A clinical analysis of 74 cases of bullous pemphigoid with stroke / 中华皮肤科杂志

Objective:To investigate differences in clinical characteristics between bullous pemphigoid (BP) patients with stroke and those without, and their relationship with the prognosis of stroke.Methods:A retrospective analysis was performed on medical records of 330 BP inpatients in the First Affiliated Hospital of Zhengzhou University from September 2012 to April 2020. These patients were divided into BP + stroke (ST) group and BP - ST group according to whether they were accompanied by stroke, and clinical manifestations and relevant laboratory examination results were compared between the two groups. According to the stroke outcome score assessed by modified Rankin Scale (mRS), patients in the BP + ST group were further divided into good-prognosis ST group (mRS ≤ 2 points) and poor-prognosis ST group (mRS > 2 points), and subgroup analysis was conducted. Correlations between measurement data (such as age, disease course and laboratory examination results) and mRS scores were analyzed.Results:In the BP - ST group (256 cases), 151 were males and 105 were females, and their age ranged from 19 to 92 (66.8 ± 13.6) years; in the BP + ST group (74 cases), 45 were males and 29 were females, and their age ranged from 48 to 92 (74.6 ± 9.6) years; Compared with the BP - ST group, the BP + ST group showed older age ( t = -5.57, P < 0.001), shorter disease course of BP ( Z = -3.07, P = 0.002), and higher anti-BP180 IgG antibody levels (215.0 [157.2, 283.1] U/ml vs. 155.0 [63.9, 279.8] U/ml; Z = -2.12, P = 0.034). The distribution of skin lesions significantly differed between the two groups ( χ2 = 10.51, P = 0.015), and the BP + ST group showed a significantly lower proportion of patients with generalized lesions ( P<0.05), but a higher proportion of patients with lesions on the limbs ( P<0.05). Subgroup analysis showed significant differences in the patients′ age, BP course, lesion distribution and anti-BP180 IgG antibody levels among the good-prognosis ST group, poor-prognosis ST group and BP - ST group ( F = 10.83, P<0.001; Z = 17.24, P<0.001; χ2 = 15.57, P = 0.026; Z = 6.29, P = 0.043, respectively). There was no significant difference in the age between the good-prognosis ST group and poor-prognosis ST group (adjusted P = 1.000), but the patients were significantly older in the two above groups than in the BP - ST group (adjusted P = 0.001, 0.007, respectively) ; the poor-prognosis ST group showed significantly shorter BP courses (adjusted P = 0.016, < 0.001, respectively) and a higher proportion of patients with lesions on the limbs (both P < 0.05) compared with the good-prognosis ST group and BP - ST group, and significantly higher serum anti-BP180 IgG antibody levels compared with the BP - ST group (226.2 [163.6, 285.8] U/ml vs. 155.0 [63.9, 279.8] U/ml; adjusted P = 0.037). There were no significant differences in the gender distribution, lesional morphology, percentages and counts of peripheral blood eosinophils, serum total IgE levels, and anti-BP230 IgG antibody levels between the BP + ST group and BP - ST group (all P > 0.05), or among the good-prognosis ST group, poor-prognosis ST group and BP - ST group (all P > 0.05). Correlation analysis in the BP + ST group showed a significantly negative correlation between the BP course and mRS scores ( r = -0.33, P = 0.004), and a significantly positive correlation between the anti-BP180 IgG antibody levels and mRS scores ( r = 0.34, P = 0.032) . Conclusion:There were differences in the patients′ age, BP course, lesion distribution, and anti-BP180 IgG antibody levels between the BP patients with stroke and those without, and the differences were more obvious between the poor-prognosis ST group and BP - ST group.

BP180-related autoimmune blistering diseases / 中华皮肤科杂志

BP180-related autoimmune blistering diseases include bullous pemphigoid,lichen planus pemphigoides,linear IgA bullous dermatosis,pemphigoid gestationis and cicatricial pemphigoid.There are multiple autoantibody-reactive sites on the extracellular region of BP180.Current studies show that there is heterogeneity in the autoimmune blistering disease-related target sites on BP 180,and different clinical manifestations of the same disease are related to the heterogeneity of target sites.However,further studies and analysis are still needed for the mechanism of the heterogeneity.

