Downregulation of aquaporin 3 in patients with pemphigus vulgaris.

Pemphigus vulgaris (PV) is an autoimmune bullous skin disease. Aquaporin 3 (AQP3) is a glycerol/water channel involved in several physiological functions. Evaluation of the tissue expression and localization of AQP3 in the skin of PV patients. Twenty-seven PV patients and 30 controls were included. The patients were subjected to history taking, clinical evaluation, Autoimmune Bullous Skin Disorder Intensity Score and 4-mm punch biopsy. The biopsies were stained using anti-human AQP3 antibody and the immunofluorescence pattern and intensity were evaluated using a scoring system and ImageJ software analysis. AQP3 was expressed in the basal epidermis in 27 (100%) and in the suprabasal epidermis in 19 PV patients (70.4%). It was expressed in all controls in basal and suprabasal layers. Intensity of AQP3 immunofluorescence was strong in 2 (7.4%), moderate in 19 (70.4%) and weak in 6 patients (22.2%) while it was strong in 18 (60%) and moderate in 12 controls (40%). AQP3 expression was significantly lower in patients than controls in the suprabasal epidermis (p = 0.001). Patients with extensive disease had significantly weaker AQP3 intensity than those with marked disease (p = 0.005) Downregulation of AQP3 in patients with PV, especially in the suprabasal layers and in extensive clinical disease, suggests a potential role of AQP3 in the pathogenesis of PV.

Pemphigus vulgaris relapse during the coronavirus disease pandemic.

Pemphigus vulgaris (PV) is the most common type of pemphigus group of autoimmune skin diseases. The treatment of PV relapse is challenging especially during the coronavirus disease (COVID-19) pandemic. In this prospective study, we aimed to evaluate the treatment of patients with relapsing PV during the pandemic. Twelve patients with PV who experienced relapse from March 2020 to January 2022 were included. The patients were asked whether they experienced COVID-19 symptoms and the pemphigus disease area index (PDAI) was measured. PCR for COVID-19, chest computed tomography, routine investigations, and electrocardiography were performed for the admitted patients. The mean PDAI of the patients during relapse was 23.6 ± 14.8 (range 5-60). Seven patients received azathioprine; one patient received mycophenolate mofetil; and six patients received 1000 mg of rituximab (RTX) twice at an interval of 14 days. None of the 12 patients had COVID-19-suggestive symptoms. Only 1 patient relapsed after receiving the COVID-19 vaccine. The six admitted patients who received RTX were negative for COVID-19 based on the PCR testing results. Out of the 12 patients, eight achieved complete remission, while four achieved partial remission. No major adverse effects were observed. In conclusion, the treatments with systemic steroids, immunosuppressive drugs, and rituximab were well tolerated by the patients with relapsing PV, provided that there was no contact with individuals with COVID-19. These treatments can then be provided to patients with PV during the pandemic with careful follow-up.

Diagnosis and management of pemphigus: Recommendations of an international panel of experts.

BACKGROUND: Several European countries recently developed international diagnostic and management guidelines for pemphigus, which have been instrumental in the standardization of pemphigus management. OBJECTIVE: We now present results from a subsequent Delphi consensus to broaden the generalizability of the recommendations. METHODS: A preliminary survey, based on the European Dermatology Forum and the European Academy of Dermatology and Venereology guidelines, was sent to a panel of international experts to determine the level of consensus. The results were discussed at the International Bullous Diseases Consensus Group in March 2016 during the annual American Academy of Dermatology conference. Following the meeting, a second survey was sent to more experts to achieve greater international consensus. RESULTS: The 39 experts participated in the first round of the Delphi survey, and 54 experts from 21 countries completed the second round. The number of statements in the survey was reduced from 175 topics in Delphi I to 24 topics in Delphi II on the basis of Delphi results and meeting discussion. LIMITATIONS: Each recommendation represents the majority opinion and therefore may not reflect all possible treatment options available. CONCLUSIONS: We present here the recommendations resulting from this Delphi process. This international consensus includes intravenous CD20 inhibitors as a first-line therapy option for moderate-to-severe pemphigus.

Recipient site preparation by cryoblebbing in melanocyte keratinocyte transplantation procedure over the fingers in vitiligo: A pilot study.

