C3d在大疱性类天疱疮患者皮损组织中的表达

目的 探讨C3d在石蜡包埋的大疱性类天疱疮患者皮损组织中的表达及临床意义。方法 免疫组化SP法在25例大疱性类天疱疮、10例大疱性表皮松解症及10例正常成人皮肤组织标本中进行C3d、IgG、IgA进行检测,并对其在大疱性类天疱疮皮损中阳性率进行比较。结果 大疱性类天疱疮皮损中C3d、IgG、IgA的阳性率分别为96%、72%、0%。C3d、IgG在BP组织中表达阳性率的差异有统计学意义（χ2 = 4.17,P < 0.05）,C3d、IgA在BP组织中表达阳性率的差异有统计学意义（χ2 = 22.04,P < 0.01）。C3d、IgG、IgA在10例EB及正常成人皮肤组织标本中表达均为阴性。结论 免疫组化方法检测C3d的表达可以协助在石蜡组织中进行大疱性类天疱疮的诊断。     

盐裂皮肤间接免疫荧光及BP180 NC16a酶联免疫吸附测定在大疱性类天疱疮诊断中的意义

目的 探讨盐裂皮肤间接免疫荧光及大疱性类天疱疮（BP）180 NC16a-酶联免疫吸附测定（ELISA）检测在BP诊断中的意义。方法 收集2015年1月至2017年8月在中国医学科学院皮肤病医院用盐裂皮肤间接免疫荧光（IIF?SSS）和BP180 NC16a?ELISA检测BP患者174例和对照组129例血清。其中25例BP患者用直接免疫荧光（DIF）进行检测并与IIF?SSS和BP180 NC16a?ELISA敏感性进行比较。结果 IIF?SSS、BP180 NC16a?ELISA的敏感性分别为93.67%、96.55%;特异性分别为100%、96.12%。IIF?SSS与BP180 NC16a?ELISA相关系数0.147,为弱相关。其中25例BP患者血清学诊断方法（IIF?SSS,BP180 NC16a?ELISA）和DIF敏感性比较差异无统计学意义。结论 BP血清学诊断方法特异性强、敏感性高,值得临床推广应用。     

C4d immunohistochemical stain is a sensitive method to confirm immunoreactant deposition in formalin-fixed paraffin-embedded tissue in bullous pemphigoid

Background:  Bullous pemphigoid (BP) is characterized clinically by the onset of pruritic urticarial plaques, vesicles and bullae in a predominantly elderly population. While the diagnosis may be suspected on routine hematoxylin and eosin histology of formalin-fixed paraffin-embedded tissue, fresh-frozen tissue must be used to show the immunologic nature of the bullous process by direct immunofluorescence (DIF). The diagnosis is further confirmed and separated from epidermolysis bullosa acquisita (EBA) by subsequent serologic studies to detect antibodies directed against BP180 and BP230 antigens and characteristic antibody deposition on salt-split skin.
Methods:  Using a polyclonal complement fragment 4d (C4d) antibody, we stained formalin-fixed paraffin-embedded skin biopsy specimens from cases of BP and controls.
Results:  We showed characteristic linear basement membrane deposition of C4d in formalin-fixed paraffin-embedded tissue in seven of nine cases diagnosed as BP vs. EBA by DIF on fresh-frozen tissue. None of the four controls for which we had adequate tissue were positive.
Conclusion:  These results indicate that formalin-fixed paraffin-embedded tissue can be stained for the immunoreactant C4d to show characteristic immunoreactant deposition, potentially obviating the need for repeat biopsy for DIF and allowing clinicians to proceed to serologic confirmation of BP.     

用石蜡切片行C3d、C4d免疫组化染色辅助诊断大疱性类天疱疮的意义

目的 探讨C3d、C4d免疫组化染色在石蜡包埋组织切片中辅助诊断大疱性类天疱疮的价值.方法 通过免疫组织化学SP法检测20例大疱性类天疱疮患者石蜡包埋组织切片中C3d、C4d的表达,并与家族性良性天疱疮、大疱性表皮松解症患者及正常皮肤进行对照.结果 20例大疱性类天疱疮患者石蜡包埋切片C3d、C4d真表皮交界基底膜处沉积率为95％(19/20),9例大疱性表皮松解症患者中真表皮交界基底膜处C3d、C4d阳性率0％(0/9),4例家族性慢性良性天疱疮患者基底膜带均为阴性.结论 石蜡包埋组织切片中,C3d、C4d免疫组化染色可以作为辅助诊断大疱性类天疱疮的方法之一.     

盐裂皮肤间接免疫荧光技术在大疱性类天疱疮诊断中的价值评估

目的:评价盐裂皮肤间接免疫荧光（IIF-SSS）在大疱性类天疱疮（BP）诊断中的价值。方法:采用单中心临床回顾性研究。纳入2013年1月至2019年1月在中国医学科学院皮肤病医院就诊的初诊BP患者163例,对照组404例,包括天疱疮161例、湿疹67例、药疹26例、多形红斑23例、结节性痒疹18例等。于患者用药前采血,...     

