Department of Medicine B and Zabludowicz Center for Autoimmune Diseases, Sheba Medical Center, Tel Hashomer, Israel.
Clin Exp Immunol. 2010 Dec;162(3):543-9. doi: 10.1111/j.1365-2249.2010.04265.x. Epub 2010 Oct 21.
Pemphigus vulgaris is a rare life-threatening autoimmune bullous disease caused by immunoglobulin G (IgG) autoantibodies directed against desmogleins 1 and 3. Previously, we showed that intravenous immunoglobulin (IVIG) ameliorates anti-desmoglein-induced experimental pemphigus vulgaris in newborn naive mice. The aim of this study was to examine the efficacy of anti-anti-desmoglein-specific IVIG in a similar model. Pemphigus-vulgaris-specific IVIG (PV-sIVIG) was affinity-purified from IVIG on a column of single-chain variable fragment (scFv) anti-desmogleins 1 and 3. The anti-idiotypic activity of PV-sIVIG was confirmed by enzyme-linked immunosorbent assay, inhibition assay. After induction of pemphigus by injection of anti-desmogleins 1 and 3 scFv to newborn mice, the animals were treated with PV-sIVIG, IVIG (low or high dose) or IgG from a healthy donor (n = 10 each). The skin was examined 24-48 h later, and samples of affected areas were analysed by histology and immunofluorescence. In vitro study showed that PV-sIVIG significantly inhibited anti-desmogleins 1 and 3 scFv binding to recombinant desmoglein-3 in a dose-dependent manner. Specificity was confirmed by inhibition assay. In vivo analysis revealed cutaneous lesions of pemphigus vulgaris in mice injected with normal IgG (nine of 10 mice) or low-dose IVIG (nine of 10 mice), but not in mice treated with PV-sIVIG (none of 10) or high-dose IVIG (none of 10). On immunopathological study, PV-sIVIG and regular IVIG prevented the formation of acantholysis and deposition of IgG in intercellular spaces. In conclusion, the PV-sIVIG preparation is more effective than native IVIG in inhibiting anti-desmoglein-induced pemphigus vulgaris in mice and might serve as a future therapy in patients with the clinical disease.
寻常型天疱疮是一种罕见的危及生命的自身免疫性大疱性疾病,由针对桥粒芯糖蛋白 1 和 3 的免疫球蛋白 G (IgG) 自身抗体引起。此前,我们表明静脉注射免疫球蛋白 (IVIG) 可改善新生无经验小鼠的抗桥粒芯糖蛋白 1 和 3 诱导的实验性寻常型天疱疮。本研究旨在检查类似模型中抗抗桥粒芯糖蛋白 1 和 3 特异性 IVIG 的疗效。通过将抗桥粒芯糖蛋白 1 和 3 的单链可变片段 (scFv) 柱亲和纯化,从 IVIG 中获得寻常型天疱疮特异性 IVIG (PV-sIVIG)。通过酶联免疫吸附试验、抑制试验证实了 PV-sIVIG 的抗独特型活性。通过向新生小鼠注射抗桥粒芯糖蛋白 1 和 3 scFv 诱导寻常型天疱疮后,将动物用 PV-sIVIG、IVIG (低或高剂量) 或来自健康供体的 IgG 治疗 (每组 10 只)。24-48 小时后检查皮肤,并通过组织学和免疫荧光分析受影响区域的样本。体外研究表明,PV-sIVIG 以剂量依赖性方式显著抑制抗桥粒芯糖蛋白 1 和 3 scFv 与重组桥粒芯糖蛋白-3 的结合。通过抑制试验证实了特异性。体内分析显示,注射正常 IgG(10 只小鼠中有 9 只)或低剂量 IVIG(10 只小鼠中有 9 只)的小鼠出现寻常型天疱疮皮肤病变,但用 PV-sIVIG(10 只小鼠中均无)或高剂量 IVIG(10 只小鼠中均无)治疗的小鼠则没有。在免疫病理学研究中,PV-sIVIG 和常规 IVIG 可防止棘层松解和 IgG 在细胞间隙中的沉积。总之,与天然 IVIG 相比,PV-sIVIG 制剂在抑制抗桥粒芯糖蛋白 1 和 3 诱导的小鼠寻常型天疱疮方面更有效,可能成为临床疾病患者的未来治疗方法。