中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
9期
853-859
,共7页
李炳宗%庄文卓%陈萍%张宏%黄海雯%傅晋翔
李炳宗%莊文卓%陳萍%張宏%黃海雯%傅晉翔
리병종%장문탁%진평%장굉%황해문%부진상
多发性骨髓瘤%间充质干细胞%T细胞%免疫调节功能%骨髓瘤骨病
多髮性骨髓瘤%間充質榦細胞%T細胞%免疫調節功能%骨髓瘤骨病
다발성골수류%간충질간세포%T세포%면역조절공능%골수류골병
Multiple myeloma%Mesenchymal stem cells%T cells%Immunoregulatory capability%Myeloma bone disease
目的 探讨多发性骨髓瘤(MM)患者骨髓间充质干细胞(MSCs)免疫调节功能及其对骨髓瘤骨病的影响.方法 分离MM患者和正常对照MSCs,流式细胞技术比较二者的免疫表型.Real-time PCR检测实验组及对照组MSCs的TGF-β1、TGF-β2、TGF-β3、IL-6、IL-3、TNF-α、FasL和NF-κB配体的受体激活物(RANKL)表达量.共培养系统与流式细胞术检测MSCs对T细胞增殖、凋亡和早期活化标志CD25和CD69表达的影响.Von kossa染色、real-time PCR和Western blot等技术检测T细胞对正常对照MSCs向成骨细胞分化的影响.结果 MM来源和正常对照MSCs具有相似的形态学和免疫表型.MM来源的MSCs表达TGF-β1、IL-6、IL-3、TNF-α和RANKL较正常对照MSCs增加,而TGF-β2、TGF-β3和FasL的表达下降.MM来源的MSCs对T细胞增殖的抑制作用明显减弱.正常对照MSCs较MM来源MSCs使更多T细胞静止于G0/G1期.正常对照MSCs明显促进T细胞凋亡,而MM来源的MSCs对T细胞的促凋亡作用减弱.正常对照MSCs明显抑制T细胞活化分子表达,而MM患者的MSCs此抑制作用明显降低.与MM患者的MSCs共培养后的T细胞以及直接来源于MM患者的T细胞均可以明显抑制正常对照MSCs向成骨细胞分化.结论 MM患者的MSCs免疫调节功能较正常对照MSCs降低,主要表现为抑制T细胞活化的功能减弱,而活化的T细胞可抑制MSCs向成骨细胞分化,此可能为骨髓瘤骨病发病机制之一.
目的 探討多髮性骨髓瘤(MM)患者骨髓間充質榦細胞(MSCs)免疫調節功能及其對骨髓瘤骨病的影響.方法 分離MM患者和正常對照MSCs,流式細胞技術比較二者的免疫錶型.Real-time PCR檢測實驗組及對照組MSCs的TGF-β1、TGF-β2、TGF-β3、IL-6、IL-3、TNF-α、FasL和NF-κB配體的受體激活物(RANKL)錶達量.共培養繫統與流式細胞術檢測MSCs對T細胞增殖、凋亡和早期活化標誌CD25和CD69錶達的影響.Von kossa染色、real-time PCR和Western blot等技術檢測T細胞對正常對照MSCs嚮成骨細胞分化的影響.結果 MM來源和正常對照MSCs具有相似的形態學和免疫錶型.MM來源的MSCs錶達TGF-β1、IL-6、IL-3、TNF-α和RANKL較正常對照MSCs增加,而TGF-β2、TGF-β3和FasL的錶達下降.MM來源的MSCs對T細胞增殖的抑製作用明顯減弱.正常對照MSCs較MM來源MSCs使更多T細胞靜止于G0/G1期.正常對照MSCs明顯促進T細胞凋亡,而MM來源的MSCs對T細胞的促凋亡作用減弱.正常對照MSCs明顯抑製T細胞活化分子錶達,而MM患者的MSCs此抑製作用明顯降低.與MM患者的MSCs共培養後的T細胞以及直接來源于MM患者的T細胞均可以明顯抑製正常對照MSCs嚮成骨細胞分化.結論 MM患者的MSCs免疫調節功能較正常對照MSCs降低,主要錶現為抑製T細胞活化的功能減弱,而活化的T細胞可抑製MSCs嚮成骨細胞分化,此可能為骨髓瘤骨病髮病機製之一.