Surgical treatment of vitiligo lesions over the fingers has poor outcome. In this intra-patient comparative study, 12 patients with stable non-segmental vitiligo (NSV) affecting the middle three fingers of one hand were included. Three variations were used in treatment of finger vitiligo lesions: minipuch grafting, melanocytes keratinocyte transplantation procedure (MKTP) preceded by cryoblebbing or full CO2 laser resurfacing of the recipient site. Liquid nitrogen was used to create blebs in one finger 24 hours before therapy. On the following day, the second finger was treated by minipunch grafting and the third finger was resurfaced by CO2 laser. A suspension was prepared and 0.1 mL was injected into each cryobleb. It was also applied to the resurfaced skin. All patients underwent topical PUVA therapy and were followed-up for 12 months. Ten cases with 52 lesions completed the follow-up period. About 4/18 lesions treated by cryoblebbing followed by MKTP showed ≥75% repigmentation while only 1/17 lesions treated by laser resurfacing + MKTP and 1/17 lesions treated by minipunch grafting showed 30% and 10% repigmentation, respectively. No complications occurred in MKTP treated lesions. Cryoblebbing of the recipient site seems to improve the outcome of MKTP in lesions over the fingers in stable NSV.

A prospective study comparing patients with early and late relapsing pemphigus treated with rituximab.

BACKGROUND: Rituximab (RTX) is an effective therapy for patients with pemphigus; however, the therapy does not prevent relapse. OBJECTIVES: To compare early relapsing patients (before 12 months) and late relapsing patients (after 24 months) following RTX therapy. METHOD: In this prospective study, 19 patients were enrolled (14 with pemphigus vulgaris and 5 with pemphigus foliaceus). The baseline disease score, autoantibody levels, and percentage of CD20+ cells of patients with pemphigus were measured. Patients received 1 cycle of RTX and were followed for 26 months. RESULTS: Among early relapsing patients (n = 5), the time to relapse was 6 to 11 months. Among late relapsing patients (n = 6), the time to relapse was 24 to 26 months. A significant difference was observed in the mean baseline anti-desmoglein 1 (DSG1) index between early relapsing (705.72) and late relapsing patients (210.4) (P = .0014). A significant negative correlation was found between the baseline anti-DSG1 index and time to relapse (r = -0.777, P = .00009). LIMITATIONS: The small number of patients with pemphigus foliaceus. CONCLUSIONS: Because patients with high baseline anti-DSG1 indices relapsed earlier, it may be important to follow these patients closely for the initial 12 months after RTX therapy. These patients may require a maintenance RTX dose during the first 12 months after RTX therapy.

Effect of combination of fractional CO2 laser and narrow-band ultraviolet B versus narrow-band ultraviolet B in the treatment of non-segmental vitiligo.

The present study was designed to evaluate the effect of combining fractional CO2 laser with narrow-band ultraviolet B (NB-UVB) versus NB-UVB in the treatment of non-segmental vitiligo. The study included 20 patients with non-segmental stable vitiligo. They were divided into two groups. Group I received a single session of fractional CO2 laser therapy on the right side of the body followed by NB-UVB phototherapy twice per week for 8 weeks. Group II received a second session of fractional CO2 laser therapy after 4 weeks from starting treatment with NB-UVB. The vitiligo lesions were assessed before treatment and after 8 weeks of treatment by VASI. At the end of the study period, the vitiligo area score index (VASI) in group I decreased insignificantly on both the right (-2.6%) and left (-16.4%) sides. In group II, VASI increased insignificantly on the right (+14.4%) and left (+2.5%) sides. Using Adobe Photoshop CS6 extended program to measure the area of vitiligo lesions, group I showed a decrease of -1.02 and -6.12% in the mean area percentage change of vitiligo lesions on the right and left sides, respectively. In group II the change was +9.84 and +9.13% on the right and left sides, respectively. In conclusion, combining fractional CO2 laser with NB-UVB for the treatment of non-segmental vitiligo did not show any significant advantage over treatment with NB-UVB alone. Further study of this combination for longer durations in the treatment of vitiligo is recommended.

Platelet-rich plasma for resistant oral erosions of pemphigus vulgaris: A pilot study.

