Comparação da população linfocitária e da expressão da selectina-e no líquen plano oral e na reação liquenóide oral

Introdução: A ausência de critérios para distinguir os aspectos clínicos e histopatológicos das lesões de líquen plano oral (LPO) e da reação liquenóide oral (RLO) já foi descrita por diversos autores. Alguns critérios foram propostos na tentativa da diferenciação entre elas, especialmente devido a possível associação do LPO com o carcinoma oral. Objetivo geral: Comparar e avaliar a população linfocitária e a expressão da selectina-E em lesões de líquen plano oral e reação liquenóide oral. Objetivos específicos: verificar a presença e quantificar a imunoexpressão, comparando os grupos no que diz respeito as células T, TCD4, TCD8, CLA CD4, CLA CD8, selectina-E no LPO e RLO; avaliar se existe correlação entre a expressão da selectina-E com a expressão do antígeno cutâneo associado a linfócitos no LPO e na RLO. Material e Métodos: estudo aprovado pelo Comitê de Ética em Pesquisa (CAAE: 47567515.1.0000.5243). Os participantes com diagnóstico clínico de LPO e RLO foram recrutados no ambulatório de Diagnóstico Oral do HUAP-UFF que necessitavam realizar biópsia. O tecido foi congelado e em seguida corado em H&E e imunomarcado para CD4/CLA, CD3/CD4, CD3/CD8, CD8/CLA, CD62E. Foram selecionadas cinco áreas na lâmina de cada participante para a imunofluorescência, sendo contadas no mínimo 150 células e avaliada no programa Image J. A análise estatística foi realizada no Programa SPSS 20.0, com nível de significância de 5% (p≤0,05). Resultados: foram atendidos 36 participantes, destes 25 com diagnóstico histopatológico de LPO e 11 de RLO. Na análise intragrupo, no GLPO a comparação entre a quantidade de linfócitos CD3+CD4+ e CD3+CD8+ foi estatisticamente significante (p=0,00); e entre os linfócitos CD4+CLA+ e CD8+CLA+ também (p=0,031). No GRLO a comparação entre a quantidade de linfócitos CD3+CD4+ e CD3+CD8+ não foi estatisticamente significante, assim como entre os linfócitos CD4+CLA+ e CD8+CLA+. Na análise intergrupos quando avaliadas as células CD3+, CD3+CD4+, CD3+CD8+, CLA+, CD4+CLA+ e CD8+CLA+ e a expressão de selectina-E nenhuma variável obteve significância estatística. Na avaliação das proporções somente CD3+CD4+/CD3+ e CD3+CD8+/CD3+ tiveram significância estatística (p=0,027 e 0,038 respectivamente). Na análise de correlação entre CD4+CLA+ e selectina-E, e CD8+CLA+ e selectina-E, ambos os grupos não apresentaram correlação. Conclusão: Os linfócitos CD3+, CD3+CD4+, CD3+CD8+, CD4+CLA+, CD8+CLA+ e a selectina-E, estão presentes em ambas as lesões. A maior proporção de células CD3+CD4+ na RLO e CD3+CD8+ no LPO sugere que podem ter uma importância no mecanismo etiopatogênico dessas lesões. Não foi observada significância estatística entre os grupos em relação ao linfócito CD3+, talvez pelo fato de que ambas as lesões possuem uma origem inflamatória onde predomina este tipo celular. No GLPO a maior presença de linfócitos CD4+CLA+ pode justificar sua importância no mecanismo etiopatogênico dessa lesão. A imunoexpressão da selectina-E não foi significante, e não houve correlação entre as células CD4+CLA+, CD8+CLA+ e a selectina-E nos grupos, sugerindo que outras moléculas de adesão podem participar da transmigração