The reliability of long-term storage of direct immunofluorescent staining slides at room temperature

BACKGROUND: Long-term preservation of immunofluorescence is important for re-examinations. We investigated whether the storage of direct immunofluorescent (DIF)-positive slides at room temperature was reliable in daily practice. METHODS: One hundred and twenty-five DIF-positive slides from the skin of 52 patients were evaluated. Sections were examined for the presence of immunoglobulin A (IgA), IgG, IgM, C3, and fibrinogen using fluorescein isothiocyanate-conjugated antisera and mounted with a ready-to-use permanent mounting medium containing an antifading reagent and sodium azide (DAKO, Glostrup, Denmark, S3023). The slides were stored at room temperature for 16-24 months. Changes in diagnostic pattern, fluorescence intensity, and the form and location of accumulation of immunoreactants and technical deformation were investigated. RESULTS: Over the entire observation period, 49.6% of the slides faded away; the median length of survival was 16 months. Before 12 months, the survival rate of slides was 92.0%, whereas after 20 months it was 28.0%. In the early faded slides, which faded away before 16 months following the first examination, C3 and IgA were the most frequently observed immunoreactants. The technical deformations did not prevent the diagnosis. CONCLUSIONS: The preservation of fluorescence in DIF-positive slides using mounting media with an antifading reagent is possible for 2 years at room temperature. However, in daily practice, storage for longer than 11 months prevents a reliable diagnosis.     

Enzyme‐linked immunosorbent assay (ELISA) and indirect immunofluorescence testing in a bullous pemphigoid and pemphigoid gestationis

Enzyme‐linked immunosorbent assay (ELISA) is an excellent tool for detection of circulating antibodies against the NC16A portion of BP180 antigen. We compared the sensitivity and specificity of a commercially available BP180‐NC16a domain ELISA with that of an indirect immunofluorescence (IIF) testing in the evaluation of bullous pemphigoid (BP) and pemphigoid gestationis (PG), and analyzed the relationship between ELISA results and the presence of IgG deposition, in an epidermal or combined pattern, on direct immunofluorescence (DIF) testing of salt‐split skin. ELISA was performed on serum from 28 patients (24 BP, 4 PG) and 50 controls. IIF testing was performed on serum from 27 patients and 98 controls. For the group of 28 patients with BP or PG, ELISA had a sensitivity of 93% and specificity of 96% (P < 0.001), while sensitivity was 74% and specificity 96% (P < 0.001) for IIF testing. In these patients, ELISA has a higher sensitivity than IIF testing, but similar specificity. Evaluation of controls who had IgG deposition on the dermal side of salt‐split skin on DIF testing showed specificity for the ELISA of 100% (all four cases negative) and 80% for IIF testing (one of five positive). Positive ELISA correlated with a diagnosis of BP or PG only in patients who had IgG at the basement membrane zone (BMZ) by DIF testing. Overall, ELISA appears to have greater sensitivity and specificity for BP or PG than does IIF testing.     

Paraneoplastic pemphigus with negative direct immunofluorescence in epidermis or mucosa but positive findings in adnexal structures

Introduction:  Paraneoplastic pemphigus (PNP) is considered an autoimmune, multiorgan disease caused by antiplakin antibodies. We present three PNP patients who had negative epithelial direct immunofluorescence (DIF) findings in one or more biopsies.
Patients:  An early lip biopsy of uninvolved oral epithelia in patient 1 was negative. A later biopsy from foreskin showed intense intercellular immunoglobulin G (IgG) deposits in the epithelia. In the early phase of the disease in patient 2, the intercellular fluorescence was negative in the epidermis, while intercellular IgG and C3 were observed in the sweat ducts. A later biopsy showed weak intercellular epidermal IgG and C3 fluorescence. Patient 3 showed intercellular IgG and/or C3 in follicular, sebaceous and sweat duct structures in several biopsies. No intercellular IgG or C3 was observed in the epithelia.
Discussion:  The presence of immunoreactants in adnexal structures suggests that desmoplakins can be more strongly expressed in adnexa than in the epidermis, facilitating visualization of antibody deposits.
Conclusions:  Negative DIF findings in epithelia do not rule out the diagnosis of PNP, and the presence of IgG and/or C3 at the intercellular level of adnexal structures can help establish this diagnosis.     

