Distinction between epidermal antigens binding pemphigus vulgaris and pemphigus foliaceus autoantibodies.
Distinction between epidermal antigens binding pemphigus vulgaris and pemphigus foliaceus autoantibodies.
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表皮抗原结合寻常型天疱疮和落叶型天疱疮自身抗体的区别。
DOI：
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发表时间：
1984
影响因子：
15.9
通讯作者：
C. Thivolet
中科院分区：
文献类型：
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作者：
J. Stanley;Leena iCoulu;C. Thivolet
寻常型天疱疮（PV）和落叶型天疱疮（PF）是自身免疫性起泡性疾病，其中抗体发展到表皮细胞的细胞表面。在这项研究中，我们试图确定抗体的抗原特异性与PV和PF患者的血清。血清从12例PV被用来免疫沉淀提取物的培养的人表皮细胞，放射性标记的14 C-氨基酸。免疫沉淀物通过SDS聚丙烯酰胺凝胶电泳（PAGE）和荧光法鉴定。所有12个PV血清沉淀蛋白质，当还原时，在SDS-PAGE上显示130，000和80，000 mol wt的链。非还原条件下的电泳鉴定出210，000-mol wt分子，推测其是由130，000和80，000-mol wt链的二硫键交联形成的。用14 C-葡糖胺标记的表皮细胞提取物的免疫沉淀物表明，130，000-mol wt链。在该免疫沉淀试验中，与PV血清同时运行的8份PF血清中有7份没有沉淀这种糖蛋白，也没有特异性沉淀任何蛋白质。为了确定是否可以识别与PF血清中的抗体反应的特定分子，我们使用正常人表皮提取物的免疫印迹分析。将这些提取物中的蛋白质还原，通过SDS-PAGE分离，并电泳转移到硝酸纤维素纸或2-氨基苯硫醚纸上。用PF血清对转移的蛋白质进行免疫过氧化物酶染色表明，8份PF血清中有4份含有染色160，000 mol wt蛋白质条带的抗体。使用正常人皮肤作为底物的间接免疫荧光表明，从该蛋白条带洗脱的IgG以细胞表面图案染色表皮。PV血清通过免疫印迹分析不特异性识别任何条带。用从细胞培养物提取物中免疫沉淀的PV抗原进行的免疫印迹表明，一旦变性用于SDS-PAGE，PV抗原不再具有免疫反应性。综上所述，这些数据表明：来自不同患者的PV血清中所含的自身抗体具有独特的分子特异性;来自大多数PF血清的自身抗体具有不同于PV自身抗体的特异性;来自不同PF患者的自身抗体可能不具有相同的抗原特异性。PV和PF血清之间抗原特异性的这些差异可能解释了这些疾病之间的临床和组织学差异。
Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune blistering diseases in which antibodies develop to the cell surface of epidermal cells. In this study we sought to determine the antigenic specificity of antibodies in the sera of patients with PV and PF. Sera from 12 patients with PV were used to immunoprecipitate extracts of cultured human epidermal cells that were radiolabeled with 14C-amino acids. Immunoprecipitates were identified by SDS polyacrylamide gel electrophoresis (PAGE) and fluorography. All 12 PV sera precipitated a protein which, when reduced, displayed chains of 130,000 and 80,000 mol wt on SDS-PAGE. Electrophoresis under nonreducing conditions identified a 210,000-mol wt molecule, which was presumably formed by disulfide crosslinking of the 130,000 and 80,000-mol wt chains. Immunoprecipitates of epidermal cell extracts that were labeled with 14C-glucosamine indicated that the 130,000-mol wt chain. Seven of eight PF sera, which were run concurrently with the PV sera in this immunoprecipitation assay, did not precipitate this glycoprotein, nor did they specifically precipitate any protein. To determine if a specific molecule which reacted with antibodies in PF sera could be identified, we used immunoblot analysis of extracts of normal human epidermis. The proteins in these extracts were reduced, separated by SDS-PAGE, and electrophoretically transferred to nitrocellulose sheets or to 2-aminophenylthioether paper. Immunoperoxidase staining of the transferred proteins with PF sera indicated that four of eight PF sera contained antibodies that stained a protein band of 160,000 mol wt. Indirect immunofluorescence, using normal human skin as the substrate, indicated that IgG that was eluted from this protein band stained the epidermis in a cell surface pattern. PV sera did not specifically recognize any bands by immunoblot analysis. Immunoblots performed with PV antigen that was immunoprecipitated from cell culture extracts suggested that, once denatured for SDS-PAGE, PV antigen is no longer immunoreactive. Taken together, these data indicate that: autoantibodies contained in PV sera from various patients have a unique molecular specificity; autoantibodies from most PF sera have a specificity different from that of PV autoantibodies; and autoantibodies from various PF patients may not have identical antigenic specificities. These differences in antigenic specificity between PV and PF sera may account for the clinical and histologic differences between these diseases.
影响因子：
158.5
作者：
ANHALT, GJ;LABIB, RS;DIAZ, LA
通讯作者：
DIAZ, LA