Immunofluorescence and immunoblot studies on the reactivity of pemphigus vulgaris and pemphigus foliaceus sera with desmoglein 3 and desmoglein 1.

Epithelial cell biology Pub Date : 1995-01-01
T Hashimoto, M Amagai, D R Garrod, T Nishikawa
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Abstract

We have investigated expression of pemphigus vulgaris (PV) antigen, desmoglein 3 (Dsg3), and pemphigus foliaceus (PF) antigen, desmoglein 1 (Dsg1), in various tissues. Immunofluorescence studies suggested that Dsg1 and Dsg3 are preferentially expressed in the upper and lower epidermis, respectively. With immunoblotting of human epidermal extracts, all PV sera reacted with Dsg3 but not with Dsg1. By contrast, only half of PF sera reacted with Dsg1 but none reacted with Dsg3. These results confirm the distinct antibody specificity between PV and PF sera. They also suggest that PV sera contain antibodies against linear epitopes present even on the denatured antigen, while some PF sera contain antibodies only against conformational epitopes present on the native antigen. With immunoblotting of bovine desmosome preparations, certain PV sera and an anti-Dsg monoclonal antibody reacted with both Dsg1 and Dsg3. Affinity-purification of these PV antibodies suggested that the simultaneous reactivity with Dsg1 and Dsg3 was produced by two different subsets of antibodies and not by cross-reactivity of single antibodies. This study indicates that pemphigus serum is a useful probe with which we learn various aspects of keratinocyte biology.

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免疫荧光和免疫印迹法研究寻常型天疱疮和叶状天疱疮血清对粘连蛋白3和粘连蛋白1的反应性。
我们研究了寻常型天疱疮(PV)抗原、桥粒蛋白3 (Dsg3)和叶状天疱疮(PF)抗原、桥粒蛋白1 (Dsg1)在不同组织中的表达。免疫荧光研究表明,Dsg1和Dsg3分别优先表达于上表皮和下表皮。对人表皮提取物进行免疫印迹检测,所有PV血清均与Dsg3有反应,但与Dsg1无反应。相比之下,只有一半的PF血清与Dsg1反应,而没有与Dsg3反应。这些结果证实PV和PF血清之间存在明显的抗体特异性。他们还表明,PV血清含有针对变性抗原上存在的线性表位的抗体,而一些PF血清只含有针对天然抗原上存在的构象表位的抗体。用牛桥粒制备的免疫印迹法,某些PV血清和抗dsg单克隆抗体能与Dsg1和Dsg3反应。这些PV抗体的亲和纯化表明,与Dsg1和Dsg3的同时反应性是由两个不同的抗体亚群产生的,而不是由单个抗体的交叉反应性产生的。本研究表明,天疱疮血清是一个有用的探针,我们了解角质细胞生物学的各个方面。
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