Análise das células dendríticas na língua, linfonodos cervicais e tonsilas palatinas de pacientes autopsiados com Síndrome da Imunodeficiência Adquirida

Orientadores: Pablo Agustin Vargas, Luiz Paulo KowalskiTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Na infecção pelo HIV, as células dendríticas (CDs) podem desempenhar vários papéis, incluindo a provável captação inicial do HIV, transporte para os linfonodos, e posterior transferência para células T, desempenhando um importante papel no sistema imune. As manifestações orais observadas em pacientes infectados pelo HIV, incluindo aquelas associadas ao HSV-1 podem estar diretamente relacionadas à injúria das CDs. A proposta deste estudo foi identificar e quantificar as CDs intersticiais na língua de pacientes autopsiados com AIDS e portadores de infecção herpética lingual (n=10), pacientes com AIDS e sem lesões linguais (n=10) e pacientes sem AIDS e sem lesões linguais (n=10) por meio de reações imunoistoquímicas. Além disso, investigamos a população de CDs nos linfonodos e tonsilas palatinas de pacientes com AIDS (n=32) e sem AIDS (n=21). Nos tecidos linguais, foram utilizados os anticorpos contra CD1a e CD83 para identificação das CDs e o anticorpo contra HSV-1 para detecção do vírus da herpes simples tipo 1. Nos linfonodos e tonsilas palatinas foi utilizados além dos anticorpos contra CD1a e CD83, o anticorpo contra fator XIIIa. Para a quantificação das CDs nos tecidos linguais foi utilizado análise histomorfométrica convencional e nos tecidos linfóides foi aplicado o método analítico Positive Pixel Count (software Image Scope). Os resultados mostraram uma intensa depleção na população de CDs em tecidos linguais e linfóides de pacientes com AIDS e a infecção lingual pelo HSV-1 não potencializou a redução de CDsAbstract: During HIV infection, dendritic cells (DCs) may play several roles, including the probable initial uptake of HIV, transport to the lymph nodes, and subsequent transfer to T cells. Oral opportunistic infections observed in HIV-infected patients, including those associated with HSV-1 may be directly related to injury of DCs. The purpose of this study was to identify and quantify the interstitial DCs in the tongue of autopsied patients with AIDS and lingual herpes (n = 10), AIDS patients with normal tongues (n = 10) and non-AIDS patients with normal tongues (n = 10) by immunohistochemistry. Furthermore, we investigated the DCs population in lymph nodes and palatine tonsils of AIDS patients (n = 32) and non-AIDS patients (n = 21). CD1a and CD83 antibodies were carried out to identify DCs in lingual tissues and HSV-1 antibody for detection of herpes simplex virus type 1. In lymphoid tissues, CD1a, CD83 and factor XIIIa antibodies were carried out to identify DCs. Interstitial DCs were measured by conventional histomorphometry whereas the lymphoid DCs were measured by Positive Pixel Count Algorithm method using ImageScope software. The results showed a decreased population of DCs in lingual and lymphoid tissues of AIDS patients independently of the presence of concomitant infection by HSV-1DoutoradoPatologiaDoutor em Estomatopatologi

Expression of Langerhans cells in the tongue of autopsied patients with advanced AIDS