The predominance of IgG4 in prodromal bullous pemphigoid

Background Prodromal bullous pemphigoid (PBP) and bullous pemphigoid (BP) demonstrate immunoglobulin G (IgG) and/or C3 deposition at the basement membrane zone (BMZ) on direct immunofluorescence. BP‐180‐specific IgG1, IgG4, and IgE antibodies have been detected in BP. However, the distribution of IgG subclasses is unknown in PBP. Objectives We will describe the role of anti‐BMZ IgG subclasses in PBP and we will correlate these findings to better understand the pathogenesis of PBP. Methods Skin biopsies and serum samples were obtained from 45 patients who had PBP. The skin tissue was processed for direct immunofluorescence studies. Sera were analyzed by indirect immunofluorescence for the presence of circulating anti‐BMZ IgG antibodies (by standard IIF) and IgG subclasses antibodies (by sandwich double antibody immunofluorescence [SDAI]). Sera were also analyzed for antibodies against BP‐180 and BP‐230 antigens by enzyme‐linked immunosorbent assay (ELISA). Results Thirty‐two patients (71%) had IgG and C3 staining at the BMZ, while 13 patients (29%) had isolated C3 staining at the BMZ on direct immunofluorescence. All patients demonstrated staining on the epidermal side of the salt‐split skin. Of the seven skin specimens that were available for C5‐9 SDAI testing, all were found to be positive along BMZ area. Standard IIF studies demonstrated the presence of circulating BMZ antibodies in 11 of the 30 patients (36.6%). When SDAI for IgG subclass differentiation was utilized, 17 of 30 (56.6%) patients were found to have circulating anti‐BMZ antibodies. All of these 17 patients had IgG4 subclass antibodies. Thirteen patients did not have detectable IgG subclass anti‐BMZ antibody on SDAI. Sixteen of 30 patients had detectable anti‐BP‐180 or anti‐BP‐230 antibodies, while 12 (40%) did not have detectable antibody against BP antigens on ELISA. Conclusions IgG4 is the initial and predominant anti‐BMZ antibody subclass detected in PBP. Demonstration of linear C5‐9 at the BMZ enhances the early diagnosis of PBP. Predominance of IgG4 and the initial presence of IgG4 on skin lesions as well as the presence of only IgG4 subclass anti‐BMZ antibody suggest that IgG4 subclass antibody could be the initial immunologic event encountered in patients with PBP.     

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.     

