Desmosomal Glycoprotein Research Articles

Revisiting pemphigus vulgaris: A case report and review of literature

Introduction: Pemphigus is an autoimmune blistering mucocutaneous disease. Production of auto-antibodies against desmosomal glycoproteins is a characteristic feature of the disease. Oral lesions of pemphigus vulgaris are many a times first sign of the disease and hence it is important for a dentist to be familiar with the clinical appearance, diagnosis, and treatment modalities. Case Report: This report describes a case of pemphigus vulgaris in a female patient who presented with ulcers on the gingiva. On the basis of clinical examination and histopathology, we arrived at a diagnosis of pemphigus vulgaris, which was confirmed by immunofluorescence. Conclusion: If left undiagnosed or untreated pemphigus vulgaris may be fatal. Early diagnosis will aid in appropriate treatment and better outcome of the disease.

An observational study to determine the role of indirect immunofluorescence and ELISA for desmogleins in the diagnosis and monitoring of autoimmune bullous disorders

BackgroundAutoimmune bullous disorder (AIBD) is a diverse group of blistering dermatoses that affects the skin and mucous membrane, characterized by the formation of autoantibodies against the desmosomal glycoproteins and adhesion molecular components of the basement membrane zone. Various immunoassay techniques for serological diagnosis are Direct Immunofluorescence (DIF), Indirect Immunofluorescence (IIF), Enzyme Linked Immunosorbent Assay (ELISA) and immunoblotting. Quantitative ELISA titer can also be used to monitor the disease activity and response to treatment. The aim of this study was to evaluate the role of IIF in diagnosing various AIBDs and the role of ELISA for desmogleins in monitoring disease activity in the pemphigus group of disorders. MethodsA total of 45 patients with freshly diagnosed AIBD were included in the study. Skin biopsies were done to establish the diagnosis by histopathology and DIF. BIOCHIP-based IIF assay was performed on the sera of the patient. The sensitivity and specificity of the IIF assay were then calculated based on the established diagnosis by Histopathological examination (HPE) and DIF. Quantitative ELISA titer was performed to measure the antibody (anti desmoglein (Dsg) 1 and 3) titre in serum samples of patients with pemphigus at baseline, 1 and 3 months follow-up after initiation of appropriate treatment therapy and results were then compared with disease activity scoring system. ResultsSensitivity and specificity of IIF for Dsg 1 for the pemphigus group were derived to be 85.7% and 98.2% respectively, while that for Dsg 3 were 100% and 98.7%, respectively. Sensitivity (90–100%) of IIF for Bullous pemphigoid was higher than specificity (85–96%). The ELISA titers of anti Dsg1 and 3 were also observed to fall sequentially so was the disease activity score during the follow-up visits at 1 and 3 months. ConclusionBiochip IIF assay can be used as a screening tool for the serological diagnosis of AIBD and quantitative ELISA for monitoring the disease activity and response to treatment.

Pemphigus Vulgaris- A Case Report

Introduction: Pemphigus is a rare and life-threatening autoimmune disease characterized by blisters and erosion of the skin and mucous membranes throughout the entire body. It mostly affects the mouth, eye, nose, throat as well as genitals. Pemphigus vulgaris is most common type of pemphigus. The epithelial lesions are caused by autoantibodies reacting with desmosomal glycoproteins found on the keratinocyte's cell surface. The binding of immunoglobulin G autoantibodies to desmoglein 3, a transmembrane glycoprotein adhesion molecule found on desmosomes, is the underlying process that causes intraepithelial lesions.
 Case Presentation: A 40-year-old male patient came to the hospital with a complaint of a mouth ulcer and a genital lesion that had been since 15 years. He is having a history of oral ulcer in oral mucosa and involvement of genital area in glans penis with a history of pain, bleeding, difficulty in swallowing. 
 Intervention: The patient was admitted to the hospital on 29/07/2021 and taken Inj. Rituximab 1gm in 500Ml Normal Saline in infusion pumps in over 6 Hours.
 Conclusion: In this case report, we mainly focus on expert dermatological management and excellent nursing care in managing the rare complicated case nicely. Early diagnosis and treatment of pemphigus Vulgaris help determine the course of the disease of the patient and is done by a dermatologist.