Oral erosions and ulcers of pemphigus vulgaris (PV) are a debilitating condition that is usually difficult to treat. The wound healing properties of platelet-rich plasma (PRP) encouraged us to evaluate its usefulness in treatment of non-healing oral PV lesions. Seven patients with chronic oral PV, resistant to conventional therapy, were treated with weekly to monthly injections of PRP of affected mucosal membranes. All recruits reported improvement in pain and mastication and 6 of 7 patients had an improvement in pemphigus disease area index scores with PRP treatment. PRP injections seems to accelerate the healing process and decrease the pain and eating discomfort associated with the oral erosions and ulcers induced by PV.

Comparison of vascular endothelial growth factor expression between lesional and non-lesional skin in patients with morphea: a dermoscopy-guided immunohistochemical study.

Morphea is an inflammatory fibrosing disease, initiated by vascular injury resulting in increased collagen formation and decreased collagen degradation. This study was designed to evaluate the role of angiogenic vascular endothelial growth factor (VEGF) in the vascular changes which are dermoscopically evident in morphea lesions, compared with that in non-lesional skin, by assessing its expression immunohistochemically on tissue blood vessels. Twenty patients with morphea were subjected to clinical and dermoscopic examinations. Two skin biopsies from lesional and non-lesional skin were obtained and stained with hematoxylin and eosin (H&E) and immunohistochemically with VEGF. Dermoscopic examination showed linear blood vessels in 90% of the lesions. No significant difference in the number of VEGF-stained and unstained blood vessels, was observed between the lesional and non-lesional skin (p = 0.475 and 0.191, respectively). A weak inverse correlation was found between the total number of blood vessels positive for VEGF and the disease duration, (r = - 0.48; p = 0.032). Significant differences were found between different stages of morphea and total number of blood vessels negative for VEGF, (p = 0.017). In conclusion, VEGF immunostaining, which represents the newly formed blood vessels, showed no difference between lesional and non-lesional skin in patients with morphea. Thus, the dermoscopically observable blood vessels in lesions compared with non-lesional skin are not due to angiogenesis, but rather due to the thinning and atrophy of the overlying epidermis in morphea cases, rendering the blood vessels more obvious.

Upregulation of Anti-Desmocollin 3 Antibodies in Pemphigus Diseases: A Case-control Study.

BACKGROUND: Pemphigus diseases are a subgroup of autoimmune bullous diseases characterized by autoantibodies against desmogleins and occasionally desmocollins. Desmocollin 3 is the main desmocollin isoform that contributes to cell adhesion in the epidermis. OBJECTIVE: To evaluate the presence and level of anti-desmocollin 3 antibodies in pemphigus diseases, and to investigate whether their presence is associated with a specific type, presentation, or clinical pattern. METHODS: Forty patients with pemphigus diseases and forty healthy controls were enrolled. Medical history, clinical examination, and pemphigus disease area index (PDAI) scoring were recorded for all patients. Serum samples were collected from both groups for assessment of anti-desmocollin 3 antibody reactivity by ELISA. RESULTS: The presence of anti-desmocollin 3 antibodies was significant among patients with pemphigus compared with controls (P=0.003). The level of anti-desmocollin 3 antibodies was also significantly higher in patients with pemphigus compared with controls (P=0.01). There was no significant relationship between the presence of anti-desmocollin 3 antibodies and any of the clinical presentations of pemphigus (type, severity, duration, activity, presence of annular pattern, or site of affection - mucosal, cutaneous, on the scalp, palmoplantar, or flexural). CONCLUSION: Anti-desmocollin 3 antibodies are upregulated in pemphigus diseases and can contribute to the pathogenesis of pemphigus. No specific clinical type, presentation, or pattern was found to be associated with the presence of anti-desmocollin 3 antibodies.

Serum and tissue levels of coenzyme Q10 in pemphigus vulgaris.

