CAJ | 학술논문

目的探讨利用携带人工合成的共刺激分子细胞毒性T淋巴细胞抗原4(CTLA-4)拮抗剂(CTLA4Ig)的真核表达载体,对自身免疫性甲状腺炎进行基因治疗的效果.方法采用辅助性T淋巴细胞1型(Th1)倾向的C57BL/6J雌性小鼠,构建实验性自身免疫性甲状腺炎(EAT)动物模型20只,随机分为基因治疗组(治疗组)和模型对照组(模型组),每组各10只,分别行甲状腺内质粒pCI/CTLA4Ig和空质粒pCI局部注射,另设非干预正常对照组(对照组)10只.获得血清和甲状腺、脾脏组织,通过实时定量PCR及酶联免疫吸附试验(ELISA)方法检测样本中甲状腺自身抗体及Th1、Th2相关细胞因子的表达.结果与模型组比较,治疗组的甲状腺组织CTLA-4的表达升高近两倍(P=0.038),Th1细胞因子干扰素γ(IFN-γ)及细胞间黏附分子1(ICAM-1)显著降低(P值分别为0.016及0.042),而Th2细胞相关因子白细胞介素4(IL-4)的表达显著升高(P=0.044).脾脏组织和小鼠血清中细胞因子水平显示了同样的改变.血清中抗甲状腺过氧化物酶抗体水平在治疗前后改变不显著.结论甲状腺局部CTLA4Ig基因的注射可通过诱导Th1优势向Th2偏移而对EAT起治疗作用.
목적 탐토이용휴대인공합성적공자격분자세포독성T림파세포항원4(CTLA-4)길항제(CTLA4Ig)적진핵표체재체,대자신면역성갑상선염진행기인치료적효과.방법 채용보조성T림파세포1형(Th1)경향적C57BL/6J자성소서,구건실험성자신면역성갑상선염(EAT)동물모형20지,수궤분위기인치료조(치료조)화모형대조조(모형조),매조각10지,분별행갑상선내질립pCI/CTLA4Ig화공질립pCI국부주사,령설비간예정상대조조(대조조)10지.획득혈청화갑상선、비장조직,통과실시정량PCR급매련면역흡부시험(ELISA)방법검측양본중갑상선자신항체급Th1、Th2상관세포인자적표체.결과 여모형조비교,치료조적갑상선조직CTLA-4적표체승고근량배(P=0.038),Th1세포인자간우소γ(IFN-γ)급세포간점부분자1(ICAM-1)현저강저(P치분별위0.016급0.042),이Th2세포상관인자백세포개소4(IL-4)적표체현저승고(P=0.044).비장조직화소서혈청중세포인자수평현시료동양적개변.혈청중항갑상선과양화물매항체수평재치료전후개변불현저.결론 갑상선국부CTLA4Ig기인적주사가통과유도Th1우세향Th2편이이대EAT기치료작용.
Objective To investigate the therapeutic effect of CTLA4Ig gene on experimental autoimmune thyroiditis (EAT) by the use of portable synthetic costimulatory molecules of cytotoxic T lymphocyte antigen 4 (CTLA-4) antagonist (CTLA4Ig) eukaryotic expression vector.Methods Thirty C57BL/6J female mice were divided into three groups,named EAT model group (EAT,n =10),CTLA4Ig-treatment group (CTLA4Ig-EAT,n =10) and control group(n =10).At 28 day after first immunization,plasmids mixture with pCI or pCI/CTLA4Ig were injected into thyroid tissues of EAT and CTLA4Ig-EAT by surgery,respectively.Serum,thyroid tissues and spleens were collected as samples.Thyroid autoantibody and expression of interleukin (Th)1,Th2 related cytokinesby were measured by real-time quantitative PCR and ELISA.Results Compared with EAT group,the expression of CTLA-4 in thyroid of CTLA4Ig-EAT group was elevated double folds (P =0.038),and the expression of Th1 cytokine interferon γ and intercellular adhesion molecule-1 decreased significantly (P =0.016,0.042).Meanwhile,Th2 cytokine IL-4 was increased after CTLA4Ig treatment (P =0.044).The same changes were seen in spleen tissues and serum.There was no significant difference in terms of TPOAb between EAT and treated group.Conclusion Local thyroid injection of CTLA4Ig gene shows the therapeutic effect to same degree on EAT through adjusting the underlying Th1/Th2 imbalance.