Oral Pemphigus Vulgaris with Cutaneous Manifestations: A Case Report

Pemphigus is a life threatening mucocutaneous disease characterized by production of auto-antibodies against desmosomal glycoproteins. This results in the formation of intra epithelial blisters. The etiology of this disease remains obscure, although the presence of autoantibodies confirms autoimmune origin. The initial manifestation is frequently the development of intraoral lesions, and later, may involve other mucous membranes and skin. As oral lesions are first to appear, dental professionals must be well acquainted with the clinical presentation of pemphigus vulgaris to ensure timely diagnosis and appropriate treatment. In this article, we report a case of 54- year-old male, who was diagnosed with pemphigus vulgaris.

Association of NFKB1 -94ATTG ins/del polymorphism (rs28362491) with pemphigus vulgaris.

Pemphigus vulgaris is a rare chronic blistering skin disease resulting from IgG autoantibodies directed against transmembrane desmosomal glycoprotein desmoglein 3 and is the most common form of pemphigus. Since interleukin-1 receptor-associated kinase (IRAK-1)/nuclear factor-kappa B (NF-kappa B) pathway plays an essential role in the pathogenesis of autoimmune diseases, the aim of the present study was to explore the role of polymorphisms in three genes, named IRAK1 (rs3027898), NFKBIA (rs696) and NFKB1 (-94ATTG insertion/deletion variant, - rs28362491), in PV susceptibility. Forty-four unrelated patients with PV (23 males) were enrolled in the study. Additionally, 77 ethnic matching healthy volunteers (45 males) with no personal or family history of chronic autoimmune or infectious diseases were studied. Strong statistical significant difference was observed between PV patients and controls for polymorphism -94 insertion/deletion ATTG in the promoter region of NFKB1 gene (P=0.00005). Additional dedicated studies in larger groups of patients of various ethnicities are needed to replicate and confirm the preliminary findings.

Oral Pemphigus Vulgaris: Case Report

Pemphigus is a potentially life threatening autoimmune disease that causes blisters and erosions of the skin and the mucous membrane. The epithelial lesions are a result of auto-antibodies that react with desmosomal glycoproteins that are present on the cell surface of the keratinocyte. The autoimmune reaction against these glycoproteins causes a loss of cell to cell adhesion, resulting in the formation of intraepithelial bullae. Eighty to ninety percent of patients with pemphigus vulgaris develop oral lesions and in 60% of cases oral lesions are the first sign. Timely recognition and therapy of oral lesion is critical as it may prevent skin involvement. If treatment is instituted during this time, the disease is easier to control and the chance for an early remission of the disorder is enhanced. This case report describes the case of a patient who complained of ulcers of the mouth and difficulty in swallowing since 20 days, who was diagnosed as having Pemphigus vulgaris. Due to early diagnosis, lower doses of medication for a shorter period of time could control the disease. Dental professionals must be sufficiently familiar with the clinical manifestations of pemphigus vulgaris to ensure early diagnosis and treatment which in turn determines the prognosis and course of the disease.

Usefulness of desmoglein 1 and 3 in serodiagnosis of pemphigus vulgaris and its correlation with disease activity - ELISA study

Background: Pemphigus is an autoimmune blistering disease with three subtypes, which includes pemphigus vulgaris (PV), pemphigus foliaceus (PF) and paraneoplastic pemphigus (PP), which are characterized by circulating autoantibodies to the desmosomal glycoproteins desmoglein 3 (Dsg 3) and Dsg 1. Detection and quantification of these antibodies can be useful in diagnosis of PV. Aims: The purpose of this study was to evaluate the practical application of ELISAs for diagnosis of PV based on immunological reactivity of Dsg 1 and Dsg3. Materials and Methods: Based on clinical presentation and histopathologic confirmation, 10 patients previously diagnosed of PV were included in the study, out of which, five were with active lesions and five were under medication and hence were at remission stages. Sera of the patients were tested for Dsg 1 and Dsg 3 titres by Desmoglien enzyme-linked immunosorbent assay (ELISA) test. Results: Desmoglien ELISA was positive in all the patients, with higher values of Dsg antibody titers in patients with active disease. Dsg 3 titers exceeded the cut off value in patients with oral lesions and titers of both Dsg 1 and Dsg 3 were greater than cut value in patients with mucocutaneous involvement. Conclusion: Thus, the Dsg 1 and Dsg 3 provided objective and quantitative data which allowed differentiation of PV, and in view of these advantages; they are likely to become a routine technique in diagnostic laboratories.

