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目的研究扶正透毒祛毒复方加味青蒿鳖甲汤对髓系微小残留白血病(MRD-L)患者骨髓CD34+细胞源树突状细胞(DC)的影响。方法用Ficoll离心法分离急性髓系MRD-L患者骨髓单个核细胞(BMNC),采用免疫磁珠分选出CD 34+细胞,并培养扩增,用不同浓度中药含药血清联合细胞因子进行体外诱导培养DC,倒置显微镜下观察DC的形态学特征,流式细胞术检测DC表面分子CD 83、CD 80、CD 86、CD 1a、HLA-DR的表达。诱导转化成熟的DC分别激发异体、自体T细胞,MTT法检测激发后的T细胞在不同效靶比时对人白血病细胞株K562的杀伤作用。结果各中药含药血清联合细胞因子培养髓系MRD-L患者CD 34+9 d后均能促进CD 34+分化为形态特征典型的DC,各中药联合细胞因子组、细胞因子组均明显上调CD 83、CD 80、CD 86、HLA-DR的表达,与胎牛血清及空白兔血清对照组比较有统计意义(P<0.01),中剂量及低剂量含药血清组能促进CD 1a的表达提高,与高剂量组、细胞因子组比较有差异(P<0.01),其余各组无明显差异。DC激发T细胞杀伤K562细胞,各中药联合细胞因子组的杀伤率均高于细胞因子组、胎牛血清组和空白兔血清组,其中中剂量和低剂量组又明显优于(P<0.01)或优于(P<0.05)细胞因子组、胎牛血清组和空白兔血清组。T细胞的杀伤作用随效靶比的增高而增强,正常T细胞与患者T细胞对K562细胞的杀伤作用无明显差异(P>0.05)。结论加味青蒿鳖甲汤能促进髓系MRD-L患者CD 34+细胞向DC转化。
Objective To study the effect of Fuzheng Tongdu Tuodudui Decoction of Modified Artemisia annua Biejia Decoction on dendritic cells (DC) of bone marrow CD34 + cells in patients with myeloid leukemia (MRD-L). The method of separation by Ficoll centrifugation MRD-L acute myeloid bone marrow mononuclear cells (the BMNC), immunomagnetic beads sorted CD 34+ cells, and expanded in culture containing serum combined with different concentrations of cytokines in vitro TCM DCs were induced to induce morphological changes of DCs under inverted microscope. Flow cytometry was used to detect the expression of CD83, CD80, CD86, CD1a and HLA-DR on DCs. Induced DCs were stimulated allogeneic, autologous T cells, MTT assay T cells challenged with different effective target ratio of human leukemia cell line K562 killing effect. Results All the Chinese medicinal serum combined with cytokines could promote the differentiation of CD34 + into typical DCs after culturing MRD-L patients with CD34 + 9 d, and all the combination of cytokines and cytokines were significantly up-regulated The expression of CD80, CD80, CD86 and HLA-DR in serum was significantly higher than that in fetal bovine serum and blank rabbit serum (P <0.01) , Compared with high-dose group and cytokine group (P <0.01), the other groups had no significant difference. DC-induced T cell killing K562 cells, the combination of Chinese medicine and cytokines killing rate were higher than the cytokine group, fetal bovine serum and blank rabbit serum group, which was significantly better than the medium dose and low dose group (P <0.01) Or better than (P <0.05) cytokine group, fetal calf serum group and blank rabbit serum group. T cell killing effect increased with the increase of target ratio, the normal T cells and patient T cells killing K562 cells had no significant difference (P> 0.05). Conclusion Modified Artemisia annua Biejia decoction can promote the transformation of CD34 + cells to DC in myeloid MRD-L patients.