Orientador: Pablo Agustin VargasDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: A língua de pacientes com AIDS é acometida freqüentemente por infecções oportunistas e neoplasias. O objetivo deste estudo foi quantificar as células de Langerhans (CL) presentes em regiões lesionais e não lesionais na língua de pacientes autopsiados com AIDS em fase avançada, correlacionando a diminuição das CL com a presença de patologias infecciosas em diferentes regiões da língua (anterior, média, posterior e lateral) e comparar estes achados com a língua de pacientes HIV negativos. Foram utilizadas neste trabalho as línguas de 40 pacientes autopsiados com AIDS divididos em 04 grupos (10 com candidose lingual, 10 com herpes lingual, 10 com leucoplasia pilosa oral e 10 sem lesões em língua) e as línguas de 23 pacientes autopsiados (grupo controle) que morreram por outras patologias não associadas à AIDS e que não apresentaram patologias em língua. Realizamos estudo imunoistoquímico com os marcadores HLA-DR, CD1a e CD83 para identificar as CL e quantificá-las por meio de análise histomorfométrica. O índice de positividade foi obtido através da leitura de 7 campos seqüenciados orientados por ocular micrométrica. As células positivas foram contadas para cada um dos anticorpos nas quatro diferentes regiões da língua e os resultados expressos em células positivas por área de epitélio e por comprimento de membrana basal. O anticorpo HLA-DR mostrou a presença média das CL na língua dos pacientes com AIDS (LA=24,28/mm2 e 3,64/mm, LM=24,60/mm2 e 3,68/mm, LP=20,95/mm2 e 3,14/mm, LL=19,84/mm2 e 2,97/mm) e no grupo controle (LA=68,18/mm2 e 10,23/mm, LM=60,73/mm2 e 9,11/mm, LP=62,94/mm2 e 9,44/mm, LL=50,24/mm2 e 7,53/mm). O anticorpo CD1a mostrou a presença média das CL na língua dos pacientes com AIDS (LA=17,30/mm2 e 2,59/mm, LM=21,11/mm2 e 3,16/mm, LP=13,48/mm2 e 2,02/mm, LL=15,55/mm2 e 2,33/mm) e no grupo controle (LA=205,38/mm2 e 30,81/mm, LM=218,36/mm2 e 32,75/mm, LP=167,29/mm2 e 25,09/mm, LL=223,60/mm2 e 33,54/mm). O anticorpo CD83 mostrou a presença média das CL na língua dos pacientes com AIDS (LA=6,19/mm2 e 0,92/mm, LM=6,34/mm2 e 0,95/mm, LP=6,82/mm2 e 1,02/mm, LL=7,14/mm2 e 1,07/mm) e no grupo controle 68,18/mm2 e 10,23/mm, LM=68,46/mm2 e 10,27/mm, LP=69,28/mm2 e 10,39/mm, LL=63,49/mm2 e 9,52/mm), sendo que foram extremamente significantes as diferenças entre ambos os grupos em todas as regiões e anticorpos estudados (p<0,001). Portanto, podemos concluir que as CL estavam degeneradas e diminuídas em número em todas as regiões da língua e em todos os grupos com AIDS em relação ao grupo controle e as lesões infecciosas orais oportunistas não influenciaram na depleção das CL nas línguasAbstract: The tongues of AIDS patients can be affected by opportunistic infections and neoplasms. Objectives: to quantify and compare the expression of Langerhans cells (LC) in lesional and non lesional areas in the tongue from patients with and without AIDS (control group), using autopsy material. Methods: we analysed the expression of CD1a, HLA-DR and CD83 using immunohistochemistry to identify and quantify LC in the tongues of AIDS patients (n=40), which were divided into 04 groups (10 lingual candidiasis, 10 lingual herpes, 10 oral hairy leukoplakia and 10 none lesions), and 23 tongues from HIV-negative controls. The immunoreactivity rate was obtained after reading at least seven fields sequenced driven ocular micrometer. The positive LC were detected in the lingual surface epithelium in four different regions (anterior, middle, posterior and lateral) and the results expressed as positive cells per area of epithelium and basement membrane length. Results: LC showed the following immunoreactivity for CD1a in the tongue of AIDS patients (LA=17.30/mm2 and 2.59/mm, LM=21.11/mm2 and 3.16/mm, LP=13.48/mm2 and 2.02/mm, LL=15.55/mm2 and 2.33/mm), and in the control group (LA=205.38/mm2 and 30.81/mm, LM=218.36/mm2 and 32.75/mm, LP=167.29/mm2 and 25.09/mm, LL=223.60/mm2 and 33.54/mm); HLA-DR (AIDS patients) (LA=24.28/mm2 and 3.64/mm, LM=24.60/mm2 and 3.68/mm, LP=20.95/mm2 and 3.14/mm, LL=19.84/mm2 and 2.97/mm), and the control group (LA=68.18/mm2 and 10.23/mm, LM=60.73/mm2 and 9.11/mm, LP=62.94/mm2 and 9.44/mm, LL=50.24/mm2 and 7.53/mm); CD83 (AIDS patients) (LA=6.19/mm2 and 0.92/mm, LM=6.34/mm2 and 0.95/mm, LP=6.82/mm2 and 1.02/mm, LL=7.14/mm2 and 1.07/mm), and the control group (LA=68.18/mm2 and 10.23/mm, LM=68.46/mm2 and 10.27/mm, LP=69.28/mm2 and 10.39/mm, LL=63.49/mm2 and 9.52/mm). The statistical analysis identified significant differences in the both groups and in all regions, and among the 3 antibodies (p<0.001). Conclusions: LC were degenerated and reduced in number in all regions of the tongue of AIDS patients in relation to the control group and the depletion of LC in the tongues of AIDS patients is not associated with oral opportunistic infectionsMestradoPatologiaMestre em Estomatopatologi