IgG Autoantibody Response against Keratinocyte Cadherins in Endemic Pemphigus Foliaceus (Fogo Selvagem)

It is well established that autoantibodies against desmoglein 3 and desmoglein 1 are relevant in the pathogenesis of pemphigus vulgaris and pemphigus foliaceus, including its endemic form, Fogo Selvagem (FS). Isolated reports have shown that in certain patients with these diseases, autoantibodies against other desmosomal cadherins and E-cadherin may also be present. The goal of this investigation was to determine if FS patients and normal individuals living in endemic areas possess autoantibodies against other desmosomal cadherins and E-cadherin. Testing a large number of FS and endemic control sera by ELISA we find a consistent and specific autoantibody response against desmoglein 1 and other keratinocyte cadherins in these individuals, which is quite different from US controls. Overall, the highest correlations among the autoantibody responses tested are in the endemic controls, followed by FS patients, and lowest in the US controls. These findings suggest that multiple, perhaps cross reactive, keratinocyte cadherins are recognized by FS patients and endemic controls.

Profile of Trypanosoma cruzi Reactivity in a Population at High Risk for Endemic Pemphigus Foliaceus (Fogo Selvagem)

Fogo selvagem (FS) is an autoimmune bullous disease with pathogenic IgG autoantibodies recognizing desmoglein 1 (Dsg1), a desmosomal glycoprotein. In certain settlements of Brazil, a high prevalence of FS (3%) is reported, suggesting environmental factors as triggers of the autoimmune response. Healthy individuals from endemic areas recognize nonpathogenic epitopes of Dsg1, and exposure to hematophagous insects is a risk factor for FS. Fogo selvagem and Chagas disease share some geographic sites, and anti-Dsg1 has been detected in Chagas patients. Indeterminate Chagas disease was identified in a Brazilian Amerindian population of high risk for FS. In counterpart, none of the FS patients living in the same geographic region showed reactivity against Trypanosoma cruzi. The profile of anti-Dsg1 antibodies showed positive results in 15 of 40 FS sera and in 33 of 150 sera from healthy individuals from endemic FS sites, and no cross-reactivity between Chagas disease and FS was observed.

Rôle des infections à staphylocoques dans la survenue de pemphigus endémique tunisien

Pemphigus is a severe autoimmune blistering disorder, in which immunoglobulin G autoantibodies are directed against desmosomal glycoproteins. The antibodies against desmogleine1 are prevalent in normal subjects living in Tunisia, that suggests the role of environment in the pathogenesis of this endemic type of pemphigus and the need for additional factors to switch from a subclinical form to a clinical form of the disease. The aim of this study was to investigate explanatory environmental factors. We tested the hypothesis that normal subjects whom presented staphylococcal infections have antibodies against desmoglein. We recruited 29 patients with staphylococcal infections from the dermatology department of the university hospital of Tunis. 29 controls healthy subjects are used to compare the prevalence of anti-desmoglein1 and 3 between these two groups. We used a specific and sensitive enzyme linked immunosorbent assay to detect autoantibodies against Dsg1. (10.3% versus 7% in healthy Tunisian blood donors: p < 0.05). Conversely, we observed a significant association with antibodies against Dsg3 (20.78% versus 3% in healthy Tunisian blood donors: p < 0.05). Subclasses analysis of anti-desmoglein antibodies showed that they were almost exclusively the IgG 1, 2 and 3 subclasses in positive normal sera. The antibodies against desmogleins binding activity of these positive sera were confirmed by indirect immunofluorescence analysis and by immunobloting using human epidermal extract. In conclusion, we detected autoantibodies in the sera of patients with staphylococcal infections and found elevated levels of autoantibodies against Dsg 3.

Desmosomal molecules in and out of adhering junctions: normal and diseased States of epidermal, cardiac and mesenchymally derived cells.

