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Archive > Volume 23 Issue 2 > 2016,23(2):188-194. DOI:10.3872/j.issn.1007-385X.2016.02.006 Prev Next
Basic Research
Efficient antitumor immune response induced by DC sensitized with heat-shocked gastric cancer cell-derived exosomes
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Abstract:
Objective:To explore response of tumor-specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DCs) sensitized with exosomes that derived from heat-shocked gastric cancer cell. Methods: exosomes from mice murine foregastric cancer (MFC) cells (Exo), heat-shocked MFC cells (Exo/HS), and lysates (Lys) of MFC cells were prepared, respectively. Morphology of the exosomes was observed by electron microscopy and protein components of the exosomes were detected by Western blotting. DCs generated from murine bone marrow were cultured and sensitiaed with Exo/HS, Exo and Lys, and the prepared DCs vaccines were termed as DC-Exo/HS, DC-Exo, and DC-Lys, respectively. Also, DCs were sensitized with exosomes from heat-shocked MFC tumor cells that transfected with HSP70 siRNA. Phenotypes of DCs were detected by flow cytometry. Mice were immunized with DC-Exo/HS, DC-Exo, DC-Ly, DC, or PBS respectively; the proliferation of splenic T cells was measured by 3H-TdR and activity of spleen CTL was measured by LDH. MFC cells-bearing mouse models were constructed to examine immunotherapy effects of the DCs vaccines. Results: exosomes were confirmed as small membranous vesicles with electron microscopy. Detection of Western blotting showed that Exo/HS contained high level of HSP70. Flow cytometry demonstrated that the exosomes from heat-shocked MFC cells could significantly up-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs, and interference with HSP70 siRNA could down-regulate expressions of MHC-Ⅱ, CD80, CD86 and CD40 molecules on the DCs sensitized with the exosomes from heat-shocked MFC tumor cells. Results of 3H-TdR showed that proliferation ability of T cells in DC-Exo/HS group was markedly enhanced as compared with those in DC-Exo, DC-Lys, DC and PBS groups (P<0.01). Results of LDH showed that higher level of CTL activity was induced in DC-Exo/HS group as compared with those in DC-Exo, DC-Lys, DC or PBS (P<0.01), and induced CTL activity in HSP70 siRNA group was significantly lower than that in control siRNA group (P<0.01). Results of the immunotherapies showed that inhibitive effect of DC-Exo/HS on tumor in the carcinoma-bearing mice was significantly better than those of all the other groups (P<001). Conclusion: DCs sensitized with exosomes that derived from heat-shocked gastric cancer cell could induce efficient antitumor immune response.
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Project Supported:
Project supported by the Natural Science Foundation of Zhejiang Province (No. LY14HY160011)
Clc Number:
