中国免疫学杂志
中國免疫學雜誌
중국면역학잡지
Chinese Journal of Immunology
2015年
10期
1320-1323
,共4页
余毅恺%冯业晨%张木勋%叶丛%涂巍
餘毅愷%馮業晨%張木勛%葉叢%塗巍
여의개%풍업신%장목훈%협총%도외
ICOSL%Graves病%TSHR抗体%基因枪
ICOSL%Graves病%TSHR抗體%基因鎗
ICOSL%Graves병%TSHR항체%기인창
ICOSL%Graves disease%TSHR autoantibody%Gene gun
目的：研究可诱导共刺激分子及配体（ICOS／ICOSL）在Graves 病（以下简称甲亢，GD）中的病理生理机制。方法：45只远交系BALB／c 小鼠随机分成三组各15只，用基因枪将不同质粒注射。A 组采用表达pCDNA3．0－mICOSL 和pCDNA3．0－hTSHR 的质粒免疫；B 组用表达pCDNA3．0－hTSHR 的质粒免疫，C 组采用空质粒pcDNA3．0免疫作为对照组。放免法测定小鼠血清游离甲状腺素（FT4）、脾脏细胞上清液IFN－γ度，ELISA 法血清促甲状腺素受体抗体（TRAb），小鼠血清和转染人hTSHR－CHO 细胞共同孵育后检测cAMP 浓度判断TRAb 自身抗体活性。结果：质粒注射后A 组小鼠血清FT4水平（0．49±0．25）pg／ml 高于C 组（q＝6．571，P ＝0．023）；成模达标率（10／15例）高于B、C 两组（χ2＝14．47，P ＝0．005）。A 组小鼠脾脏原代培养上清液IFN－γ的产量（1．88±0．41）pmol／L 明显高于B、C 两组（F ＝17．85，P ＝0．003）。A 组小鼠血清TRAb 活性188．3（179．7～260．2）％高于B、C 组（P ＝0．027）。结论：外源性导入pCDNA3．0－mICOSL 到GD 小鼠，能刺激其脾脏淋巴细胞分泌IFN－γ，增加自身抗体TRAb 活性，从而上调GD 动物体内免疫应答。
目的：研究可誘導共刺激分子及配體（ICOS／ICOSL）在Graves 病（以下簡稱甲亢，GD）中的病理生理機製。方法：45隻遠交繫BALB／c 小鼠隨機分成三組各15隻，用基因鎗將不同質粒註射。A 組採用錶達pCDNA3．0－mICOSL 和pCDNA3．0－hTSHR 的質粒免疫；B 組用錶達pCDNA3．0－hTSHR 的質粒免疫，C 組採用空質粒pcDNA3．0免疫作為對照組。放免法測定小鼠血清遊離甲狀腺素（FT4）、脾髒細胞上清液IFN－γ度，ELISA 法血清促甲狀腺素受體抗體（TRAb），小鼠血清和轉染人hTSHR－CHO 細胞共同孵育後檢測cAMP 濃度判斷TRAb 自身抗體活性。結果：質粒註射後A 組小鼠血清FT4水平（0．49±0．25）pg／ml 高于C 組（q＝6．571，P ＝0．023）；成模達標率（10／15例）高于B、C 兩組（χ2＝14．47，P ＝0．005）。A 組小鼠脾髒原代培養上清液IFN－γ的產量（1．88±0．41）pmol／L 明顯高于B、C 兩組（F ＝17．85，P ＝0．003）。A 組小鼠血清TRAb 活性188．3（179．7～260．2）％高于B、C 組（P ＝0．027）。結論：外源性導入pCDNA3．0－mICOSL 到GD 小鼠，能刺激其脾髒淋巴細胞分泌IFN－γ，增加自身抗體TRAb 活性，從而上調GD 動物體內免疫應答。
목적：연구가유도공자격분자급배체（ICOS／ICOSL）재Graves 병（이하간칭갑항，GD）중적병리생리궤제。방법：45지원교계BALB／c 소서수궤분성삼조각15지，용기인창장불동질립주사。A 조채용표체pCDNA3．0－mICOSL 화pCDNA3．0－hTSHR 적질립면역；B 조용표체pCDNA3．0－hTSHR 적질립면역，C 조채용공질립pcDNA3．0면역작위대조조。방면법측정소서혈청유리갑상선소（FT4）、비장세포상청액IFN－γ도，ELISA 법혈청촉갑상선소수체항체（TRAb），소서혈청화전염인hTSHR－CHO 세포공동부육후검측cAMP 농도판단TRAb 자신항체활성。결과：질립주사후A 조소서혈청FT4수평（0．49±0．25）pg／ml 고우C 조（q＝6．571，P ＝0．023）；성모체표솔（10／15례）고우B、C 량조（χ2＝14．47，P ＝0．005）。A 조소서비장원대배양상청액IFN－γ적산량（1．88±0．41）pmol／L 명현고우B、C 량조（F ＝17．85，P ＝0．003）。A 조소서혈청TRAb 활성188．3（179．7～260．2）％고우B、C 조（P ＝0．027）。결론：외원성도입pCDNA3．0－mICOSL 도GD 소서，능자격기비장림파세포분비IFN－γ，증가자신항체TRAb 활성，종이상조GD 동물체내면역응답。
Objective:To study the pathological mechanism of the inducible co-stimulator molecular and ligand ( ICOS/ICOSL) in Graves disease animal.Methods:45 out-bred BALB/c mice were randomly divided into three groups with 15 rats in each group;using gene gun to deliver different plasmid injection.Group A was delivered with pCDNA3.0-mICOSL and pCDNA3.0-hTSHR, Group B with pCDNA3.0-hTSHR and null pCDNA3.0 with Group C for immunization as the control group.The concentration of serum free thyroxine immunization was deter mined with immunoassay and serum thyrotropin receptor antibody ( TRAb ) with ELISA, supernatant of IFN-γconcentration in mouse spleen cells was measured with radioimmunoassay,and hTSHR transected CHO cells were incubated to detect the concentration of cAMP to deter mine autoantibody TRAb activity.Results: After plasmid injection serum FT4 level in Group A (0.49±0.25) pg/ml ( q=6.571,P=0.023) was higher than that in Group C,the standard rate was higher than Group B and C (χ2=14.47,P=0.005).IFN-γconcentration of mice spleen cultured supernatant in Group A (1.88±0.41) pmol/L was significantly higher than the other two groups.The activity of autoantibody TRAb in Group A 188.3 (179.7-260.2) %was higher than that in the other two groups ( P=0.027 ) .Conclusion: Exogenous delivery of pCDNA3.0-mICOSL plasmid in GD mice could stimulate the spleen lymphocytes to secrete more IFN-γ,increase the activity of TRAb autoantibodies and might lead to upregulation of immune response in Graves animal model in vivo.
