盐裂皮肤直接和间接免疫荧光联合应用鉴别诊断表上大疱病的…

本文探讨盐裂皮肤ＤＩＦ和盐裂皮肤ＩＩＦ联合应用鉴别诊断表皮下大疱病的价值。２０例两种盐裂皮肤免疫荧光均阳性的病例比较表明，盐裂皮肤ＩＩＦ示抗基底膜带体结合表皮侧或真皮侧与盐裂皮肤ＤＩＦ示免疫反应物（ＩｇＧ、ＩｇＡ、Ｃ３等）沉积于表皮侧或真皮侧的部位基本一致。提示多数情况下盐裂皮肤ＤＩＦ或ＩＩＦ均可韵用于表皮下大将民病诊断，两者对比可以提高诊断率。     

儿童获得性大疱表皮松解症(附4例报道)

儿童获得性大疱表皮松解症（ＥＢＡ）是一种自身免疫性表皮下大疱病,比较少见。本文报道４例儿童ＥＢＡ,年龄为１、４、７、１４岁,均为男性。临床上似营养不良性大疱表皮松解症２例,儿童型线形ＩｇＡ大疱病２例。３例有口腔损害,伴血疱,萎缩性瘢痕和栗丘疹。病理为表皮下水疱。ＤＩＦ均示ＩｇＧ和Ｃ３呈线状分布于基底膜带,３例还见ＩｇＡ,１例见ＩｇＭ沉积于基底膜带。盐裂ＤＩＦ示３例ＩｇＧ沉积于真皮侧。ＩＩＦ示３例Ｉ     

盐裂皮损围围皮肤直接免疫荧光检查诊断表皮下水疱病

本文报告了应用盐裂皮损周围皮肤直接免疫荧光检查（ＤＩＦ）诊断２２例表皮下水疱病的结果，所有２２例表皮下水疱病盐裂皮损周围皮肤ＤＩＦ均显示ＩｇＧ免疫复合物沉积在表皮侧、真皮侧或表皮真皮两侧，阳性率为１００％。其中表皮侧１７例、表皮真皮两侧均有２例，此１９例均为大疱性类天疱疮（Ｂｐ）；真皮侧３例，２例为获得性大疱性在皮松解症（ＥＢＡ）、１例为大疱性系统性红斑狼疮（ＢＳＬＥ）。３例天疱疮和４例正常人皮肤均阴性。此法简便，易行，准确可靠，重复性好，值得推广使用。     

盐裂皮损围围皮肤直接免疫荧光检查诊断表皮下水泡病

本文报告了应用盐裂皮损周围皮肤直接免疫荧光检查（ＤＩＦ），诊断２２例表皮下水疱病的结果，所有２２例表皮下水泡病盐裂皮损周围皮肤ＤＩＦ均显示ＩｇＧ免疫复事物沉淀在表皮侧，真皮侧或表皮真皮两侧，阳性率为１００％，其中表皮侧１７例，表皮真皮两侧均有２例，此１９例的均为大泡性类天疱疮（Ｂｐ），真皮侧３例，２例为获得性大疱性表皮性松解症（ＥＢＡ），１例大疱性系统性红斑狼疮（ＢＳＬＥ），３例天疱疹和４例正常人     

1M NaCl分离皮肤的免疫荧光法在表皮下大疱疹诊断中的应用

该文介绍了１Ｍ ＮａＣｌ分离皮肤免疫荧光法诊断表皮下大疱病的原理和优越性。同时介绍了盐裂正常人皮肤ＩＩＦ法以及盐裂皮损周围皮肤的ＤＩＦ法对表皮下大疱病诊断与鉴别诊断的价值。     

表皮下大疱病的鉴别诊断和抗原表达区域性差别的研究

通过间接免疫荧光和盐裂皮损周围皮肤直接免疫荧光（简称盐裂ＤＩＦ），分别研究正常人皮肤、类天疱疮（ＢＰ）及获得性大疱性表皮松解症（ＥＢＡ）抗原表达的区域性差别和表皮下大疱病鉴别诊断。 窝、肘窝、上背、下背、股内侧和下腹部皮肤ＢＰ抗原表达率较高；膝、阳窝、足背、肘、肘窝和下腹部皮肤ＥＢＡ抗原表达率较高。皮肤ＤＩＦ显示２５例表皮下大疱病中１６例（６４％）基底膜带有Ｃ３或ＩｇＧ或伴Ｃ３和ＩｇＡ沉积；盐裂ＤＩＦ表明２５例（１００％）均有ＩｇＧ或伴Ｃ３和ＩｇＡ沉积在表皮侧或真皮侧。结果提示，ＢＰ抗原高表达率与皮损好发部位相一致；ＥＢＡ抗原高表达率一部分与皮损好发部位一致。盐裂ＤＩＦ不仅提高ＤＩＦ阳性率，而且根据免疫反应物沉积部位可以鉴别出ＢＰ与ＥＢＡ以及大疱性系统性红斑狼疮。     