de células no mecanismo etiopatogênico das lesõesIntroduction: Several authors have described the lack of association between clinical and histopathological aspects of oral lichen planus lesions (OLP) and oral lichenoid reactions (OLR). Some criteria have been proposed in attempt to differentiate them, especially due to the possible association of OLP with the development of oral carcinoma. Main objective: To evaluate and compare lymphocyte population and the expression of E-selectin in OLP and OLR lesions. Specific objectives: verify the presence and quantify the immunoexpression, and as well as to compare OLP and OLR groups with respect to T cells, TCD4, TCD8, CLA CD4, CLA CD8, and E-selectin. Evaluate if there is a correlation between the expression of E-selectin and the expression of cutaneous lymphocytes antigen in OLP and OLR lesions. Material and Methods: this study was approved by ethic committee (CAAE: 47567515.1.0000.5243). The participants with clinical diagnosis of OLP and OLR were selected in Oral Diagnosis Clinic of HUAP-UFF for biopsy confirmation. Frozen tissue sections were obtained and stained by H&E and immunostained for CD4/CLA, CD3/CD4, CD3/CD8, CD8/CLA, and CD62E. Five areas were selected at the fragment specimens of each immunostain in both OLP and OLR groups, a minimum of 150 cells was counted for immunofluorescence evaluation, and the Image J Program was used for the analysis. Statistical analysis was performed using SPSS 20.0, considered significance level of 5% (p ≤ 0,05). Results: a total of 36 patients were evaluated, 25 had histopathological diagnosis of OLP and 11 of OLR. In intragroup analysis the comparison between CD3+CD4+ and CD3+CD8+ lymphocytes was statistically significant (p=0.00) in OLPG; and CD4+CLA+ and CD8+CLA+ lymphocytes was as well (p=0.031). In OLRG, the comparison between CD3+CD4+ and CD3+CD8+ lymphocytes were not statistically significant, as well as between CD4+CLA+ and CD8+CLA+ lymphocytes. In the intergroup analysis for CD3+, CD3+CD4+, CD3+CD8+, CLA+, CD4+ CLA+ and CD8+CLA+ cells and E-selectin expression were not statistically significant. However, the proportions of CD3+CD4+/CD3+ and CD3+CD8+/CD3+ were statistical significant (p=0.027 and 0.038 respectively). In the correlation, analysis between CD4+CLA+ and E-selectin, and CD8+CLA+ and E-selectin for both groups showed no correlation. Conclusion: CD3+, CD3+CD4+, CD3+CD8+, CD4+CLA+, CD8+CLA+ lymphocytes and E-selectin are present in both OLP and OLR lesions. The higher proportion of CD3+CD4+ cells in OLR and CD3+CD8+ cells in OLP lesions suggest that they may play an important etiopathogenic mechanism for these lesions. It is not observed statistical significance between the groups with respect to CD3+ lymphocytes may be because both are inflammatory lesions with the predominance of this type of cells. The higher presence of CD4+CLA+ lymphocytes in OLPG may justify its importance in the etiopathogenic mechanism. E-selectin immunoexpression was not significant and correlations between CD4+CLA+ cells and E-selectin, CD8+CLA+ and E-selectin cells are not observed in both group, suggesting that other adhesion molecules may play a role in cells transmigration in the etiopathogenic mechanism of both lesions104f