Expression of BP180 antibody in sera of patients with bullous pemphigoid and nervous system / 中华皮肤科杂志

Objective To measure the expression of BP180 antibody in sera of patients with bullous pemphigoid (BP) and/or nervous system diseases (ND),and to explore the relationship between BP and ND.Methods Clinical data were collected from some inpatients and outpatients with BP in Department of Dermatology,as well as some inpatients with ND in Department of Neurology,the Second Hospital of Jilin University between March 2012 and September 2013.Peripheral blood samples were obtained from 20 BP patients without a medical history of ND (BP group),20 patients with ND alone (ND group),20 BP patients with a medical history of ND (BP + ND group),and 20 healthy controls (healthy control group) separately.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum level of BP180 antibody in the above groups.Statistical analysis was carried out with SPSS17.0 software by using chi-square test for comparing enumeration data,analysis of variance for comparing the anti-BP180 antibody titer or ages among the above 4 groups,and Games-Howell test for multiple comparisons.Results No anti-BP180 antibody was detected in any of the 20 subjects in the healthy control group.All of the patients in the BP group and BP + ND group were positive for anti-BP180 antibody,and the antibody titers were 128.347 ± 54.678 and 143.482±72.568 respectively.Of the 120 patients in the ND group,7 were positive for anti-BP180 antibody,including 4 with cerebral hemorrhage and 3 with cerebral thrombosis,and the highest anti-BP180 antibody titer of 39.638 was recorded in 1 patient with cerebral infarction.There was a significant difference in the anti-BP180 antibody titer among the 4 groups (F =55.624,P ＜ 0.05).In addition,there was no significant difference in the anti-BP180 antibody titer between the BP group and BP + ND group (P =0.878),while the anti-BP180 antibody titers significantly differed between the BP group and ND group,between the BP group and healthy control group,between the BP + ND group and ND group,between the BP + ND group and healthy control group,and between the ND group and healthy control group (P ＜ 0.05).Conclusion BP may occur in ND patients with an increased anti-BP180 antibody titer.

There is no correlation between the serum level of interleukin-21 and bullous pemphigoid / 中华皮肤科杂志

Objective To investigate the correlations of the serum level of interleukin-21 (IL-21) with anti-BP 180 antibodies,peripheral eosinophil level,bullous pemphigoid disease area index (BPDAI)in patients with bullous pemphigoid (BP).Methods A total of 31 patients with bullous pemphigoid and 31 healthy controls were enrolled into this study.Blood samples were collected from all the subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to detect the serum level of IL-21 and serum titer of anti-BP180 antibodies.BPDAI was assessed among 13 patients,and the correlation of the IL-21 level with anti-BP180 antibody titers,eosinophil count and BPDAI were analyzed.Results No significant difference in the serum level of IL-21 was observed between the patients (M [P25-P75]:255.28 [156.19-361.59] ng/L) and healthy controls (150.47 [72.4-268.73] ng/L,P =0.14).There was no correlation between the serum level of IL-21 and serum titer of anti-BP180 antibodies (82.98 [26.82-132.92] U/ml,rs =0.18,P =0.332).Among the 27 BP patients who underwent blood cell count,no correlation was found between the serum levels of IL-21 and whole-blood eosinophil counts (rs =0.104,P =0.606).Among the 13 BP patients who underwent assessment of BPDAI,there was no correlation between the serum levels of IL-21 and BPDAI scores (rs =0.233,P =0.464).Conclusion The IL-21 level is uncorrelated with anti-BP 180 antibody level,peripheral eosinophil level or BPDAI in patients with BP.

Evaluation of the combination of BP180-NC16a enzyme-linked immunosorbent assay and BP230 enzyme-linked immunosorbent assay in the diagnosis of bullous pemphigoid.

Background: Bullous pemphigoid (BP) is an acquired autoimmune subepidermal blistering disease characterized by circulating IgG autoantibodies directed against BP180 and BP230 hemidesmosomal proteins. Previous studies have demonstrated that antibodies against the NC16a domain of BP180 mediate BP pathogenesis, while antibodies against BP230 enhance the inflammatory response. Recently, commercial BP180-NC16a enzyme-linked immunosorbent assay (ELISA) and BP230 ELISA kits were developed to detect anti-BP180 and anti-BP230 autoantibodies in human BP sera. Aims: To evaluate the efficacy of BP180-NC16a ELISA and BP230 ELISA in the initial diagnosis of BP. Methods: Sera from 62 BP patients and 62 control subjects were tested by BP180-NC16a ELISA and BP230 ELISA and compared with findings from indirect immunofluorescence (IIF) and immunoblotting (IB) to determine the sensitivity and specificity of these assays. Results: The sensitivities of BP180-NC16a ELISA and BP230 ELISA were 87.1% (54/62) and 56.5% (35/62), respectively, and the specificities of both were 100% (62/62). Using both ELISAs for diagnosis increased the sensitivity to 95.2% (59/62) and was statistically comparable with IB sensitivity. Conclusions: ELISA is a convenient, effective, and reliable method for serodiagnosis of BP, and combined use of BP180-NC16a ELISA and BP230 ELISA can increase the sensitivity of this diagnostic approach.

