国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2011年
10期
750-756
,共7页
曾宪升%于宝丹%任敦强%王君丽%何臣%罗永峰%刘明%郑丽霞%陈敏慧%汪延生%付志萍%徐军
曾憲升%于寶丹%任敦彊%王君麗%何臣%囉永峰%劉明%鄭麗霞%陳敏慧%汪延生%付誌萍%徐軍
증헌승%우보단%임돈강%왕군려%하신%라영봉%류명%정려하%진민혜%왕연생%부지평%서군
肺纤维化%自然杀伤T细胞%间充质干细胞%免疫调节
肺纖維化%自然殺傷T細胞%間充質榦細胞%免疫調節
폐섬유화%자연살상T세포%간충질간세포%면역조절
Pulmonary fibrosis%Natural killer T cell%Mesenchymal stem cell%Immunomodulation
目的 研究骨髓间充质干细胞在间质性肺纤维化中的抗损伤治疗作用机制.方法 通过体外诱导的自然杀伤T细咆(NKT细胞)模拟间质性肺纤维化炎症损伤特点.以干扰素γ(IFN-γ),CD3,白介素2刺激培养外周血单个核细胞,得到含高比例CD3+ CD56+ NKT细胞,将这种培养的CD3+ CD56+ NKT细胞与人骨髓间充质干细胞共培养,留取培养基上清液,流式细胞仪鉴定NKT表型.用ELISA方法检测上清液中转化生长因子γ(TGF-γ)和干扰素诱导蛋白10(IP-10)细胞因子水平,并用液相芯片技术检测共培养上清中IFN-γ、肿瘤坏死因子α(TNF-α)等炎性因子水平.CD3+ CD56+NKT细胞与人骨髓间充质干细胞共作用后,再与16HBE共作用4 h,以CCK8法检测16HBE细胞增殖活力.结果 体外培养可见间充质干细胞对包括CD3+ CD56+ NKT细胞在内的多种T细胞亚群具有免疫调节作用,能够降低外周血单个核细胞中高比例的CD3+ CD56+ NKT细胞,诱导调节性T细胞的分化,下调IFN-γ、TNF-α等多种炎性因子水平,间充质干细胞作用后的CD3+ CD56+ NKT细胞由(20.33±1.05)%降为(15.17±1.75)%(P＜0.05).能够分泌高水甲的TGF-β1及IP-10是其具有免疫调节作用的重要分子基础,间克质干细胞减轻了NKT细胞对肺上皮细胞的杀伤作用.结论 间充质干细胞对包括CD3+ CD56+ NKT细胞在内的多种T细胞亚群具有免疫调节作用,对NKT细胞杀伤肺上皮细胞具有保护作用.
目的 研究骨髓間充質榦細胞在間質性肺纖維化中的抗損傷治療作用機製.方法 通過體外誘導的自然殺傷T細咆(NKT細胞)模擬間質性肺纖維化炎癥損傷特點.以榦擾素γ(IFN-γ),CD3,白介素2刺激培養外週血單箇覈細胞,得到含高比例CD3+ CD56+ NKT細胞,將這種培養的CD3+ CD56+ NKT細胞與人骨髓間充質榦細胞共培養,留取培養基上清液,流式細胞儀鑒定NKT錶型.用ELISA方法檢測上清液中轉化生長因子γ(TGF-γ)和榦擾素誘導蛋白10(IP-10)細胞因子水平,併用液相芯片技術檢測共培養上清中IFN-γ、腫瘤壞死因子α(TNF-α)等炎性因子水平.CD3+ CD56+NKT細胞與人骨髓間充質榦細胞共作用後,再與16HBE共作用4 h,以CCK8法檢測16HBE細胞增殖活力.結果 體外培養可見間充質榦細胞對包括CD3+ CD56+ NKT細胞在內的多種T細胞亞群具有免疫調節作用,能夠降低外週血單箇覈細胞中高比例的CD3+ CD56+ NKT細胞,誘導調節性T細胞的分化,下調IFN-γ、TNF-α等多種炎性因子水平,間充質榦細胞作用後的CD3+ CD56+ NKT細胞由(20.33±1.05)%降為(15.17±1.75)%(P＜0.05).能夠分泌高水甲的TGF-β1及IP-10是其具有免疫調節作用的重要分子基礎,間剋質榦細胞減輕瞭NKT細胞對肺上皮細胞的殺傷作用.結論 間充質榦細胞對包括CD3+ CD56+ NKT細胞在內的多種T細胞亞群具有免疫調節作用,對NKT細胞殺傷肺上皮細胞具有保護作用.
