摘要
目的:建立CD45RBhighCD4+幼稚T细胞诱导严重联合免疫缺陷型(SCID)小鼠慢性结肠炎模型,并探讨该结肠炎模型的免疫病理特征。方法:采用流式细胞仪分离BALB/c小鼠(H-2d基因表型)脾脏CD45RBhighCD4+幼稚T细胞,经尾静脉或腹腔注射植入21只同基因型SCID小鼠体内,观察慢性结肠炎的发生。8周后处死SCID小鼠,分离结肠黏膜固有层内CD4+T淋巴细胞,体外刺激,分别采用ELISA法和RT-PCR法检测炎症结肠黏膜组织内炎症细胞因子(IFN-γ、IL-2、IL-4、IL-5、TNF-α)及其mRNA的表达水平。以16只正常BALB/c小鼠作对照。结果:CD45RBhighCD4+幼稚T细胞植入SCID小鼠(4.1±0.3)周后,小鼠体重开始下降,大便变软;(7.4±0.6)周后出现腹泻、体重明显下降、脱肛等慢性结肠炎症状;组织病理检查发现结肠黏膜层、黏膜下层、肌层及浆膜层有大量的淋巴细胞浸润,以黏膜层和黏膜下层明显,并出现肠上皮细胞炎性增生、黏膜溃疡形成。与正常BALB/c小鼠比较,CD45RBhighCD4+幼稚T细胞诱导后,SCID小鼠结肠黏膜固有层CD4+T细胞IFN-γ、IL-2、TNF-α分泌水平增高(P<0.005),结肠黏膜组织内IL-2、IFN-γ、TNF-αmRNA水平亦升高(P<0.005)。结论:CD45RBhighCD4+幼稚T细胞可诱导SCID小鼠发生慢性结肠炎。
Aim： To establish severe combined immunodeficient （SCID） mice induced by CD45^high CD4^＋ naive T cells chronic colitis model, and investigate the modelg immuno-pathological features. Methods：CD45^high CD4^＋ naive T cells isolated from BALB/c mice （H-2d） were sorted through flow cytometry and then transferred intravenously or intraperitoneally into syngenetic SCID mice. Mice were sacrificed at 8 weeks after cell transfer and CD4^＋ T cells were isolated from the inflamed colonic lamina propria. Proinflammatory cytokines （IFN-γ, IL-2, TNF-α, IL-4, IL-5 ） were analyzed by ELISA after in vitro stimulation with anti-CD3, and mRNA levels of these cytokines in inflamed colonic mucosa were also determined by quantitative PCR. Results： SCID mice appeared weight decrease and soft stool at （4.1 ± 0. 3 ） weeks after the reconstitution, and developed colitis at （7.4 ±0. 6） weeks thereafter, characterized by diarrhea, weight sharp decrease, prolapse and large number of lymphocyte infiltration in mucosa and submucosa, intestinal epithelial cell hyperplasia and ulceration. Compared with healthy BALB/c mice, lamina propfia CD4^＋ T cells from the inflamed colon secreted significantly higher levels of IFN-γ, IL-2 and TNF-α（ P 〈 0. 005 ） after anti-CD3 stimulation in vitro, and mRNA levels of IFN-γ, IL-2 and TNF-α also increased （P 〈0. 005）. Conclusion： CD45RB^high CD4^＋ T cells can induce chronic colitis in SCID mice.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2006年第5期815-818,共4页
Journal of Zhengzhou University(Medical Sciences)
基金
国家自然科学基金资助项目30571751
教育部"新世纪优秀人才支持计划"基金资助项目NCET-05-0609
