IL-2/GM-CSF 体外处理的外周血单个核细胞对 再生障碍性贫血小鼠T 淋巴细胞亚群 及细胞因子的影响

Effects of peripheral blood mononuclear cells treated with IL-2 and GM-CSF in vitro on T lymphocyte subsets and cytokines in aplastic anemia mice

摘要:

目的 探讨白细胞介素-2（IL-2）/ 粒细胞- 巨噬细胞集落刺激因子（GM-CSF）体外处理的 外周血单个核细胞输注对再生障碍性贫血（以下简称再障）小鼠T 淋巴细胞亚群和细胞因子的影响及其促造 血作用。方法 选取清洁级雌性BALB/c 小鼠并复制免疫介导再障小鼠模型，设立对照组、模型组及干预组。 干预组取正常同源小鼠外周血单个核细胞，在IL-2、GM-CSF等作用下培养48 h 后进行尾静脉输注（模型 复制第1、2、4 及6 天），模型组输注等体积生理盐水。全部小鼠分别于模型复制第7、14、21 及28 天断尾 采血检测外周血白细胞、血红蛋白和血小板计数；于模型复制第28 天或濒死时处死小鼠，计数骨髓有核细胞 （粒系、红系、淋巴细胞比例及巨核细胞数），检测外周血CD3+CD4+（T 辅助细胞、Th 细胞）、CD3+CD8+（T 抑制细胞、Ts 细胞）及CD4+CD25+CD127-（调节性T 细胞、T-regs）细胞比例，检测血清肿瘤坏死因子-α （TNF-α）、γ 干扰素（IFN-γ）水平。结果 模型组和干预组外周血WBC 较对照组低（P <0.05），干预 组在第21 和28 天较模型组高（P <0.05）。模型组和干预组外周血PLT 较对照组低（P <0.05），干预组在第 21 和28 天较模型组高（P <0.05）。模型组和干预组外周血Hb 较对照组低（P <0.05），干预组在第28 天较模 型组高（P <0.05）。模型组和干预组粒系比例、红系比例及巨核细胞数较对照组低，淋巴细胞比例较对照组 高（P <0.05），干预组骨髓粒系比例、红系比例及巨核细胞数较模型组高，淋巴细胞比例较模型组低（P <0.05）。 模型组Th 细胞比例、T-regs 细胞比例较对照组低（P <0.05），Ts 细胞比例较对照组高（P <0.05）。干预组Th 细胞比例、T-regs 细胞比例较模型组高，Ts 细胞比例较模型组低（P <0.05）。模型组外周血TNF-α、IFN-γ 水平较对照组高（P <0.05），干预组较模型组低（P <0.05）。结论 IL-2/GM-CSF 体外处理的外周血单个核细 胞输注有助于调节再障小鼠T 淋巴细胞亚群紊乱，下调造血负调控细胞因子，从而改善骨髓造血功能。

Abstract:

Objective To explore the effects of peripheral blood mononuclear cells treated with IL-2 and GM-CSF in vitro on T lymphocyte subsets and cytokines in aplastic anemia mice. Methods Models of immune-mediated aplastic anemia were established according to literature methods, and a control group, a model group and an intervention group were established. Peripheral blood mononuclear cells were isolated from normal homologous mice and cultured for 48 hours under the effects of IL-2 and GM-CSF and administered intravenously to mice in the intervention group (on the 1st d, 2nd d, 4th d and 6th d). The model group was given equal volume of normal saline. The counts of WBC, PLT, and Hb level in peripheral blood were tested in all mice on the 7th d, 14th d, 21st d, and 28th d. All mice were sacrificed on the 28th d or the brink of death. The numbers of bone marrow nucleated cells (granulocytes, erythrocytes, lymphocytes, megakaryocytes) were counted. The proportions of CD3+CD4+(T helper cells, Th cells), CD3+CD8+(T suppressor cells, Ts cells), CD4+CD25+CD127?(regulatory T cells, T-regs) cells in peripheral blood were measured, and the levels of TNF-α and IFN-γ in serum were measured. Results The WBC of the model group and the intervention group was lower than that of the control group (P < 0.05), and the WBC of the intervention group was higher than that of the model group on the 21st and 28th d (P < 0.05). PLT in the model group and the intervention group was lower than that in the control group (P < 0.05), and higher in the intervention group on the 21st and 28th day (P < 0.05). The Hb of model group and intervention group was lower than that of control group (P < 0.05), and the Hb of intervention group was higher than that of model group on the 28th day (P < 0.05). The proportion of granulocytes, erythrocytes and megakaryocytes in the model group and the intervention group were lower than those in the control group, and the proportion of lymphocytes in the intervention group was higher than that in the control group (P < 0.05). The proportion of Th cells and T-regs cells in the model group was lower than that in the control group (P < 0.05), and the proportion of TS cells in the model group was higher than that in the control group (P < 0.05). The proportion of Th cells and T-regs cells in the intervention group was higher than that in the model group, and the proportion of TS cells in the intervention group was lower than that in the model group (P < 0.05). The levels of TNF-α and IFN-γ in the model group were higher than those in the control group (P < 0.05), and the levels in the intervention group were lower than those in the model group (P < 0.05). Conclusions Infusion of peripheral blood mononuclear cells treated with IL-2/GM-CSF in vitro can help to regulate T lymphocyte subsets disorder and reduce the hematopoietic negative cytokines, so as to improve the bone marrow hematopoietic function in aplastic anemia mice.