大鼠免疫功能恢复与白细胞介素-10基因转染的关系

Relationship between the recovery of immunological function and interleukin-10 gene transfection

目的:通过建立实验性自身免疫性甲状腺炎(experimentalautoimmunethyroiditis,EAT)动物模型的基础上,探索通过基因转染方法,以纠正EAT大鼠免疫功能异常状态。方法:选用雌性Wistar大鼠建立EAT动物模型。成模大鼠分为4组:A组5只(PBS+PLL),B组5只(pORF质粒+PLL)、C组10只(pORFmIL10质粒+PLL)及D组5只(pORFmIL10质粒+MEM)。使用放射免疫方法测定TgAb,TmAb滴度。采用Charreive法计算甲状腺内淋巴细胞浸润指数。脾脏淋巴细胞培养,采用犤3H犦-TdR标记方法测定cpm值并计算淋巴细胞增殖指数。结果:C组与A,B,D组比较,第4,6,8周TgAb,TmAb滴度显著降低(P均<0.001,F=42.66,32.65,9.66;F=22.25,81.35,14.84)。C组甲状腺淋巴细胞浸润指数为1.10±0.18,显著低于其他各组(F=4.39,P<0.01)。此外,淋巴细胞增殖试验结果显示,C组PTg刺激的淋巴细胞增殖指数显著低于A,B组(F=2.78,P<0.05)。结论:通过大鼠甲状腺组织内局部注射pORFmIL10质粒使甲状腺滤泡上皮细胞表达白细胞介素10基因,并且能够清除甲状腺内浸润的淋巴细胞,降低自身抗体水平和针对抗原反应的T淋巴细胞增殖反应。多聚赖氨酸可以延长目的基因表达时间,更有效地治疗EAT。

AIM:To explore the effect of gene transfection on redressing the immunological functional abnormality of experimental autoimmune thyroiditis(EAT) rats on the basis of establishing EAT animal models. METHODS:Wistar female rats were selected to establish EAT animal models.The ra ts were divided into four groups:group A(PBS+PLL,n=5),group B(pORF+PLL,n=5),g roup C[pORFm interleukin(IL)-10 +PLL,n=10], group D(pORFmIL10+MEM,n=5).The me thod of radio-immunity was used to determine the titre of TgAb and that of Char reive to calculate the infiltrative index of thyroid endolymphatic cell.Spleen l ymphocytes were cultured,and -TdR marking method was used to determine the value of cpm and calculate the proliferation index. RESULTS:The titre of TgAb and TmAb in group C at the 4,6,8 weeks decreased sig nificantly than those in group A,B and D(P< 0.001,F=42.66,32.65,9.66,22.25,81.3 5,14.84). The infiltrative index of thyroid endolymphatic cell in group C was ( 1.10±0.18), which was significantly lower than those in other 3 groups(P< 0.01 ,F=4.39).Besides,the result of lymphocytes proliferation index test showed that proliferation index of the cells,which were stimulated by PTg,in group C were s ignificantly lower than those in group A and B(P< 0.05,F=2.78).