BACKGROUND: Pemphigus vulgaris (PV) is a debilitating autoimmune blistering disease of the skin and mucous membranes. It occurs due to the action of autoantibodies against various keratinocyte self-antigens. Anti-mitochondrial autoantibodies are detected in patients with PV. Coenzyme Q10 (CoQ10) is a member of the mitochondrial respiratory chain involved in cellular metabolism, including apoptosis. This study aimed to assess the serum and tissue levels of CoQ10 of patients with PV and healthy controls to determine its relevance to the disease pathogenesis. METHODS: In this case-control study, 20 patients with PV and 20 healthy controls were included. Blood and skin samples were collected for the measurement of CoQ10 levels using ELISA. RESULTS: CoQ10 was significantly lower in both serum and tissue of patients with PV compared with controls (p = 0.001). Similar results were found when gender subgroups were separately compared. A significant positive correlation was found between serum and tissue CoQ10 levels in controls (p = 0.019, r = 0.521), but not in patients with PV. CONCLUSION: CoQ10 appears to be one of the parameters affected by the autoimmune response in PV, which may contribute to the tissue damage caused by autoantibodies. The absence of a significant correlation between CoQ10 level and disease severity or duration may be caused by the complex pathophysiological process in PV with multiple autoantibodies against different keratinocyte antigens.

Assessment of tissue E-cadherin and its proteolytic serum fragment in pemphigus vulgaris before and after remission: A case-control study.

BACKGROUND: E-cadherin is a classic cadherin that mediates keratinocyte adhesion. AIMS: To assess the tissue expression of E-cadherin and its proteolytic serum fragment (soluble E-cadherin) in pemphigus vulgaris (PV) before and after clinical remission compared with controls. PATIENTS: Thirty-seven PV patients and thirty controls were enrolled. Pemphigus disease area index (PDAI) was calculated for patients at baseline and after remission. Punch biopsy specimens were taken from patients before, and after remission, and from controls for assessment of tissue E-cadherin by immunofluorescence. Similarly, serum samples were collected for assessment of serum soluble E-cadherin by ELISA. RESULTS: Presence, intensity, and mean intensity of tissue E-cadherin were significantly reduced in PV patients before treatment compared with controls (p < 0.001). Detected E-cadherin showed mainly a basal and suprabasal distribution with cell surface and a cytoplasmic expression. Serum E-cadherin was significantly higher in patients before treatment compared with controls (p = 0.006). With remission, tissue E-cadherin presence, intensity, mean intensity, and serum E-cadherin showed statistically significant improvement (p = 0.003, <0.001, <0.001, and 0.003 respectively). Tissue E-cadherin presence and serum E-cadherin level reached values equivalent to the controls (p = 0.49 and 0.44, respectively). CONCLUSIONS: Disruption of tissue E-cadherin and upregulation of serum soluble E-cadherin can contribute to the pathogenesis of PV. Clinical remission of PV is associated with normalization of tissue and serum E-cadherin.

How Do Experts Treat Patients with Bullous Pemphigoid around the World? An International Survey.

Many treatments are currently proposed for treating patients with bullous pemphigoid (BP). We assessed treatment modalities of BP depending on the different countries, BP extent, and patients' comorbidities. We surveyed worldwide experts about how they treat patients with BP. A total of 61 experts from 27 countries completed the survey. Severe and moderate BP were treated with oral prednisone (61.4 and 53.7%, respectively) or superpotent topical corticosteroids (CSs) (38.6 and 46.3%, respectively). Conventional immunosuppressants were more frequently combined with oral prednisone (74.5%) than with superpotent topical CS (37.5%) in severe BP. Topical CSs were mainly used in Europe in mild (81.1%), moderate (55.3%), and severe (54.3%) BP. In the United States of America and Asia, systemic CSs were mainly proposed for treating severe (77.8 and 100%, respectively), moderate (70 and 77.8%, respectively), and also mild (47.1 and 33.3%, respectively) BP. Most experts reduced the initial dose of oral CS in patients with diabetes mellitus (48.1%) or cardiac insufficiency (40.2%) but rarely changed BP treatment in patients with neurological disorders or neoplasia. This survey showed major differences in the way patients with BP are treated between AmeriPac countries (United State of America, Latin America, and Australia) and Asia on the one hand and Europe and the Middle East on the other hand.

Serum levels of soluble PD1 in pemphigus vulgaris: A useful marker for disease severity.

