Memar O M, Rajaraman S, Thotakura R, Tyring S K, Fan J L, Seetharamaiah G S, Lopez A, Jordon R E, Prabhakar B S
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, 77555-1019, USA.
J Invest Dermatol. 1996 Feb;106(2):261-8. doi: 10.1111/1523-1747.ep12340663.
The development of an animal model for studying the pathogenesis of pemphigus vulgaris (PV) has been hampered by the unavailability of the purified full-length autoantigen desmoglein 3 (Dsg 3).Therefore, we expressed Dsg 3 using a baculovirus expressed system. The expressed protein was identified as Dgs 3 by its reactivity with a pan-cadherin anti-serum, an anti-serum to a Dsg 3 synthetic peptide, or patient serum, and by amino-terminal sequencing. Carbohydrate analysis showed that recombinant Dsg 3 was glycosylated. While a majority of the recombinant protein was cell associated, by immunoprecipitation, some Dsg 3 was demonstrated in the medium. The Dgs 3 could adsorb out blister-causing antibodies from patient sera. Rabbit anti- Dsg 3 antibodies induced by the recombinant Dsg 3 showed specific binding to intercellular spaces of monkeys esophagus by indirect immunofluorescence. Moreover, these antibodies induced PV-like blisters in neonatal mice and weakly bound perilesional epidermis. Availability of large quantities of relatively pure Dsg 3 should now facilitate studies aimed at understanding Dsg 3 structure and pathogenesis of PV, with implications for developing specific immunotherapies.
由于无法获得纯化的全长自身抗原桥粒芯糖蛋白3(Dsg 3),寻常型天疱疮(PV)发病机制研究的动物模型开发受到了阻碍。因此,我们使用杆状病毒表达系统表达Dsg 3。通过其与泛钙黏蛋白抗血清、Dsg 3合成肽抗血清或患者血清的反应性以及氨基末端测序,将表达的蛋白鉴定为Dgs 3。碳水化合物分析表明重组Dsg 3被糖基化。虽然大多数重组蛋白与细胞相关,但通过免疫沉淀在培养基中发现了一些Dsg 3。Dgs 3可以从患者血清中吸附出导致水疱的抗体。重组Dsg 3诱导的兔抗Dsg 3抗体通过间接免疫荧光显示与猴食管的细胞间空间特异性结合。此外,这些抗体在新生小鼠中诱导出PV样水疱,并与病损周围表皮弱结合。大量相对纯的Dsg 3的可获得性现在应该有助于旨在了解Dsg 3结构和PV发病机制的研究,这对开发特异性免疫疗法具有重要意义。