睾丸内胰岛移植对CD4^+ CD25^+调节性T细胞分布的影响  

Distribution of CD4～+ CD25～+ regulatory T cells following islets transplantation into the testis

作　　者：许宝华[1] 万福生[1,2] 李华[2] 刘丽乔[2] 戴振华[3] 

机构地区：[1]南昌大学实验动物科学部,江西省南昌市330006 [2]南昌大学医学院生物化学与分子生物学教研室,江西省南昌市330006 [3]美国德克萨斯大学医学中心,tylertxusa70758

出　　处：《中国组织工程研究与临床康复》2008年第B12期10407-10410,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30660179)~~

摘　　要：背景:CD4+CD25+调节性T细胞是维持机体免疫耐受的重要调控者,参与了多种移植免疫耐受的诱导。目的:拟观察小鼠睾丸内胰岛移植后CD4+CD25+调节性T细胞分布的特点。设计、时间及地点:观察对照动物实验,2007-04/2008-01在江西省实验动物中心完成。材料:成年Balb/c小鼠。方法:分离小鼠胰岛细胞,采用胰管内注射胶原酶水浴消化及Ficoll400不连续密度梯度离心法纯化,以双硫腙染色,计算胰岛细胞纯度,以体外葡萄糖刺激胰岛素分泌试验判定胰岛细胞功能。将胰岛移植至小鼠睾丸或肾包膜下,每只移植300～400个胰岛。在胰岛移植24h后麻醉处死小鼠,取脾脏、睾丸及淋巴结,制成细胞悬液,免疫磁珠法分离CD4+CD25+T细胞,通过流式细胞仪分析计数。主要观察指标:胰岛细胞的纯度及功能,CD4+CD25+调节性T细胞的分布。结果:纯化后每只胰腺获得(478±53)个胰岛细胞,纯度为(81.5±12.3)%,纯化后细胞形态完好,活度大于90%。睾丸内胰岛移植时,睾丸、淋巴结及脾脏中CD4+CD25+调节性T细胞均显著增多(P<0.05～0.01)。结论:睾丸内胰岛移植能明显上调睾丸、淋巴结及脾脏中CD4+CD25+调节性T细胞数量。BACKGROUND： CD4^＋ CD25^＋ regulatory T cells are the important regulators for the immune tolerance of the human body. And they participate in several kinds of transplantative immune tolerance induction. OBJECTIVE： To observe the distribution characteristics of CD4^＋ CD25^＋ regulatory T cells after islets transplantation in mouse testis. DESIGN, TIME AND SETTING： A controlled observational animal experiment was performed at Medical Laboratory Animal Center in Jiangxi Province from August 2007 to January 2008. MATERIALS： Adult Balb/c mice were selected in the experiment. METHODS： Islet cells isolated from mice were digested with collagenase injected into pancreatic duct, followed by incubation in water bath. Then, the cells were purified by Ficol1400 discontinuous density gradient centrifugation. The purity of islet cells was evaluated by dithizone staining. The function of islet cells was determined by glucose-stimulated insulin release test. The 300-400 islet cells were transplanted into the testis or subrenal capsule of each mouse. Under anesthesia, all mice were sacrificed 24 hours after transplantation. The spleens, testises and lymph nodes were separated and made cell suspension. The CD4^＋ CD25^＋ regulatory T cells were isolated by immunomagnetic bead separation. The flow cytometry were used to count CD4^＋ CD25^＋ regulatory T cells. MAIN OUTCOME MEASURES： Purity and function of islet cells, and distribution ofCD4^＋ CD25^＋ regulatory T cells were observed. RESULTS： After purification, the quantity of islet cells were （478±53） per pancreas, the average purity was （81.5±12.3） %, and the viability of islets was more than 90 %. The quantity ofCD4^＋ CD25^＋ regulatory T cells was increased significantly in the testis, lymph node and spleen after islet transplantation into the testis （P 〈 0.05-0.01）. CONCLUSION： Islets transplantation in the testis can increase the quantity of CD4^＋ CD25^＋ regulatory T cells in the testis, lymph node, and spleen obviously.

关 键 词：胰岛移植 CD4^+CD25^+T细胞 睾丸 免疫赦免 细胞移植 

分 类 号：R617[医药卫生—外科学]