Characterization of the Cytokine Pattern of Porcine Bone Marrow‐Derived Cells Treated with 1α,25(OH)2D3
Characterization of the Cytokine Pattern of Porcine Bone Marrow‐Derived Cells Treated with 1α,25(OH)2D3
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1α,25(OH)2D3 处理的猪骨髓来源细胞的细胞因子模式表征
DOI：
10.1111/j.1439-0442.2005.00755.x
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发表时间：
2005
期刊：
影响因子：
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通讯作者：
P. Pietschmann
中科院分区：
文献类型：
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作者：
W. Sipos;J. Duvigneau;F. Schmoll;B. Exel;G. Hofbauer;G. Baravalle;R. Hartl;M. Dobretsberger;P. Pietschmann
最近，维生素 D(3) [1α,25(OH)(2)D(3)] 的生物活性形式不仅被描述为影响骨代谢，而且还发挥免疫调节活性，这也可能对骨形成/吸收产生影响。在本研究中，我们分析了 1alpha,25(OH)(2)D(3) 对 1-3 周龄仔猪骨髓来源细胞细胞因子模式的影响。培养1周后，测定破骨细胞的数量，其中抗酒石酸酸性磷酸酶（TRAP）阳性的多核细胞被认为是破骨细胞。对培养的骨髓细胞衍生的 mRNA 进行半定量 RT-PCR，对一组猪细胞因子（IL-1α、IL-6、IL-8、IL-10 和 TNF-α）具有特异性。此外，还使用特异针对 IL-1β、IL-2、IL-4、IL-6、IL-12、TNF-α 和 IFN-γ 的抗猪单克隆抗体进行了免疫荧光分析。为了证明 NF-κB 配体受体激活剂 (RANKL) 的猪同源物的存在，使用鼠 RANKL 特异的引物对通过 RT-PCR 研究了骨髓细胞以及猪白细胞来源的 mRNA。通过设计用于定量人 RANKL 的 ELISA 分析细胞培养物上清液中的可溶性 RANKL。通过RT-PCR方法，在有和没有1α,25(OH)(2)D(3)培养的细胞中均发现IL-1α、IL-6、IL-8、IL-10和TNF-α mRNA的表达。免疫荧光分析显示，基质细胞和破骨细胞均产生 IL-1、IL-6 和 TNF-α。除了其已知的破骨作用外，1α,25(OH)(2)D(3) 倾向于下调各自的细胞因子，但显着上调 RANKL 表达。猪RANKL特异性序列与相应的人RANKL序列之间的同源性为79%。发现的数据支持这样的观点：猪骨髓细胞培养物可能为人类骨学研究中的鼠类系统提供合适的替代品。
The biologically active form of vitamine D(3) [1alpha,25(OH)(2)D(3)] has recently been described not only to influence bone metabolism but also to exert immunomodulating activities, which may have an impact on bone formation/resorption as well. In this study, we analysed the effects of 1alpha,25(OH)(2)D(3) on the cytokine pattern of porcine bone marrow-derived cells from piglets aged 1-3 weeks. After culture for 1 week, the number of osteoclasts was determined, with tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated cells being considered osteoclasts. Cultured bone marrow cell-derived mRNA was subjected to semiquantitative RT-PCR specific for a panel of porcine cytokines (IL-1alpha, IL-6, IL-8, IL-10, and TNF-alpha). In addition, an immunofluorescence analysis using anti-porcine mAbs specific for IL-1beta, IL-2, IL-4, IL-6, IL-12, TNF-alpha, and IFN-gamma was performed. In order to prove the existence of a porcine homologue of the receptor activator of NF-kappaB ligand (RANKL) bone marrow cell- as well as porcine white blood cell-derived mRNA was investigated by RT-PCR using primer pairs specific for murine RANKL. Cell culture supernatant was analysed for soluble RANKL by means of an ELISA designed for quantification of human RANKL. By means of RT-PCR, expression of IL-1alpha, IL-6, IL-8, IL-10 and TNF-alpha mRNA could be found in cells cultured with and without 1alpha,25(OH)(2)D(3). Immunofluorescence analysis revealed that IL-1, IL-6, and TNF-alpha were produced by both stromal cells and osteoclasts. Besides its known osteoclastogenic effects, 1alpha,25(OH)(2)D(3) tended to downregulate the respective cytokines, but significantly upregulated RANKL expression. The homology between the porcine RANKL-specific sequence and the corresponding human RANKL sequence was 79%. The data found support the idea that porcine bone marrow cell cultures may provide a suitable alternative to murine systems in human osteological research.
甲状旁腺激素刺激 TRANCE 并抑制小鼠骨髓培养物中骨保护素信使核糖核酸的表达：与破骨细胞样细胞形成的相关性。
DOI：
10.1210/endo.140.8.6887
发表时间：
1999
期刊：
Endocrinology.
影响因子：
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作者：
Lee,SK;Lorenzo,JA
通讯作者：
Lorenzo,JA