Oral pigmented lesions: clinicopathologic features and review of the literature

Diagnosis of pigmented lesions of the oral cavity and perioral tissues is challenging. Even though epidemiology may be of some help in orientating the clinician and even though some lesions may confidently be diagnosed on clinical grounds alone, the definitive diagnosis usually requires histopathologic evaluation. Oral pigmentation can be physiological or pathological, and exogenous or endogenous. Color, location, distribution, and duration as well as drugs use, family history, and change in pattern are important for the differential diagnosis. Dark or black pigmented lesions can be focal, multifocal or diffuse macules, including entities such as racial pigmentation, melanotic macule, melanocytic nevus, blue nevus, smoker's melanosis, oral melanoacanthoma, pigmentation by foreign bodies or induced by drugs, Peutz-Jeghers syndrome, Addison's disease and oral melanoma. The aim of this review is to present the main oral black lesions contributing to better approach of the patients

Hybrid ameloblastoma and central giant cell lesion : challenge of early diagnosis

Hybrid lesions encompass the occurrence of different entities in one lesion. A 67-year-old woman was referred to the Oral and Maxillofacial Surgery Service for treatment of mandibular Central Giant Cell Lesion (CGCL) previously diagnosed. Intraoral examination revealed edentulism and a painless swelling extending from the alveolar ridge to the buccal vestibule with hard consistency covered by normal mucosae, with unknown duration. Panoramic radiograph revealed a large, multilocular and well-defined radiolucent lesion extending from the region of left mandibular lateral incisor teeth to right mandibular first molar with no evidence of osseous perforation. Initially, a treatment with intralesional injection of corticosteroids was performed. After 18 months of treatment, an increase in size of the osteolytic lesion was noted. An incisional biopsy was carried out and the microscopic examination revealed a unicystic ameloblastoma associated to CGCL. It was performed marsupialization and later the enucleation of residual lesion. The follow-up remains being performed

Clinicopathologic analysis of 14 cases of odontogenic myxoma and review of the literature

Odontogenic myxoma is a rare benign neoplasm that originates from odontogenic ectomesenchyme. There is no standard of care and recurrences are frequent after conservative surgical procedures. A retrospective study conducted at a single cancer center, with analysis of medical records of all patients diagnosed with odontogenic myxoma from 1980 to 2010, along with a literature review. There were 14 patients with diagnosis of odontogenic myxoma (OM). Most patients were female (78.6%) and Caucasian (100%), with ages ranging from 7 to 51 years (21.6 ± 11.6 years). The time period between the first symptom and first consultation ranged from 0 to 60 months (19.4 ± 19.97 months). The most frequent complaints were increased local volume or failure to tooth eruption. The most common tumor site was the mandible (11 cases, 78.5%). About radiological findings, most lesions were multilocular (9 cases, 64.3%) and with imprecise limits (12 cases, 85.7%). Surgery was performed in all cases and curettage was the most applied technique (10 cases, 71.4%). Three patients underwent mandibulectomy and complex reconstructions including iliac crest microvascular flap. Three patients had postoperative complications and 4 had local recurrences of the tumor. The follow up time ranged from 12 to 216 months (112 ± 70.8 months). All patients are without clinical and radiographic evidence of disease. OM is a locally aggressive and rare tumor. There is no gold standard surgical management and the therapeutic decision should be individualized taking into account the characteristics and extension of the tumor