목적 탐토다발성골수류(MM)환자골수간충질간세포(MSCs)면역조절공능급기대골수류골병적영향.방법 분리MM환자화정상대조MSCs,류식세포기술비교이자적면역표형.Real-time PCR검측실험조급대조조MSCs적TGF-β1、TGF-β2、TGF-β3、IL-6、IL-3、TNF-α、FasL화NF-κB배체적수체격활물(RANKL)표체량.공배양계통여류식세포술검측MSCs대T세포증식、조망화조기활화표지CD25화CD69표체적영향.Von kossa염색、real-time PCR화Western blot등기술검측T세포대정상대조MSCs향성골세포분화적영향.결과 MM래원화정상대조MSCs구유상사적형태학화면역표형.MM래원적MSCs표체TGF-β1、IL-6、IL-3、TNF-α화RANKL교정상대조MSCs증가,이TGF-β2、TGF-β3화FasL적표체하강.MM래원적MSCs대T세포증식적억제작용명현감약.정상대조MSCs교MM래원MSCs사경다T세포정지우G0/G1기.정상대조MSCs명현촉진T세포조망,이MM래원적MSCs대T세포적촉조망작용감약.정상대조MSCs명현억제T세포활화분자표체,이MM환자적MSCs차억제작용명현강저.여MM환자적MSCs공배양후적T세포이급직접래원우MM환자적T세포균가이명현억제정상대조MSCs향성골세포분화.결론 MM환자적MSCs면역조절공능교정상대조MSCs강저,주요표현위억제T세포활화적공능감약,이활화적T세포가억제MSCs향성골세포분화,차가능위골수류골병발병궤제지일.
Objective To deplore the immunoregulatory function changes of mesenchymal stem cells(MSCs)from multiple myeloma(MM)patients and its effects on the pathogenesis of myeloma bone disease.Methods MSCs from MM patients and normal controls were isolated and the immunophenotype was detected.Real-time PCR was performed to detect the expressions of TGF-β1,TGF-β2,TGF-β3,IL-6,IL3,TNF-α,FasL and RANKL of MSCs.Transwell coculture systems were performed between MSCs and T cells.Lymphocyte proliferative assay was employed to detect the effect of MSCs on T cell proliferation.The effect of MSCs on T cell cycle and T cell activation markers CD25 and CD69 expression were analyzed by flow cytometry.Cleaved caspase 3 protein by western blot and hoechst 33258 staining were employed to detect the apotosis of T cells.Influence of T cells on the osteogenesis potential of MSCs were detected by Von kossa stain,real-time PCR and Western blot.Results MSCs from both MM patients and normal controls possessed similar morphology and immunophenotypes.MM derived MSCs exhibited increased expressions of TGF-β1,IL-6,IL-3,TNF-α and RANKL and decreased expression of TGF-β2,TGF-β3 and FasL.The inhibitory effect of MM derived MSCs on T cell proliferative ability was attenuated compared to control MSCs.MSCs from normal controls silence more T cells in Go/G1 phase than those from MM patients.The daupening effect of MM derived MSCs on activation-induced T apoptosis seemed to be enhanced.Expression of T cell activation markers were significantly inhibited by MSCs from normal controls.Both T cells cocultured with MM deprived MSCs and T cells directly from MM patients inhibited osteogenesis potential of MSCs from normal controls.Conclusion MSCs from MM patients showed impaired immunoregulatory capability on T cells.The activated T cells,in turn,inhibited the osteogenesis potential of MSCs.This may participate in the pathogenesis of myeloma bone disease.