목적 연구골수간충질간세포재간질성폐섬유화중적항손상치료작용궤제.방법 통과체외유도적자연살상T세포(NKT세포)모의간질성폐섬유화염증손상특점.이간우소γ(IFN-γ),CD3,백개소2자격배양외주혈단개핵세포,득도함고비례CD3+ CD56+ NKT세포,장저충배양적CD3+ CD56+ NKT세포여인골수간충질간세포공배양,류취배양기상청액,류식세포의감정NKT표형.용ELISA방법검측상청액중전화생장인자γ(TGF-γ)화간우소유도단백10(IP-10)세포인자수평,병용액상심편기술검측공배양상청중IFN-γ、종류배사인자α(TNF-α)등염성인자수평.CD3+ CD56+NKT세포여인골수간충질간세포공작용후,재여16HBE공작용4 h,이CCK8법검측16HBE세포증식활력.결과 체외배양가견간충질간세포대포괄CD3+ CD56+ NKT세포재내적다충T세포아군구유면역조절작용,능구강저외주혈단개핵세포중고비례적CD3+ CD56+ NKT세포,유도조절성T세포적분화,하조IFN-γ、TNF-α등다충염성인자수평,간충질간세포작용후적CD3+ CD56+ NKT세포유(20.33±1.05)%강위(15.17±1.75)%(P＜0.05).능구분비고수갑적TGF-β1급IP-10시기구유면역조절작용적중요분자기출,간극질간세포감경료NKT세포대폐상피세포적살상작용.결론 간충질간세포대포괄CD3+ CD56+ NKT세포재내적다충T세포아군구유면역조절작용,대NKT세포살상폐상피세포구유보호작용.
Objective To explore the mechanism of anti-injury effects of human bone marrow mesenchymal stem cells on interstitial pulmonary fibrosis. Methods Natural killer T cell (NKT cell) was induced in vitro to simulate the inflammatory characteristics of interstitial pulmonary fibrosis. Peripheral blood mononuclcar cells (PBMC) were cultured with addition of interferon-γ (IFN-γ),CD3 antibody and interleukin-2 to contain high proportion of CD3+ CD56+ NKT cell,which was collected and cultured with human bone marrow mesenchymal stem cells. The cultured supernatants were harvested and NKT phenotype was identified by flow cytometry. The levels of transforming growth factor-β1 (TGF-β1 ) and interferon-inducible protein-10 (IP-10) in supernatants were detected by enzyme linked immunosorbent assay. The levels of IFN-γ and tumor necrosis factor α (TNF-α) were detected by liquid chip technique. CD3+ CD56+ NKT cell was cultured with human bone marrow mesenchymal stem cells and then co-cultured with 16HBE for four hours. The cell survival counting of 16HBE was detected by CCK8 assay. Results Mesenchymal stem cells had immunomodulatory effects on T lymphocyte subsets including CD3+ CD56+ NKT cell,reduced CD3+ CD56+ NKT cell in peripheral blood monouclear cells,induced differentiation of regulatory T cell,and reduced the levels of IFN-γ,TNF-α and other inflammatory cytokines. After co-cultured with mesenchymal stem cells,CD3+ CD56+ NKT cell decreased from (20. 33±1. 05)%to (15.17±1.75)%( P ＜0. 05). Secreting high levels of TGF-β1 and IP-10 was an important molecular basis of immunomodulation of mesenchymal stem cells. Mesenchymal stem cells relieved NKT cell-mediated damage of lung epithelial cells. Conclusions Mesenchymal stem cells have immunomodulatory effects on T lymphocyte subsets including CD3+ CD56+ NKT cell,and play a protective role in killing lung epithelial cells mediated by NKT cell.
