ABSTRACT
Introducción El liquen escleroso (LiE) es una enfermedad crónica escleroatrófica que afectará generalmente el área anogenital y ocasionalmente a localizaciones extragenitales. Las células dendríticas dérmicas CD34 positivas (DDC) contribuyen al mantenimiento de la microarquitectura dérmica y a la modulación de la respuesta inmunitaria. El p53 es un gen supresor de tumores importante para la regulación del ciclo celular y de la apoptosis. De manera similar a lo que ocurre en la morfea (una condición escleroatrófica estrechamente relacionada con el LiE), la esclerosis dérmica, las alteraciones de las DDC y de la microvasculatura dérmica pueden ser mecanismos patogénicos subyacentes importantes en el LiE. Objetivos Examinar el perfil de las DDC positivas para el CD34, la densidad de microvasos (MVD) y la proteína p53 en el LiE. Materiales y métodos Se evaluaron los perfiles inmunohistológicos de las DDC, de la MVD y del p53 en 19 casos de LiE y en la piel normal de pacientes emparejados por edad y sexo (10 muestras), utilizando los anticuerpos contra el CD34 y el p53. Resultados Hubo una marcada disminución de los recuentos (1,7±0,5/mm2) o pérdida completa de DDC CD34+en el LiE en comparación con su elevada expresión en la piel normal (23,4±2,1/mm2, p =0,000). La MVD estaba notablemente aumentada en las lesiones de LiE (20±0,47) en comparación con la de la piel normal (5,50±0,20, p =0,000). Se observó una tinción nuclear discontinua, de células aisladas, débilmente positiva para el p53, localizada en los queratinocitos de las capas basales epidérmicas de la piel sana y de la piel afectada por el LiE. Conclusiones Hasta donde tenemos conocimiento, este es el primer estudio que analiza los perfiles de las DDC, de la MVD y del p53 de manera simultánea en el LiE. Los hallazgos sugirieron que las alteraciones de las DDC y de la MVD tienen papeles en la patogénesis del LiE (AU)
Background Lichen sclerosus (LiS) is a chronic scleroatrophic condition that usually affects the anogenital area and occasionally the extragenital sites. CD34-positive dermal dendritic cells (DDCs) contribute to the maintenance of the dermal microarchitecture and modulation of the immune response. p53 is a tumor suppressor gene important for the regulation of the cell cycle and apoptosis. Similar to morphea (a LiS-closely related scleroatrophic condition), dermal sclerosis, alterations of DDCs, and dermal microvasculature may be important underlying pathogenetic mechanisms in LiS. Objectives To examine the profile of CD34-positive DDCs, microvessel density (MVD), and p53 protein in LiS. Materials and methods The immunohistological profiles of DDCs, MVD, and p53 were examined in 19 cases of LiS and their age- and sex-matched normal skin (10 specimens), using antibodies against CD34 and p53. Results There was a markedly decreased counts (1.7±0.5/mm2) or complete loss of CD34-positive DDCs in LiS against their abundance in the normal skin (23.4±2.1/mm2, p=0.000). MVD was markedly increased in LiS lesions (20±0.47) as compared to normal skin (5.50±0.20, p=0.000). Discontinuous single-cell p53 weakly positive nuclear staining was seen in the epidermal basal cell keratinocytes in normal skin and LiS lesions. Conclusions To the best of this author's knowledge, this is the first study analyzing DDCs, MVD, and p53 profiles together in LiS. The findings suggest that alterations of DDCs and MVD have roles in the pathogenesis of LiS (AU)
Subject(s)
Humans , Male , Female , Middle Aged , Aged , Lichen Sclerosus et Atrophicus/pathology , Dendritic Cells/pathology , Antigens, CD34 , Vulvar Lichen Sclerosus/pathology , Tumor Suppressor Protein p53/metabolism , Microvessels/pathology , Retrospective Studies , ImmunohistochemistryABSTRACT
