RESUMO
Immunological tests may represent valuable tools for the diagnosis of human tegumentary leishmaniasis (TL) due to their simple execution, less invasive nature and potential use as a point-of-care test. Indeed, several antigenic targets have been used with the aim of improving the restricted scenario for TL-diagnosis. We performed a worldwide systematic review to identify antigenic targets that have been evaluated for the main clinical forms of TL, such as cutaneous (CL) and mucosal (ML) leishmaniasis. Included were original studies evaluating the sensitivity and specificity of immunological tests for human-TL, CL and/or ML diagnosis using purified or recombinant proteins, synthetic peptides or polyclonal or monoclonal antibodies to detect Leishmania-specific antibodies or antigens. The review methodology followed PRISMA guidelines and all selected studies were evaluated in accordance with QUADAS-2. Thirty-eight original studies from four databases fulfilled the selection criteria. A total of 79 antigens were evaluated for the detection of antibodies as a diagnostic for TL, CL and/or ML by ELISA. Furthermore, three antibodies were evaluated for the detection of antigen by immunochromatographic test (ICT) and immunohistochemistry (IHC) for CL-diagnosis. Several antigenic targets showed 100% of sensitivity and specificity, suggesting potential use for TL-diagnosis in its different clinical manifestations. However, a high number of proof-of-concept studies reinforce the need for further analysis aimed at verifying true diagnostic accuracy in clinical practice.
Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/sangue , Leishmania/imunologia , Leishmaniose Tegumentar Difusa/diagnóstico , Leishmaniose Mucocutânea/diagnóstico , Antígenos de Protozoários/classificação , Antígenos de Protozoários/imunologia , Cromatografia de Afinidade/normas , Ensaio de Imunoadsorção Enzimática/normas , Humanos , Imuno-Histoquímica/normas , Leishmaniose Tegumentar Difusa/imunologia , Leishmaniose Tegumentar Difusa/parasitologia , Leishmaniose Mucocutânea/imunologia , Leishmaniose Mucocutânea/parasitologia , Testes Imediatos/normas , Guias de Prática Clínica como Assunto , Sensibilidade e EspecificidadeRESUMO
Cutaneous leishmaniasis (CL) is one of the most frequent parasitic zoonoses in Panama. Currently, conventional, molecular and histopathological tests are performed to diagnose CL. Immunohistochemistry (IHC) has proven to be a valuable tool to facilitate the diagnosis of leishmaniasis and to study the cellular immune response developed during the infection. Therefore, considering the absence of IHC in the diagnostic routine in Panama, the objective of this study is to demonstrate the usefulness of this test as a complementary diagnostic tool for improving the sensitivity of histopathology (HP) and helping to study the cellular immune response of patients. Samples from patients with suspected CL were analysed by intradermal reaction of Montenegro (IDRM), smears, culture, PCR (Viannia, Hsp-70), HP and IHC. According to the diagnostic criteria, 95.8% of patients were positive for Leishmania sp., that was characterized as Leishmania (V.) panamensis by PCR-HSP70/RFLP. From positive samples, Leishmania was detected by the tested diagnostic methods in the following degrees: 100% by IDRM, 60% by smears, 93.3% by culture, 100% by kDNA PCR, 78.3% by PCR Hsp-70, 50% by HP and 73.9% by IHC. Although IHC had a poor correlation (kâ¯=â¯0.191) with the diagnostic criteria, the sensitivities of both HP (76.1%) and smears (89.1%) were improved by combining them with IHC. IHC considerably improved the detection of the Leishmania parasites in the histopathological sections, supporting the need to implement this diagnostic tool in Panama. In addition, immunohistochemistry allows evaluation of the patient's immune response and thus provides new guidelines for the treatment and control of CL in Panama.
Assuntos
Imuno-Histoquímica/normas , Leishmania/isolamento & purificação , Leishmaniose Cutânea/diagnóstico , Pele/patologia , Adulto , Idoso , Antígenos de Protozoários/imunologia , Biópsia , DNA de Protozoário/genética , Feminino , Técnicas Histológicas , Humanos , Imunidade Celular , Leishmania/genética , Masculino , Pessoa de Meia-Idade , Panamá , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade , Pele/parasitologia , Adulto JovemRESUMO
BACKGROUND: Despite the very low or absent parasitism in the lungs, the interstitial pneumonitis is a common lesion found in humans and dogs with visceral leishmaniasis. The lung is a neglected organ in the study of dogs and humans with visceral leishmaniasis, but interstitial pneumonitis represents an important lesion characterized by thickening of the alveolar septum due to fibrosis and inflammatory exudate, and its pathogenesis is still uncertain. In this study, the polymerase chain reaction (PCR) was used to detect Leishmania infantum in paraffin-embedded lung biopsies from naturally infected dogs from an endemic area in Minas Gerais State, Brazil; PCR was compared to histological and immunohistochemical techniques for detecting Leishmania. RESULTS: Eighteen dogs in which leishmaniasis had been diagnosed by serological tests - indirect immunofluorescence assay (IFAT), enzyme-linked immunosorbent assay (ELISA) and complement fixation tests (CFT) - were classified as asymptomatic, oligosymptomatic or symptomatic. Nine of the 18 dogs studied had a positive PCR (50%) but parasites were not detected by histopathological and immunocytochemistry methods. CONCLUSIONS: These data indicate that PCR on DNA extracted from paraffin-embedded tissue is a valuable method for detecting Leishmania infantum parasites in lungs of naturally infected dogs, despite the apparent absence of parasites from standard HE (hematoxylin and eosin) stained slides and of labeled parasites from immunocytochemical preparations.
