Salamero J, Remy J J, Michel-Bechet M, Charreire J
Eur J Immunol. 1987 Jun;17(6):843-8. doi: 10.1002/eji.1830170617.
Monoclonal autoantibodies obtained using the native porcine thyroglobulin (Tg) molecule for immunization were shown to react against a defined epitope of the Tg molecule. Its biochemical characterization was undertaken, using trypsin treatment of Tg, followed by gel filtration, affinity chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and high pressure liquid chromatography analysis. Immunoreactivity tested by immunoblotting with appropriate monoclonal anti-Tg antibody allowed the detection of one peptide fragment of 5-10 kDa, after limited proteolysis with trypsin. We demonstrated that this peptide reacts exclusively with monoclonal anti-Tg antibodies which block in vitro proliferative T cell response; moreover, the 5-10-kDa tryptic fragments from porcine Tg were able to induce thyroiditis when injected into normal CBA mice, while porcine Tg depleted from these fragments was not able to do so.
使用天然猪甲状腺球蛋白（Tg）分子进行免疫获得的单克隆自身抗体被证明可与Tg分子的一个特定表位发生反应。对其进行了生化特性分析，先用胰蛋白酶处理Tg，然后进行凝胶过滤、亲和色谱、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和高压液相色谱分析。在用胰蛋白酶进行有限的蛋白水解后，通过用适当的单克隆抗Tg抗体进行免疫印迹测试免疫反应性，可检测到一个5 - 10 kDa的肽片段。我们证明，该肽仅与阻断体外增殖性T细胞反应的单克隆抗Tg抗体发生反应；此外，将来自猪Tg的5 - 10 kDa胰蛋白酶片段注射到正常CBA小鼠中时能够诱发甲状腺炎，而去除这些片段的猪Tg则不能。
