Identification of Alternatively Spliced Forms of human OSCAR in Osteoclasts
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
연구목적: 파골세포는 다핵 세포로서 골흡수를 유발한다. 생쥐에서 OSCAR는 파골세포 분화에서 중요한 역할을 함이 밝혀졌다. 인간 OSCAR의 분비형이 클로닝되었고 이들의 파골세포 분화에서의 역할을 분석하고자 하였다. 연구방법: 파골세포 분화는 M-CSF와 RANKL의 처리에 의해 유도하였으며 TRAP 염색법과 pit 형성도를 측정하였다. 발현 정도는 유세포분석, Northern, 및 Western 분석 방법을 사용하였다. 결 과: 인간 OSCAR는 파골세포에서 발현하였으며 PBMC로부터 유도되는 파골세포 형성에 관여하였다. 선택적 이어맞추기에 의해 만들어진 두 종류의 인간 OSCAR가 확인되었다. 이들은 막관통영역이 없는 구조로서 파골세포로부터분비되는 형태로 존재하였다. 이러한 분비형의 인간 OSCAR는 파골세포 분화 및 골흡수를 저해하였다. 결 론: 인간 OSCAR는 최소 5종류의 구조적아형을 갖고 있으며, 이들은 서로 다른 역할을 하고 있다. 특히 분비형인간 OSCAR는 파골세포 분화에서 음성 조절자로서 역할을 한다.
Abstract
Objectives: Osteoclasts are multinucleated giant cells which can resorb bone and differentiated from hematopoietic cells. We have previously reported murine osteoclast-associated receptor (OSCAR) may be an important bone-specific regulator of osteoclast differentiation. We have cloned soluble form of human OSCAR (hOSCAR) and examined the role of hOSCAR on osteoclast differentiation. Methods: Osteoclast differentiation was induced by treatment with macrophage colony-stimulating factor (M-CSF)and receptor activator of nuclear factor kappa B ligand (RANKL) and tartrate-resistant acid phosphatase (TRAP)staining and pit formation were performed. Expression was measured by flow cytometry analysis, Northern and Western blot analysis. Results: hOSCAR is expressed in osteoclast cells and involved in the differentiation of osteoclasts from peripheral blood mononuclear cells (PBMC). Two alternatively spliced forms (soluble hOSCAR [hOSCAR-S]) of hOSCAR were identified from osteoclasts complementary deoxyribonucleic acid (cDNA) library derived from PBMC. Putative transmembrane domain was not found in hOSCAR-S forms and it suggested that these forms might be secreted from osteoclast cells. These secreted forms of hOSCAR attenuated RANKL-induced osteoclast formation and bone resorption. Conclusions: Human osteoclasts express at least five different OSCAR messenger ribonucleic acid (mRNA) isoforms which could play different regulatory roles for differentiation. The secreted forms of hOSCAR might be a negative regulator of membrane-bounded forms of OSCAR.
참고문헌 (16)
- Walsh MC, "Osteoimmunology: Interplay between the immune system and bone metabolism", ANNUAL REVIEW OF IMMUNOLOGY, 24, pp.33~63 (2006), ANNUAL REVIEWS
- Marks SC, Jr., "Osteopetrosis, a new recessive skeletal mutation on chromosome 12 of the mouse", J Hered, 67, pp.11~18 (1976)
- -related activation-induced cytokine), a new TNF family member predominantly expressed in T cells, is a dendritic cell-specific survival factor, "J Exp Med", 1997, 2075~2080
- Anderson DM, "A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function", Nature, 390, pp.175~179 (1997)
- Yasuda H, "Osteoclast differentiation factor is a ligand for osteoprotegerin", osteoclastogenesisinhibitory factor and is identical to TRANCE, Proc Natl Acad Sci U S A, pp.1998, 95[DOI]
- Lacey DL, "Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation", Cell, 93, pp.165~176 (1998)
- Suda T, "Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families", Endocr Rev, 20, pp.345~357 (1999)
- Koga T, "Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis", Nature, 428, pp.758~763 (2004)
- Ishida N, "Large scale gene expression analysis of osteoclastogenesis in vitro and elucidation of NFAT2 as a key regulator", J Biol Chem, 277, pp.41147~41156 (2002)
- Ishikawa S, "Involvement of FcRgamma in signal transduction of osteoclast-associated receptor (OSCAR)", Int Immunol, 16, pp.1019~1025 (2004)
- Herman S, "Induction of osteoclast-associated receptor, a key osteoclast costimulation molecule, in rheumatoid arthritis", Arthritis Rheum, 58, pp.3041~3050 (2008)
- Merck E, "Fc receptor gamma-chain activation via hOSCAR induces survival and maturation of dendritic cells and modulates Toll-like receptor responses", Blood, 105, pp.3623~3632 (2005)
- Merck E, "OSCAR is an FcRgamma-associated receptor that is expressed by myeloid cells and is involved in antigen presentation and activation of human dendritic cells", Blood, 104, pp.1386~1395 (2004)
- Merck E, "Ligation of the FcR gamma chain-associated human osteoclast-associated receptor enhances the proinflammatory responses of human monocytes and neutrophils", J Immunol, 176, pp.3149~3156 (2006)
- Sallusto F, "Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte", macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha, 1994, pp.179, 1109~1118
- Kim N, "A novel member of the leukocyte receptor complex regulates osteoclast differentiation", J Exp Med, 195, pp.201~209 (2002)