Current cell biology textbooks mention only two kinds of cell-to-cell adhering junctions coated with the cytoplasmic plaques: the desmosomes (maculae adhaerentes), anchoring intermediate-sized filaments (IFs), and the actin microfilament-anchoring adherens junctions (AJs), including both punctate (puncta adhaerentia) and elongate (fasciae adhaerentes) structures. In addition, however, a series of other junction types has been identified and characterized which contain desmosomal molecules but do not fit the definition of desmosomes. Of these special cell-cell junctions containing desmosomal glycoproteins or proteins we review the composite junctions (areae compositae) connecting the cardiomyocytes of mature mammalian hearts and their importance in relation to human arrhythmogenic cardiomyopathies. We also emphasize the various plakophilin-2-positive plaques in AJs (coniunctiones adhaerentes) connecting proliferatively active mesenchymally-derived cells, including interstitial cells of the heart and several soft tissue tumor cell types. Moreover, desmoplakin has also been recognized as a constituent of the plaques of the complexus adhaerentes connecting certain lymphatic endothelial cells. Finally, we emphasize the occurrence of the desmosomal transmembrane glycoprotein, desmoglein Dsg2, out of the context of any junction as dispersed cell surface molecules in certain types of melanoma cells and melanocytes. This broadening of our knowledge on the diversity of AJ structures indicates that it may still be too premature to close the textbook chapters on cell-cell junctions.

Recognition, calcium and the control of desmosome formation.

Since desmosome formation requires the participation of two adjacent cells, a crucial initiating event must be recognition between desmosomal adhesion molecules. Studies of mutual desmosome formation between different cell types suggest that the recognition mechanisms are highly conserved between different tissues and different species of animals. A further requirement for desmosome formation is an adequate extracellular concentration of Ca2+ (greater than 0.1 mM). Keratinocytes, MDCK cells and MDBK cells all show Ca2+-induced desmosome formation. The desmosomes of these cells also show variable stability to reduction in [Ca2+] and Ca2+ chelation. Desmosome formation at low [Ca2+] is triggered by tunicamycin in keratinocytes, suggesting that the carbohydrate moieties of desmosomal glycoproteins may be involved in the Ca2+ control mechanism. The desmosomal glycoproteins appear to bind Ca2+, while the desmosomal adhesion molecules known as desmocollins, like other Ca2+-dependent adhesion molecules, yield a soluble fragment on trypsinization in the presence of Ca2+. For desmocollins the soluble fragment has a relative molecular mass of 42,000.

Developmental potential of bovine oocytes cultured in different maturation and culture conditions

Diverse groups of chemicals in culture media are needed for successful bovine oocyte maturation and embryo development during which dramatic cytoplasmic and nuclear reprogramming events take place. In vitro embryo production (IVP) procedures frequently include supplements such as serum and/or co-culture with various types of somatic cells. However, the presence of undefined serum in culture media introduces a variation from batch to batch, increases viral or prion contamination risk, and leads to problems during fetal development. The aim of the present study was to investigate the possibility of using chemically defined-synthetic serum substitute (SSS) in place of fetal calf serum (FCS) during maturation and long-term culture to stimulate in vitro maturation (IVM), fertilization (IVF) and subsequent embryo development. In Experiment I, the effect of the protein source on in vitro maturation was tested by maturing oocytes in culture media supplemented with 10% FCS (Control Group), 10% SSS (Group I) and 10% SSS + 10 ng/ml epidermal growth factor (EGF) (Group II). In Experiment II, effects of SSS on both oocyte maturation and embryo development during in vitro culture (IVC) were tested by maturing oocytes in media supplemented with 10% FCS (FCS Group) or 10% SSS + 10 ng/ml EGF (SSS Group), followed by IVF and IVC in SOF media supplemented with 10% FCS and 10% SSS on day 4 for FCS and SSS Groups, respectively. Even though rates for cleavage and development to blastocyst stage were not different, blastocyst cell numbers were higher in Group II containing SSS and EGF. The SSS supplementation group had higher apoptotic nuclei as compared to the FCS Group in Experiment II. Transcripts for heat shock protein 70 (Hsp70), interferon tau (IF-τ), DNA methyltransferase 3a (Dnmt3a), desmosomal glycoprotein desmocollin III (DcIII) and insulin-like growth factor II receptor (Igf-2r) were altered in different culture conditions in Experiment I. However, only glucose transporter-1 (Glut-1) mRNA was different in the SSS and FCS Groups in the second experiment. In summary, SSS and EGF in maturation medium and replacement of FCS with SSS alone in culture medium on day 4 of IVC support oocyte maturation and embryo development in vitro. However, significance of culture condition induced changes on the genome-wide abundance of messenger ribonucleic acid and the significance of the apoptotic nuclei during fetal development still remain to be determined.

