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目的 探讨重组白细胞介素2(rIL 2)激活的骨髓(ABM)抗骨髓瘤活性的免疫学机制。方法 应用rIL 2体外分别激活多发性骨髓瘤(MM)患者骨髓及正常对照骨髓,同时设未激活骨髓(NBM)为对照。用MTT比色分析法测定正常对照组的ABM对U266细胞的杀伤活性,应用流式细胞仪检测培养72h后MM组骨髓瘤残余病变标志(CD45-CD38+CD138+细胞百分率)的变化,ELISA法检测MM组及正常对照组不同时间段ABM、NBM培养液中TNF α、IFN γ的水平,并进行比较和相关性分析。结果 正常对照组NBM 72h对U266细胞的杀伤率为( 43. 20±12. 39 )%, 24h杀伤率为(26. 53±5. 48)%,ABM 72h对U266细胞的杀伤率为( 69. 70±26. 57 )%, 24h杀伤率为( 34. 25±11. 93)%, 24h及72h的ABM细胞杀伤活性均较NBM增高,其中72hABM与NBM对U266细胞杀伤率的差异有统计学意义(P<0. 05);MM患者骨髓经rIL 2激活72h后CD45-CD38+CD138+细胞的百分率明显下降[激活前为(8. 46±3. 66)%,激活后为(4. 79±1. 56)%, P<0. 05];正常骨髓及MM患者骨髓经rIL 2激活后,随着培养时间的延长,培养液中TNF α、IFN γ的含量增加,无论24h或72h,两组ABM中TNF α、IFN γ的水平均较NBM明显增高(P<0. 05);正常对照组ABM 24h及72h培养液中TNF α、IFN γ的水平分别与本组同时相的细胞杀伤
Objective To investigate the immunological mechanism of recombinant interleukin-2 (IL-2) -activated bone marrow (ABM) against myeloma activity. Methods The bone marrow of patients with multiple myeloma (MM) and normal control bone marrow (BMMNC) were activated by rIL 2 in vitro. Inactivated NBM was used as control. The killing activity of ABM on U266 cells in normal control group was determined by MTT colorimetric assay. The changes of residual myeloid markers (CD45-CD38 + CD138 + cells) in MM group were detected by flow cytometry MM group and normal control group at different time periods ABM, NBM medium TNFα, IFN γ levels, and compared and correlation analysis. Results The killing rate of U266 cells in NBM 72h group was (43.20 ± 12.39%) in NBM group, (26.53 ± 5.58)% in 24h group, and (69 .70 ± 26.57%), the killing rate of ABM cells was (24.45 ± 11.93)% at 24 h and 72 h, respectively, higher than that of NBM at 72 h, and statistical difference was found between 72 h ABM and NBM on U266 cell killing rate (P <0.05). The percentage of CD45-CD38 + CD138 + cells in bone marrow of MM patients significantly decreased after activated by rIL 2 for 72 h [(8.66 ± 3.66)% before activation and (4) 79 ± 1.56)%, P <0.05]. After activation of bone marrow cells in normal bone marrow and MM patients, the levels of TNFα and IFNγ in the culture fluid increased with the prolongation of culture time, no matter 24h or 72h , The levels of TNFα and IFNγ in ABM of both groups were significantly higher than that of NBM (P <0.05); the levels of TNFα and IFNγ in ABM 24h and 72h of normal control group were significantly higher than that of NBM Kill