盐裂皮肤间接免疫荧光检测抗基底膜带抗体的意义

以１ｍｏｌ／Ｌ盐裂正常人皮肤为底物，间接免疫荧光（ⅡＦ）检测３１例表皮下大疱病患者血清，２６例（８３．９％）检出抗基底膜带（ＢＭＺ）抗体，而常规ⅡＦ仅１１例（３５．５％）检出此种抗体。结果提示，盐裂皮肤ⅡＦ较常规ⅡＦ敏感，并可根据免疫球蛋白结合于盐裂皮肤表皮侧或真皮侧，诊断和鉴别诊断表皮下大疱病。     

疱液在诊断表皮下自身免疫性大疱病中的意义

目的：探讨表皮下自身免疫性大疱病疱液中抗基底膜带（ＢＭＺ）自身抗体的情况及在诊断中的意义。方法：应用非盐裂皮肤及盐裂皮肤间接免疫荧光技术（ＩＦ）检测３８例表皮下自身免疫性大疱病患者疱液和血清中ＩｇＧ、ＩｇＡ、ＩｇＭ、ＩｇＥ及ＩｇＧ亚型ＩｇＧ１～ＩｇＧ４抗ＢＭＺ抗体及滴度，并检测疱液和血清中结合补体Ｃ３的特异性抗ＢＭＺ抗体。结果：（１）疱液和血清中抗ＢＭＺ抗体及滴度和结合补体Ｃ３的特异性抗ＢＭＺ抗体均无显著性差异。（２）大疱性类天疱疮患者的疱液和血清中ＩｇＧ抗ＢＭＺ抗体的亚型分布相同，主要为ＩｇＧ１和ＩｇＧ４。结论：疱液可作为ＩＩＦ检测表皮下大疱病抗ＢＭＺ抗体的另一个有价值的标本来源。     

自身免疫性表皮下大疱病基底膜带自身抗体的检测

目的 比较免疫印迹（ＩＢ）和盐裂皮肤间接免疫荧光（ＩＩＦ）检测自身免疫性表皮下大疱病（ＳＡＢＤ）基底膜带自身抗体（ＢＭＺ－Ａｂ）的敏感性和特异怀。方法 分另应用ＩＩＦ和ＩＢ技术对９７例ＳＡＢＤ患者血清中ＩｇＧ型或ＩｇＡ型ＢＭＺ－Ｚｂ进行检测。结果 ＩＩＦ法阳性率７５．３％,免疫印迹法阳性率７９．４％,两者在检测灵敏度上无显著性差异。结论 二者均可用于ＳＡＢＤ中ＢＭＺ－Ａｂ的检测,二者联用对于提高Ｂ     

1M NaCl分离皮肤的免疫荧光法在表皮下大疱病诊断中的应用

该文介绍1M NaCl分离皮肤免疫荧光法诊断表皮下大疱病的原理和优越性。同时介绍了盐裂正常人皮肤IIF法以及盐裂皮损周围皮肤的DIF法对表皮下大疱病诊断与鉴别诊断的价值。     

病情与外周血嗜酸粒细胞计数平行的大疱性类天疱疮一例

报告1例病情与外周血嗜酸粒细胞计数相平行的大疱性类天疱疮.患者男,65岁.全身泛发红斑、水疱1个月,加重15天.皮肤科检查:皮损泛发头面颈、躯干、四肢及会阴部,基本损害为正常皮肤或红斑基础上出现大小不等的水疱,疱液为草黄色,散在红色糜烂面,尼氏征阴性,部分皮损表面敷以灰褐色痂片,以胸背部、四肢尤为严重.组织病理、直接免疫荧光、间接免疫荧光及盐裂皮肤直接免疫荧光检查结果符合大疱性类天疱疮.入院后依据病情变化,先后给予3次糖皮质激素冲击治疗,治疗过程中,定期监测血常规,提示外周血嗜酸粒细胞计数随病情波动.     