Ionizing radiation–induced pemphigus foliaceus

To the Editor: Pemphigus foliaceus (PF) is a chronic autoimmune bullous disorder, with primarily IgG4 subclass autoantibodies against desmoglein-1, a 106 kDa desmosomal glycoprotein of the cadherin family, which has been described in association with ultraviolet (UV) and ionizing radiation. Reports of PF in association with ionizing radiation are uncommon. A 92-year-old female with a history of intra-ductal carcinoma in situ of the left breast underwent wide local excision followed by radiation with 5040 cGy without complication.

Antibodies against desmoglein 1 in healthy subjects in endemic and nonendemic areas of pemphigus foliaceus (fogo selvagem) in Peru

Endemic pemphigus foliaceus or fogo selvagem is an autoimmune skin disease characterized by the presence of subcorneal superficial blisters and antibodies of the immunoglobulin G4 (IgG4) class specific for the desmosomal glycoprotein, desmoglein 1. In Peru, no studies have been published on the seroprevalence of antibodies against desmoglein 1 in healthy subjects from endemic foci. This was a cross-sectional study. The sample included 82 healthy subjects, 41 from the Pueblo Libre community, a focus of endemic pemphigus foliaceus, and 41 from a nonendemic urban area in Pucallpa City. Enzyme-linked immunosorbent assay (ELISA) was used to determine the presence of antibodies against desmoglein 1. Samples were processed and tested at the Department of Dermatology and Cutaneous Surgery, School of Medicine, University of Miami, Miami, Florida. It was found that 31.7% of healthy individuals (13 subjects) from the endemic focus had anti-desmoglein 1 antibodies. A statistically significant association was found between the distance from the endemic focus and the presence of antibodies against desmoglein 1 in subjects living within the endemic focus [Mantel-Haenszel odds ratio (OR), 3.34; P = 0.03; 95% confidence interval (CI), 1.06-10.48]. Agriculture as an occupation showed a statistically significant association with the presence of antibodies against desmoglein 1 (Mantel-Haenszel OR, 7.84; P < 0.001; 95% CI, 2.47-24.87). Antibodies against desmoglein 1 are present in healthy subjects exposed to an endemic focus of pemphigus foliaceus (fogo selvagem). Agriculture is associated with a high risk of development of antibodies against desmoglein 1 in the endemic focus of the Pueblo Libre community.

265 DEVELOPMENT AND DYNAMICS OF GENE EXPRESSION OF BOVINE EMBRYOS CULTURED IN VITRO IN THREE DIFFERENT MEDIA

Dramatic reprogramming of gene expression occurs during embryonic genome activation (EGA), an essential event initiating as early as the 1-cell zygotic stage in the bovine and increasing gradually as embryonic development advances. It is this reprogramming of gene expression that sets the stage for later development. Expression of embryonic genes is altered in different culture conditions and this may influence developmental potential both during pre-implantation and during fetal development. The objective of this study was to define some most commonly used embryo culture media (KSOMaa, CR1aa, and SOFaa) based on their ability to support embryonic development to the blastocyst stage, mean cell number, percentages of apoptotic cells, and the expression patterns of a panel of developmentally important genes. Oocytes with several layers of cumulus cells obtained from an abattoir were matured in TCM 199 (supplemented with 0.25 mM pyruvate, 0.5 μg/mL FSH, 5 μg/mL LH, 100 U/mL penicillin, 100 μg/mL streptomycin, and 10% FCS) for 24 h and in vitro-fertilized (Day 0) using frozen bull semen. Presumptive zygotes were transferred into three different media (KSOMaa, CR1aa, and SOFaa) 16–18 h post-insemination, supplemented with 10% FCS on Day 4, and cultured until Day 8 at which time they were fixed or frozen for further analysis. Mean cell numbers and percentages of apoptotic cells in blastocysts were determined using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL). Real-time quantitative PCR was performed to assess gene transcripts of glucose transporter-1 (Glut-1), heat shock protein 70.1 (Hsp70), interferon-tau (IF-tau), insulin-like growth factor II receptor (Igf-2r), desmosomal glycoprotein desmocollin III (DcIII), and DNA methyltransferase 3a (Dnmt3a). Gene expression data were analyzed relative to transcripts of housekeeping gene glyceraldehyde 3-phosphate dehydrogenase (Gapdh). In three separate trials, a total of 538, 518, and 503 oocytes were used for KSOMaa, CR1aa, and SOFaa groups, respectively. Cleavage rates were 79.2%, 77.5%, and 80.2%; and rates of development to the blastocyst stage were 22.2%, 23.4%, and 32.9% for KSOMaa, CR1aa, and SOFaa groups, respectively. The blastocyst rate of the SOFaa group was significantly higher than those of the KSOMaa and CR1aa groups (P &lt; 0.05). Mean cell numbers were 109.3, 101.0, and 114.0; and the percentages of apoptotic cell numbers per blastocyst were 1.25, 1.91, and 1.87 for KSOMaa, CR1aa, and SOFaa groups, respectively. There was no difference among groups in terms of mean cell numbers and percentages of apoptotic cells per blastocyst. The expressions of Glut-1 and DcIII genes did not differ among the groups. However, expressions of Hsp70, IF-tau, and Dnmt3a genes were all significantly up-regulated in the CR1aa group as compared to the SOFaa and KSOMaa groups (P &lt; 0.05). In conclusion, SOFaa supports higher development to the blastocyst stage than KSOMaa and CR1aa, and culture conditions influence gene expression.

