Study of dendritic cells co-cultured with cytokine induced killer cells sensitized by cholangiocarcinoma antigen on antitumor activity

doi:10.3877/cma.j.issn.1674-0793.2017.04.001

Abstract

Abstract:

Objective

To investigate the morphological changes and proliferation of cholangiocarcinoma antigen sensitized DC-CIKs and to detect its antitumor activity.

Methods

DC and CIK were generated by culturing peripheral blood mononuclear cells (PBMCs) of healthy blood donors. Adherent cells were collected to generate DC, and non-adherent cells were collected to generate CIK. After being matured with rhGM-CSF, rhIL-4 and rhTNF-α, DC were sensitized with RBE tumor antigen lysates. CIK was matured with rhIL-2, IL-1β. DC and CIK were co-cultured at the ratio of 1:10, 120, 140. The DC mature phenotype were analyzed by flow cytometry. The anti-tumor effect of the co-culture sensitized DC and CIK was evaluated by luciferase. The secretion of IL-12, IFN-γ were assayed by ELISA.

Results

The growth and proliferation of antigen-sensitized DC was more potent than non- antigen-sensitized DC and pre-induced DC cells. The antitumor activity of the antigen-sensitized DC-CIK group was not significantly higher than that of the non-sensitized DC-CIK group at the effective target ratio of 101. At the effective target ratio of 201 and 401, the antitumor activity of antigen-sensitized DC-CIK group was significantly higher than that of non-sensitized DC-CIK group (P<0.05). The expression of cytokines (IL-12, IFN-γ) in antigen-sensitized DC-CIK group was significantly higher than that in non-sensitized DC-CIK group (P<0.05) and CIK group (P<0.05).

Conclusions

Allogeneic cholangiocarcinoma antigen can effectively sensitize DC cell of healthy human and promote proliferation ability, maturation and phenotype. The antigen-sensitized DC-CIK had the stronger cytotoxic activity against RBE than that of non-sensitized DC-CIK and CIK.

Key words: Bile duct neoplasms, Dendritic cells, Cytokine-induced killer cells, Tumor suppressor

Xiaofeng Jiang, Xiaopeng Yuan, Dawei Zhang, Haiwu Lu, Zilong Wen, Qiang Zheng, Songhang Liu, Ping Xue. Study of dendritic cells co-cultured with cytokine induced killer cells sensitized by cholangiocarcinoma antigen on antitumor activity[J]. Chinese Archives of General Surgery(Electronic Edition), 2017, 11(04): 217-221.

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URL: https://zhptwkxwx.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0793.2017.04.001

https://zhptwkxwx.cma-cmc.com.cn/EN/Y2017/V11/I04/217

Figures/Tables 5

References 14

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组别	CD3⁺CD8⁺双阳性细胞率	CD3⁺CD56⁺双阳性细胞率
CIK组	49.49±3.79	14.35±2.46
未致敏DC-CIK组	73.66±3.62^a	14.29±1.53
致敏DC-CIK组	85.41±2.61^a	56.78±2.38^a
统计值	16.27	13.82
P值	0.001	0.001

组别	CD3⁺CD8⁺双阳性细胞率	CD3⁺CD56⁺双阳性细胞率
CIK组	49.49±3.79	14.35±2.46
未致敏DC-CIK组	73.66±3.62^a	14.29±1.53
致敏DC-CIK组	85.41±2.61^a	56.78±2.38^a
统计值	16.27	13.82
P值	0.001	0.001

组别	10∶1	20∶1	40∶1
CIK组	18.94±2.65	26.88±2.97	42.44±3.25
未致敏DC-CIK组	19.58±1.80	37.56±2.54	55.48±3.44
致敏DC-CIK组	25.28±2.08	55.50±3.65	70.88±7.36
统计值	6.64	7.82	9.21
P值	0.01	0.01	0.01

组别	10∶1	20∶1	40∶1
CIK组	18.94±2.65	26.88±2.97	42.44±3.25
未致敏DC-CIK组	19.58±1.80	37.56±2.54	55.48±3.44
致敏DC-CIK组	25.28±2.08	55.50±3.65	70.88±7.36
统计值	6.64	7.82	9.21
P值	0.01	0.01	0.01

组别	IL-12（μg/L）	INF-γ（ng/L）
CIK组	13.92±10.24	207.82±62.78
未致敏DC-CIK组	74.35±37.26	503.42±108.26
致敏DC-CIK组	94.67±39.13	552.21±138.21
统计值	11.34	16.27
P值	0.01	0.001

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