Tumoricidal effect of dendritic cells pulsed with Ar-He cryoablation on glioma cells

2013 
Objective To explore the effect of Ar-He cryoablation on maturation of bone marrow-derived dendritic cells (BMDCs) in C57BL/6 mouse,and the mechanism of sensitized dendritic cells (DCs) induced tumor killing effect.Methods Ar-He cryoablation of GL261 glioma cells was used as experimental group,only inserting probe to GL261 glioma cells as negative control group and Ar-He cryoablation on the same amount of PBS as blank control group.The protein concentration of freezing and thawing products was measured.DCs were induced from BMDCs in vitro,and after rumor cell lysate was added,morphological characteristics of DCs were observed.The expressions of surface markers CD80,CD86,major histocompatibility complex-Ⅰ (MHC-Ⅰ) and MHC-Ⅱ were analyzed by flow cytometry.The IL-12p70 concentration in the supematants was detected by enzyme-linked immunosorbent assay (ELISA).T cells were extracted and co-cultured with DCs,and then,the expressions ofT cell phenotypes CD3,CD4 and CD8 were analyzed by flow cytometry.The killing rate ofT cells (choosing from sensitized DCs) to GL261 glioma cells was detected using CCK-8 method at different effect-target ratios (10:1,50:1 and 100:1).Results The protein concentration of the experimental group ([1071 ±240.94] μg/mL) was higher than that of negative control group([91.42±55.75]μg/mL) and blank control group (0 μg/mL,P<0.05).The experimental group showed typical characteristics of mature DCs,and the expressions ofDCs phenotypes CD80,CD86,MHC-Ⅱ and MHC-Ⅱ were higher than those in the negative control group and blank control group (P<0.05); IL-12p70 secreted by experimental group DCs was higher than that by control group (P<0.05).There were significantly more CD8+ T cells and less CD4+ T cells in the experimental group than negative control group and blank control group (P<0.05).The killing rate of CTLs to GL261 glioma cells in the experimental group was significantly higher than that in the negative control group and blank control group at different effect-target ratios (P<0.05).Conclusions A large amount of tumor antigen can be obtained from GL261 glioma cells after Ar-He cryoablation which can induce maturation of DCs.After being pulsed with tumor lysate,these DCs can produce anti-tumor effect and its mechanism is related to up-regulation of DCs surface molecules CD80,CD86,MHC-Ⅰ,MHC-Ⅱ and induce T cells into cytotoxic T cells. Key words: Ar-He cryoablation;  Glioma;  Dendritic cell;  Cytotoxic T lymphocyte
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