Ovarian Carcinoma cell line lysates conditionated with Heat Shock are potent stimulus to maturate DCs towards finding an immunotherapy against Ovarian Cancer.
Jimena
P.
Gatica Arenas1, 2, 3,
Felipe
E.
Falcón Beas1, 2, 3,
Iván
E.
Flores Colmann1, 2, 3,
Cristián
A.
Falcón Beas1, 2, 3,
María
A.
Gleisner1, 2, 3,
Flavio
A.
Salazar Onfray1, 2, 3 and
Mercedes
N.
Lopez1, 2, 3, 4*
-
1
Faculty of Medicine, University of Chile, Chile
-
2
Institute of Biomedical Sciences, Faculty of Medicine, Chile
-
3
Millennium Institute on Immunology and Immunotherapy, Chile
-
4
Clinical Hospital, University of Chile, Chile
Introduction: Ovarian epithelial carcinoma is the most prevalent ovarian malignancy with a high mortality on advanced stages, where conventional treatments are not effective. Dendritic cells (DCs) are professional antigen presenting cells that are able to induce a strong immune response against tumor cells. Immature DCs have the capability to acknowledge endogenous danger signals (DAMPs) and phagocyte tumoral antigens, presenting them to T cells on a HLA context initiating an antitumor response. Previous results of our laboratory on immunotherapy have shown that DCs treated with a heat shocked melanoma cell lysate called TRIMEL can mature and induce a strong antitumor response. These have been used in more than 120 stage III and IV melanoma patients with positive outcomes. According with this results we propose that DCs derived from monocytes from peripheral blood (PBMC) can mature and activate T cells into a proinflamatory profile using cell lysates from 3 distinct ovarian carcinoma cell lines that underwent heat shock (TRIO).
Methods: Monocytes were obtained from PBMC from healthy donors through a density gradient (FICOLL-Hypaque). Then, DCs were generated using the established protocol of our laboratory. Briefly, we differentiated monocytes into immature DCs using IL-4 and GM-CSF. Then, Heat-shock conditioned ovarian cells lysate (TRIO) were used as maturation stimulus. We evaluated the expression of coestimuatory molecules using flow cytometry (FACS). The quantification of the secreted cytokines by the DCs was assayed from the supernatant of the different DCs, cultured with a NIH3T3 CD40L+ cell line. The secretion of proinflamatory cytokines was measured using the CBA kit (BD bioscience). Also, DCs and allogenic PBL were cultured for 5 days and INF-γ, IL-4, IL-17 and CD4 expression were analyzed by flow cytometry.
Results: Trio induces maturation of DCs based on an increased expression of CD83 and CD86 in a dose dependent manner, showing maximum effect between 100ug/ml and 200ug/ml of protein. Interestingly we did not find significant differences comparing this condition with TRIMEL. Afterwards, we further studied a wider panel of maturation molecules on DCs stimulated using 100-200ug/ml conditions. Our results showed that TRIO is capable of inducing a higher expression of CD80, CD40, HLAI, CCR7 and an increase on TNF-α and IL-6 secretion compared to tolerogenic and immature DCs and similar to TRIMEL DCs levels. When allogenic lymphocytes were cocultured with TRIO DCs, we obtained a higher proportion of INFγ + and IL-17 + T cells, in comparison to coculturing with tolerogenic DCs.
Discussion: These results suggest that DCs stimulated with TRIO can develop a proinflamatory profile, behaving similarly to TRIMEL DCs, which make them a good candidate for a future DCs based vaccination treatment for advance staged ovarian carcinoma patients, though further studies are needed in order to evaluate their clinical efficacy.
Acknowledgements
Funded by grants FONDEF-D11I1036; FONDECYT-1130324; IMII P09/016-F.
Keywords:
Immunotherapy,
ovarian cancer,
Dendritic Cells,
T lymphocytes,
cancer lysate
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:
Gatica Arenas
JP,
Falcón Beas
FE,
Flores Colmann
IE,
Falcón Beas
CA,
Gleisner
MA,
Salazar Onfray
FA and
Lopez
MN
(2015). Ovarian Carcinoma cell line lysates conditionated with Heat Shock are potent stimulus to maturate DCs towards finding an immunotherapy against Ovarian Cancer..
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.2015.05.00083
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Received:
15 May 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
MD, PhD. Mercedes N Lopez, Faculty of Medicine, University of Chile, Santiago, Chile, melopez@me.com