Dendritic cells transfected with allel-specific polyepitope DNA-constructs stimulate cytotoxic response mononuclear cells of breast cancer patients in vitro
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1
Federal State Budgetary Institution «Research Institute of Clinical Immunology» under the Russian Academy of Medical Sciences Siberian Branch, Russia
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2
Municipal Clinical Hospital number 1, Russia
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3
Avaxis Bio Co.Ltd, Russia
Breast cancer (BC) has the highest frequency of women's cancers. From 10 to 35% of breast cancers are characterized by the enhanced expression of HER2 protein often associated with more aggressive tumors and poor outcome. HER2 over-expression was also found in other cancers including ovarian, colon, uterine, gastric, prostate, and adenocarcinoma of the lung. HER2/neu is a member of the family of epidermal growth factor receptors, regulating cell proliferation, differentiation, migration, and adhesion. Upregulation of HER2 leads to suppression of apoptosis and active proliferation and can lead to cancer. Peptides predicted as HLA-A*0201-binders were selected to construct HLA-A*0201-specific immunogen. HLA-A*0201 allele was chosen as the most frequently found HLA class I allele all over the world. We investigated the efficacy of DC transfected polyepitope constructs containing HLA-A0201-associated peptide CTL epitopes of different tumor-associated antigens for breast cancer to stimulate the cytotoxic response in culture mononuclear cells of patients with BC.
Monocytes from mononuclear cells (MNCs) of peripheral blood from BC-patients were cultured in the presence of rhIL-4, rhGM-CSF and rhTNF-a for generation of mature DC. Delivery of DNA-construct containing HLA-A0201-associated peptide CTL epitopes, in mature DCs was performed using magnetic transfection. To stimulate the cytotoxic activity was carried co-culture of DC and MNCs in several parallel cultures at meeting the same culture conditions - the ratio of DC: MNC 1:10 and stimulate Th1-polarized by rhIL-12 and rhIL-18. Autologous tumor cells and cell lines MCF-7 we used as target cells. DNA-transfected mature DC stimulate the cytotoxic potential of MNCs against autologous tumor cells, and against MCF-7. Addition of IL-12 and IL-18 at cocultivation MNCs and transfected DC is an effective method of stimulation the specific cytotoxic response against tumor cells. The use of autologous tumor cells as a target cell results in a more pronounced cytotoxic response, compared with the cell line MCF-7.
Thus, the use polyepitope DNA constructs for antigenic load of DC and stimulation of MNCs culture with transfected DC leads to the stimulation of cytotoxic response directed at tumor cell lysis.
Acknowledgements
Work was supported by the Federal Program "Research and scientific-pedagogical personnel of innovative Russia in 2009-2013." (Agreement number 8265), the grant of the President of the Russian Federation № 16.120.11.3396-MC.
References
Antonets DV, Maksiutov AZ (2010) [TEpredict: software for T-cell epitope prediction]. Molekuliarnaia biologiia 44, 130-9.
Harari D, Yarden Y (2000) Molecular mechanisms underlying ErbB2/HER2 action in breast cancer. Oncogene 19, 6102-14.
Keywords:
Dendritic Cells,
Cytotoxicity,
breast cancer,
DNA vaccine,
Transfection
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Adaptive Immunity
Citation:
Shevchenko
YA,
Sennikov
SV,
Khantakova
YN,
Kurilin
VV,
Lopatnikova
YA,
Sidorov
SV,
Maksyutov
AZ,
Maksyutov
RA,
Gavrilova
EV and
Zaytsev
SA
(2013). Dendritic cells transfected with allel-specific polyepitope DNA-constructs stimulate cytotoxic response mononuclear cells of breast cancer patients in vitro.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00165
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Received:
11 Mar 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Miss. Yulia A Shevchenko, Federal State Budgetary Institution «Research Institute of Clinical Immunology» under the Russian Academy of Medical Sciences Siberian Branch, Novosibirsk, Russia, shevcen@ngs.ru