1 Department of Cardiac, Thoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany. 2 German Center for Lung Research, Germany. 3 Institute of Pathology, Hannover Medical School, Hannover, Germany. 4 Department of Hematology, Hannover Medical School, Hannover, Germany. 5 Department of Experimental Pneumology, Hannover Medical School, Hannover, Germany.
Transplantation. 2015 Mar;99(3):482-91. doi: 10.1097/TP.0000000000000632.
Bronchiolitis obliterans syndrome is caused by a fibroproliferative process in lung allografts resulting in irreversible damage. In this study, we induced obliterative bronchiolitis and studied the contribution of regulatory T cells to its development in immune-deficient mice receiving heterotopic porcine bronchus transplants, and major histocompatibility complex-mismatched porcine peripheral blood mononuclear cell. Furthermore, we aimed to corroborate our findings in a humanized mouse model.
Heterotopic bronchus transplantation was performed in 33 NOD.rag(−/−)γc(−/−) mice, using miniature pigs as tissue donors.The recipient mice then either received saline (negative control), unsorted MHC-mismatched PBMC (positive control), PBMC enriched with CD4(+)CD25(high) cells or PBMC depleted of CD4(+)CD25(high) cells for reconstitution. The results were validated in 28 NOD.rag(−/−)γc(−/−) mice undergoing heterotopic human bronchus transplantation and reconstitution with allogeneic human PBMC.
Histological lesions similar to those typical for obliterative bronchiolitis developed in vivo after reconstitution with allogeneic PBMC and were more severe in animals engrafted with PBMC depleted of CD4(+)CD25(high) cells. In contrast, the group reconstituted with PBMC enriched with CD4(+)CD25(high) cells showed well-preserved histology. The results of the humanized model confirmed those obtained in the porcinized model.
In conclusion, both porcinized and humanized mouse models of heterotopic subcutaneous bronchus transplantation imitate the in vivo development of bronchiolitis obliterans syndrome-like lesions and reveal its sensitivity to T-cell regulation.
闭塞性细支气管炎综合征是由肺移植物中的纤维增生过程引起的,导致不可逆转的损伤。在这项研究中,我们在接受异位猪支气管移植和主要组织相容性复合体错配猪外周血单个核细胞的免疫缺陷小鼠中诱导闭塞性细支气管炎,并研究调节性 T 细胞对其发展的贡献。此外,我们旨在在人源化小鼠模型中证实我们的发现。
在 33 只 NOD.rag(-/-)γc(-/-)小鼠中进行异位支气管移植,使用小型猪作为组织供体。然后,受体小鼠接受生理盐水(阴性对照)、未分选的 MHC 错配 PBMC(阳性对照)、富含 CD4(+)CD25(high)细胞的 PBMC 或耗尽 CD4(+)CD25(high)细胞的 PBMC 进行重建。结果在 28 只接受异位人支气管移植和同种异体人 PBMC 重建的 NOD.rag(-/-)γc(-/-)小鼠中得到验证。
在用同种异体 PBMC 重建后,体内出现类似于闭塞性细支气管炎的组织学病变,在用耗尽 CD4(+)CD25(high)细胞的 PBMC 移植的动物中更为严重。相比之下,用富含 CD4(+)CD25(high)细胞的 PBMC 重建的组显示出保存完好的组织学。人源化模型的结果证实了猪源化模型的结果。
总之,异位皮下支气管移植的猪源化和人源化小鼠模型模拟了闭塞性细支气管炎综合征样病变的体内发展,并揭示了其对 T 细胞调节的敏感性。