Murphy-Ullrich J E, Oberley T D, Mosher D F
Am J Pathol. 1986 Oct;125(1):182-90.
It is demonstrated that BALB/c/DUB mice receiving chronic intraperitoneal injection of low doses of murine native dimeric or denatured multimeric or monomeric serum fibronectin in the absence of adjuvant produce a humoral autoimmune response to mouse fibronectin and have significant levels of circulating immune complexes. With the use of the Western immunoblotting technique, sera of mice immunized with native or sodium dodecyl sulfate (SDS)-treated fibronectin reacted with native, reduced and alkylated, and heat-and SDS-treated fibronectin. Sera from mice injected with reduced and alkylated fibronectin, however, reacted preferentially with reduced and alkylated fibronectin. In a competitive inhibition ELISA, the reactivity of immune sera from mice immunized with reduced and alkylated fibronectin to insolubilized reduced and alkylated fibronectin could be inhibited (66%) by preincubation with soluble reduced and alkylated fibronectin, but not by preincubation with native or SDS-treated fibronectin (less than 25%). Glomerular mesangial electron-dense deposits were observed in animals given injections of all three forms of fibronectin, but not in saline-injected animals. Indirect immunoperoxidase studies suggested that these dense deposits were at least partially immune in nature, because the authors were able to demonstrate binding in vivo of mouse IgG.
结果表明,在无佐剂情况下,慢性腹腔注射低剂量鼠源天然二聚体、变性多聚体或单体血清纤连蛋白的BALB/c/DUB小鼠,会对小鼠纤连蛋白产生体液性自身免疫反应,并具有显著水平的循环免疫复合物。运用Western免疫印迹技术,用天然或经十二烷基硫酸钠(SDS)处理的纤连蛋白免疫的小鼠血清,可与天然、还原烷基化以及加热和SDS处理的纤连蛋白发生反应。然而,注射还原烷基化纤连蛋白的小鼠血清,优先与还原烷基化纤连蛋白发生反应。在竞争性抑制酶联免疫吸附测定中,用还原烷基化纤连蛋白免疫的小鼠免疫血清与不溶性还原烷基化纤连蛋白的反应性,可通过与可溶性还原烷基化纤连蛋白预孵育而受到抑制(66%),但与天然或SDS处理的纤连蛋白预孵育则无此效果(小于25%)。在注射所有三种形式纤连蛋白的动物中均观察到肾小球系膜电子致密沉积物,而注射生理盐水的动物中未观察到。间接免疫过氧化物酶研究表明,这些致密沉积物至少部分具有免疫性质,因为作者能够证明小鼠IgG在体内的结合。