancer 86(6):963–970. doi:10.1038/sj.bjc.6600188 PubMedCrossRef

44.

Exton JH (1994) Phosphatidylcholine breakdown and signal transduction. Biochim Biophys Acta 1212(1):26–42PubMedCrossRef

45.

Liao YP, Wang CC, Butterfield LH, Economou JS, Ribas A, Meng WS, Iwamoto KS, McBride WH (2004) Ionizing radiation affects human MART-1 melanoma antigen processing and presentation by dendritic cells. J Immunol 173(4):2462–2469PubMed

Title: High lipid content of irradiated human melanoma cells does not affect cytokine-matured dendritic cell function
Authors: Gabriela A. Pizzurro
Florencia P. Madorsky Rowdo
Luciana M. Pujol-Lereis
Luis A. Quesada-Allué
Andrea M. Copati
María P. Roberti
Jean-Luc Teillaud
Estrella M. Levy
María M. Barrio
José Mordoh
Publication date: 01-01-2013
Publisher: Springer-Verlag
Published in: Cancer Immunology, Immunotherapy / Issue 1/2013
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-012-1295-4

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2013

Low-dose cyclophosphamide administered as daily or single dose enhances the antitumor effects of a therapeutic HPV vaccine

T cells from indolent CLL patients prevent apoptosis of leukemic B cells in vitro and have altered gene expression profile

T cell responses against microsatellite instability-induced frameshift peptides and influence of regulatory T cells in colorectal cancer

Increased prevalence of tumor-infiltrating regulatory T cells is closely related to their lower sensitivity to H2O2-induced apoptosis in gastric and esophageal cancer

Phase I trial of a multi-epitope-pulsed dendritic cell vaccine for patients with newly diagnosed glioblastoma

“Tumor Immunology Meets Oncology (TIMO) VIII” from May 4 to 5, 2012, in Halle/Saale, Germany

Keynote webinar | Spotlight on antibody–drug conjugates in cancer