LIGIA MARIA ICHIMURA FUKUMORI
Índice h a partir de 2011
3
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Instituto Central, Hospital das Clínicas, Faculdade de Medicina
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article Correlation between Histopathological Findings and Results of Direct Immunofluorescence in the Diagnosis of Canine Localized Cutaneous Chronic Lupus Erythematosus(2017) ODAGUIRI, Juliana; AOKI, Valeria; FUKUMORI, Ligia Maria Ichimura; PERIGO, Alexandre Marques; MICHALANY, Nilceo Scwery; FONTANA, Isabella; LARSSON, Carlos EduardoBackground: Localized chronic cutaneous lupus erythematosus (CCLE), also known as discoid lupus erythematosus, is one of the most prevalent canine autoimmune skin diseases. Histopathology is considered the gold standard for the diagnosis of CCLE and the accuracy of which can be increased by use of direct immunofluorescence (DIF). This study aimed to investigate the fluorescence pattern revealed by DIF in cases of canine localized CCLE and to establish and compare its effectiveness with that obtained from histopathology. Materials, Methods & Results: Eleven dogs suspicious to localized CCLE, i.e., those animals that presented mucocutaneous lesions as erythema, leucoderma, erosive-ulcerative lesions, and loss of nasal planum architecture and its transition to the haired skin underwent medical physical and laboratory examinations (blood count, platelet count, determination of alanine transaminase, alkaline phosphatase, total protein, serum albumin, urea, creatinine). Only those animals that proved to be normal across both the physical and laboratorial evaluation were included in Group I. Animals belonging to this group were submitted to general anesthesia to biopsy two samples of lesioned skin from nasal planum to histopathologic examination and DIF test. Five dogs with no skin lesions were included in Group II as negative control to the DIF assay. Two samples of no lesioned skin from nasal planum were biopsied to histopathologic and DIF evaluation. The kappa (k) coefficient was used to determine the degree of agreement and reliability of the results of both tests. A P-value < 5% was considered to be statistically significant. In Group I, all animals were normal across both the physical and laboratorial evaluation. A diagnosis of canine CCLE was established in 81.8% (9/11) of the animals based on histopathology analysis. Two dogs, numbers 10 and 11, respectively, had histopathologic findings suggestive of cutaneous leishmaniasis and compatible with canine demodicosis. The DIF assay confirmed the diagnostic of canine CCLE in 100% (11/11) of the cases, including dogs numbers 10 and 11. Direct immunofluorescence positivity for canine localized CCLE was determined by the presence of homogeneous green fluorescence in the basement membrane zone, with IgM and C3 immunoreagents being found at a significantly higher frequency (P = 0.007) than IgA and IgG (P = 0.017 and P = 0.037, respectively). In Group II, no alterations from histopathologic examination and unspecific fluorescence from DIF reaction were shown in all animals belonging to this group. A substantial and significant degree of agreement or reliability (k = 0.738 and P = 0.002, respectively) was established between the results of both tests. Discussion: In veterinary medicine, there is a lack of studies about DIF around autoimmune dermatoses. This is the first manuscript comparing histopathological and direct immunofluorescence findings in the diagnostic of dogs with localized CCLE. In the present study, the positivity to the DIF assay for canine localized CCLE was established by the presence of green fluorescence of a homogeneous morphologic pattern, especially for IgM and C3, located in the BMZ and cutaneous annexes. Direct immunofluorescence excluded the suspicion of cutaneous leishmaniasis determined by the histopathological findings and confirmed the diagnostic of canine localized CCLE in the dog number 10. Statistically, a substantial degree of agreement and reliability was shown between DIF and histopathology. Therefore, we concluded that the DIF assay is a highly effective and useful complementary examination that improves our ability to diagnose canine CCLE.- How can immunohistochemistry improve the diagnosis of pemphigus foliaceus?