白血病患儿骨髓及分离培养MSC的IL-6水平

The level of IL-6 in bone marrow and mesenchymal stem cell culture in a leukemic patient

目的研究不同白血病患儿骨髓上清的白介素-(IL-6)含量研究其在白血病患者中的分布特点,研究不同患者MSCs培养液上清IL-6的含量研究不同患者MSCs上清的分泌特点。方法白血病患儿骨髓中分离单个核细胞,体外培养间充质干细胞,用流式细胞仪鉴定MSC表面CD105、CD73、CD90、CD45、CD34、CD14、CD19、HLA-DR抗原鉴定,酶联免疫法检测非白血病和白血病儿童骨髓上清及骨髓分离培养的MSC上清中IL-6的含量。结果 MSC阳性表达CD90、CD105、CD73,而CD34、CD45、HLA-DR、CD14、CD19、CD79a、CD11b则呈阴性。不同性别、年龄儿童骨髓上清IL-6水平的差异无统计学意义;非白血病患儿与处于完全缓解期的白血病患儿骨髓IL-6水平的差异无显著性;初诊及骨髓未完全缓解的白血病患儿骨髓上清IL-6水平明显高于非白血病组及化疗后处于完全缓解期的白血病患儿(P<0.05);初诊及骨髓未达完全缓解白血病患儿中急性髓细胞白血病患儿骨髓IL-6水平高于急性淋巴细胞白血病患儿(P<0.05)。处于完全缓解期的不同类型白血病患儿骨髓IL-6水平无显著差异,骨髓缓解及未缓解的低、中、高危患儿IL-6水平均无统计学差异。结论急性白血病患儿的骨髓IL-6异常升高,与其白血病类型及肿瘤负荷有关联。

Objective To investigate the different levels of IL-6 in the hone marrow of an acute leukemic pediatric patient and the resulting distribution of IL-6 ; to investigate the different levels of IL-6 in the mesenchymal stem cell culture supematant and its resulting distribution in the mesenchymal stem cells of different patients. Methods Mononuclear cells were isolated from bone marrow. Mesenchymal stem cells were cultured in vitro, conditioned medium of mesenchymal stem cell was prepared. Flow cytometer was used to identify the phenotype CD105, CD73, CD90, CD45, CD34, CD14, CDI9 and HLA-DR of MSC. IL-6 levels in bone marrow supernatant of children with non-leukemia or leukemia and MSC cultured medium separated from hone marrow were detected by ELISA assay. Results MSC expressed positive in CD90, CD105 and CD73, while negative in CD34, CD45, HLA-DR, CD14, CD19, CD79a and CD11b. There was no significant differences in IL-6 levels between different genders and ages. No significant difference was reported as well between non-leukemic children and leukemic children in the stage of complete remission （CR）. Patients who were in the stage of preliminary diagnosis as well as those in non-remission had higher IL-6 levels than those with no leukemia and those in CR （P 〈 0.05 ）. Among the patients in the stage of preliminary diagnosis and non-remission, the IL-6 levels in acute myeloid leukemia were higher than those in acute lympboblasfic leukemia （P 〈 0.05）. No differences between low risk and high to middle risk patient were found. No significant differences in the IL-6 levels in MSC conditioned medium were found between the children with leukemia and without leukemia. Neither were significant differences found between different genders, ages and types of leukemia. Conclusion The acute lymphoblastic leukemic patient had a slightly higher level of IL-6, which might possibly indicate the relationship between IL-6 and the types of leukemia and the tumor burden.