166 EFFECTS OF LEPTIN AND IGF-1 ON PRE-IMPLANTATION DEVELOPMENT, DNA FRAGMENTATION, AND GENE EXPRESSION OF BOVINE EMBRYOS CULTURED IN VITRO

Adequate regulatory proteins, growth factors, and hormones in in vitro embryo culture systems are important for improving the quality of embryos to a level similar to that in vivo conditions. The objective of this study was to define the effects of leptin, insulin-like growth factor-1 (IGF-1), and their combination on embryonic development, apoptosis, and expression profiles of a panel of developmentally important genes. Presumptive zygotes (16–18 h post-insemination) were randomly assigned and cultured in control (no supplementation), 5 ng/mL leptin (Group I), 100 ng/mL IGF-1 (Group II), and 5 ng/mL leptin and 100 ng/mL IGF-1 (Group III), all supplemented with 10% FCS on Day 4. On Day 8, the embryos reaching blastocyst stage were randomly either fixed for determination of DNA-fragmented nuclei by using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) or frozen for real-time relative quantitative RT-PCR analysis. The RT-PCR was performed to assess gene transcripts of glucose transporter-1 (Glut-1), heat shock protein 70.1 (Hsp70.1), interferon tau (IF-tau), insulin-like growth factor II receptor (IGF-IIr), desmosomal glycoprotein desmocollin III (DcIII), and DNA methyltransferase 3a (Dnmt3a). A total of 349, 322, 347, and 360 zygotes were used for the control group and Groups I, II, and III, respectively. Data were analyzed with a randomized complete block design and arcsine square root transformation of the dependent variables consisting of four treatments and six replicates. Cleavage rates were 79.5, 84.2, 87.3, and 82.4% for the control group and Groups I, II, and III, respectively, and only Group II was different from the control (P &lt; 0.05). The percentages of embryos developed beyond the 8–16 cell stage were 44.2, 48.2, 49.0, and 50.7 for the control group and Groups I, II, and III, respectively, and Group III was different from the control (P &lt; 0.05). Percentages of blastocyst development were 26.7, 29.6, 31.5, and 29.8, and the mean blastocyst cell numbers were 96.6, 98.6, 104.4, and 104.1 for the control group and Groups I, II, and III, respectively. The percentage of nuclei with fragmented DNA were 4.2, 3.3, 2.5, and 1.9 for the control group and Groups I, II, and III, respectively. Addition of IGF-1 and/or combination with leptin (Groups II and III) decreased the number of nuclei with fragmented DNA (P &lt; 0.01) as compared to the control group. Although the expression of Glut1, DcIII, and Igf2r did not change among the groups, IF-tau and Dnmt3a were down-regulated in Group II. Hsp70 and IF-tau were up regulated in Group III. Results indicate that addition of IGF-I in culture media improved the cleavage rate; combination with leptin also improved the development rates to 8–16-cell-stage embryos, decreased the TUNEL-positive nuclei, and altered expression of some of the developmentally important genes.