Purification and identification of anti-BP180 NC16A antibodies from the sera of patients with bullous pemphigoid or herpes gestationis / 中华皮肤科杂志

Objective To develop a method to purify and identify anti-BP180 NC16A antibodies from the sera of patients with bullous pemphigoid (BP) or herpes gestationis.Methods The GST/NC16A fusion protein was expressed in a prokaryotic expression vector pGEX-2TBP180NC16A,and then crosslinked to glutathione sepharose beads.Anti-BP180 NC16A antibodies were isolated from the sera of 3 patients with BP and 2 patients with herpes gestationis by affinity chromatography,and analyzed by immunofluorescence,Western blot and enzyme linked immunosorbent assay (ELISA).Results The GST/NC16A fusion protein with a relative molecular mass of 37 000 was successfully expressed by the prokaryotic vector pGEX-2TBP180NC16A.Purified anti-BP180 NC16A antibodies were obtained from the sera of patients by the affinity chromatography,and ELISA revealed that the concentration of anti-BP180 NC16A was 2.4 mg/ml.The purified antibody could bind to the basement membrane zone of human skin,suggesting a strong biological activity of the antibodies.Western blot showed a single band corresponding to the expected molecular mass for anti-BP180 NC16A antibodies,indicating a high purity of the isolated antibodies.Conclusion The anti-BP180 NC16A antibodies purified by microbead-based affinity chromatography from the sera of patients with BP or herpes gestationis are highly active and specific.

Usefulness of Enzyme-linked Immunosorbent Assay Using Recombinant BP180 and BP230 for Serodiagnosis and Monitoring Disease Activity of Bullous Pemphigoid

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune subepidermal bullous disease associated with autoantibodies against BP180 and BP230. Enzyme-linked immunosorbent assay (ELISA) is a sensitive tool for the detection of immunoglobulin G (IgG) anti-BP180 and anti-BP230 autoantibodies. OBJECTIVE: The aim of this study was to evaluate the usefulness of ELISA for diagnosing and monitoring the disease activity of BP. METHODS: We evaluated serum IgG levels of anti-BP180 and anti-BP230 autoantibodies in 47 BP patients, 16 epidermolysis bullosa aquisita patients, and 15 healthy volunteers using ELISA. Through retrospective review of the medical records, the clinical characteristics of BP including disease activity, duration, pruritus severity and peripheral blood eosinophil counts were assessed. RESULTS: The sensitivity of BP180 ELISA was 97.9%, BP230 ELISA 72.3%, and a combination of the two was 100%. The specificity of BP180 ELISA was 90.3%, BP230 ELISA 100%, and a combination of the two was 90.3%. BP180 ELISA scores showed strong associations with disease activity, pruritus severity, peripheral blood eosinophil counts, and disease duration, whereas BP230 ELISA scores did not. CONCLUSION: BP180 and BP230 ELISAs are highly sensitive methods for the diagnosis of BP, and BP180 ELISA, in particular, is a sensitive tool for monitoring the disease activity of BP.

A multi-center, randomized, double-blind and controlled study of BP180NC16a enzyme-linked immu-nosorbent assay (BP180NC16a-ELISA) in the diagnosis of bullous pemphigoid / 中华皮肤科杂志

Objective To evaluate the sensitivity and specificity of BP180NC16a-ELISA in the diagnosis of bullous pemphigoid (BP). Methods A multi-center, randomized, double-blind, parallel-controlled study was conducted. Sera were collected from 106 patients with clinically confirmed active BP and 106 control subjects including patients with non-BP bullous diseases, scleroderma, psoriasis or systemic lupus erythematosus,late pregnant women and healthy blood donors. BP180NC16a-ELISA was performed on these sera. The IgG antibody levels measured by ELISA kit were compared with those measured by indirect immunofluorescence (IIF) test. Results Of the 106 BP sera, 81 were positive for BP180NC16a-ELISA with a sensitivity of 76.4%,83 for ⅡF test with a sensitivity of 78.3%. Among the 106 control serum samples, 95 were negative for BP180NC16A-ELISA with a specificity of 89.6%, and 102 for ⅡF test with a specificity of 96.2%. There was no significant difference between the two tests in dignostic sensitivity and specificity for BP (both P ＞ 0.05).Conclusion BP180NC16A-ELISA may serve as an adjuvant tool for the diagnosis of BP.

