Pemphigus vulgaris IgG causes a rapid depletion of desmoglein 3 (Dsg3) from the Triton X-100 soluble pools, leading to the formation of Dsg3-depleted desmosomes in a human squamous carcinoma cell line, DJM-1 cells
Pemphigus vulgaris IgG causes a rapid depletion of desmoglein 3 (Dsg3) from the Triton X-100 soluble pools, leading to the formation of Dsg3-depleted desmosomes in a human squamous carcinoma cell line, DJM-1 cells
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DOI：
10.1046/j.1523-1747.1999.00463.x
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发表时间：
1999-01-01
影响因子：
6.5
通讯作者：
Kitajima, Y
中科院分区：
文献类型：
--
作者：
Aoyama, Y;Kitajima, Y
在这项研究中，我们检查桥粒芯糖蛋白（Dsg）3和其他桥粒分子后，寻常天疱疮（PV）-免疫球蛋白G（IgG）结合DJM-1细胞，人鳞状细胞癌细胞系细胞表面上的Dsg 3。在细胞与PV-IgG孵育不同的时间段后，（0、5、10、20、30、60分钟或30小时），将细胞分级成磷酸盐缓冲盐水可溶性（胞质溶胶）、磷酸盐缓冲盐水不溶性-Triton X-100可溶性（膜）和Triton X-100不溶性（细胞骨架）组分，并使用针对Dsg 1、Dsg 3、斑珠蛋白、桥粒斑蛋白1和细胞角蛋白的抗体进行免疫印迹和免疫荧光显微术。用PV-IgG的免疫印迹分析显示，在PV-IgG处理后20分钟，Dsg 3已经从膜级分中急剧耗尽，而通过免疫印迹检测，在细胞骨架级分中没有检测到Dsg 3的减少。然而，与PV-IgG孵育30小时，引起Dsg 3的显着消失，但不是其他桥粒分子，从细胞骨架组分。此外，双染色免疫荧光显微镜显示，Dsg 3从桥粒耗尽，而Dsg 1，桥斑蛋白1，斑珠蛋白，角蛋白丝被绑定到桥粒。这些结果为更好地理解PV中起泡的机制提供了新的解释;即，PV-IgG产生异常桥粒形成的可能性，所述异常桥粒在Dsg 3中是缺乏的，而不是其他桥粒成分。
In this study, we examined desmoglein (Dsg) 3 and other desmosomal molecules after pemphigus vulgaris (PV)-immunoglobulin G (IgG) binding to the Dsg3 on the cell surface in DJM-1 cells, a human squamous cell carcinoma cell line. After cells were incubated with PV-IgG for various time periods (0, 5, 10, 20, 30, 60 min, or 30 h), cells were fractionated into phosphate-buffered saline soluble (cytosol), phosphate-buffered saline insoluble-Triton X-100 soluble (membrane), and Triton X-100 insoluble (cytoskeleton) fractions, and subjected to immunoblotting and immunofluorescence microscopy using antibodies against Dsg1, Dsg3, plakoglobin, desmoplakin 1, and cytokeratins. Immunoblot analysis with PV-IgG revealed that Dsg3 was already dramatically depleted from the membrane fraction 20 min after PV-IgG treatment, whereas no reduction of Dsg3 was detected in the cytoskeleton fraction as examined by immunoblotting. A 30 h incubation with PV-IgG, however, caused a marked disappearance of Dsg3, but not other desmosomal molecules, from cytoskeleton fractions. Furthermore, double-staining immunofluorescence microscopy revealed that Dsg3 was depleted from the desmosomes whereas Dsg1, desmoplakin 1, plakoglobin, and keratin filaments were bound to desmosomes. These results provide a novel interpretation for a better understanding of mechanisms for blistering in PV; i.e., a possibility that PV-IgG generates the formation of aberrant desmosomes, which are lacking in Dsg3, but not other desmosomal constituents.