BACKGROUND: Pemphigus vulgaris (PV) is a life-threatening autoimmune blistering disease targeting the skin and mucous membranes. Programmed cell death protein 1 (PD1) is an immune checkpoint which binds to two ligands, PDL1 and PDL2 resulting in negative regulation of antigen receptor signaling, thus, play a critical role in the immune regulation of autoimmune diseases. AIM: In this work we aimed to assess serum levels of soluble PD1 (sPD1) in patients with active PV and in patients in remission in an attempt to evaluate its effect on disease severity. METHODS: In this case-control study, 60 pemphigus vulgaris patients (30 clinically active and 30 in a clinical remission) and 30 age matched healthy control subjects were included. Severity of PV was assessed using pemphigus disease area index (PDAI) score. Serum levels of sPD1 were measured by ELISA for both patients and healthy control. RESULTS: Serum levels of sPD1 were significantly lower in PV patients than in controls (P < .001) and significantly lower in patients with active disease than in those in remission (P < .001). Serum sPD1 correlated negatively with the severity of the disease (P < .001, r = -0.4). CONCLUSION: A defect in PD1 pathway is suggested in PV patients, and this defect plays a substantial role in determining the severity of the disease. Thus, sPD1 could be considered a useful marker for disease severity and targeting PD1 pathway could be a potential aim for future therapies of PV.

Evaluation of the pathogenicity of anti-desmoglein antibodies of pemphigus vulgaris patients using human organ culture assay.

Anti-desmoglein (anti-Dsg) antibodies are key players in the pathogenesis of pemphigus vulgaris (PV) disease. We aimed to evaluate the pathogenicity of anti-Dsg antibodies of PV patients using human organ culture assay and to assess the correlation between the pathogenicity and the disease score. In this cross-sectional study, sera from 37 PV patients were included. The organ culture acantholysis index (OCAI) was calculated as (width of blister/total width of the specimen) × 100. The sera were analyzed using Dsg ELISAs and ethylenediaminetetraacetic acid (EDTA) treated ELISAs. OCAI ranged from 0 to 100%, median = 50%. There was a moderate significant correlation between OCAI and the disease severity, r = 0.503, p = 0.002. There was a moderate significant correlation between OCAI and non-Ca2+-dependent anti-Dsg3 and anti-Dsg1 antibodies, p values were 0.01 and 0.021, respectively. The OCAI was assessed along the disease time course of four patients. The OCAI fluctuated parallel to the disease severity along the time course of the four patients. Human organ culture assay is an objective tool that detects the pathogenicity of anti-desmoglein antibodies. It can be used before stopping systemic steroids especially in patients in remission with high titer or if the Dsg ELISA is not available.

Assessment of the quality of life of Egyptian and Tunisian autoimmune bullous diseases' patients using an Arabic version of the autoimmune bullous disease quality of life and the treatment of autoimmune bullous disease quality of life questionnaires.

BACKGROUND: The Autoimmune Bullous Disease Quality of Life (ABQOL) and the Treatment of Autoimmune Bullous Disease Quality of Life (TABQOL) questionnaires proved to be reliable tools that measure the disease and treatment burden. OBJECTIVES: We aimed to assess the ABQOL and TABQOL in the Arabic population. METHODS: The English questionnaires were translated into the Arabic language by a certified translation agency. Eighty autoimmune bullous disease (AIBD) patients were included in this study. Patients were asked to answer 2 questionnaires. After 1 week the same patients were asked to answer the same questionnaires again. RESULTS: The age of the patients ranged from 19 to 81 years (mean=46), 19 males, 61 females. The ABQOL ranged from 0-37 (mean=16.4±9.2). The TABQOL ranged from 2-43 (mean=21.5±9.4). Test-retest reliability was acceptable, Cronbach's alpha was 0.76 for ABQOL and 0.74 for TABQOL. There was no significant correlation between the age of the patients and ABQOL, r =-0.2, p value was 0.183. There was a significant negative correlation between the age of the patients and the TABQOL, r=-0.2, p value was 0.039. There was a significant negative correlation between the education of the patients and the TABQOL, r=-0.3, p value was 0.007. STUDY LIMITATIONS: Small sample size of some AIBDs and patients with severe disease. CONCLUSION: Objective and valuable measurements such as ABQOL and TABQOL are now available to help physicians understand their patient's distress and should be used in every patient with AIBD. Younger and less educated patients appear to have more effects on their QOL from the treatments.