Anatomical distribution and immunological characteristics of epidermolysis bullosa acquisita antigen and bullous pemphigoid antigen

A Japanese patient with epidermolysis bullosa acquisita (EBA) was autopsied, and direct immunofluorescence (DIF) testing was performed. Using this patient's serum (EBA serum) and three bullous pemphigoid (BP) sera, the anatomical distribution and immunological characteristics of EBA antigen and BP antigen were investigated by indirect immunofluorescence (IIF). EBA antigen showed the same anatomical distribution as BP antigen in DIF and IIF studies; both antigens were limited to the skin, tongue, oesophagus, trachea, cornea and bladder. EBA antigen was located on the dermal side of both NaCl and PBS-separated skin, whereas BP antigen was limited to the epidermal side. Ethanol fixation abrogated the antigenic stability of BP antigen, but not that of EBA antigen. No difference was found when acetone or formalin fixation was used. The separation methods and prefixation in ethanol could be useful techniques applicable to the classification of the bullous disorders which manifest circulating anti-BMZ antibodies.     

DIFFERENTIATION OF BULLOUS PEMPHIGOID FROM EPIDERMOLYSIS BULLOSA ACQUISITA ON FROZEN SKIN BIOPSIES

Patients with bullous pemphigoid and epidermolysis bullosa acquisita may have similar clinical, histologic, and routine immunohistologic features. These two diseases can be distinguished by routine diagnostic studies either on a patient's serum tested by indirect immunofluorescence on salt-split normal skin or by obtaining a fresh perilesional skin biopsy, inducing a split at the lamina lucida, and testing for the site of IgG deposition by direct immunofluorescence. Often the serum studies are negative, while direct immunofluorescent studies yield the characteristic linear IgG staining of the basement membrane zone. To eliminate the need for a repeat biopsy to make a laboratory differential diagnosis, we studied the efficacy of salt-splitting perilesional skin biopsies that had been previously submitted and frozen for routine direct immunofluorescent studies. The biopsies were thawed, salt-split, and processed for direct immunofluorescence. Three epidermolysis bullosa acquisita biopsies and seven bullous pemphigoid biopsies examined demonstrated IgG staining at sites consistent with their respective diagnoses. The IgG appeared in the dermal side of the split biopsies in epidermolysis bullosa acquisita and predominantly, or exclusively, in the epidermal side in bullous pemphigoid. Thus the direct immunofluorescent study of previously frozen and subsequently salt-split skin biopsies may be used for the differential diagnosis of bullous pemphigoid from epidermolysis bullosa acquisita. In most cases, it may eliminate the need for a repeat biopsy.     

Bullous pemphigoid in Liguria: A 2-year survey

BACKGROUND: The epidemiology of bullous pemphigoid (BP) is not clear because of the heterogeneity of the disease, and its possible association with internal malignancies has been under debate for many years. We report the findings of a 2-year study on incident BP cases in the Liguria region of Italy. SUBJECTS AND METHODS: Thirty-two patients with BP were collected over the 2-year period. Diagnosis was made based on clinical findings and confirmed by histology, direct immunofluorescence (DIF) and indirect immunofluorescence (IIF) with salt-split skin and monkey oesophagus, and immunoblotting (IB). All patients were thoroughly investigated for possible malignancies and all were followed up for 6 months to monitor the response to treatment. RESULTS: DIF showed linear deposits at the dermoepidermal junction in all but one patient. IIF gave positive findings for 15 sera tested with monkey oesophagus and 20 tested with salt-split skin. IB gave positive findings in 19 cases. There was a malignancy in six cases, but no clinical or immunological features that could be considered to predict this occurrence. CONCLUSIONS: The findings of this study are in accordance with most of the data found in the literature, including the fact that IgG serum levels did not predict the course of the disease. Contrary to previous indications, IgE levels were not indicative of disease course either. Mucosal lesions, erythema multiform-like lesions, negative IIF findings and antibodies to AgPB2 were not a prediction for the development of malignancy.     

Frequency of IgA antibodies in pemphigus, bullous pemphigoid and mucous membrane pemphigoid

Circulating and bound IgA antibodies can be found in the autoimmune blistering diseases, but their prevalence, clinical relevance and target antigens remain unknown. Thirty-two patients with pemphigus, 73 with bullous pemphigoid and 28 with mucous membrane pemphigoid were studied retrospectively. Direct immunofluorescence (DIF) analysis of IgG, IgA, IgM and C3 was carried out for all cases. Sera were studied by standard indirect immunofluorescence, indirect immunofluorescence on salt-split skin, immunoblotting for bullous pemphigoid and mucous membrane pemphigoid and ELISA for pemphigus. With DIF, we found IgA autoantibodies in 22 of all 133 cases. Circulating IgA antibodies to skin were detected in 2 of 3 IgA-DIF-positive patients with pemphigus, in 3 of 6 with bullous pemphigoid, and in 6 of 13 with mucous membrane pemphigoid. We confirm that the IgA reactivity is more frequently associated with mucous membrane involvement, especially in cases without critical involvement (5/8). The role of IgA and its antigenic specificity in these diseases remain unclear.     

