骨髓移植小鼠急性移植物抗宿主病早期弥漫性肺损伤中TH17细胞的作用

目的 探讨骨髓移植小鼠急性移植物抗宿主病(aGVHD)早期肺损伤中TH 17细胞的作用和机理.方法 Balb/c小鼠为受鼠,经致死量全身照射(TBI)后,输注C57BL/6小鼠来源的骨髓细胞和脾细胞,建立aGVHD模型.实验分为3组:TBI组小鼠仅接受TBI,异基因骨髓移植(BMT)组小鼠TBI后输注供者骨髓细胞和脾细胞,常山酮(HF)组小鼠TBI后输注骨髓细胞和脾细胞,并注射HF.动态观察小鼠GVHD的表现,并进行肺组织病理学、T淋巴细胞亚群及相关细胞因子的检测.结果 移植后小鼠出现典型GVHD的表现.移植后6d时HF组肺组织病理学评分为(2.00±0.35)分,BMT组为(0.67±0.07)分.BMT组TH 1细胞占CD4+T淋巴细胞的比例为(5.53±0.11)％,TH 17细胞占(1.04±0.34)％;HF组TH1细胞占(8.61±0.21)％,TH 17细胞占(0.49±0.07)％;组间比较,差异有统计学意义(P＜0.05).两组均未检测到TH2细胞.移植后6d,BMT组白细胞介素(IL)-17A为(2.81±0.19)pg/ml,γ干扰素(IFN-γ)为(42.97±0.23) pg/ml; HF组IL-17A＜0.8 pg/ml,IFN-γ为(9.89±0.51)pg/ml;组间比较,差异有统计学意义(P＜0.05).两组均未检测到IL-10.结论 在异基因造血干细胞移植早期阻断TH17细胞及其细胞因子的分泌,导致炎症因子分泌失衡,加重肺损伤.

The function and mechanism of TH 17 cells in aGVHD early diffuse lung injury

Objective To assess the functional role of TH 17 cells in allogeneic hematopoietic stem cell transplantation (allyHSCT).Methods Bone marrow monocytes and splenic T cells were enriched from C57/BL6 donors.Recipient Balb/c mice were irradiated with 7.5 Gy total body irradiation (TBI) and injected with 5 105 splenic T cells and 5 106 bone marrow monocytes.Survival was monitored daily,clinical graft-versus-host disease (GVHD) was assayed three times a week,and detailed histopathologic analyses of lung were performed at day six after Allo-HSCT.Flow cytometry analysis was performed using CD3-FITC,CD4-PE,CD45-PerCP-CyS.5 monoclonal antibodies.Cells were stained for intracellular cytokines using mouse TH 1/TH2/TH 17 cytokine kit.Results All the experimental animals showed GVHD manifestations on the day 6 after transplantation.Animals from BMT and HF groups were scarified and histological analysis of lung was performed.Absence of TH 17 cells induced severe pathologic pulmonary lesions.The histopathology of the lung tissue was characterized by disorganization,epithelia cell damage,interstitial fibroplasias,and monocytes infiltration.The proportion of TH1 and TH 17 in BMT group was (5.53 ± 0.11 ) ％ and ( 1.04 ± 0.34)％ respectively,both significantly different from that in HF group.The levels of IL-17A and IFN-γin BMT group were (2.81 ±0.19) and (42.97 ± 0.23) pg/mL respectively.IL-17A could not be detected in HF group,yet the level of IFN-y was only (9.89 ± 0.51 ) pg/mL.IL-10 in both HF and BMT groups was not detectable.Conclusion Lung is on target of aGVHD.IL-17A may play a key role in the lung injury after transplantation.