Polymorphic eruption of pregnancy

A 29-year-old woman developed a pruritic papular eruption that appeared symmetrically on the lateral part of her arms and elbows and on the media part of her thighs and knees in the 35th week of gestation (Fig. 1). The papules were erythematous, firm, flat, with a slightly elevated border, and measured 2–5 mm in diameter. A few papules quickly coalesced to form urticarial plaques. There were no lesions on the abdomen. In the 40th week of gestation, she delivered a healthy girl and a week later the eruption cleared up spontaneously. Two years later, in the 37th week of gestation of her second pregnancy, the same patient developed an itchy papular eruption, clinically comparable with that of the previous pregnancy, which spontaneously disappeared a few days after delivery of a healthy boy. During both pregnancies routine laboratory investigations, hormonal tests, and tests to detect cholestasis and viral hepatitis (A,B,C) were performed. A biopsy of lesional and perilesional skin was taken for histologic and direct immunofluorescence (DIF) examination. Serum was tested by indirect Immunofluorescence (IIF) staining for IgG, IgM, and IgA, and by a complement fixation test. For histologic examination, biopsy specimens were fixed in 10% buffered formalin, embedded in paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin. Direct immunofluorescence was performed on cryostat sections from biopsy specimens, washed in phosphate-buffered saline (PBS) and incubated with fluorescein isothiocyanate (FITC) labeled antibodies (anti-IgG, IgA, IgM, C3, fibrinogen) (DAKO, Glostrup, Denmark). The slides were mounted in buffered glycerol and examined by fluorescent microscopy. An immunohistochemical study was also performed on cryostat sections using the alkaline phosphatase antialkaline phosphatase (APAAP) technique with monoclonal antibodies (anti-CD45RO, CD3, CD4, CD8, CD25, CD22, CD1a, HLA-DR, CD54). The results of routine laboratory and hormonal tests and tests for cholestasis and viral hepatitis (A,B,C) were normal or negative. The histopathologic examination of a papule showed mild spongiosis in the epidermis, slight oedema, and a moderate perivascular infiltrate of lymphocytes and monocytes in the superficial dermis (Fig. 2). Examination by DIF on lesional skin showed a dense granular-linear deposit of IgM and C3 along the basement membrane zone (BMZ) of the epidermis and hair follicles (Fig. 3). After healing of the dermatosis, repeated DIF near the previously examined location was negative. An IIF study revealed no circulating anti-BMZ IgG, IgM, or IgA autoantibodies. The complement fixation test was also negative. The immunohistochemical study showed a perivascular infiltrate in the dermis consisting mainly of T lymphocytes (CD4+, DRH+, CD25+).     

系统性红斑狼疮患者皮损中C_3沉积与病情活动的关系

目的探讨系统性红斑狼疮(SLE)患者皮损中C3的沉积与血清中抗体的阳性率、补体C3水平及病情活动之间的关系。方法用直接免疫荧光法(DIF)检测皮损部位的免疫反应物;常规方法检测患者血清中自身抗体及补体;根据临床表现和实验室检查指标,进行系统性红斑狼疮病情活动性评分(SLEDAI)。结果 65例SLE患者中,37例(56.92%)真表皮交界处有一种或几种抗体和/或补体沉积,即DIF阳性,将65例患者按皮损中C3的沉积情况分成3组:DIF阴性组28例(43.08%),C3沉积组21例(32.31%)和仅有其他抗体沉积而无C3沉积组16例(24.62%)。各组间血清抗体ANA,dsDNA,SSA,RNP和Sm的阳性率差异均无统计学意义(P>0.05)。而C3沉积组的血清C3水平明显低于其余两组(P<0.05),且C3沉积组的病情活动性明显高于其余两组(P<0.05)。结论 C3单独及与其他抗体在皮损部位的沉积,比DIF阴性组和仅有其他抗体而无C3沉积组具有更低的血清C3水平及更高的病情活动性。     

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 Study of Pemphigus from North India

Both serum studies by indirect immunofluorescence (IIF) tests and skin biopsy examination by direct immunofluorescence (DIF) were performed on 22 cases of pemphigus with clinically active lesions. Twenty cases had pemphigus vulgaris and two, pemphigus foliaceus. The majority of cases (70%) were between 21 and 60 years old; the mean age was 39.5 ± 12.7 years. There was no sex predilection. DIF showed the positive fluorescence of intercellular cement substance (ICS) of the epidermis in all 22 cases (100%). IgG was positive in 77.27%, followed sequentially by C3C (50%), IgA (45.45%), and IgM (36.36%). Six cases (27.27%) also showed granular immunoglobulin and/or complement deposits at the dermoepidermal junction. IIF detected serum antiepithelial antibodies in 18 cases (81.81%) using human esophagus as substrate. Most of these cases (88.88%) showed IgG type of antibodies; the other 11.11% exhibited IgA and IgM in low concentrations. 1+ titer positivity was observed in 15 cases. This study demonstrates the value of DIF for a definitive diagnosis of pemphigus. However, it is also important to appreciate that immunofluorescence is not a substitute for histopathology, but rather complementary to it.     

