wcjdwjgnet.com 世界华人消化杂志 2009年12月8日; 17(34): 3507-3513 ISSN 1009-3079 CN 14-1260/R基础研究 BASIC RESEARCHTIM1与TIM4对小鼠食物过敏模型中CD4+CD25+调节性T 细胞功能的影响王新亭, 郑鹏远, 罗 予, 刘志强, 张利利王新亭, 郑鹏远, 刘志强, 张利利, 郑州大学第二附属医院 消化内科 郑州大学医学微生态学研究所 河南省郑州市 450014 罗予, 河南省医药科学研究院 河南省郑州市 450052 王新亭, 在读硕士, 主要从事消化系疾病的研究. 国家自然科学基金资助项目, No. 30772028 作者贡献分布: 王新亭与郑鹏远对此文贡献均等; 此课题由王新 亭、郑鹏远及罗予设计; 研究过程由王新亭、张利利及刘志强 完成; 研究所用新试剂和分析工具由郑鹏远与罗予提供; 数据分 析由王新亭完成; 本论文写作由王新亭、郑鹏远及刘志强完成. 通讯作者: 郑鹏远, 教授, 主任医师, 博士生导师, 450014, 河南 省郑州市经八路2号, 郑州大学第二附属医院消化内科, 郑州大 学医学微生态学研究所. medp7123126.com 电话: 0371-65261035 传真: 0371-63934118 收稿日期: 2009-09-14 修回日期: 2009-11-13 接受日期: 2009-11-16 在线出版日期: 2009-12-08TIM4 and TIM1 modulate the function of CD4+CD25+ T regulatory cells in mice with food allergy Xin-Ting Wang, Peng-Yuan Zheng, Yu Luo, Zhi-Qiang Liu, Li-Li ZhangXin-Ting Wang, Peng-Yuan Zheng, Zhi-Qiang Liu, Li-Li Zhang, Department of Gastroenterology, the Second Affili- ated Hospital of Zhengzhou University; Institute of Medical Microecology, Zhengzhou University, Zhengzhou 450014, Henan Province, China Yu Luo, Henan Academy of Medical and Pharmaceutical Sciences, Zhengzhou 450052, Henan Province, China Supported by: National Natural Science Foundation of China, No. 30772028 Correspondence to: Professor Peng-Yuan Zheng, Depart- ment of Gastroenterology, the Second Affiliated Hospital of Zhengzhou University; Institute of Medical Microecology, 2 Jingba Road, Zhengzhou 450014, Henan Province, China. medp7l23126.com Received: 2009-09-14 Revised: 2009-11-13 Accepted: 2009-11-16 Published online: 2009-12-08Abstract AIM: To evaluate the function of T regulatory (Treg) cells and determine the role of T cell immunoglobulin and mucin domain-containing protein 4 (TIM4) and TIM1 in modulating the function of Treg cells in mice with food allergy.METHODS: Thirty-two BALB/c mice fed an ovalbumin (OVA)-free diet were randomly and equally divided into four groups: normal saline (NS) group, staphylococcal enterotoxin B (SEB) plus OVA group, anti-TIM1 antibody plus SEB and OVA group, and anti-TIM4 antibody plus SEB and OVA group. Mice in the four groups were sensitized by intraperitoneal injections of NS, SEB/OVA, anti-TIM1/SEB/OVA, and anti- TIM4/SEB/OVA on days 0, 3 and 9, respec- tively. All mice (except the NS group) were chal- lenged by intraperitoneal injections of SEB/OVA on days 7 and 14, respectively. The expression of forkhead box P3 (FOXP3) mRNA in the jejunum and spleen and TIM4 mRNA in the jejunum was measured by reverse transcription-polymerase chain reaction (RT-PCR). The levels of trans- forming growth factor 1 (TGF-1) and inter- leukin-10 (IL-10) in the serum were analyzed by enzyme-linked immunosorbent assay (ELISA). The expression of TGF-1 and IL-10 proteins in the jejunum was detected by immunohistochemistry.RESULTS: Compared with the NS group, the expression levels of FOXP3 mRNA in the jeju- num and spleen significantly decreased (0.401 0.145 vs 0.732 0.162 and 0.407 0.082 vs 0.691 0.145, respectively; both P 0.05), the expres- sion level of TIM4 mRNA increased significantly (P 0.05), and the levels of TGF-1 in the serum and jejunum decreased significantly (7859.853 126.704 ng/L vs 8342.814 488.461 ng/L and 108.834 9.634 ng/L vs 156.298 12.002 ng/L, respectively; both P 0.05) in the SEB/OVA group. In contrast, the expression levels of FOXP3 mRNA in the jejunum and spleen and TGF-1 in the serum and jejunum were signifi- cantly higher in the anti-TIM1/SEB/OVA and anti-TIM4/SEB/OVA groups than in the SEB/ OVA group (all P 0.05).CONCLUSION: Treg cells in SEB/OVA-sensi- tized mice are dysfunctional. Pretreatment with anti-TIM1 or anti-TIM4 can restore the function of Treg cells, suggesting that the TIM4-TIM1 pathway may play a key role in the develop- ment of food allergy.Key Words: Food allergy; CD4+CD25+ T regulatory www.wjgnet.com背景资料 FA在世界上广泛 存在, 最近几十 年其发病率在全 球范围内大大提 高. 然而FA的发 病机制尚不清楚, 最近几年有关FA 领域研究发展迅 速. 口服耐受受 损是致FA的重要 因素之一, 维持 口服耐受的复杂 免疫调节网络成 为研究热点.同行评议者 范建高, 教授, 上 海交通大学医学 院附属新华医院 消化内科cells; Oral tolerance; TIM proteinWang XT, Zheng PY, Luo Y, Liu ZQ, Zhang LL. TIM4 and TIM1 modulate the function of CD4+CD25+ T regulatory cells in mice with food allergy. Shijie Huaren Xiaohua Zazhi 2009; 17(34): 3507-3513摘要 目的: 探讨在食物过敏小鼠模型中CD4+CD25+ Treg细胞的功能状态及TIM4与TIM1对其的 影响, 分析食物过敏的发生机制. 方法: 无受试蛋白喂养BALB/c小鼠32只, 随 机分为4组: 空白对照组、金黄色葡萄球菌肠 毒素B(SEB)+卵清蛋白(OVA)共同作用组、 TIM1抗体干预组及TIM4抗体干预组, 分别于 0、3、9 d ip生理盐水, SEB和OVA, TIM1抗 体+SEB+OVA, TIM4抗体+SEB+OVA, 并于第 7、14天给予SEB+OVA(空白对照组以生理盐 水ig)ig. RT-PCR检测空肠及脾脏Foxp3 mRNA 表达、空肠TIM4 mRNA表达, ELISA法测定 血清TGF-1与IL-10的表达. 免疫组织化学法 检测空肠黏膜TGF-1与IL-10表达. 结果: 与空白对照组相比, SEB+OVA共同作 用组小鼠空肠及脾脏Foxp3 mRNA表达明 显下降(0.4010.145 vs 0.7320.162; 0.407 0.082 vs 0.6910.145, 均P0.05), TIM4 mRNA表达明显增高(P0.05), 血清和空肠黏 膜TGF-1表达显著降低(7859.853126.704 ng/L vs 8342.814488.461 ng/L; 108.834 9.634 ng/L vs 156.29812.002 ng/L, 均 P0.05); 与SEB+OVA组相比, TIM1和TIM4 抗体干预组小鼠空肠及脾脏Foxp3 mRNA 及血清和空肠黏膜