负载抗原DC联合CIK对肝癌免疫微环境的影响

田大广; 于恒海; 魏晓平

doi:10.12259/j.issn.2095-610X.S20210127

负载抗原DC联合CIK对肝癌免疫微环境的影响

doi: 10.12259/j.issn.2095-610X.S20210127

昆明医科大学第二附属医院肝胆胰外科一病区云南昆明　650101

基金项目: 云南省教育厅科学研究基金资助项目（2019J1264）

详细信息

作者简介:
田大广（1970～），男，江苏无锡人，医学硕士，主任医师，主要从事肝胆外科临床工作

通讯作者:
魏晓平，E-mail：florakm@163.com

中图分类号: R730.54；R320.2445
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出版历程
- 收稿日期: 2020-10-11
- 网络出版日期: 2021-01-26
- 刊出日期: 2021-01-26

Effect of Tumor Antigen Loaded DC-CIK on Microenvironment of Hepatocellular Carcinoma

The First Department of HBP Surgery，The 2nd Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650101，China

摘要

摘要: 目的探讨负载肝癌抗原树突状细胞（dendritic cell，DC）联合细胞因子诱导杀伤细胞（cytokine-induced killer，CIK）对肝细胞性肝癌（hepatocellular carcinoma，HCC）微环境的影响。方法采用手术切除HCC组织制备HCC抗原，采集健康志愿者单个核细胞，诱导并分离出DC，CIK，按是否加入HCC抗原及共培养将细胞分为DC、Ag-Dc、CIK、Ag-DC-CIK四组，流式细胞仪测定各组细胞表型后，ELISA检测IL-6、IL-10及IFN-γ水平；免疫细胞化学SP法检测血管内皮生长因子（vascular endothelial growth factor，VEGF）的表达；RT-PCR检测基质金属蛋白酶（matrix metalloproteinase，MMP）-2，MMP-3，MMP-9mRNA表达水平。结果负载抗原后DC组CD80及CD86表达阳性率，Ag-DC-CIK组CD3⁺CD56⁺、CD8⁺CD56⁺双阳性细胞均显著升高（P < 0.05）； Ag-DC-CIK组IL-6及IFN-γ浓度显著低于其余各组；而IL-10浓度最高（P < 0.05）；Ag-DC-CIK组VEGF及MMP-2、MMP-3、MMP-9mRNA表达水平均显著下降（P < 0.05）。结论在DC负载自身抗原的基础上，与CIK共培养，可显著增强DC及CIK活性，有效降低HePG2微环境中血管形成，炎症反应及纤维化相关因子表达。
- 肝细胞肝癌 /
- 微环境 /
- 树突状细胞 /
- 细胞因子诱导杀伤细胞
Abstract: Objective To study the effect of tumor antigen loaded DC-CIK on the microenvironment of hepatocellular carcinoma. Methods The antigen of HCC coming from resected HCC and peripheral blood mononuclear cells were isolated from healthy donors. The cells were divided into 4 groups as DC, Ag-DC, CIK and Ag-DC-CIK according to the loading of HCC antigen or coculture of DC-CIK or not. Cell phenotype of 4 groups were evaluated by FCM. Furthermore, levels of IL-6, IL-10 and IFN-γ; expression of VEGF and MMP-2、MMP-3、MMP-9mRNA were observed. Results The contents of CD80, CD86 in group Ag-DC and CD3⁺CD56⁺, CD8⁺CD56⁺ double positive cells in group Ag-DC-CIK elevated significantly（P < 0.05）. The level of IL-6 and IFN-γdecreased significantly in group Ag-DC-CIK（P < 0.05）. Expression of VEGF and MMP-2, MMP-3, MMP-9mRNA also reduced significantly in group Ag-DC-CIK（P < 0.05）. Conclusion Coculture of DC and CIK on the basis of loading HCC antigen shows the stronger activity and reduces the expression of relating factors about inflammation, angiopoiesis and fibrosis in HePG2 microenvironment.
- Hepatocellular carcinoma /
- Microenvironment /
- Dendritic cell /
- Cytokine-induced killer

HTML全文

图 1 各组IL-6、IL-10及IFN-γ水平

与NC组比较，* P < 0.05；** P < 0.01。

Figure 1. Level of IL-6、IL-10 and IFN-γ in all groups

下载: 全尺寸图片幻灯片

图 2 各组VEGF表达水平

与NC组比较，* P < 0.05；** P < 0.01。

Figure 2. Level of VEGF in all groups

下载: 全尺寸图片幻灯片

图 3 各组MMP-2、MMP-3、MMP-9mRNA表达水平

与NC组比较，* P < 0.05；** P < 0.01。

Figure 3. Level of MMP-2、MMP-3 and MMP-9mRNA in all groups

下载: 全尺寸图片幻灯片

表 1 两组DC表面标志分子表达比较（$\bar x \pm s $）

Table 1. Comparison of surface marker expression between DC and Ag-DC（$\bar x \pm s $）

表面标记物	分组		t	P
表面标记物	DC	Ag-DC	t	P
CD80	52.07 ± 0.78	63.47 ± 0.93	−134.5	0.020
CD86	43.20 ± 0.95	61.33 ± 0.78	−150.7	0.016

下载: 导出CSV

表 2 各组CIK细胞表型比较（$\bar x \pm s $）

Table 2. Comparison of immune phenotypes in different CIK groups（$\bar x \pm s $）

细胞表型	分组			F	P
细胞表型	CIK	DC-CIK	Ag-DC-CIK	F	P
CD3⁺CD8⁺	48.37 ± 2.09	59.35 ± 1.28	76.27 ± 3.32	407.29	0.013
CD3⁺CD56⁺	16.43 ± 1.21	35.51 ± 2.03	57.73 ± 2.67	356.19	0.021

下载: 导出CSV

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