Análise da expressão do antígeno cutâneo associado ao linfócito e da expressão da selectina-e nas lesões de líquen plano oral através da imunofluorescência

O Líquen Plano Oral (LPO) é uma doença autoimune inflamatória crônica, afeta aproximadamente 0,1 a 2% da população, na maioria mulheres, entre a quinta e sexta décadas de vida. Acredita-se que os linfócitos T CD8+ desempenhem um papel fundamental na manifestação da doença. Os antígenos cutâneos associados aos linfócitos (CLA) migram preferencialmente para a pele através da interação com a selectina-E. Possivelmente nas lesões de LPO a maior expressão do CLA está diretamente relacionada a maior expressão da selectina-E. Assim, como a pele é afetada por doenças crônicas autoimunes como o líquen plano, é interessante determinar se os linfócitos T CLA+ são expressos na mucosa oral assim como na pele. Dessa forma, esse trabalho se propõe a analisar a expressão do CLA nas áreas de líquen plano oral e verificar se existe correlação com a expressão da selectina-E através da imunofluorescência. Os pacientes foram selecionados do ambulatório de Diagnóstico Oral do Hospital Universitário Antônio Pedro através de uma abordagem direta, foram informados por escrito dos objetivos do estudo, seus riscos e benefícios e assinaram um termo de consentimento para a participação. Foi preenchido um questionário de anamnese. Todos os pacientes tinham mais de 18 anos de idade e preencheram os critérios clínicos e histopatológicos para LPO. Foram excluídos gestantes, lactantes e os que não preencheram um dos critérios clínicos ou histopatológicos para LPO. Foi realizada uma única biópsia em cada paciente em mucosa jugal, englobando uma área de LPO (GLPO) e uma área clinicamente livre de lesão (GPL). O material obtido foi congelado a -80oC em OCT e posteriormente, obtidas secções de tecido com 6 μm de espessura. A coloração pela hematoxilina e eosina (H&E) foi realizada para o diagnóstico histopatológico bem como avaliar a intensidade do infiltrado inflamatório. Foi realizada a imunofluorescência utilizando os seguintes anticorpos: CD3, CD4, CD8, CD62E e CLA. Foi utilizado o programa Image J para análise da imunomarcação CD3, CD4, CD8 e CLA, e uma contagem visual para CD62E. A análise estatística foi realizada através do programa SPSS 17.0 e análise descritiva para os dados demográficos e características clínicas. Foram selecionados 11 pacientes do total de 37, na maioria mulheres (90,9%), idade média 52,2 anos, e 54,2% eram pardos. Os sítios mais acometidos foram mucosa jugal (100%) e língua (80%). Os padrões clínicos mais comuns foram o reticulado (100%) e o placa (90%). A histopatologia demonstrou em todos os casos infiltrado inflamatório em faixa, degeneração hidrópica da camada basal e ausência de displasia. A imunofluorescência demonstrou no GLPO em relação ao GC um predomínio de células CD3+CD8+ (p=0,003), CD8+CLA+ (p=0,004) e de CD62E (p=0,003). No GLPO houve um predomínio de CD3+CD8+ (p=0,01) e ligeiro predomínio de CD8+CLA+(p>0,05). As correlações entre CD3+ ou CLA e o CD62E foi fraca (p>0,05). A correlação entre o CLA e o CD62E foi fraca, não apresentando significância estatística. Os resultados indicam que tanto o CLA como a selectina-E parecem estar envolvidas e são importantes no mecanismo etiopatogênico do LPO porém uma outra molécula de adesão vascular pode ter importância na emigração destes linfócitosOral lichen planus (OLP) is a chronic inflammatory and autoimmune disease, affecting approximately 0,1 to 2% of the population, mainly females, and most frequently during the fifth and sixth decades of life. It is believed that the T CD8+ lymphocytes play a fundamental role in its manifestation. The cutaneous lymphocyte-associated antigen (CLA) appears to migrate preferentially into skin by interacting with E-selectin on vascular endothelium. In lichen planus it is possible a directly relationship with a higher expression of CLA and a higher expression of E-selectin. Since the mouth may be affected by chronic immune-inflammatory dermatoses such as lichen planus it is interesting to investigate if the CLA is expressed on oral mucosa as it is expressed in the skin. The aim of this work is to analyze the CLA expression in oral lichen planus area and to verify if there is a correlation with the E-selectin expression by using immunofluorescence stain. The samples were selected from Oral Diagnostic Clinic of Hospital Universitario Antonio Pedro. All patients were informed about the study, their risks and benefits and a consent form were signed in order to participate of the study. A questionnaire was filled out with all patients’ data. All patients were over 18 years old and clinical and histopathological criteria for OLP were considered. Pregnant and nursering mothers were excluded and those who clinical and histopathological criteria for OLP were not found. A biopsy was obtained from each patient from buccal mucosa, an area with OLP (GOLP) and without OLP (GPL). The obtained samples were frozen at -80oC in OCT for a 6 μm cut. Samples were stained for Hematoxylin and Eosin (HE) for inflammatory intensity evaluation and for histopathologic diagnosis. An immunofluorescence stain was performed using CD3, CD4, CD8, CD62E and CLA antibodies. An Image J program was used for immune stain analysis of CD3, CD4, CD8 and CLA and a visual count was performed for CD62E stain. Statistical analysis was performed for SPSS 17.0 package and descriptive analysis for clinical features and demographic data were performed. It was selected 11 patients from 37, the majority was female (90,9%), median age 52,2 years old, and 54,2% were brown. Buccal mucosa (100%) and the tongue (80%) were the sites most affected. Reticular (100%) and plaque (80%) pattern were the most common. In all cases histopathology demonstrated a band inflammatory infiltrate, basal layer hidropic degeneration and no dysplasia. The comparison of GOLP with CG an immunofluorescence demonstrated a predominance of CD3+CD8+ (p=0,003), CD8+CLA+ (p=0,004) and CD62E (p=0,003) in favor of GOLP. In GOLP was observed a predominance of CD3+CD8+ (p=0,01) and slightly predominance with regard to CD8+CLA+(p>0,05). The correlation among CD3+ or CLA and CD62E were weak. The correlation between CLA and CD62E was weak with no statistical significance was found. The results indicated that both CLA and E-selectin seems to envolved and are importants in the etiopatogenic mechanism of OLP however other vascular adhesion molecules could be enrolled and important into these lymphocytes emigration136 f