BACKGROUND: Lichen sclerosus (LiS) is a chronic scleroatrophic condition that usually affects the anogenital area and occasionally the extragenital sites. CD34-positive dermal dendritic cells (DDCs) contribute to the maintenance of the dermal microarchitecture and modulation of the immune response. p53 is a tumor suppressor gene important for the regulation of the cell cycle and apoptosis. Similar to morphea (a LiS-closely related scleroatrophic condition), dermal sclerosis, alterations of DDCs, and dermal microvasculature may be important underlying pathogenetic mechanisms in LiS. OBJECTIVES: To examine the profile of CD34-positive DDCs, microvessel density (MVD), and p53 protein in LiS. MATERIALS AND METHODS: The immunohistological profiles of DDCs, MVD, and p53 were examined in 19 cases of LiS and their age- and sex-matched normal skin (10 specimens), using antibodies against CD34 and p53. RESULTS: There was a markedly decreased counts (1.7 ± 0.5/mm2) or complete loss of CD34-positive DDCs in LiS against their abundance in the normal skin (23.4 ± 2.1/mm2, p = 0.000). MVD was markedly increased in LiS lesions (20 ± 0.47) as compared to normal skin (5.50 ± 0.20, p = 0.000). Discontinuous single-cell p53 weakly positive nuclear staining was seen in the epidermal basal cell keratinocytes in normal skin and LiS lesions. CONCLUSIONS: To the best of this author's knowledge, this is the first study analyzing DDCs, MVD, and p53 profiles together in LiS. The findings suggest that alterations of DDCs and MVD have roles in the pathogenesis of LiS.
ABSTRACT
BACKGROUND: Lichen sclerosus (LiS) is a chronic scleroatrophic condition that usually affects the anogenital area and occasionally the extragenital sites. CD34-positive dermal dendritic cells (DDCs) contribute to the maintenance of the dermal microarchitecture and modulation of the immune response. p53 is a tumor suppressor gene important for the regulation of the cell cycle and apoptosis. Similar to morphea (a LiS-closely related scleroatrophic condition), dermal sclerosis, alterations of DDCs, and dermal microvasculature may be important underlying pathogenetic mechanisms in LiS. OBJECTIVES: To examine the profile of CD34-positive DDCs, microvessel density (MVD), and p53 protein in LiS. MATERIALS AND METHODS: The immunohistological profiles of DDCs, MVD, and p53 were examined in 19 cases of LiS and their age- and sex-matched normal skin (10 specimens), using antibodies against CD34 and p53. RESULTS: There was a markedly decreased counts (1.7±0.5/mm2) or complete loss of CD34-positive DDCs in LiS against their abundance in the normal skin (23.4±2.1/mm2, p=0.000). MVD was markedly increased in LiS lesions (20±0.47) as compared to normal skin (5.50±0.20, p=0.000). Discontinuous single-cell p53 weakly positive nuclear staining was seen in the epidermal basal cell keratinocytes in normal skin and LiS lesions. CONCLUSIONS: To the best of this author's knowledge, this is the first study analyzing DDCs, MVD, and p53 profiles together in LiS. The findings suggest that alterations of DDCs and MVD have roles in the pathogenesis of LiS.
ABSTRACT
BACKGROUND: Nevi of special sites (NOSS) are benign melanocytic lesions that occur at particular sites. Although the histological features of NOSS have been described, their immunophenotypic features have not been fully characterized. AIMS: To present the clinicopathological characteristics of a case series of NOSS and to characterize their immunohistochemical profile. MATERIALS AND METHODS: Thirty-five NOSS were assessed using immunoperoxidase staining techniques for the melanocytic (S100, Melan-A, and HMB45) and proliferation (Ki-67) markers RESULTS: All of the cases of NOSS showed concerning architectural changes (prominent lentiginous melanocytic proliferation, irregularities, crowdedness, and dyhesiveness of the nests), and cytological atypia (large nevomelanocytes with vesicular nuclei, clear cytoplasm, and dusty melanin pigment) that can lead to a misdiagnosis of atypical nevi or even melanomas. All of the cases of NOSS showed diffuse expression of S100 and Melan-A proteins. Ki-67 labeling index of the nevomelanocytes was extremely low. HMB45 protein expression was limited to the junctional and superficial dermal nevomelanocytes. CONCLUSIONS: NOSS can show histological features that can easily mimic atypical nevi or melanomas and this diagnostic consideration should be kept in mind to avoid their misdiagnosis. The expression of HMB45 protein in NOSS indicates that their nevomelanocytic cells have an activated phenotype. The decreased HMB45 protein expression following a gradient from junctional to deeper dermal localization in NOSS is indicative of their immunohistochemical maturation.
