Shanxi Medical University, Taiyuan, China; Department of Gastroenterology, The First Hospital of Shanxi Medical University, Taiyuan, China.
Department of Gastroenterology, The Second Hospital of Shanxi Medical University, Taiyuan, China.
Biochem Biophys Res Commun. 2019 Apr 23;512(1):14-21. doi: 10.1016/j.bbrc.2019.02.147. Epub 2019 Mar 7.
Regulatory T cell (Treg)-based therapy can effectively control autoimmune hepatitis (AIH). Hepatic stellate cells (HSC) can selectively stimulate allogeneic Treg proliferation following liver transplantation. This study tested the therapeutic effect and potential mechanisms underlying the action of HSC-stimulated Tregs on AIH in a mouse model of Concanavalin A (ConA)-induced AIH. HSC were isolated from BALB/c mice and characterized. Splenic CD4CD25 Tregs were isolated from C57BL/6 mice by immunomagnetic beads. The cells were co-cultured with primary (HSC-0), the second generation of HSC (HSC-2) for 72 h. The proliferation of Tregs was determined by flow cytometry. Similarly, the Tregs were co-cultured with HSC in transwell plates to determine the potential cell-cell contact dependent. The CD4CD25 effector T cells (CFSE-Teffs) were co-cultured with Teff or Tregs in the presence or absence of HSC to determine the suppressive capacity of Tregs. The effects of Tregs or HSC-stimulated Tregs on AIH severity and the frequency of splenic Tregs and Th17 cells were examined in mice. Co-culture with HSC-2 significantly promoted Treg proliferation in a dose- and cell-cell contact-dependent manner, and allogeneic HSC enhanced the suppressive activity of Tregs to inhibit the proliferation of Teff in vitro. Adoptive transfer of Tregs, particularly of HSC-stimulated Tregs, significantly reduced liver injury, inflammation and Ishak modified histology activity index in AIH mice, which were associated with improving the balance of Treg and Th17 cell responses. Our data indicated that mature HSC stimulated allogeneic Treg proliferation in a dose and cell-cell contact-dependent manner, and HSC enhanced the suppressive activity of Tregs to inhibit the proliferation of Teff. Adoptive transfer of HSC-stimulated Tregs significantly reduced liver injury in AIH mice by modulating the balance of Treg and Th17 cell responses.
调节性 T 细胞（Treg）为基础的治疗可有效控制自身免疫性肝炎（AIH）。肝星状细胞（HSC）在肝移植后可选择性地刺激同种异体 Treg 的增殖。本研究在伴刀豆球蛋白 A（ConA）诱导的 AIH 小鼠模型中，检测了 HSC 刺激的 Treg 对 AIH 的治疗作用及其潜在机制。从 BALB/c 小鼠中分离 HSC 并进行鉴定。用免疫磁珠从 C57BL/6 小鼠中分离脾 CD4CD25Treg。将细胞与原代（HSC-0）、第二代 HSC（HSC-2）共培养 72h。通过流式细胞术测定 Treg 的增殖。同样，将 Treg 与 Transwell 板中的 HSC 共培养，以确定潜在的细胞-细胞接触依赖性。将 CD4CD25效应性 T 细胞（CFSE-Teffs）与 Teff 或 Treg 在有无 HSC 的情况下共培养，以确定 Treg 的抑制能力。在小鼠中检测 Treg 或 HSC 刺激的 Treg 对 AIH 严重程度以及脾 Treg 和 Th17 细胞频率的影响。与 HSC-2 共培养以剂量和细胞-细胞接触依赖性方式显著促进 Treg 的增殖，同种异体 HSC 增强了 Treg 的抑制活性，以抑制 Teff 在体外的增殖。过继转移 Treg，特别是 HSC 刺激的 Treg，可显著减轻 AIH 小鼠的肝损伤、炎症和 Ishak 改良组织学活动指数，这与改善 Treg 和 Th17 细胞反应的平衡有关。我们的数据表明，成熟的 HSC 以剂量和细胞-细胞接触依赖性方式刺激同种异体 Treg 的增殖，HSC 增强了 Treg 的抑制活性，以抑制 Teff 的增殖。过继转移 HSC 刺激的 Treg 通过调节 Treg 和 Th17 细胞反应的平衡，显著减轻 AIH 小鼠的肝损伤。
