UNASSIGNED: Immunophenotyping of human peripheral blood and liver Tregs was performed with flow cytometry, Vectra-6 multicolor immunohistochemistry (IHC), and real-time polymerase chain reaction. Measured outcomes of Treg augmentation with the interleukin-2 monoclonal antibody JES6-1/interleukin-2 in the RRV-induced mouse model of BA included survival, direct bilirubin, IHC, and liver flow cytometry.
UNASSIGNED: Patients with BA had decreased peripheral blood Treg frequency and lack of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) upregulation despite a highly activated, effector Treg phenotype. IHC revealed decreased liver Treg frequency and Treg CTLA-4 expression. Treg augmentation in the murine model led to increased survival, decreased direct bilirubin levels and liver inflammation, and expansion of resident macrophages. In addition to the M2 phenotype of resident macrophages, these cells adopted an inflammatory M1 phenotype in response to RRV infection, which was inhibited with Treg augmentation.
UNASSIGNED: Patients with BA have Treg deficiencies associated with lack of sufficient CTLA-4 expression that is necessary for cell-cell contact inhibition of inflammatory responses. Treg augmentation therapy in murine BA protected from disease. Future treatment trials for BA should include agents that enhance Treg number or function, mimic CTLA-4 function, and promote anti-inflammatory M2 macrophage phenotypes.
■用流式细胞术对人外周血和肝脏Tregs进行免疫分型,Vectra-6多色免疫组织化学(IHC),和实时聚合酶链反应。在RRV诱导的BA小鼠模型中,用白介素-2单克隆抗体JES6-1/白介素-2增强Treg的测量结果包括存活,直接胆红素,IHC,和肝脏流式细胞术。
BA患者外周血Treg频率降低,缺乏细胞毒性T淋巴细胞相关抗原-4(CTLA-4)上调,尽管高度活化,效应物Treg表型。IHC显示肝脏Treg频率和TregCTLA-4表达降低。小鼠模型中的Treg增加导致存活率增加,直接胆红素水平下降和肝脏炎症,和常驻巨噬细胞的扩增。除了常驻巨噬细胞的M2表型,这些细胞在对RRV感染的反应中采用了炎性M1表型,被Treg增强抑制。
■BA患者具有与缺乏足够的CTLA-4表达相关的Treg缺陷,所述CTLA-4表达是细胞-细胞接触抑制炎症反应所必需的。鼠BA中的Treg增强疗法可预防疾病。BA的未来治疗试验应包括增强Treg数量或功能的药物,模拟CTLA-4功能,并促进抗炎M2巨噬细胞表型。