Kelly R H, Rao K N, Harvey V S, Lombardi B
Acta Hepatogastroenterol (Stuttg). 1979 Aug;26(4):302-9.
Feeding a choline-deficient diet containing 0.5% DL-ethionine induces an acute hemorrhagic pancreatitis in 100% of young female mice. Evidence for deposition of the third component of complement (C3) on acinar cell plasma membranes was sought, during the inductive stages, by a sandwich immunofluorescence technique. Such a localization could not be demonstrated even though the method is capable of detecting less than 8 x 10(-5) microgram of protein/mm2 of cell membrane. Artifactual binding of immunoglobulin reagents was encountered when goat antisera, with high levels of circulating immune complexes, formed the middle layer in the sandwich technique. This was attributed to the appearance of Fc receptors on the plasma membrane of degenerating acinar cells, and could be avoided by ultracentrifuging acinar cells, and could be avoided by ultracentrifuging the goat antisera prior to sue. In view of the fact that C3 cleavage represents an amplification loop in both the calssical and alternate pathways of complement activation, the lack of demonstrable C3 staining in tbe present experiments strongly suggests that complement plays no role in acinar cell necrosis in this model of pancreatitis.
给年轻雌性小鼠喂食含0.5% DL-乙硫氨酸的胆碱缺乏饮食会导致100%的小鼠发生急性出血性胰腺炎。在诱导阶段，通过夹心免疫荧光技术寻找补体第三成分（C3）在腺泡细胞质膜上沉积的证据。尽管该方法能够检测到每平方毫米细胞膜上少于8×10⁻⁵微克的蛋白质，但仍未证实有这样的定位。当含有高水平循环免疫复合物的山羊抗血清在夹心技术中形成中间层时，会出现免疫球蛋白试剂的人为结合。这归因于退化腺泡细胞质膜上Fc受体的出现，通过对腺泡细胞进行超速离心可以避免这种情况，并且在使用前对山羊抗血清进行超速离心也可以避免。鉴于C3裂解在补体激活的经典途径和替代途径中均代表一个放大环，本实验中缺乏可证实的C3染色强烈表明补体在该胰腺炎模型的腺泡细胞坏死中不起作用。