(2018) MIYAMOTO, D.; MARUTA, C.; SANTI, C.; ZOROQUIAIN, P.; DIAS, A. B.; FUKUMORI, L.; PERIGO, A.; AOKI, V.; BURNIER, M.Purpose Pemphigus foliaceus (PF) is a rare, autoimmune blistering disorder characterized by the production of autoantibodies against desmoglein 1. The mainstay for diagnosis is the demonstration of immune complex deposition by direct immunofluorescence (DIF) in fresh tissue samples. Immunohistochemistry (IHC) recognizes autoantibodies in formalin-fixed paraffin-embedded specimens, but studies regarding its use in PF are scarce. This study aims to evaluate immunoglobulin and C3 deposition using IHC in patients with confirmed PF by DIF and indirect immunofluorescence (IIF). Material and methods Six biopsies obtained from five patients with PF and six healthy individuals were included in this study. Anti-C3c, -IgG, -IgM, and -IgA antibodies were used for DIF and automated IHC. After digitalizing the slides, staining was classified as negative (0) or positive (1 = mild/2 = intense). Results DIF revealed intraepidermal intercellular deposition of IgG and C3c (n = 6), without deposits in dermal structures. IHC was positive in the intercellular spaces between keratinocytes with anti-IgG (n = 6) and anti-C3c antibodies (n = 6); no intercellular immune complexes deposition was observed in healthy individuals. In patients with PF, inflammatory cells were tagged by anti-IgG and anti-C3c (n = 6), anti-IgM (n = 1), and anti-IgA (n = 1); and immune complexes at vessel walls were detected with anti-C3c, anti-IgG, anti-IgA (n = 6), and anti-IgM (n = 5) antibodies. Adnexal positivity occurred with anti-C3c and anti-IgG (n = 6), anti-IgM (n = 1), and anti-IgA (n = 3). Healthy individuals also presented positivity in inflammatory cells with anti-IgG and anti-C3c (n = 4), anti-IgM (n = 1), and anti-IgA (n = 3); vessels were stained with anti-IgG and anti-C3c (n = 5), anti-IgM and anti-IgA (n = 4); adnexa were not represented in all samples obtained from healthy individuals. Conclusion IHC may serve as a reliable method to assess PF diagnosis. Immune deposits in dermal structures suggest their participation in autoimmune/inflammatory processes in PF. IHC may contribute to evaluate disease mechanisms, prognostic factors, and target-oriented treatment in PF. © 2017 The Authors
- Clinical and immunological profile of umbilical involvement in pemphigus vulgaris and pemphigus foliaceus(2013) OLIVEIRA JUNIOR, J. V.; MARUTA, C. W.; SOUSA JR., J. X.; SANTI, C. G.; VALENTE, N. Y. S.; ICHIMURA, L. M. F.; PERIGO, A. M.; AOKI, V.Background. Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune vesicobullous disorders with IgG autoantibodies directed against desmoglein (Dsg)1 and 3, which lead to intraepidermal acantholysis. Aim. To characterize the clinical and immunological profile of patients with PF or PV with umbilical involvement. Methods. In total, 10 patients (7 women, 3 men; age range 2470 years, disease duration 316 years) diagnosed with either PV (n = 5) or mucocutaneous PF (n = 5) were assessed according to their clinical features, histopathology and immunological findings [direct and indirect immunofluorescence (DIF and IIF) and ELISA with recombinant Dsg1 and Dsg3]. Results. Erythema, erosions, crusts and vegetating skin lesions were the main clinical features of the umbilical region. DIF of the umbilical region gave positive results for intercellular epidermal IgG and C3 deposits in eight patients and for IgG alone in the other two. Indirect immunofluorescence with IgG conjugate showing the typical pemphigus pattern was positive in all 10 patients, with titres varying from 1 : 160 to 1 : 2560. ELISA with recombinant Dsg1 gave scores of 24266 in PF and 0270 in PV. Reactivity to recombinant Dsg3 was positive in all five patients with PV (ELISA 2298) and was negative in all PF sera. Conclusions. All 10 patients with pemphigus with umbilical presentation had the clinical and immunopathological features of either PF or PV. This peculiar presentation, not yet completely elucidated, has rarely been reported in the literature. A possible explanation for this unique presentation may be the presence of either novel epitopes or an association with embryonic or scar tissue located in the umbilical-cord region.