Proteomic analysis of heat-conditioned cancer cell lysates used for anti-melanoma DC-based immunotherapy
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1
Millennium Institute on Immunology and Immunotherapy, University of Chile, Chile
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2
Department of Medical Biochemistry and Biophysics, Karolinska Institute, Sweden
Autologous dendritic cells (DCs) loaded with tumor-associated antigens (TAAs) from cancer cell lysates, have emerged as a promising immunological tool for cancer therapy due to their abilities to stimulate the antitumor response and trigger the generation of immunological memory in cancer patients. We have recently shown that DCs loaded with an allogeneic cancer cell lysate (TRIMEL) can induce a potent immunological response in melanoma patients. Our results have shown that TRIMEL not only works as a source of TAAs but also contains other molecules, specific damage associated molecular patterns (DAMPs), which cooperate in DC maturation improving their capacity as antigen presenting cells. Our aim was to characterize the proteomic profile of complete TRIMEL and its three cell lines separately with and without heat activation, in order to identify new DAMPs from biologically relevant protein candidates. Five independently produced batches of TRIMEL, Mel1, Mel2 and Mel3 cell lines with and without heat activation at 42°C for 1 hour were prepared. Samples were digested and a high-throughput proteome analysis to over >5000 proteins was performed. The raw data acquired was analyzed using MaxQuant software, version 1.3.0.5, for peptide and protein identification. Our result showed 71 up-regulated proteins from TRIMEL, 137 from Mel1, and 106 from Mel2 under heat-shock treatment. Some of the up-regulated proteins were: Eukaryotic translation initiation factor family proteins, haptoglobin, and UTP--glucose-1-phosphate uridylyltransferase, among others. Some of these proteins will be considered as putative DAMPs for further immunological analysis. Identification of new DAMPs from cancer cells, will allow a better understanding of DCs activation during antitumor response and, in turn assist the development of improved and more effective approaches in cancer immunotherapy.
Acknowledgements
Funded by grant FONDECYT 11130607
Keywords:
Cancer Vaccines,
Dendritic Cells,
DAMPs,
Tumor immunotherapy,
Melanoma
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Poster Presentation
Topic:
Immunotherapy
Citation:
González
F,
Pereda
C,
Chernobrovkin
A,
Fuentes
C,
López
J,
Salazar-Onfray
F and
Zubarev
R
(2015). Proteomic analysis of heat-conditioned cancer cell lysates used for anti-melanoma DC-based immunotherapy.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00086
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Received:
30 May 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
PhD. Fermín González, Millennium Institute on Immunology and Immunotherapy, University of Chile, Santiago, Chile, fgonzalez@uchile.cl