Alteration of serum and tissue tumor necrosis factor alpha levels: A possible mechanism of action of oral pulse steroids in the treatment of alopecia areata.

BACKGROUND: Alopecia areata (AA) is a multifactorial disease in which tumor necrosis factor alpha (TNF-α) plays an important role. OBJECTIVE: To study the effect of oral pulse steroids on both serum and tissue levels of TNF-α in AA patients. METHODS: Skin biopsies and serum samples were collected from 20 patients with patchy AA before and after treatment (oral prednisolone for two consecutive days every week for 3 months) for determination of the levels of TNF-α levels using ELISA technique. RESULTS: Both serum and tissue levels of TNF-α in AA patients were significantly higher than in controls before (P < 0.001) as well as after treatment (P = 0.0169 and P = 0.3051), respectively. The duration of disease negatively correlated with tissue TNF-α before treatment (P < 0.0001). Serum and tissue levels of TNF-α dropped significantly after treatment (P < 0.0001). The percentage of reduction of both tissue and serum TNF-α levels correlated positively with the percentage of clinical improvement (r = 0.682, P = 0.0009; r = 0.567, P = 0.009, respectively). CONCLUSION: TNF-α plays an important role in the evolution of AA lesions, and alteration in both serum and tissue levels of TNF-α could be considered one of the important mechanisms of action of systemic oral pulse steroids in the treatment of AA.

Downregulation of aquaporin 3 in bullous pemphigoid patients.

Bullous pemphigoid (BP) is a chronic autoimmune skin disease. Aquaporin 3 (AQP 3) has a possible role in the pathogenesis of many dermatological diseases. In this work, we aimed to evaluate the expression of AQP 3 in BP. Perilesional skin biopsies were taken from 24 BP patients and 13 controls. The biopsies were stained by direct immunofluorescence using rabbit anti-human AQP 3 FITC antibody. The expression of AQP 3 was weak in 5 patients (20.8%), moderate in 18 patients (75%), strong in 1 patient (4.2%) in the suprabasal layers. It was negative in 4 patients (16.7%), weak in 18 patients (75%), moderate in 2 patients (8.3%) and no strong fluorescence was seen in the basal layers. In the controls, the expression was strong in ten controls (76.9%), moderate in three controls (23.1%) and no controls showed weak fluorescence in the suprabasal layer. The basal layer showed strong fluorescence in 11 controls (84.6%), moderate in 2 controls (15.4%) and no controls showed mild or no fluorescence. There was a statistically significant difference in the expression of AQP 3 between basal and suprabasal layers of BP patients but not of the controls. There was statistically significant difference in the expression of AQP 3 between patients and controls in both the basal (P value < 0.001) and the suprabasal layers (P value < 0.001). In conclusion, AQP 3 was downregulated in BP patients especially in the basal cell layer. This suggests that AQP 3 plays a role in the pathogenesis of BP.

Autologous platelet-rich plasma and triamcinolone acetonide intralesional injection in the treatment of oral erosions of pemphigus vulgaris: a pilot study.

Oral erosions of pemphigus vulgaris (PV) represent a therapeutic challenge. In this work we compared intralesional injection of steroids (ILS) and autologous platelet-rich plasma (PRP) treatment of oral erosions of PV. In this split-mouth randomized double blind study, one side of the buccal mucosa of 11 PV patients was injected with 10 mg/ml triamcinolone and the opposite side was injected with 1 millilitre of autologous PRP every 14 days for 3 months. The percentage of clinical improvement, pemphigus disease area index and pain score were recorded before and after therapy. Out of 11 recruited patients, only 9 completed the study. Seven out of the nine patients (78%) showed improvement in PDAI and/or pain score. The mean oral PDAI of the steroid injected sides decreased from 2.3 to 0.9. The mean oral PDAI of the PRP injected sides decreased from 2.6 to 1.0. No significant difference was detected in clinical improvement, PDAI and pain score between autologous PRP and ILS in the treatment of oral erosions of PV. Autologous PRP can be used for the treatment of resistant oral erosions of PV when ILS is contraindicated.