Abediankenari Saeid, Yousefzadeh Yousef, Azadeh Hossein, Vahedi Mohammad
Department of Microbiology and Immunology, Mazandaran University of Medical Sciences, Sari, Iran.
Iran J Immunol. 2010 Jun;7(2):83-7.
Dendritic cells (DCs) are professional antigen presenting cells that have an important role in the initiation of immune response. The use of maturation factors in dendritic cell differentiation provides a promising approach in immunotherapy.
In this study, we compared tumor necrosis factor-α, polyribocytidylic acid, lipopolysacharide and CpG oligonucleotides in inducing dendritic cell maturation.
We generated immature dendritic cells with GM-CSF in combination with IL-4 from peripheral blood mononuclear adherent cells and used tumor necrosis factor-α, polyribocytidylic acid, lipopolysacharide and CpG for the induction of dendritic cell maturation. CD83 maturation marker on the dendritic cells was analyzed by flowcytometry after 7 days. In addition, mixed leukocyte reaction between dendritic cells and T cells was performed by MTT proliferation assay.
Flow cytometry results demonstrated a comparable high level of CD83 expression on the mature dendritic cells generated by TNF-α, CpG, Poly I:C, and LPS treatment of the immature dendritic cells. However, a significantly poorer proliferation of lymphocytes cocultured with the Poly I:C-treated DCs was observed compared to the CpG-treated DCs in mixed leukocyte reaction (p=0.026). Conversely, a significantly stronger proliferation of lymphocytes was observed when cocultured with TNF-α-treated DCs compared to the LPS-treated DCs (p=0.025).
Our results indicated that all of studied maturation inducing factors can be used in DC maturation but TNF-α and CpG were the preferred in vitro maturation factors. It is concluded that maturation of dendritic cells by CpG motif and TNF-α can be used to regulate immune responses.
树突状细胞（DCs）是专业的抗原呈递细胞，在免疫反应的启动中起重要作用。在树突状细胞分化过程中使用成熟因子为免疫治疗提供了一种有前景的方法。
在本研究中，我们比较了肿瘤坏死因子-α、聚肌胞苷酸、脂多糖和CpG寡核苷酸在诱导树突状细胞成熟方面的作用。
我们从外周血单核贴壁细胞中用粒细胞-巨噬细胞集落刺激因子（GM-CSF）联合白细胞介素-4生成未成熟树突状细胞，并使用肿瘤坏死因子-α、聚肌胞苷酸、脂多糖和CpG诱导树突状细胞成熟。7天后通过流式细胞术分析树突状细胞上的CD83成熟标志物。此外，通过MTT增殖试验进行树突状细胞与T细胞之间的混合淋巴细胞反应。
流式细胞术结果表明，用肿瘤坏死因子-α、CpG、聚肌胞苷酸（Poly I:C）和脂多糖处理未成熟树突状细胞所产生的成熟树突状细胞上CD83表达水平相当高。然而，在混合淋巴细胞反应中，与用CpG处理的树突状细胞相比，观察到与用聚肌胞苷酸处理的树突状细胞共培养的淋巴细胞增殖明显较差（p = 0.026）。相反，与用脂多糖处理的树突状细胞相比，与用肿瘤坏死因子-α处理的树突状细胞共培养时观察到淋巴细胞增殖明显更强（p = 0.025）。
我们的结果表明，所有研究的成熟诱导因子均可用于树突状细胞成熟，但肿瘤坏死因子-α和CpG是体外成熟的首选因子。得出结论，CpG基序和肿瘤坏死因子-α诱导树突状细胞成熟可用于调节免疫反应。