Endemic pemphigus foliaceus (fogo selvagem) and pemphigus vulgaris: immunoglobulin G heterogeneity detected by indirect immunofluorescence

Pemphigus are autoimmune intraepidermal blistering diseases in which immunoglobulin G (IgG) autoantibodies are directed against desmosomal glycoproteins. The aim of this study was to determine the IgG subclass profile of endemic pemphigus foliaceus (fogo selvagem) and pemphigus vulgaris utilizing indirect immunofluorescence. Twenty-five patients with pemphigus vulgaris, 25 with endemic pemphigus foliaceus (fogo selvagem), and 25 healthy controls were analyzed by indirect immunofluorescence for circulating autoantibodies (total IgG and its subclasses). Our data revealed a significant correlation (P <.05) of disease activity and autoantibody levels in both forms of pemphigus, i.e., negative titers related to clinical remission, whereas positive results related to active disease. Immunoglobulin G subclass analysis in fogo selvagem demonstrated that in patients in remission, 56% showed positive immunoglobulin G4; in active disease, immunoglobulin G4 was the predominant subclass (100% positive in all cases). The IgG subclass profile in pemphigus vulgaris showed that in patients in remission, only 10% were positive for immunoglobulin G4; in active disease, positivity for immunoglobulin G4 was present in 78% to 88% of the cases. Subclass characterization of immunoglobulin G autoantibodies is a useful tool for pemphigus follow-up, since immunoglobulin G4 (IgG4) is the subclass that is closely related to recognition of pathogenic epitopes, and consequently with disease activity. Careful monitoring should be performed for fogo selvagem in clinical remission with a homogeneous IgG4 response, since this may indicate more frequent relapses.

Pathogenicity and Epitope Characteristics of Anti-Desmoglein-1 from Pemphigus Foliaceus Patients Expressing Only IgG1 Autoantibodies

Pemphigus foliaceus (PF) is an antibody-mediated autoimmune disorder with IgG1 and IgG4 as the predominant subclasses of autoantibodies against a desmosomal glycoprotein, desmoglein-1 (Dsg1). Previously, we found that the IgG4 anti-Dsg1 autoantibodies only recognize a conformational epitope(s), whereas the IgG1 autoantibodies recognize both conformational and linear epitopes but do not display pathogenicity in the passive transfer animal model. The purpose of this study was to analyze the epitopes recognized by autoanti-bodies from a subset of PF patients who only express anti-Dsg1 of the IgG1 isotype throughout the course of their diseases and to further characterize the pathogenicity of their IgG1 anti-Dsg1. We found that IgG1 auto-antibodies in this subset of PF patients, similar to IgG4 autoantibodies from other PF patients, are able to bind both human and mouse skin and induce the experimental PF in mice. Moreover, a detailed epitope mapping reveals that the conformational epitopes recognized by IgG1 autoantibodies from these PF patients are restricted to the first 161 amino acids of Dsg1, whereas the linear epitopes are spread throughout the entire ectodomain. In conclusion, our study reveals that the isotype of IgG does not necessarily determine the epitopes and pathogenicity of pemphigus autoantibodies.

Polymorphisms within the tumor necrosis factor and lymphotoxin-alpha genes and endemic pemphigus foliaceus--are there any associations?

The purpose of this study was to analyze the possible influence of the TNF and LTA loci polymorphisms on the susceptibility/resistance to endemic pemphigus foliaceus, also named fogo selvagem (FS), an autoimmune disease characterized by blisters due to acantholysis of the superficial-most epidermal cells. Autoantibodies, mainly of the IgG4 subclass, are directed against a desmosomal glycoprotein known as desmoglein 1. FS shares clinical, histological and immunological features with nonendemic pemphigus foliaceus. Most residents of the endemic regions do not develop the disease, and familial clustering has been documented, suggesting that host factors play a role in susceptibility. In fact, strong positive and negative associations with HLA class II genes have been reported. The TNF and LTA genes are located in the class III region of the Human Major Histocompatibility Complex. Their location, the function of their products, which are cytokines and pluripotent immunomodulators, as well as their genetic variability make them candidate genes for complex diseases with an altered immune response. A total of 162 patients and 191 controls were enrolled in this study. No significant associations were found with any one of the three LTA single nucleotide polymorphisms (SNP) analyzed (at nucleotides 249, 365, 720), nor with the TNF SNP located at positions -863 and -308. The frequency of allele TNF*238A was slightly decreased in patients (OR = 0.45). In conclusion, the results of this study indicate that genetic variability of the TNF and LTA genes does not play a major role in susceptibility/resistance to pemphigus foliaceus.