CHRONIC DISCOID LUPUS ERYTHEMATOSUS IN THAILAND: DIRECT IMMUNOFLUORESCENCE STUDY

Background. Studies of chronic discoid lupus erythematosus (DLE) lesions by direct immunofluorescence (DIF) were heterogeneous with respect to classes of immunoglobulins and sites where these were deposited. Most of the studies were done in the USA and European countries. Materials and Methods. To obtain representative data from Asiatic countries, we analyzed the direct immunofluorescent abnormalities of 100 DLE lesions in Thai patients who were diagnosed on the basis of clinical and histologic criteria. Results. Granular deposits at the dermoepidermal junction (DEJ) were detected in 90% of cases. The common immunoreactants at the DEJ were IgG (63%) and IgM (47%). The deposits were usually combinations of various classes of immunoglobulins, mostly IgG (53%) and IgM (41%). Deposits of IgG and IgM alone at the DEJ were observed in 12% and 8%, respectively. Deposits at colloid bodies, dermal blood vessel walls, and epidermal nuclei were sometimes also seen. Conclusions. The DIF test of skin biopsy specimens is diagnostically significant in chronic DLE. Our study in Thai patients showed that the most common deposit was a combination of various classes of immunoglobulins, mostly IgG and often IgM as well as C3, and occurred at the DEJ of the involved area.     

Immunofluorescence testing in the diagnosis of autoimmune blistering diseases: overview of 10-year experience

BACKGROUND

Immunofluorescence testing is an important tool for diagnosing blistering diseases.

OBJECTIVE

To characterize the immunofluorescence findings in patients diagnosed with autoimmune blistering skin diseases.

METHODS

We retrospectively analyzed immunofluorescence results encompassing a 10-year period.

RESULTS

421 patients were included and divided into 2 groups: group 1- intraepidermal blistering diseases (n=277) and 2- subepidermal blistering diseases (n=144). For group 1, positive DIF findings demonstrated: predominance of IgG intercellular staining (ICS) and C3 for pemphigus foliaceus-PF (94% and 73% respectively), pemphigus vulgaris-PV (91.5%-79.5%) and paraneoplastic pemphigus-PNP (66%-33%); ICS IgA in 100% of IgA pemphigus cases, and IgG deposits in the basement membrane zone (BMZ) along with ICS in one Hailey-Hailey patient. The IIF findings revealed mean titers of 1:2.560 for PV and 1:1.280 for PF. For paraneoplastic pemphigus, IIF was positive in 2 out of 3 cases with rat bladder substrate. In group 2, positive DIF findings included multiple deposits at basement membrane zone for epidermolysis bullosa acquisita-EBA (C3-89%,IgG-79%,IgA-47%,IgM-21%) mucous membrane pemphigoid-MMP (C3,IgG,IgA,IgM-80%) and bullous pemphigoid-BP (C3-91%,IgG-39%,IgA-11%,IgM-6%), and IgA at basement membrane zone for IgA linear disease (99%) and dermatitis herpetiformis-DH (dermal papillae in 84.6%). For lichen planus pemphigoides, there was C3 (100%) and IgG (50%) deposition at basement membrane zone. indirect immunofluorescence positive findings revealed basement membrane zone IgG deposits in 46% of BP patients, 50% for EBA, 15% for IgA linear dermatosis and 50% for LPP. Indirect immunofluorescence positive results were higher for BP and EBA with Salt-Split skin substrate.

CONCLUSION

Our results confirmed the importance of immunofluorescence assays in diagnosing autoimmune blistering diseases, and higher sensitivity for indirect immunofluorescence when Salt-split skin technique is performed.     

A Cross-sectional Study of Direct Immunofluorescence in the Diagnosis of Immunobullous Dermatoses

Background:

Autoimmune blistering diseases are a group of bullous disorders characterized by pathogenic antibodies directed at the target antigens, which are components of the desmosomes or adhesion complex at the dermoepidermal junction. Direct immunofluorescence (DIF) is invaluable in the diagnosis of these lesions.

Aim:

The aim of this study was to evaluate the sensitivity of DIF in immunobullous dermatoses and to study the pattern of DIF. The study also aims to correlate DIF with clinical and histologic findings and to analyze discrepancies.

