中华医学杂志(英文版)
中華醫學雜誌(英文版)
중화의학잡지(영문판)
CHINESE MEDICAL JOURNAL
2002年
2期
202-205
,共4页
许志祥%徐颖%朱剑昆%施勤%李颖%张学光
許誌祥%徐穎%硃劍昆%施勤%李穎%張學光
허지상%서영%주검곤%시근%리영%장학광
重组人Flt3配体(rhFL) 造血形成 人血管内皮细胞(ECV)
重組人Flt3配體(rhFL) 造血形成 人血管內皮細胞(ECV)
중조인Flt3배체(rhFL) 조혈형성 인혈관내피세포(ECV)
recombinant human soluble Flt3 ligand%hematopoiesis%human microvascular endothelial cell line
目的 研究重组人Flt3配体（rhFL）对造血的直接作用，如造血集落形成、树突状细胞的增殖等，并研究rhFL刺激内皮细胞（ECV）增殖，对造血的间接作用。
方法 采用甲基纤维素半固体集落形成试验，观察rhFL单独或与其它细胞因子联合对脐血造血细胞体外集落形成的影响，计数自脐血细胞产生的造血集落数；用含有100ng/mlrhFL等多种细胞因子混合的方法，自人脐血单个核细胞中诱导产生树突状细胞（DC）。用流式细胞仪测定人ECV表面Flt3受体的表达，用MTT法分析rhFL对ECV的促增殖作用。同时还测定了ECV培养上清中FL、IL-6、IL-8、 G-CSF 和GM-CSF的水平。
结果 rhFL可显著促进人脐带血造血细胞体外集落的形成，与其它细胞因子合用时，作用明显增强；当rhFL与其它诱导DC的细胞因子联合使用时，诱导的树突状细胞数增加2.5倍。ECV表达Flt3受体，rhFL促进ECV的增殖。另外，ECV能够分泌FL、IL-6、IL-8、 G-CSF 和GM-CSF，TNFα和rhFL促进ECV分泌FL。
结论 rhFL增强自脐血单个核细胞诱导产生造血集落和DC。FL不仅能够刺激ECV的增殖，而且ECV可分泌FL。FL在机体的造血过程中可能起直接和简接作用。
目的 研究重組人Flt3配體（rhFL）對造血的直接作用，如造血集落形成、樹突狀細胞的增殖等，併研究rhFL刺激內皮細胞（ECV）增殖，對造血的間接作用。
方法 採用甲基纖維素半固體集落形成試驗，觀察rhFL單獨或與其它細胞因子聯閤對臍血造血細胞體外集落形成的影響，計數自臍血細胞產生的造血集落數；用含有100ng/mlrhFL等多種細胞因子混閤的方法，自人臍血單箇覈細胞中誘導產生樹突狀細胞（DC）。用流式細胞儀測定人ECV錶麵Flt3受體的錶達，用MTT法分析rhFL對ECV的促增殖作用。同時還測定瞭ECV培養上清中FL、IL-6、IL-8、 G-CSF 和GM-CSF的水平。
結果 rhFL可顯著促進人臍帶血造血細胞體外集落的形成，與其它細胞因子閤用時，作用明顯增彊；噹rhFL與其它誘導DC的細胞因子聯閤使用時，誘導的樹突狀細胞數增加2.5倍。ECV錶達Flt3受體，rhFL促進ECV的增殖。另外，ECV能夠分泌FL、IL-6、IL-8、 G-CSF 和GM-CSF，TNFα和rhFL促進ECV分泌FL。
結論 rhFL增彊自臍血單箇覈細胞誘導產生造血集落和DC。FL不僅能夠刺激ECV的增殖，而且ECV可分泌FL。FL在機體的造血過程中可能起直接和簡接作用。
목적 연구중조인Flt3배체（rhFL）대조혈적직접작용，여조혈집락형성、수돌상세포적증식등，병연구rhFL자격내피세포（ECV）증식，대조혈적간접작용。
방법 채용갑기섬유소반고체집락형성시험，관찰rhFL단독혹여기타세포인자연합대제혈조혈세포체외집락형성적영향，계수자제혈세포산생적조혈집락수；용함유100ng/mlrhFL등다충세포인자혼합적방법，자인제혈단개핵세포중유도산생수돌상세포（DC）。용류식세포의측정인ECV표면Flt3수체적표체，용MTT법분석rhFL대ECV적촉증식작용。동시환측정료ECV배양상청중FL、IL-6、IL-8、 G-CSF 화GM-CSF적수평。
결과 rhFL가현저촉진인제대혈조혈세포체외집락적형성，여기타세포인자합용시，작용명현증강；당rhFL여기타유도DC적세포인자연합사용시，유도적수돌상세포수증가2.5배。ECV표체Flt3수체，rhFL촉진ECV적증식。령외，ECV능구분비FL、IL-6、IL-8、 G-CSF 화GM-CSF，TNFα화rhFL촉진ECV분비FL。
결론 rhFL증강자제혈단개핵세포유도산생조혈집락화DC。FL불부능구자격ECV적증식，이차ECV가분비FL。FL재궤체적조혈과정중가능기직접화간접작용。
Objective To investigate the direct effects of the Flt3 ligand (FL) on hematopoiesis, such as the stimulation of the formation of hematopoietic colonies and the proliferation of dendritic cells, as well as the indirect stimulation of hematopoiesis, especially via the proliferation of endothelial cells.
Methods Mononuclear cells from human cord blood were plated in methylcellulose medium containing different cytokines to induce hematopoietic colony formation. Dendritic cells (DCs) were induced from the mononuclear cells with a cytokine cocktail with or without recombinant human soluble FL (rhFL; 100?ng/ml). The Flt3 receptors on the surface of a human microvascular endothelial cell line (ECV) were analyzed by flow cytometry. The proliferation of ECV stimulated by rhFL was measured with the microculture tetrazolium assay. The levels of FL, IL-6, IL-8, G-CSF and GM-CSF in the supernatant of ECV cultures were measured by enzyme linked immunoabsorbent assay (ELISA).
Results rhFL stimulates colony formation from cord blood when used as a sole stimulant. FL in combination with other cytokines increased colony formation significantly. The number of DCs was approximately 2.5 times higher when rhFL was used. rhFL stimulates the proliferation of ECV on which Flt3 receptors are expressed. Furthermore, ECV secretes FL, IL-6, IL-8, G-CSF and GM-CSF, which were augmented by tumor necrosis factor-α and rhFL.
Conclusions rhFL enhances hematopoietic colony formation and DC proliferation from human cord blood cells. FL not only stimulates the proliferation of ECV, but is also secreted by ECV. FL may exert direct and indirect effects on hematopoiesis.