Atypical oral candidiasis in a psoriatic patient during targeted immunotherapy with an interleukin 17 inhibitor (secukinumab)

Abstract Background Secukinumab is a human monoclonal antibody immunoglobulin that neutralises interleukin (IL)-17A, and as such, is effective in the treatment of psoriasis. However, as IL-17A is essential in protection against fungal infections, patients treated with this drug may develop candidiasis. This report presents a case of atypical oral candidiasis occurring during targeted drug immunotherapy with an interleukin 17 (IL-17) inhibitor (secukinumab), with the aim of emphasisinge the necessity of periodical oral health assessment and monitoring. It provides a rational clinical approach to therapeutic protocol in the treatment of side effects associated with novel medications for autoimmune diseases. Case presentation Symptomatic tongue lesions were observed in a 50-year-old female patient on a monthly systemic treatment of 300 mg of secukinumab, which appeared after 60 days of using the medication. Two inconclusive biopsies and an unsuccessful application of oral corticosteroids made the diagnostic process challenging. Papillae on the back of the tongue were atrophied, forming a well-defined erythema and white non-detachable plaques on the lateral border of the tongue. Cytopathological and histopathological exam results were compatible with a diagnosis of oral candidiasis. Topical antifungal medication led to subsequent regression of the tongue lesions. During asymptomatic period and follow up for 7 months, a reduced monthly dose 150 mg of secukinumab was administered. Conclusions Patients undergoing treatment with IL-17 blockers, such as secukinumab, should be carefully monitored in order to avoid oral side effects resulting from the use of this medication

Evaluation of sclerotherapy with different dilutions of ethanolamine oleate in the treatment of oral varicose veins

ABSTRACT Objective: This study aim to evaluate the effectiveness of sclerotherapy protocols with different dilutions of ethanolamine oleate in the treatment of oral varicose veins. Methods: Clinical data and images of 14 cases treated with sclerotherapy were analyzed and descriptive analyses were performed. Results: Females (58%) and white skin color (83%) prevailed. Age varied between 14 and 79 years, with 47 years on mean (SD = 19 years). The most common anatomical locations were the buccal mucosa and lower lip. The final volume of the sclerosing agent (Ethamolin®) ranged from 0.4 to 4.3ml and the concentration ranged from 5% to 100%. The number of sessions ranged from 1 to 12 and the number of points per application was 1 to 7 points. Pain and edema were seen in 43% and 29% of patients, respectively. Conclusion: Sclerotherapy with monoethanolamine oleate diluted in anesthetic is a safe and effective option for the treatment of this lesion, regardless of concentration. However, edema and pain seem to be directly associated with increased drug concentration