Kraehenbuhl J P, Racine L, Galardy R E
Ann N Y Acad Sci. 1975 Jun 30;254:190-202. doi: 10.1111/j.1749-6632.1975.tb29169.x.
The different steps in the processing of secretory IgA (sIgA) of the mammary gland of lactating rabbits have been investigated with immunoelectron microscopy. The method employed consists of an indirect localization sequence, in which the immunologic reagents are allowed to diffuse into the fixed tissue prior to embedding and thin sectioning. In the first step, goat F(ab')2 fragments directed against sIgA, secretory component (SC), or alpha chain were used. To visualize the location of these first-step antibody fragments, a second Fab fragment directed against goat F(ab')2 antibody was prepared and coupled to a small heme octapeptide that possessed peroxidatic activity, the reaction product of which is visible by electron microscopy. Both the epithelial cell and the local plasmacytes of the mammary gland contain alpha chain, whereas SC is exclusively located in the epithelial cells. In the plasmacytes, both the cisternae of the rough endoplasmic reticulum and the saccules and vesicles of the Golgi complex contain alpha chain. In the epithelial cells, the elements of the Golgi complex and large apical vacuoles situated in the apical region of the cell. Based on these results, a model for processing of sIgA is proposed and discussed.
利用免疫电子显微镜研究了泌乳兔乳腺分泌型IgA(sIgA)加工过程中的不同步骤。所采用的方法包括间接定位序列,即在包埋和超薄切片之前,使免疫试剂扩散到固定组织中。第一步,使用针对sIgA、分泌成分(SC)或α链的山羊F(ab')2片段。为了可视化这些第一步抗体片段的位置,制备了针对山羊F(ab')2抗体的第二种Fab片段,并将其与具有过氧化物酶活性的小血红素八肽偶联,其反应产物可通过电子显微镜观察到。乳腺的上皮细胞和局部浆细胞都含有α链,而SC仅位于上皮细胞中。在浆细胞中,粗面内质网的池以及高尔基体复合体的扁平囊和小泡都含有α链。在上皮细胞中,高尔基体复合体的成分和位于细胞顶端区域的大顶端空泡。基于这些结果,提出并讨论了sIgA加工的模型。