Pemphigoid nodularis: a case report

Several variants of bullous pemphigoid have been reported including pemphigoid nodularis. Patients with pemphigoid nodularis have clinical features of prurigo nodularis in combination with clinical or immunologic characteristics of bullous pemphigoid. We report the case of a 71-year-old woman with pemphigoid nodularis. The diagnosis was suspected clinically and established by positive indirect immunofluorescence (IIF) findings characteristic of pemphigoid. Results of direct immunofluorescence (DIF) testing were negative, which emphasizes the importance of conducting both DIF and IIF when pemphigoid nodularis is suspected.     

Rat bladder epithelium: a sensitive substrate for indirect immunofluorescence of bullous pemphigoid

Serological diagnosis of bullous pemphigoid is based on immunoblotting or indirect immunofluorescence on normal human salt-split skin. These methods are expensive or time-consuming and not available as a routine test in all laboratories. We used rat bladder epithelium as substrate for indirect immunofluorescence and compared it with other substrates and with immunoblotting. Twenty-nine bullous pemphigoid sera were studied on rat bladder epithelium, monkey oesophagus, salt-split skin and with immunoblotting on human keratinocyte cultures. Indirect immunofluorescence on rat bladder epithelium proved to be more sensitive (72%) than on monkey oesophagus alone (45%) and less sensitive than on salt-split skin (97%). Rat bladder epithelium, when tested on 41 sera of a control group, showed a very high specificity: 2/41 (95%). In combination with immunoblotting on keratinocyte extracts, indirect immunofluorescence on rat bladder epithelium allowed 93% of sera to be recognized, a value close to the salt-split skin alone. Rat bladder epithelium appears to be a more sensitive substrate than monkey oesophagus for the diagnosis of bullous pemphigoid and, although less specific, it is easier and faster than using salt-split skin, which remains indispensable to distinguish bullous pemphigoid from epidermolysis bullosa acquisita.     

A comparative study of antibody titers of blister fluid and serum in patients with subepidermal immunobullous diseases

BACKGROUND: Subepidermal autoimmune bullous diseases (SABD) comprise several disorders, such as bullous pemphigoid (BP), cicatricial pemphigoid (CP), epidermolysis bullosa acquisita (EBA), herpes gestationis (HG), and linear immunoglobulin A (IgA) dermatosis (LAD), and are characterized by antibody production against the basement membrane structures of the skin and mucosa. Although indirect immunofluorescence (IIF) on serum is a routine test for the detection of basement membrane zone antibodies, there have only been a few studies related to IIF on blister fluid. Aim To perform IIF on blister fluid and to compare the results with those of serum. METHODS: IIF on salt-split skin was performed on the serum and blister fluid of 35 patients with SABD (25 bp, three EBA, three HG, three LAD, and one bullous systemic lupus erythematosus) with conjugated IgG, IgA, and C3. RESULTS: Twenty-eight of the 35 patients showed IIF-positive blister fluid with a titer similar or less than that of serum. In 25 patients with BP, the most common disease in this study, 23 cases (92%) had positive IIF on serum, 23 cases (92%) on blister fluid, and 24 cases (96%) on either serum or blister fluid. Immunoreactant titers in BP blister fluid and serum did not show significant differences (P > 0.05). Epidermal binding of immunoreactants was the most prevalent staining pattern of IIF on salt-split skin (92%) in BP. CONCLUSIONS: From the findings of this study, the blister fluid of patients with SABD can be used for IIF. Although IIF sensitivity on blister fluid is no more than that on serum, the performance of this test on blister fluid in addition to serum may reduce the number of false negative results of IIF found using either of these two substrates alone.     

Immunofluorescence in dermatology

The accurate diagnosis of bullous and other immune diseases of the skin requires evaluation of clinical, histologic, and immunofluorescence findings. Immunofluorescence testing is invaluable in confirming a diagnosis that is suspected by clinical or histologic examination. This is especially true in subepidermal bullous diseases that often have overlap in the clinical and histologic findings. Direct immunofluorescence is performed on perilesional skin for patients with bullous diseases and lesional skin for patients with connective tissue diseases and vasculitis.