Immunoregulatory properties of cell-free DNA in plasma of celiac disease patients - A pilot study
Immunoregulatory properties of cell-free DNA in plasma of celiac disease patients - A pilot study
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DOI：
10.1080/08916934.2019.1608965
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发表时间：
2019-02-17
期刊：
影响因子：
3.5
通讯作者：
Dankova, Pavlina
中科院分区：
文献类型：
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作者：
Brynychova, Iva;Zinkova, Alzbeta;Dankova, Pavlina
血浆游离 DNA (cfDNA) 浓度升高被多次报道与炎症过程相关。尽管健康个体的 cfDNA 能够调节免疫反应，但尚未研究活动性（新诊断的）乳糜泻患者 (CD) 血浆 cfDNA 的定性和定量特征。我们通过定量 PCR 测定了 CD（n=10）和健康年龄和性别匹配对照（HC，n=10）血浆 cfDNA 中端粒序列的总浓度和相对含量。为了获得所观察到的生物效应仅由 cfDNA 分子引起的证据，我们应用 DNase 处理配对血浆样本。使用配对血浆样本（未经处理/天然和经 DNase 处理），我们分析了 cfDNA 对 THP1 单核细胞系中 TLR9 和 TNF-α mRNA 表达激活的贡献。血浆 cfDNA 的数量和端粒序列的相对含量没有显着差异。当我们比较用天然 CD 和 HC 血浆样品刺激后 THP1 细胞中的 TNF-α mRNA 表达水平时，我们发现用 CD 样品刺激后表达显着升高 (p=.031)。我们还记录了 CD 血浆样本中所含 cfDNA 刺激 TLR9 mRNA 产生的能力。从 CD 血浆样本中去除 cfDNA 后，TLR9 mRNA 表达水平显着降低 (p=.014)。我们的实验设计使我们能够研究 cfDNA 的影响，而无需将其从血浆中分离出来。 CD 血浆样本中所含的 cfDNA 的免疫调节能力与 HC 血浆中的 cfDNA 存在显着差异。这些差异既不是由血浆样品中 cfDNA 浓度不同引起的，也不是由端粒序列相对丰度不同引起的。需要进一步的研究来阐明血浆 cfDNA 在乳糜泻发病机制中的作用。
The elevated plasma cell-free DNA (cfDNA) concentrations were repeatedly reported in association with the process of inflammation. The qualitative and quantitative characteristics of plasma cfDNA in active (newly diagnosed) celiac disease patients (CD) have not yet been studied despite the fact that cfDNA of healthy individuals is able to regulate immune response. We determined the total cfDNA concentration and relative content of telomeric sequences in plasma cfDNA in CD (n=10) and healthy age- and sex-matched controls (HC, n=10) by quantitative PCR. To obtain the evidence that the observed biological effects are caused solely by cfDNA molecules, we applied the treatment of paired plasma samples with DNase. Using paired samples of plasma (non-treated/native and treated by DNase), we analyzed the contribution of cfDNA to the activation of TLR9 and TNF-alpha mRNA expression in THP1 monocytic cell line. There were no significant differences in the quantities of plasma cfDNA and relative contents of telomeric sequences in their pools. When we compared the levels of TNF-alpha mRNA expression in THP1 cells achieved after stimulation with native CD and HC plasma samples, we found significantly (p=.031) higher expression after stimulation with CD samples. We documented also the ability of cfDNA contained in CD plasma samples to stimulate the production of TLR9 mRNA. The TLR9 mRNA expression levels were significantly (p=.014) lowered after cfDNA removal from CD plasma samples. The design of our experiments allowed us to study the effects of cfDNA without its isolation from plasma. cfDNA contained in CD plasma samples differs significantly in its immunoregulatory capacity from cfDNA in HC plasma. The differences are caused neither by different concentrations of cfDNA in plasma samples nor by different relative abundance of telomeric sequences. Further studies are needed to elucidate the role of plasma cfDNA in celiac disease pathogenesis.