盐裂间接免疫荧光技术在大疱性类天疱疮鉴别诊断中的应用研究

目的:探讨盐裂皮肤间接免疫荧光(IIF)技术在大疱性类天疱疮(BP)鉴别诊断中的作用.方法:应用盐裂IIF技术检测78例常规方法诊断为BP的患者血清.结果:43例血清IgG沉积于表皮侧,7例IgG沉积于双侧,11例IgG沉积于真皮侧,另有17例双侧均未见抗体沉积.结论:盐裂IIF仅能用于BP的初步鉴别诊断.     

Childhood bullous pemphigoid – report of a case with dermal fluorescence on salt-split skin

Summary Bullous pemphigoid (BP) is an acquired bullous disorder which predominantly affects the elderly. It is rare in children, and may be clinically indistinguishable from other immunobullous disorders. As routine histology may be non-specific, a definitive diagnosis of childhood BP usually depends on the results of direct and indirect immunofluorescence investigations.
We report a 5-year-old girl who developed bullous pemphigoid, associated with atypical immunofluorescence findings. Indirect immunofluorescence on split-skin showed a pure dermal pattern of IgG binding. This is usually suggestive of epidennolysis bullosa acquisit, but Western immunoblotting was positive with epidermal extracts, confirming a diagnosis of BP. Dermal binding on split-skin occurs in about 5% of adult cases of BP, and has not been reported previously in childhood BP.     

Decrease in eosinophils infiltrating into the skin of patients with dipeptidyl peptidase‐4 inhibitor‐related bullous pemphigoid

Bullous pemphigoid (BP) is an acquired autoimmune blistering disease in which autoantibodies against epitopes in the basement membrane zone of the skin such as BP180 or BP230 are produced. Dipeptidyl peptidase (DPP)‐4 inhibitors have become commonly used to treat diabetes. As DPP‐4 inhibitors are more commonly prescribed for diabetes, BP related to DPP‐4 inhibitors has been reported and has attracted attention. Therefore, we retrospectively investigated patients who were diagnosed with BP in order to examine characteristics of DPP‐4 inhibitor‐related BP (nine patients; median age, 85 years) in comparison with non‐DPP‐4 inhibitor‐related BP (21; median age, 85 years). There was no significant difference in Bullous Pemphigoid Disease Area Index between DPP‐4 inhibitor‐related BP patients and non‐DPP‐4 inhibitor‐related BP patients, except for erosions/blisters score in mucosa. Laboratory tests revealed no significant differences between DPP‐4 inhibitor‐related BP patients and non‐DPP‐4 inhibitor‐related BP patients in total white blood cell count, eosinophil count, neutrophil count and the titer of anti‐BP180 antibody. The number of eosinophils infiltrating into the skin was significantly lower in patients with DPP4 inhibitor‐related BP than in patients with non‐DPP4 inhibitor‐related BP. Our results showed that DPP‐4 inhibitor‐related BP has some distinct pathological characteristics from BP not associated with DPP‐4 inhibitor.     

Eosinophil localization to the basement membrane zone is autoantibody‐ and complement‐dependent in a human cryosection model of bullous pemphigoid

Bullous pemphigoid (BP) is an autoimmune blistering disease characterized by antibodies (IgG and IgE) targeting cell‐substrate adhesion proteins. A variety of BP models suggest that autoantibody‐dependent neutrophil degranulation is essential for blister formation. However, lesional biopsies reveal a predominance of eosinophils and few neutrophils. Our goal was to evaluate the role of antibodies and complement in eosinophil localization, degranulation and split formation at the dermo‐epidermal junction (DEJ) utilizing a human skin cryosection model of BP paired with a human eosinophilic cell line, 15HL‐60. Expression of receptors for IgG (FcγRII), IgE (FcεRI) and complement (CR1 and CR3) was confirmed on 15HL‐60 cells using flow cytometry. 15HL‐60 expression of granule protein [eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO)] mRNA and their degranulation in vitro was confirmed using RT‐PCR and ELISA, respectively. For cryosection experiments, BP or control sera or IgG and IgE antibodies purified from BP sera were utilized in combination with 15HL‐60 cells ± fresh complement. Both BP serum and fresh complement were required for localization of 15‐HL60 cells to the DEJ. Interestingly, eosinophil localization to the DEJ was dependent on IgG, but not IgE, and complement. However, no subepidermal split was observed. Additionally, the 15HL‐60 cells did not degranulate under any experimental conditions and direct application of cell lysate to cryosections did not result in a split. Our observation that eosinophil localization to the DEJ is dependent on IgG mediated complement fixation provides additional insight into the sequence of events during the development of BP lesions.