CAJ | 학술논문

目的考察自体树突状细胞(DC)活化的骨髓细胞对慢性粒细胞白血病(CGL)细胞的毒性作用。方法用免疫磁珠从2例缓解的CGL患者骨髓单个核细胞(BMMNC)分离DC，另取BMMNC分成3份，分别加入长期培养基，建立Dexter培养体系。第1份为对照，第2份加重组人白细胞介素2(rhIL-2)，第3份于培养第4天加入收获的DC，培养10 d后收集非贴壁细胞。以上述3份非贴壁细胞为效应细胞，对照非贴壁细胞为靶细胞，采用双色流式细胞技术进行细胞毒性实验，同时用流式细胞技术检测BMMNC的P210表达。结果 DC活化的效应细胞比rhIL-2活化的效应细胞对靶细胞有更大的杀伤作用，靶细胞中的死细胞比例例1分别为63.12%和42.59%，例2分别为61.60%和21.46%。另外，DC活化的效应细胞与靶细胞混合孵育后，效应细胞本身的死细胞比例明显增加，而rhIL-2活化的效应细胞无此现象。BMMNC经培养后非贴壁细胞中P210阳性细胞比例明显减少，以含DC者最为显著，含rhIL -2者次之。结论自体DC活化的骨髓细胞能对CGL细胞产生明显的细胞毒性作用，DC的作用胜过rhIL-2。这些经DC活化的骨髓回输后可能在体内发挥移植物抗白血病(GVL)效应。
목적고찰자체수돌상세포(DC)활화적골수세포대만성립세포백혈병(CGL)세포적독성작용。방법용면역자주종2례완해적CGL환자골수단개핵세포(BMMNC)분리DC，령취BMMNC분성3빈，분별가입장기배양기，건립Dexter배양체계。제1빈위대조，제2빈가중조인백세포개소2(rhIL-2)，제3빈우배양제4천가입수획적DC，배양10 d후수집비첩벽세포。이상술3빈비첩벽세포위효응세포，대조비첩벽세포위파세포，채용쌍색류식세포기술진행세포독성실험，동시용류식세포기술검측BMMNC적P210표체。결과 DC활화적효응세포비rhIL-2활화적효응세포대파세포유경대적살상작용，파세포중적사세포비례례1분별위63.12%화42.59%，례2분별위61.60%화21.46%。령외，DC활화적효응세포여파세포혼합부육후，효응세포본신적사세포비례명현증가，이rhIL-2활화적효응세포무차현상。BMMNC경배양후비첩벽세포중P210양성세포비례명현감소，이함DC자최위현저，함rhIL -2자차지。결론자체DC활화적골수세포능대CGL세포산생명현적세포독성작용，DC적작용성과rhIL-2。저사경DC활화적골수회수후가능재체내발휘이식물항백혈병(GVL)효응。
Objective　To investigate the activity of bone marrow cells activated by autologous dendrit ic cells(DC) to mediate cytotoxicity against chronic granulocytic leukemia(CGL) cells. Methods　DC were separated from bone marrow mononuclear cells(BMMNC) of two CGL patients in hematological remission and harvested after 3 d ays of culture in IMDM containing autologous plasma, rhGM-CSF and rhTNFα at 37 ℃, 5% CO2 humidified atmosphere. BMMNC obtained from the patients were divi ded into 3 groups to set up Dexter systems: the control group, rhIL-2 contain ing, and the third group having DC added at day 4. After 10 days of culture, non -adherent cells were harvested and the changes of immunological phenotype and t he percentage of P210 positive cells were analyzed. The cytotoxicity were assaye d with two-colour flow cytometry. The non-adherent cells from all the 3 system s served as effector cells, those from control system as target cells. Res u lts　The cytotoxic activity against target cells was greater in the DC- activated effector cells than that in rhIL-2-activated ones. The percentages of death cells in target cells were 63.12% versus 42.59%(case 1) and 61.60% ver sus 21.46% (case 2), respectively. In addition, there was a marked increase in the death cell percentage in the DC-activated effector cells themselves after i ncubation with target cells. This phenomenon was not found in the rhIL-2-activated effector cells. The percentage of P210 positive cells was significantly lower in non-adherent cells after 10 days of culture in Dexter s ystem, comparing with that in non-cultured BMMNC. The least P210 positive cells were found in those cultured with DC and the less in those with rhIL-2. Conclusion　Autologous DC were able to activate bone marrow cells to generate cytotoxicity against CGL cells. Their effect was greater than that of rhIL-2. These activated bone marrow cells might mediate graft versus leukemia e ffect in vivo.