Schiltz J R, Michel B
J Invest Dermatol. 1976 Aug;67(2):254-60. doi: 10.1111/1523-1747.ep12513454.
Normal human skin was maintained in organ cultures for several days in Ham's F-10 medium with good preservation of the epidermal cells. When the partially purified IgG fraction from the pooled sera of patients with pemphigus vulgaris or pemphigus foliaceous was added to this culture system, after 24 hr some evidence of epidermal acantholysis was seen. By 72 hr, extensive suprabasilar epidermal acantholysis had occurred in which the acantholytic cells were indistinguishable histologically from the acantholytic cells in biopsies from skin lesions of patients with pemphigus vulgaris. In the control cultures (i.e., F-10 medium or F-10 medium + normal human serum IgG), none of these changes was seen. Direct immunofluorescent staining of these explants using fluorescein-labeled goat antihuman IgG showed that by 6 hr binding of the pemphigus IgG had occurred in the intercellular cement substance of the epidermis. The staining intensity was maximal by 18 to 20 hr. When the pemphigus serum was fractionated by DEAE-cellulose column chromatography, three major IgG-containing peaks (presumably IgG) were eluted which bound to the epidermoid intercellular substance and caused acantholysis in culture. The complement system did not play a role in the antibody-induced acantholysis since complement was not included in this system and heating the reconstituted F-10 + pemphigus IgG for 1 hr at 58 degrees C did not destroy the acantholytic activity. Autoradiographic experiments showed that after about 2 days in culture the rates of incorporation of RNA and protein precursors in the suprabasilar cells in the presence of pemphigus IgG were reduced to less than 10% of the normal IgG controls, whereas these synthetic activities of the basal cells were only slightly affected. These observations lead to the proposal that it is the interaction of the pemphigus autoantibody(s) with the suprabasilar epidermal cell which initiates and possibly substains the process(es) of acantholysis.
正常人类皮肤在含Ham's F - 10培养基的器官培养中维持数天，表皮细胞保存良好。当将寻常型天疱疮或落叶型天疱疮患者混合血清中的部分纯化IgG组分添加到该培养系统中时，24小时后可见一些表皮棘层松解的迹象。到72小时时，发生了广泛的基底上层表皮棘层松解，其中棘层松解细胞在组织学上与寻常型天疱疮患者皮肤病变活检中的棘层松解细胞无法区分。在对照培养物（即F - 10培养基或F - 10培养基 + 正常人血清IgG）中，未观察到这些变化。使用荧光素标记的山羊抗人IgG对这些外植体进行直接免疫荧光染色显示，到6小时时，天疱疮IgG已在表皮的细胞间黏合物质中发生结合。染色强度在18至20小时时达到最大。当天疱疮血清通过DEAE - 纤维素柱色谱法分级分离时，洗脱了三个主要的含IgG峰（可能为IgG），它们与表皮样细胞间物质结合并在培养中引起棘层松解。补体系统在抗体诱导的棘层松解中不起作用，因为该系统中未包含补体，并且将重构的F - 10 + 天疱疮IgG在58℃加热1小时不会破坏棘层松解活性。放射自显影实验表明，培养约2天后，在存在天疱疮IgG的情况下，基底上层细胞中RNA和蛋白质前体的掺入率降至正常IgG对照的不到10%，而基底细胞的这些合成活性仅受到轻微影响。这些观察结果提示，天疱疮自身抗体与基底上层表皮细胞的相互作用启动并可能维持了棘层松解过程。
