Differences in the anti-basement membrane zone antibodies in ocular and pseudo-ocular cicatricial pemphigoid
Differences in the anti-basement membrane zone antibodies in ocular and pseudo-ocular cicatricial pemphigoid
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DOI：
10.3109/02713689609000763
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发表时间：
1996-05-01
影响因子：
2
通讯作者：
Ahmed, AR
中科院分区：
文献类型：
--
作者：
Bhol, K;Mohimen, A;Ahmed, AR
目的。眼部瘢痕性类天疱疮(OCP)是一种慢性自身免疫性瘢痕性疾病，影响结膜和其他鳞状上皮，导致瘢痕形成过程。在一些使用眼药水治疗青光眼的患者中观察到的一个类似的过程，仅限于结膜，被称为假眼瘢痕疙瘩(P-OCP)。免疫荧光研究表明，免疫球蛋白和补体成分沉积在结膜的基底膜区(BMZ)，患者血清中有抗基底膜区抗体。OCP与MHC-II类基因DQB1*0301显著相关。本研究的目的是在免疫印迹试验中确定OCP和P-OCP血清与不同裂解物结合的一些差异，以试图部分表征OCP和P-OCP抗原。此外，我们想要确定P-OCP的MHC-II类基因关联是否与OCP.Methods的相似。我们研究了11例活动期眼部瘢痕性类天疱疮患者和7例假性眼部瘢痕性类天疱疮患者和对照组的血清。以猴食道和盐裂正常人皮肤为底物，进行间接免疫荧光(IIF)研究。以正常人表皮、真皮和结膜为底物，建立了一种灵敏的免疫印迹试验(IBA)。应用斑点杂交技术对8例假眼瘢痕性类天疱疮患者进行了MHC-II类基因分型，并与38例匹配对照进行了比较。在用猴食道进行的IIF试验中，10份眼瘢痕性类天疱疮血清中有9份和7份假性瘢痕性类天疱疮血清中有5份基底膜带染色较弱。以盐裂的人皮肤为底物，11份眼部瘢痕性类天疱疮血清中有10份与裂解的表皮侧结合滴度低，结合力弱。假眼瘢痕性类天疱疮血清未见一致的染色模式。在11份眼瘢痕性类天疱疮血清中，有10份与IBA中的230、205、160和85 kDa蛋白结合。当裂解产物首先与BP血清反应，然后与眼瘢痕性类天疱疮血清免疫印迹时，230、160和86 kDa条带消失，只有205 kDa条带保留。7例假眼瘢痕性类天疱疮患者中有5例的血清与表皮和结膜中的290、230、205、180、97和85 kDa蛋白结合。然而，当裂解产物第一次与BP和眼部瘢痕性类天疱疮血清反应时，230、205、180和85 kDa的蛋白质被耗尽。在真皮裂解物中，假眼瘢痕性类天疱疮血清识别400、290、150和45 kDa的蛋白质。这些药物都不能被BP、眼部瘢痕性类天疱疮、寻常型天疱疮或获得性大疱性表皮松解症血清吸收。在真皮和表皮中鉴定出的290 kDa蛋白质是不同的。3种裂解物与对照血清均无结合。经统计学处理，斑点杂交分析未发现MHC DQB1*0301基因频率显著增加。眼瘢痕性类天疱疮和假性瘢痕性类天疱疮患者会产生多种自身抗体。然而，它们之间既有相似之处，也有不同之处。假性眼部瘢痕性类天疱疮相关的MHC-II类基因不同于眼部瘢痕性类天疱疮。这为研究特发性和药物相关器官特异性自身免疫中的免疫异常提供了新的模型系统。
Purpose. Ocular cicatricial pemphigoid (OCP) is a chronic autoimmune cicatrizing disease which affects the conjunctiva and other squamous epithelium, resulting in a scarring process. A similar process, limited only to the conjunctiva, observed in some patients using eye drops for the treatment of glaucoma, is called pseudo-ocular cicatricial pemphigoid (P-OCP). Immunofluorescence studies demonstrate deposition of immunoglobulins and complement components in the basement membrane zone (BMZ) of the conjunctiva and an anti-basement membrane zone antibody in the serum of patients. A striking association between OCP and MHC class II gene DQB1*0301 has been observed. The purpose of this study was to determine some of the differences in the binding of OCP and P-OCP sera to different lysate in an immunoblot assay, in an attempt to partially characterize the OCP and P-OCP antigens. Furthermore, we wanted to determine if the MHC class II gene association of P-OCP is similar to that of OCP.Methods. We studied sera from 11 patients with active ocular cicatricial pemphigoid and seven patients with pseudo-ocular cicatricial pemphigoid and controls. Indirect immunofluorescence (IIF) studies were done using monkey esophagus and salt split normal human skin as substrate. A sensitive immunoblot assay (IBA) was developed using normal human epidermis, dermis and conjunctiva as substrate. Typing for MHC class II genes was performed on eight pseudo-ocular cicatricial pemphigoid patients by dot-blot analysis and compared to 38 matched controls.Results. Weak staining of the basement membrane zone was observed in nine of ten ocular cicatricial pemphigoid sera and five of seven pseudo-ocular cicatricial pemphigoid sera in the IIF assay using monkey esophagus. Using salt split human skin as substrate, ten of eleven ocular cicatricial pemphigoid sera demonstrated low titer weak binding to the epidermal side of the split. No consistent pattern of staining was seen with pseudo-ocular cicatricial pemphigoid sera. Ten of the 11 ocular cicatricial pemphigoid sera demonstrated binding to 230, 205, 160 and 85 kDa proteins in the IBA using normal human epidermis and conjunctiva lysates. When the lysates were first reacted with BP sera and then immunoblotted with ocular cicatricial pemphigoid sera, the 230, 160, and 86 kDa bands disappeared, and only the 205 kDa band persisted. The sera of five of seven pseudo-ocular cicatricial pemphigoid patients bound to 290, 230, 205, 180, 97, and 85 kDa proteins in the epidermis and conjunctiva. However, the 230, 205, 180, and 85 kDa proteins are depleted when the lysates are first reacted with BP and ocular cicatricial pemphigoid sera. In the dermal lysate, the pseudo-ocular cicatricial pemphigoid sera recognize 400, 290, 150 and 45 kDa proteins. None of these are absorbed by BP, ocular cicatricial pemphigoid or pemphigus vulgaris or epidermolysis bullosa acquisita sera. The 290 kDa proteins identified in the dermis and epidermis are distinct from each other. No binding was seen with control sera with the 3 lysates. Statistically, dot-blot analysis did not demonstrate a significant increase in the frequency of the MHC DQB1*0301 gene.Conclusions. Patients with ocular cicatricial pemphigoid and pseudo-ocular cicatricial pemphigoid produce several autoantibodies. However, there are similarities and differences between them. The MHC class II genes associated with pseudo-ocular cicatricial pemphigoid are different from those with ocular cicatricial pemphigoid. This provides a new model system to study the immune abnormalities in idiopathic and drug-related organ specific autoimmunity.