Cytokine mRNA and protein expression in primary-culture and repeated-passage synovial fibroblasts from patients with rheumatoid arthritis.
Cytokine mRNA and protein expression in primary-culture and repeated-passage synovial fibroblasts from patients with rheumatoid arthritis.
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DOI：
10.1186/ar391
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Sack U
中科院分区：
文献类型：
--
作者：
Hirth A;Skapenko A;Kinne RW;Emmrich F;Schulze-Koops H;Sack U
在类风湿性关节炎（RA）滑膜成纤维细胞（SFB），分离自原代培养物或通过反复传代衍生的组成性mRNA表达和促炎和抗炎细胞因子的分泌进行了比较分析;正常皮肤成纤维细胞用作对照。首次传代RA-SFB（n = 3）分泌大量IL-6（15，800 ± 2，110 pg/ml;平均值± SEM），但仅分泌少量肿瘤坏死因子（TNF）-α（22.1 ± 1.1 pg/ml）或IL-10（35.7 ± 34.2 pg/ml; 3份样本中仅1份为阳性）。IL-1β、IL-15和IL-18在蛋白水平上未检测到，并且通过半定量RT-PCR显示出非常低的mRNA水平。在重复传代RA-SFB（第10代）中，IL-6（初始水平的二十分之一）和TNF-α（三分之二）的蛋白分泌显著降低，IL-10显著降低（四分之一，三个样本中仅一个呈阳性）。虽然IL-10蛋白从第1代至第10代的减少被相应的mRNA减少所抵消，但IL-6和TNF-α的相对mRNA水平实际上增加（分别为20倍和300倍），表明这些细胞因子的转录后和/或翻译后调节。然而，由于个体患者之间的高度可变水平，在原代培养和重复传代RA-SFB（第九代）之间未观察到任何细胞因子mRNA的显著差异。同样，在RA-SFB和正常皮肤成纤维细胞（原代培养和重复传代）之间没有检测到显著差异。通过产生大量的IL-6和有限量的TNF-α，RA-SFB可能有助于炎症关节中促炎和抗炎细胞因子的平衡。
Constitutive mRNA expression and secretion of proinflammatory and anti-inflammatory cytokines was comparatively analyzed in rheumatoid arthritis (RA) synovial fibroblasts (SFB), isolated from primary culture or derived by repeated passage; normal-skin fibroblasts were used as controls. First-passage RA-SFB (n = 3) secreted large amounts of IL-6 (15,800 ± 2,110 pg/ml; mean ± SEM), but only limited amounts of tumor necrosis factor (TNF)-α (22.1 ± 1.1 pg/ml) or IL-10 (35.7 ± 34.2 pg/ml; only one of three samples was positive). IL-1β, IL-15, and IL-18 were not detectable at the protein level and showed very low mRNA levels by semiquantitative RT-PCR. In repeated-passage RA-SFB (tenth passage), protein secretion was significantly lower for IL-6 (one-twentieth of the initial level) and TNF-α (two-thirds), and markedly reduced for IL-10 (one-quarter, with only one of three samples positive). While the decrease of IL-10 protein from first to tenth passage was paralleled by a corresponding decrease of mRNA, the relative mRNA levels for IL-6 and TNF-α were actually increased (20-fold and 300-fold, respectively), indicating post-transcriptional and/or post-translational regulation of these cytokines. Due to highly variable levels among individual patients, however, no significant differences were observed for any cytokine mRNA between primary-culture and repeated-passage RA-SFB (ninth passage). Likewise, no significant differences were detectable between RA-SFB and normal-skin fibroblasts (primary-culture and repeated-passage). By producing high amounts of IL-6 and limited amounts of TNF-α, RA-SFB may contribute to the (im)balance of proinflammatory and anti-inflammatory cytokines in the inflamed joint.
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DOI：
10.1186/ar109
发表时间：
2000
期刊：
Arthritis research
影响因子：
--
作者：
Kontoyiannis D;Kollias G
通讯作者：
Kollias G
影响因子：
4
作者：
BURMESTER, GR;LOCHER, P;MOHR, W
通讯作者：
MOHR, W
DOI：
10.1073/pnas.86.16.6171
发表时间：
1989-08-01
影响因子：
11.1
作者：
ELIAS, JA;REYNOLDS, MM;KERN, JA
通讯作者：
KERN, JA
影响因子：
5.5
作者：
BOMBARA, MP;WEBB, DL;BLUE, ML
通讯作者：
BLUE, ML
DOI：
10.1186/ar86
发表时间：
2000
期刊：
Arthritis research
影响因子：
--
作者：
Kinne RW;Bräuer R;Stuhlmüller B;Palombo-Kinne E;Burmester GR
通讯作者：
Burmester GR