Subject(s)
Melanoma , Nevus, Epithelioid and Spindle Cell , Nevus, Pigmented , Nevus , Skin Neoplasms , Humans , Melanoma/diagnosis , Nevus, Pigmented/diagnosis , Skin Neoplasms/diagnosisABSTRACT
INTRODUCTION: The α-chain variant Hb Q-India (c.193G>C) is caused by a point mutation GACâCAC at codon 64 of the α1 globin gene and is clinically silent. Point mutations can be diagnosed easily by many simple polymerase chain reaction (PCR) techniques including PCR-restriction digest, but for Hb Q-India the restriction digest has never been described. In this work we aimed to develop a restriction enzyme digestion assay for DNA diagnosis of Hb Q-India, in order to increase the panel of restriction enzymes used in DNA diagnosis of haemoglobinopathies and also as a simple cheap alternative to the ARMS-PCR method. METHODS: A restriction enzyme digestion assay was designed for diagnosis of Hb Q-India using the restriction enzyme EaeI enzyme as the Hb Q-India mutation abolishes the recognition site of this enzyme. Patients were screened for an abnormal haemoglobin by high performance liquid chromatography (HPLC) and those had an abnormal peak with a retention time between 4.7 and 4.8 minutes were selected for diagnosis at the molecular level. The α1 globin gene was amplified in 12 cases with a presumed diagnosis of Hb Q-India by HPLC and isoelectric focusing (IEF), and the amplified products were subjected to the EaeI digestion. RESULTS: All the 12 cases were diagnosed positive (100%) for Hb Q-India by the EaeI restriction enzyme digest. They were heterozygotes for the mutation. CONCLUSION: EaeI restriction enzyme digestion can be used as a simple and robust alternative method to ARMS-PCR for DNA diagnosis of Hb Q-India. The EaeI restriction enzyme can be added to the panel of restriction enzymes used in the DNA diagnosis of the abnormal Hb variants. Concomitant use of HPLC and IEF can be used efficiently for presumed diagnosis of this rare variant.
Subject(s)
Deoxyribonucleases, Type II Site-Specific , Hemoglobinopathies/diagnosis , Hemoglobins, Abnormal/genetics , Genetic Testing , Heterozygote , Humans , Mutation/genetics , Restriction Mapping , alpha-Globins/geneticsABSTRACT
INTRODUCTION: Haemoglobin (Hb) G-Philadelphia mutation is a common alpha-globin chain variant [α68(E17)Asn > Lys]. Combined high performance liquid chromatography (HPLC) and isoelectric focusing (IEF) can be used in a presumptive diagnosis of Hb G-Philadelphia, but there are other α-chain variants with a similar phenotype that cannot be excluded. Our aim was to develop a novel StyI restriction enzyme assay to diagnose the common Hb G-Philadelphia mutation and to identify any other variants with a similar phenotype by DNA sequencing. METHODS: Thirty-one cases given a presumptive diagnosis as Hb G-Philadelphia by HPLC and IEF were subjected to DNA analysis by restriction enzyme digestion using StyI. Negative cases were then subjected to DNA sequencing. RESULTS: Twenty-two cases (78.6%) of 28 cases amplified were tested positive for Hb G-Philadelphia by StyI restriction digestion. Sequencing of the six negative cases revealed two cases of Hb G-Philadelphia with CâA mutation in codon 68 in α2 globin gene, plus one case each of Hb G-Norfolk Hb Stanleyville-II, Hb Matsue-Oki and Hb Mizushi. CONCLUSION: A novel StyI restriction enzyme can be used to confirm the commonest type of Hb G-Philadelphia. DNA sequencing identified four other α-chain variants with a similar HPLC and IEF phenotype.
