Identification of specific expressed genes of inflammatory Osteoclasts in joints of reheumatoid arthritis patients

类风湿性关节炎患者关节炎症破骨细胞特异性表达基因的鉴定

• 批准号: 16390447
• 负责人: SUZUKI Ryuji
• 金额: $ 8.9万
• 依托单位: Clinical Research Center for Rheumatology and Allergy, Sagamihara National Hospital, National Hospital Organization
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390447/
• 关键词: Rheumatoid arthritis; Osteoclast; Nurse cell; Inflammation; Gene-analysis; 骨破壊; DNAチップ

Objective : We have previously shown in vitro differentiation of monocytes into osteoclast-like multinucleated cells by granulocyte-macrophage colony-stimulating factor (GM-CSF), well-known as the potent inhibitor of osteoclastogenesis. We compared GM-CSF-induced multicleated cells with those induced by receptor activator of nuclear factor kappa B ligand (RANKL) to elucidate the attributes of osteoclast-like cells generated under the pathgenic condition of rheumatoid arthritis (RA).Methods : The precursors were produced by the co-culture of monocytes with synovial nurse cells derived fro RA joints (RA-SNCs) and then subjected to the formation of multinucleated cells. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to measure the expression of matrix metalloproteinase (MMP)-12 and six osteoclast markers ; tartrate-resistant acid phosphatase (TRAP), crbonic anhydrase, cathepsin K, calcitonin receptor, integirin αV and MMP-9. TRAP and MMP-12 in RA joints specimen were evaluated histochemically.Results : The mononuclear cells co-cultivated with RA-SNCs expressed ostoclast markers except for MMP-9, and maintained the ability of osteoclast-like cell formation induced by RANKL, RANKL-induced cells expressed MMP-9 as well as other ostoclast markers, but not MMP-12. On the other hand, by thestimulation of GM-CSF, the mononuclear cells also differentiated into osteoclast-like multinuleated cells expressing MMP-9 and MMP-12. Farthermore, the multinucleated cells producing MMP-12 were found in the erosive bone area of rheumatoid joints.Conclusion : These findings suggest the presence of two distinct types of TRAP-positive multinucleated cells in RA joints. GM-CSF-induced cells, distinguishable from RANKL-induced cells by the expression of MMP-12, should be involved in the pathology of RA. The co-culture with RA-SNCs is very useful system to investigate mechanism of the differentiation of synovial macrophages and their roles in the pathogenesis of RA.

目的：我们以前已经显示了在体外分化的单核细胞成破骨细胞样多核细胞的粒细胞-巨噬细胞集落刺激因子（GM-CSF），众所周知的破骨细胞生成的有效抑制剂。为探讨类风湿关节炎（RA）发病条件下破骨细胞样细胞的特性，我们将GM-CSF诱导的多核细胞与核因子κ B配体受体激活剂（RANKL）诱导的多核细胞进行比较。采用逆转录-聚合酶链反应（RT-PCR）检测基质金属蛋白酶（MMP）-12和6种破骨细胞标志物：抗酒石酸酸性磷酸酶（TRAP）、骨酸酐酶、组织蛋白酶K、降钙素受体、整合素αV和MMP-9的表达。结果：RA-SNCs共培养的单个核细胞表达破骨细胞标志物（MMP-9除外），并保持RANKL诱导的破骨细胞样细胞形成能力，RANKL诱导的细胞表达MMP-9及其他破骨细胞标志物，但不表达MMP-12。另一方面，通过GM-CSF的刺激，单个核细胞也分化为表达MMP-9和MMP-12的破骨细胞样多核细胞。结论：RA关节内存在两种不同类型的TRAP阳性多核细胞，其表达的MMP-12与RA关节内的TRAP阳性多核细胞表达的MMP-12呈负相关。GM-CSF诱导的细胞与RANKL诱导的细胞通过MMP-12的表达而区别，应该参与RA的病理过程。RA SNCs与滑膜巨噬细胞共培养是研究RA滑膜巨噬细胞分化机制及其在RA发病机制中的作用的一个非常有用的系统。

项目成果

Production of interleukin-6 and interleukin-8 by nurse-like cells from rheumatoid arthritis patients after stimulation with monocytes.

类风湿性关节炎患者的护士样细胞在单核细胞刺激后产生白细胞介素 6 和白细胞介素 8。

• 发表时间: 2006
• 期刊: Mod Rheumatol. 15
• 作者: Tanaka K;Mori T;Juji T;Suzuki S;Watanabe J;Goto A;Shiobara N;Yamane S;Fukui N;Suzuki R;Ochi T
• 通讯作者: Ochi T

Production of interleukin-6 and interleukin-8 by nurse-like cells from rheumatoid arthritis patients after stimulation with monocytes

类风湿性关节炎患者的护士样细胞在单核细胞刺激后产生白细胞介素 6 和白细胞介素 8

• 发表时间: 2005
• 期刊: Mod Rheumatol 15
• 作者: Tanaka K;Mori T;Juji T;Suzuki S;Watanabe J;Goto A;Shiobara N;Yamane S;Fukui N;Suzuki R;Ochi T.
• 通讯作者: Ochi T.

Methods for isolation of Osteoclast precursor cells and inducing their differentiation in to Osteoclasts

分离破骨细胞前体细胞并诱导其分化为破骨细胞的方法

• 发表时间: 2000

Evidence for existence of oligoclonal tumor-infiltrating lymphocytes and predominant production of T helper 1/T cytotoxic 1 type cytokines in gastric and colorectal tumors.

胃和结直肠肿瘤中存在寡克隆肿瘤浸润淋巴细胞和 T 辅助细胞 1/T 细胞毒性 1 型细胞因子的主要产生的证据。

• 发表时间: 2004
• 期刊: International Journal of Oncology 25・1
• 作者: Yamazaki;K.;Shimada;S.;Kato-Nagoka;N.;Soga;H.;Itoh;T.;Nanno;M.;Kei Yaguchi;Takaji Matsutani
• 通讯作者: Takaji Matsutani

Limited VH gene usage in B-cell clones established with nurse-like cells from patients with rheumatoid arthritis

使用来自类风湿关节炎患者的护士样细胞建立的 B 细胞克隆中 VH 基因的使用有限

• 发表时间: 2006
• 期刊: Rheumatology (Oxford) 45・5
• 作者: Abe J;Hosokawa H;Sawada Y;Matsumura K;Kobayashi S;Shino Nakamura-Kikuoka
• 通讯作者: Shino Nakamura-Kikuoka