Expression of chemokines and their receptors in GVHD target organs and therapeutic target

移植物抗宿主病(GVHD)是异基因造血细胞移植后最重要的临床问题。GVHD发病过程中产生的趋化因子可促进T细胞向GVHD靶器官的迁移，从而加重GVHD的严重程度。因此，我们分析了趋化因子在GVHD靶器官中的表达，并尝试通过趋化因子拮抗剂及其受体进行治疗。在本研究中，我们使用了急性GVHD模型小鼠：C57BL/6(供体)和(C57BL/6 X DBA/2)F1(B6D2F1)(受体)。B6D2F1小鼠接受13Gy射线照射。在GVHD发生发展过程中，Th1相关趋化因子(Mig/CXCL9和IP-10/CXCL10)、Th2相关趋化因子(TARC和MDC)、MIP-1α和-1β、RANTES和KC在肝脏中的表达增加。肠组织中Th1相关趋化因子、巨噬细胞炎性蛋白-Lα和-β、单核细胞趋化蛋白-1和-3、黏附分子LTn均明显增加。根据这一发现，在GVHD诱导的受体小鼠的肝脏和肠道中观察到Th1相关趋化因子及其受体CXCR3的高水平表达。接下来，我们尝试使用IP-10拮抗剂治疗急性移植物抗宿主病。与对照组相比，IP-10拮抗剂显著改善了肝脏和肠道的损伤。与接受CD4^+CD25^+Foxp3^+Foxp3^+T细胞(Treg细胞)的受体小鼠相比，接受CD4^+CD25^+Foxp3^+CXCR3-Treg细胞(CXCR3-Treg细胞)的受体小鼠肝脏和肠道的GVHD改变显著改善。这是由于CXCR3-Treg细胞的迁移更加明显，它们在TM相关趋化因子表达器官中定位的时间更长，导致了更强的抑制活性。因此，我们成功地制备了表达趋化因子受体的Treg细胞，并证明了它们在靶器官积聚后能够改善疾病进展的能力。该方法可能为急性移植物抗宿主病的器官损害提供一种新的治疗途径。

Graft-versus-host disease (GVHD) is the most significant clinical problem that arises after allogeneic hematopoietic cell transplantation. Chemokines induced during the development of GVHD promote T-cell migration into GVHD target organs and contribute to severity of GVHD. Therefore, we analyzed the expression of chemokines in GVHD target organs in an animal model and tried the therapy through chemokine antagonists and their receptors.In this study, we used acute GVHD model mouse: C57BL/6 (donor) and (C57BL/6 X DBA/2)F1 (B6D2F1) (recipient). The B6D2F1 mice were treated by 13 Gy irradiation. During the development of GVHD, expressions of Th1-associated chemokines (Mig/CXCL9 and IP-10/CXCL10), Th2-associated chemokines (TARC and MDC), MIP-1α and-1β, RANTES, and KC were increased in liver. In contrast, Th1-associated chemokines, MIP-lα and-β, MCP-1 and-3, and LTN were increased in gut. From this finding, high levels of expression of Th1-associated chemokines and their receptor CXCR3 were observed in the liver and intestines of GVHD-induced recipient mice. Next, we tried the treatment of acute GVHD by using IP-10 antagonist. Consiquently, IP-10 antagonist ameliorated the damage significantly in liver and gut, compared with control mice.Recipient mice that had undergone transfer of CD4^+CD25^+Foxp3^+ CXCR3-transfected T cells (CXCR3-Treg cells) showed significant amelioration of GVHD changes in the liver and intestines in comparison with recipient mice that had received CD4^+CD25^+Foxp3^+ T cells (Treg cells). This was due to more pronounced migration of CXCR3-Treg cells and their localization for a longer time in TM-associated chemokine-expressing organs, resulting in stronger suppressive activity. Thus we succeeded in preparing chemokine receptor-expressing Treg cells and demonstrated their ability to ameliorate disease progression upon accumulation in target organs. This method may provide a new therapeutic approach for organ damage in acute GVHD.