Phillips Kate L E, Chiverton Neil, Michael Anthony L R, Cole Ashley A, Breakwell Lee M, Haddock Gail, Bunning Rowena A D, Cross Alison K, Le Maitre Christine L
Arthritis Res Ther. 2013;15(6):R213. doi: 10.1186/ar4408.
The aims of these studies were to identify the cytokine and chemokine expression profile of nucleus pulposus (NP) cells and to determine the relationships between NP cell cytokine and chemokine production and the characteristic tissue changes seen during intervertebral disc (IVD) degeneration.
Real-time q-PCR cDNA Low Density Array (LDA) was used to investigate the expression of 91 cytokine and chemokine associated genes in NP cells from degenerate human IVDs. Further real-time q-PCR was used to investigate 30 selected cytokine and chemokine associated genes in NP cells from non-degenerate and degenerate IVDs and those from IVDs with immune cell infiltrates (‘infiltrated’). Immunohistochemistry (IHC) was performed for four selected cytokines and chemokines to confirm and localize protein expression in human NP tissue samples.
LDA identified the expression of numerous cytokine and chemokine associated genes including 15 novel cytokines and chemokines. Further q-PCR gene expression studies identified differential expression patterns in NP cells derived from non-degenerate, degenerate and infiltrated IVDs. IHC confirmed NP cells as a source of IL-16, CCL2, CCL7 and CXCL8 and that protein expression of CCL2, CCL7 and CXCL8 increases concordant with histological degenerative tissue changes.
Our data indicates that NP cells are a source of cytokines and chemokines within the IVD and that these expression patterns are altered in IVD pathology. These findings may be important for the correct assessment of the ‘degenerate niche’ prior to autologous or allogeneic cell transplantation for biological therapy of the degenerate IVD.
这些研究的目的是确定髓核(NP)细胞的细胞因子和趋化因子表达谱,并确定NP细胞细胞因子和趋化因子产生与椎间盘(IVD)退变期间所见特征性组织变化之间的关系。
使用实时定量聚合酶链反应(q-PCR)cDNA低密度阵列(LDA)研究来自退变人IVD的NP细胞中91种细胞因子和趋化因子相关基因的表达。进一步使用实时q-PCR研究来自非退变和退变IVD以及具有免疫细胞浸润(“浸润”)的IVD的NP细胞中30种选定的细胞因子和趋化因子相关基因。对四种选定的细胞因子和趋化因子进行免疫组织化学(IHC),以确认并定位人NP组织样本中的蛋白质表达。
LDA鉴定出众多细胞因子和趋化因子相关基因的表达,包括15种新的细胞因子和趋化因子。进一步的q-PCR基因表达研究确定了来自非退变、退变和浸润IVD的NP细胞中的差异表达模式。IHC证实NP细胞是白细胞介素-16(IL-16)、趋化因子配体2(CCL2)、趋化因子配体-7(CCL7)和趋化因子配体-8(CXCL8)的来源,并且CCL2、CCL7和CXCL8的蛋白质表达与组织学退变组织变化一致增加。
我们的数据表明NP细胞是IVD内细胞因子和趋化因子的来源,并且这些表达模式在IVD病理学中发生改变。这些发现对于在自体或异体细胞移植用于退变IVD的生物治疗之前正确评估“退变微环境”可能很重要。