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目的探讨猪甲状腺球蛋白(porcine thyroglobulin,pTg)免疫诱导C57BL6小鼠Graves病模型的效果及适宜剂量。方法 8周龄雌性C57BL6小鼠29只,随机分为对照组9只,免疫1组10只,免疫2组10只,对照组仅给予蒸馏水+弗氏佐剂,免疫1组和免疫2组分别给予pTg 25、50μg/鼠+弗氏佐剂进行免疫,10周龄时加强免疫1次,之后每周加强免疫1次,共8次。19周龄时,3组小鼠采用放射免疫法测定血清游离甲状腺素(free thyroxine,FT4)水平,采用ELISA法测定甲状腺刺激性抗体(thyroid stimulating antibody,TSAb)水平,并行甲状腺组织病理学检查。结果免疫2组小鼠血清FT4[(55.61±22.30)pmol/L]、TSAb[(2 546.76±606.06)μIU/L]水平明显高于对照组[(15.11±5.69)pmol/L、(1 805.18±213.48)μIU/L](P<0.05);免疫1组小鼠血清FT4[(24.74±17.40)pmol/L]、TSAb[(2 157.10±536.92)μIU/L]与对照组比较差异无统计学意义(P>0.05);免疫2组小鼠血清FT4水平高于免疫1组(P<0.05),TSAb水平与免疫1组比较差异无统计学意义(P>0.05);免疫1组4只和免疫2组7只小鼠甲状腺组织病理检查示甲状腺弥漫性肿大,甲状腺滤泡上皮细胞高度增加,且甲状腺未见明显淋巴细胞浸润;免疫2组Graves病成模率(70%)高于免疫1组(40%)(P<0.05);免疫2组TSAb与FT4呈明显正相关(r=0.863,P=0.006)。结论利用pTg诱导C57BL6小鼠Graves病模型可行,且50μg/鼠可能是最适宜的免疫抗原剂量。
Objective To investigate the effect of porcine thyroglobulin (pTg) immunization on Graves disease induced by porcine thyroid carcinoma (C57BL6) mice and its suitable dosage. Methods Twenty-nine female C57BL6 mice aged 8 weeks were randomly divided into control group (n = 9), immune group (n = 10) and immune group (n = 10). Control group was given distilled water and Freund’s adjuvant only Immunizations were given at pTg 25, 50 μg / mouse + Freund’s adjuvant, boosted at 10 weeks of age and then boosted once per week for a total of 8 times. At 19 weeks of age, free thyroxine (FT4) levels were measured by radioimmunoassay in 3 groups of mice. Thyroid stimulating antibody (TSAb) levels were measured by ELISA, and histopathological examination of thyroid was performed. Results The levels of serum FT4 (55.61 ± 22.30 pmol / L) and TSAb (2 546.76 ± 606.06) μIU / L in the two groups were significantly higher than those in the control group [(15.11 ± 5.69) pmol / L, ± 21.48) μIU / L] (P <0.05). There was no significant difference in serum FT4 [(24.74 ± 17.40) pmol / L and TSAb [(2 157.10 ± 536.92) μIU / L] (P> 0.05). The level of serum FT4 in the two immunized groups was significantly higher than that in the immunized group (P <0.05), while the level of TSAb was not significantly different from that in the immunized group (P> 0.05) Immunohistochemistry and immunohistochemistry showed that the pathological examination of thyroid in 7 mice showed that the thyroid was diffusely enlarged, the thyroid follicular epithelial cells were highly increased and there was no obvious lymphocyte infiltration in the thyroid gland. The rate of Graves’ disease (70% Immunization group 1 (40%) (P <0.05). There was a significant positive correlation between TSAb and FT4 in immunization group 2 (r = 0.863, P = 0.006). Conclusion It is feasible to use pTg to induce Graves’ disease model in C57BL6 mice, and 50μg / mouse may be the most suitable immunization antigen dose.