Liu W, Gao C, Zhou B-G, Li W-M
The First Affiliated Hospital, Harbin Medical University, Department of Cardiology, Heilongjiang, 150001, PR China.
Autoimmunity. 2006 Mar;39(2):83-92. doi: 10.1080/08916930500507870.
To explore the therapeutic alliance effects of adenovirus vector-mediated gene transfer of ICOSIg and CTLA4Ig fusion protein on experimental autoimmune myocarditis (EAM).
Expression vector pAdeno-CTLA4Ig and pAdeno-ICOSIg was constructed and transfected into HEK293 cells. Adenovirus expresses CTLA4Ig and ICOSIg was produced. Ad-CMV-GFP was used as controls. EAM was induced in Lewis rats by injection of procine cardiac myosin. All the immunized rats were divided into four groups. Group A (n = 15) received adenovirus containing CTLA4Ig and ICOSIg from day 14-28; group B (n = 15), group C (n = 15) and group D (n = 15) received adenovirus containing CTLA4Ig, ICOSIg and GFP, respectively. Group E (n = 10) was normal controls never received immunization. On day 28, all the rats were killed after echocardiography examination. Histopathological examination was used to observe inflammation in the myocardium. Western blot was used to detect CTLA4, ICOS, ICOSL and competitive RT-PCR for B7-1, B7-2 expression. T lymphocyte proliferation assay was performed and ELISPOT was used to detect the Th1 and Th2 production.
Alliance application of CTLA4Ig and ICOSIg exerts therapeutic effects on EAM. After a treatment duration of 14 days, cardiac function and myocardial inflammation improved significantly compared to group D. Expression of CTLA-4, ICOS and ICOSL, B7-1 was statistically decreased in group A, B and C compared with group D. T-cell proliferation was inhibited by costimulatory blockade in a dose-dependent style. ICOSIg blockade significantly augments IL-4 and IL-10 production while diminished IFN-gamma production.
Blockade of costimulatory pathway with alliance therapy of CTLA4Ig and ICOSIg alleviated autoimmune damage in EAM and improved cardiac function. The mechanisms may be downregulation of costimulatory molecules and anti-inflammation.
探讨腺病毒载体介导的ICOSIg和CTLA4Ig融合蛋白基因转移对实验性自身免疫性心肌炎（EAM）的治疗性联盟效应。
构建表达载体pAdeno-CTLA4Ig和pAdeno-ICOSIg并转染至HEK293细胞。产生表达CTLA4Ig和ICOSIg的腺病毒。使用Ad-CMV-GFP作为对照。通过注射猪心肌肌凝蛋白在Lewis大鼠中诱导EAM。所有免疫大鼠分为四组。A组（n = 15）从第14天至28天接受含CTLA4Ig和ICOSIg的腺病毒；B组（n = 15）、C组（n = 15）和D组（n = 15）分别接受含CTLA4Ig、ICOSIg和GFP的腺病毒。E组（n = 10）为未接受免疫的正常对照。在第28天，所有大鼠在超声心动图检查后处死。采用组织病理学检查观察心肌炎症。使用蛋白质免疫印迹法检测CTLA4、ICOS、ICOSL，并采用竞争性逆转录聚合酶链反应检测B7-1、B7-2的表达。进行T淋巴细胞增殖试验，并使用酶联免疫斑点法检测Th1和Th2的产生。
CTLA4Ig和ICOSIg联合应用对EAM具有治疗作用。治疗14天后，与D组相比，心功能和心肌炎症明显改善。与D组相比，A组、B组和C组中CTLA-4、ICOS、ICOSL、B7-1的表达在统计学上降低。共刺激阻断以剂量依赖方式抑制T细胞增殖。ICOSIg阻断显著增加IL-4和IL-10的产生，同时减少IFN-γ的产生。
CTLA4Ig和ICOSIg联合治疗阻断共刺激途径可减轻EAM中的自身免疫损伤并改善心功能。其机制可能是共刺激分子的下调和抗炎作用。
