胰岛素对脐血来源树突状细胞功能状态的影响及其对树突状细胞诱导的 CTL 对胰腺癌细胞系的细胞毒性作用。

Effect of insulin on functional status of cord blood-derived dendritic cells and on dendritic cell-induced CTL cytotoxicity against pancreatic cancer cell lines.

机构信息

Department of Surgery, First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.

出版信息

Hepatobiliary Pancreat Dis Int. 2009 Oct;8(5):529-34.

PMID:19822498

Abstract

BACKGROUND

Dendritic cells (DCs) are the most important antigen-presenting cells in the human body, and DCs with different mature status possess different or even opposite functions. This study was designed to explore the influence of insulin on the functional status of cord blood-derived DCs and on DC-induced cytotoxic T lymphocyte (CTL) activity against pancreatic cancer cell lines.

METHODS

Mononuclear cells were isolated from fresh cord blood. Interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were used to induce or stimulate the mononuclear cells. Insulin at different concentrations served to modify DCs, and then DC morphology, number, and growth status were assessed. The DC immunophenotype was detected with a flow cytometer. The IL-12 in DC supernatant was determined by ELISA. DC functional status was evaluated by the autologous mixed lymphocyte reaction. T lymphocytes were induced by insulin-modified DCs to become CTLs. The CTL cytotoxicity against pancreatic cancer cell lines was determined.

RESULTS

Mononuclear cells from cord blood can be differentiated into DCs by cytokine induction and insulin modification. With the increase in insulin concentration (2.5-25 mg/L), the expression of DC HLA-DR, CD1alpha, CD80, and CD83 was significantly increased, the DC ability to secrete IL-12 was significantly improved, DC function to activate autologous lymphocytes was significantly enhanced, and the cytotoxicity of CTLs induced by insulin-modified DCs against pancreatic cancer cell lines was significantly strengthened.

CONCLUSIONS

Insulin may facilitate DC induction and maturation, and improve the reproductive activity of autologous lymphocytes. The cytotoxicity of CTLs induced by insulin-modified DCs against pancreatic cancer cell lines was significantly enhanced. Insulin may serve as a factor modifying DCs and inducing CTLs in vitro in insulin biotherapy.

摘要

背景

树突状细胞（DC）是人体内最重要的抗原呈递细胞，不同成熟状态的 DC 具有不同甚至相反的功能。本研究旨在探讨胰岛素对脐血来源的 DC 功能状态以及 DC 诱导的抗胰腺癌细胞系细胞毒性 T 淋巴细胞（CTL）活性的影响。

方法

从新鲜脐血中分离出单个核细胞。白细胞介素-4（IL-4）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）用于诱导或刺激单核细胞。用不同浓度的胰岛素修饰 DC，然后评估 DC 形态、数量和生长状态。用流式细胞仪检测 DC 免疫表型。用 ELISA 法测定 DC 上清液中 IL-12 的含量。通过自体混合淋巴细胞反应评估 DC 功能状态。用胰岛素修饰的 DC 诱导 T 淋巴细胞成为 CTL。测定 CTL 对胰腺癌细胞系的细胞毒性。

结果

通过细胞因子诱导和胰岛素修饰，脐血中的单个核细胞可分化为 DC。随着胰岛素浓度（2.5-25 mg/L）的增加，DC HLA-DR、CD1alpha、CD80 和 CD83 的表达显著增加，DC 分泌 IL-12 的能力显著提高，DC 激活自身淋巴细胞的功能显著增强，胰岛素修饰的 DC 诱导的 CTL 对胰腺癌细胞系的细胞毒性显著增强。