CD1a+and CD83+Langerhans cells are reduced in lower lip squamous cell carcinoma

BACKGROUND : Actinic cheilitis (AC) is a potentially malignant lesion diagnosed in the lip of patients chronically exposed to the sun that may give rise to a fully invasive lower lip squamous cell carcinoma (LLSCC). It is known that ultraviolet radiation causes dendritic cells (DCs) depletion in the epidermis, but the role of this cellular population in lip cancer progression remains uncertain. Therefore, this study investigated the distribution of DCs in normal, dysplastic and neoplastic tissues of the lower lip. METHODS : Thirteen cases of lower lip mucocele, 42 of ACs and 21 of LLSCC were retrieved and original diagnoses confirmed by two oral pathologists, who further classified ACs as low- and high-risk lesions. Immunoreactions against CD1a and CD83 identified immature and mature DCs, respectively. RESULTS : Immature CD1a+ Langerhans cells (LCs) were significantly decreased in LLSCC when compared to morphologically normal (P 0.05), but ACs demonstrated a lower concentration of CD1a+ LCs than normal epithelium (P 0.05). CONCLUSION : These results suggest that depletion of epithelial LCs, but not IDCs in the connective tissue, would represent an important step for lip cancer development.http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1600-07142017-07-31hb2016Oral Pathology and Oral Biolog

Different methods of cell quantification can lead to different results : a comparison of digital methods using a pilot study of dendritic cells in HIV-positive patients

Although new digital pathology tools have improved the positive cell quantification, there is a heterogeneity of the quantification methods in the literature. The aim of this study was to evaluate and propose a novel dendritic cells quantification method in squamous cell carcinoma comparing it with a conventional quantification method. Twenty-six squamous cell carcinomas HIV-positive cases affecting the oropharynx, lips and oral cavity were selected. Immunohistochemistry for CD1a, CD83, and CD207 was performed. The immunohistochemical stains were evaluated by automated examination using a positive pixel count algorithm. A conventional quantification method (unspecific area method; UA) and a novel method (specific area method; SA) were performed obtaining the corresponding density of positive dendritic cells for the intratumoral and peritumoral regions. The Mann-Whitney U test was used to verify the influence of the quantification methods on the positive cell counting according to the evaluated regions. Data were subjected to the ANOVA and Student?s t-test to verify the influence of the tumour location, stage, histological grade, and amount of inflammation on the dendritic cells density counting. The cell quantification method affected the dendritic cells counting independently of the evaluated region (P-value <0.05). Significant differences between methods were also observed according to the tumour features evaluations. The positive cell quantification method influences the dendritic cells density results. Unlike the conventional method (UA method), the novel SA method avoids non-target areas included in the hotspots improving the reliability and reproducibility of the density cell quantification

Immunohistochemical expression of RANKL in oral giant cell lesions is predictive of aggressiveness

Abstract The aim of this study was to evaluate the immunohistochemical expression of receptor activator of nuclear factor kappa-B ligand (RANKL) and of osteoprotegerin (OPG), important proteins correlated with osteoclastogenesis, in central giant cell lesions (CGCL) and peripheral giant cell lesions (PGCL) and to compare their expression with the histological and clinical parameters for quantification of multinucleated giant cells (MGC) and their nuclei, lesion size, and recurrences. Twenty cases of each lesion type were selected to quantify the number of MGCs and nuclei/mm2 of connective tissue. The immunoreactivity of RANKL and OPG was expressed as a percentage of the marked area in the stroma. Clinical data were collected from pathoanatomical and medical reports. No statistical differences were found for the number of MGCs (p = 0.24) between PGCL and CGCL, but the number of nuclei within the MGCs was higher in CGCL (p = 0.01). RANKL expression was higher in CGCL than in PGCL (p = 0.04) and all recurrent lesions showed higher RANKL and OPG expressions than nonrecurrent lesions. We report higher RANKL expression and a greater number of nuclei in CGCL, which may explain the difference in clinical behaviour between these lesions and their pathogenesis