Materials and Methods:

Total 100 biopsies received over a period of 2 years in the Department of Pathology were analyzed. DIF, histopathology and clinical data were reviewed.

Results:

Out of 100, 89 cases showed DIF patterns concordant with clinical/histologic diagnosis. The sensitivity of DIF was 94.44% (51/58) in the pemphigus and 84% (21/25) in the bullous pemphigoid (BP) group, 100% each in dermatitis herpetiformis (2/2) and linear IgA disease (1/1). A total of 11 histologically proven cases of immunobullous lesions were DIF negative-four (three of pemphigus vulgaris and one of BP) due to having no epidermis, three (cases of BP) owing to sampling/technical errors and the remaining four (cases of pemphigus vulgaris) due to being on treatment.

Conclusion:

Immunofluorescence helps confirm the diagnosis of bullous lesions in which there is clinical and the histopathologic overlap. Sampling errors contributed to false negative (FN) results.     

抗核抗体阴性的大疱性系统性红斑狼疮1例

报告１例抗核抗体阴性的大疱性系统性红斑狼疮。在整个病程中共查过７次抗核抗体均阴性。ＤＩＦ、ＩＩＦ，免疫印迹均无阳性发现。肾脏穿刺活检免疫荧光见ＩｇＧ颗粒状沉积。并讨论其诊断依据和大疱性系统性红斑狼疮抗核抗体为阴性的可能原因。     

Childhood bullous pemphigoid: a clinicopathologic study and review of the literature

Bullous pemphigoid (BP) is an acquired bullous disorder that predominantly affects the elderly. It is rare in children but when it occurs, there is considerable clinical and histologic overlap with other acquired or congenital blistering disorders. A definitive diagnosis of childhood BP requires direct immunofluorescence and, in some cases, characterization of the target antigen. Three cases of childhood BP are presented, with their histologic and immunofluorescence findings. The first was a 5-month-old male infant who presented with erythema and bullae of the palms and soles and was found to have linear deposition of IgG and C3 along the dermoepidermal junction on direct immunofluorescence (DIF). Histopathologic examination revealed a subepidermal blister containing eosinophils. Type IV collagen was demonstrated along the floor of the blister cavity by a direct immunoperoxidase technique. The second case was an 8-month-old female infant who presented with a blistering eruption of her palms and soles that then became widespread. Direct immunofluorescence showed linear IgG and C3 at the dermoepidermal junction, with laminin deposition at the base of the blister. The third case was a 7-year-old female with bullae and erosions on the vulva and vaginal mucosa. A subepidermal blister was seen on microscopic examination whereas immunofluorescence demonstrated linear IgG and C3 deposition at the basement membrane zone (BMZ). A literature review uncovered 50 cases of childhood BP confirmed by direct or indirect immunofluorescence, or both, and often with evidence of autoantibodies against either the 180 kD or the 230 kD human bullous pemphigoid antigens (BP180 or BP230). This review was used to delineate characteristics of childhood BP, including the newly proposed subtypes: infantile BP and childhood localized vulval BP. Infantile BP presents within the first year of life and is characterized by BP-like lesions on erythematous or normal acral skin. Localized vulval BP is a self-limited, nonscarring BP-like process that involves only the vulva. Both subtypes are normally self-limited and respond well to either topical or systemic steroids, if treatment is initiated before the disease becomes widespread.     

A case of antiepiligrin cicatricial pemphigoid with nephrotic syndrome

We report a 71-year-old woman with antiepiligrin cicatricial pemphigoid associated with nephrotic syndrome. She presented with a six-month history of pruritic blisters over her trunk and legs. She also had episodes of recurrent painful oral erosions. A skin biopsy showed a subepidermal bulla, and a direct immunofluorescence (DIF) study revealed linear deposition of IgG and C3 at the basement membrane zone (BMZ). Indirect immunofluorescence (IIF) staining of 1 M NaCl-split skin demonstrated circulating IgG autoantibodies reactive with the dermal side. Immunoprecipitation studies of the patient's serum disclosed IgG autoantibodies directed against a set of polypeptides that corresponded to laminin 5 (beta 3 gamma 2). Based upon the long-standing edema of her legs and her hypoproteinemia, she was diagnosed with nephrotic syndrome. To our knowledge, the association of antiepiligrin cicatricial pemphigoid with nephrotic syndrome has not been reported previously.     