Localized Bullous Pemphigoid on Both Upper Extremities in a Hemiplegic Patient / 대한피부과학회지

Bullous pemphigoid is a chronic autoimmune blistering disease characterized clinically by tense bullae that develop on normal or erythematous skin. Bullous pemphigoid is associated with autoantibodies to two hemidesmosomal proteins, BPAG1 (230 kD) and BPAG2 (180 kD). The localized form of BP is an unusual variant that occurs in 5~30% of the patients. A 58-year-old man who had been suffering from right hemiplegia since 2006, presented with multiple tense bullae localized on both arms and hands. Direct immunofluorescence test showed linear deposition of IgG and C3 along the basement membrane zone. The antibodies against the recombinant NC16a-domain of BP180 were positive by ELISA and immunoblotting using epidermal extract of normal human foreskin demonstrated that the patient's serum reacted with only BP180 antigen. Here, we report a case of localized bullous pemphigoid on both upper extremities in a hemiplegic patient predominantly on the opposite side to the hemiplegia.

Pemphigoid gestationis: clinical and laboratory evaluation

INTRODUCTION: Pemphigoid gestationis, also known as herpes gestationis, is a rare autoimmune blistering disease associated with pregnancy. It usually occurs during the second or third trimester, but it may be present at any stage of pregnancy or the puerperium. The clinical, histologic, and immunopathological features of pemphigoid gestationis are similar to those of the pemphigoid group of disorders. METHODS: We hereby report seven patients who were diagnosed with pemphigoid gestationis and followed at the Autoimmune Blistering Disease Clinic in the Department of Dermatology of the University of Sao Paulo Medical School between 1996 and 2008. DISCUSSION: Demographic and clinical findings, such as median age, sites of involvement, and gestational age of onset of our patients, coincide with those described in previous reports. The majority of patients (85 percent) exhibited complement C3 or C3 and immunoglobulin G (IgG) deposition along the basement membrane zone (BMZ) on immunofluorescence. Herpes gestationis factor (HG) factor was postitive in four out of six patients (67 percent), and three out of five patients recognized the bullous pemphigoid recombinant antigen (BP180) by ELISA. CONCLUSION: This study revealed a good outcome of the newborns from pemphigoid gestationis affected mothers, based on the absence of pemphigoid gestationis cutaneous lesions, mean birth weight, and normal Apgar scores and gestational age at birth.

Immunoblot Analysis Using Epidermal Extract and 180kD Bullous Pemphigoid Antigen NC16a Domain in the Diagnosis of Subepidermal Bullous Diseases / 대한피부과학회지

BACKGROUND: The 180kD bullous pemphigoid antigen (BP180) is known to be recognized by sera from patients with bullous pemphigoid (BP), herpes gestationis (HG), and cicatricial pemphigoid (CP). A series of previous studies using BP180 recombinant proteins has shown that most sera from patients with BP and HG react with the NC16a domain of BP180. OBJECTIVE: This study was performed to compare the immunoblotting results by using human epidermal extracts and by using recombinant protein of BP180 NC16a domain in the diagnosis of subepidermal bullous skin diseases. METHOD: We observed the reactivity with the epidermal extract of normal human foreskin and recombinant protein of BP180 NC16a domain in immunoblotting assay. RESULTS: In immunoblotting with epidermal extract, 7(78%) and 6(67%) of the 9 BP sera reacted with BP230 and BP180, respectively. 1(50%) of the 2 HG and 2(100%) of the 2 CP sera reacted with BP180. In immunoblotting with recombinant protein of BP180 NC16a domain, 8(89%) of the 9 BP sera and 2(100%) of the 2 HG sera were immunoreactive. But, none of the CP and pemphigus sera reacted with the recombinant protein. CONCLUSION: The immunoblotting analysis using BP180 NC16a domain is a highly sensitive method without ambiguity in the diagnosis of subepidermal bullous skin diseases, and also is useful for the differential diagnosis of BP and HG from CP.