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.     

The usefulness of immunofluorescent tests in pemphigus patients in clinical remission

Direct and indirect immunofluorescent studies (DIF, IIF) were performed on 24 pemphigus vulgaris patients who were in a state of clinical remission. The tests were repeated after an interval of 6 months. All the patients were on maintenance therapy with oral prednisone. The DIF in eight patients showed negative results among whom seven remained negative. Six patients out of 24 showed weakly positive fluorescence and ten patients showed strong positive fluorescence. The IIF was negative in 17 patients and positive in seven patients who also showed positive DIF. During a follow-up period of 20 months, one of eight patients with negative DIF relapsed compared with two of six patients with weak positive DIF and five of 10 patients with strong DIF. Five patients with strong DIF for IgG also had C3, of whom three relapsed, compared with five of 19 patients who were negative for C3. Four of seven patients with positive IIF relapsed compared with four of 17 with negative IIF. It is suggested that repeated DIF tests in pemphigus patients, who are in clinical remission, may serve as an indicator for the immunological activity and be of help in the management of these cases.     

间接免疫荧光技术对类天疱疮的循环抗基底膜抗体的研究

运用正常人皮肤及分离正常人皮肤为底物的两种间接免疫荧光方法，对２０例大疱性类天疱疮血中循环抗基底膜抗体进行研究。发现传统的ＩＩＦ法阳性率为６５％（１３／２０），分离皮肤ＩＩＦ法阳性率为１００％（２０／２０）。分离皮肤法不单敏感性明显高于传统方法，且能知道抗体在基底膜的结合部位，比传统方法更有助于类天疱疮和几种表皮下大疱病的鉴别诊断。     

IgE and its related phenomena in bullous pemphigoid

This study was designed to analyse IgE and its related phenomena in bullous pemphigoid (BP). We analysed 17 BP sera by indirect immunofluorescenee (IIF) and immunoblotting (IB) using a monoclonal antibody to IgE. In addition, inflammatory cells in lesional skin from 11 patients with BP were analysed by the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique using monoclonal antibodies to IgE and Fc?RII/CD23. IgE class anti-basement membrane zone (BMZ) autoantibody was detected in nine of 17 sera (52.9%) by IIF. IgG class anti-BMZ antibody could block the BMZ-binding reactivity of IgE class antibody. Titres of IgE class autoantibody in the sera ranged from 1:40 to 1:320, and statistically correlated with serum IgF levels. Two of 11 sera contained an IgE class autoantibody which recognized a 230-kDa BP antigen by IB. By radio-allergosorbent test (RAST), IgE-specific antibodies to an extended series of common inhalant and food allergens were detectable in six sera with higb concentrations of total IgF(over 3300 IU/ml). IgE-bearing and Fc?RII-expressing cells were demonstrated in the upper dermis and along the BMZ in seven of 11 biopsy specimens by tbe APAAP technique. The distribution and number of IgE-bearing cells in the lesions were similar to those of the FcEERII-expressing cells. Tbese results suggest that botb IgE-mediated immune responses and autoimmunity characterize BP as distinctive features.     

大疱性类天疱疮合并绒癌1例

患者女性,３８岁,因发现腹部包块５个月,全身皮肤起大疱２个月入院,皮肤科检查可见躯干,四肢大量紧张性水疱,大疱,尼氏征阴性,皮肤组织理学检查可见表皮下水疱形成伴嗜酸性粒细胞浸润,直接免疫荧光可见基底膜带ＩｇＧ和Ｇ３呈线性沉积,间接免疫荧光检查血清抗类天疱疹抗体阴性。盆腔术后病理诊断为绒癌侵及卵巢。