Assuntos
DNA de Protozoário/análise , Doenças do Cão/diagnóstico , Leishmaniose Visceral/veterinária , Pulmão/parasitologia , Animais , Brasil , Doenças do Cão/parasitologia , Doenças do Cão/patologia , Cães , Imuno-Histoquímica/normas , Imuno-Histoquímica/veterinária , Leishmania infantum/genética , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/patologia , Pulmão/química , Reação em Cadeia da Polimerase/normas , Reação em Cadeia da Polimerase/veterináriaRESUMO
Accessibility to immunohistochemistry (IHC) is invaluable to proper diagnosis and treatment of pediatric patients with malignant neoplasms. Whereas IHC is widely available in anatomic pathology laboratories in high-income countries, access to it in anatomic pathology laboratories of low- and middle-income countries remains a struggle, with many limitations. To advance the quality of the pathology service offered to children with cancer in areas with limited resources, a 5-day pathology training workshop was offered to pathologists and histotechnologists from various countries of the Central American and Caribbean region. An initial assessment of the workshop participants' current laboratory capacities was performed, and a regional training center was selected. Didactic and hands-on activities were offered, and review and evaluation of the IHC slides produced during the training course were compared with original slides from the participants' sites. This model of intensive 5-day training appears to be effective and can potentially be used in other budget-constrained regions. Moreover, it can serve as a continuing education activity for pathologists and histotechnologists, and as part of validations and quality improvement projects to build capacity and develop IHC assay proficiency in low- and middle-income countries.
Assuntos
Imuno-Histoquímica , Neoplasias/diagnóstico , Neoplasias/epidemiologia , Assistência ao Paciente , Região do Caribe/epidemiologia , América Central/epidemiologia , Criança , Gerenciamento Clínico , Humanos , Imuno-Histoquímica/métodos , Imuno-Histoquímica/normas , Gradação de Tumores , Estadiamento de Neoplasias , Neoplasias/terapiaRESUMO
Therapy for non-small cell lung carcinoma (NSCLC) is currently determined by histologic subtype and the presence or absence of actionable mutations. Accurate subclassification is therefore essential for appropriate selection of cases for molecular studies and guiding treatment. The gold standard for subclassification of NSCLC is identification of differentiating morphologic features in correlation with diagnostic immunohistochemistry (IHC) in cases of poorly differentiated carcinoma. Whereas Napsin A, TTF1, and p40 antibodies have been used individually for the subtyping of NSCLC, few studies have examined the 3 in cocktail form. Using a novel triple IHC antibody cocktail (TNP) composed of TTF1 (brown nuclear), Napsin A (red granular cytoplasmic), and p40 (red nuclear), a randomized, double-blinded subclassification was performed on a representative histologic section of 32 previously resected primary NSCLCs. TNP results were then compared with the gold-standard diagnosis. TNP accurately identified all (100%, 10/10) squamous cell carcinomas (SCCs) (p40+/TTF1-/Napsin A-) and 89% (16/18) of adenocarcinomas (ADCs) (p40-/TTF1+/Napsin A+). TNP was negative in 7 (20%) tumors (p40-/TTF1-/Napsin A-), including 2 mucinous ADCs. TNP showed no overlapping or discordant immunostaining. Using traditional IHC with p63, CK5/6, and TTF1, all TNP (-) cases remained unclassifiable. With the exception of mucinous ADC, which was TNP negative, all TNP cases correlated with gold-standard diagnosis; 78% of tumors were also definitively classified as either ADC or SCC and required only a single slide for classification.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/patologia , Imuno-Histoquímica/métodos , Imuno-Histoquímica/normas , Anticorpos Monoclonais/metabolismo , Humanos , Reprodutibilidade dos TestesRESUMO
Cancer stem cells (CSC) have been investigated as prognostic markers in oral squamous cell carcinoma (OSCC). However, an assessment of the reporting quality of these studies has not been performed yet. The aim of this study was to describe the reporting quality of prognostic studies involving CSCs and OSCC, focusing mainly on the immunohistochemical reproducibility. By means a systematic review, 34 articles were selected. Analyses of both general reporting quality and immunohistochemistry technique were performed by using checklists for multiple aspects related to study reproducibility. A total of 21 different CSC markers were cited in the selected studies, evaluated by means of a wide range of antibodies, most of them (40.3%) without clone description. Discrepancies in intracellular immunolabeling were noted for some markers. The mean global score for general quality assessment revealed limits in the quality of the articles. The main problems were related to lack of report on OSCC characteristics and treatment, sample size rationale, and sensitivity analysis or internal validation of the markers. Although there was a high frequency of studies having "good or very good" score for immunohistochemistry reproducibility, the frequency of articles with "poor or very poor" score for individual items was expressive, mainly for description of immunolabeling analysis (38.2% of the studies were poorly described). In conclusion, although there is a significant range of CSC markers with promising results for prognosis of OSCC, the inadequate reporting of important sections in the published studies, including immunohistochemistry technique, may limit the quality of the investigation.
Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/diagnóstico , Imuno-Histoquímica/normas , Neoplasias Bucais/diagnóstico , Células-Tronco Neoplásicas/patologia , Regulação Neoplásica da Expressão Gênica , Humanos , Prognóstico , Garantia da Qualidade dos Cuidados de Saúde , Reprodutibilidade dos TestesRESUMO
INTRODUCTION: Heat-induced epitope retrieval (HIER) of formalin-fixed paraffin-embedded tissues is now a standard practice in immunohistochemistry (IHC). In this study, we aimed to test the effect of altering HIER temperature on IHC staining quality at high altitude, the hypothesis being that lower HIER temperatures would result in improved staining patterns. MATERIALS AND METHODS: In a laboratory at high altitude (Aurora, CO), we used a platform with automated onboard epitope retrieval, and systematically tested 3 different HIER temperatures (100°C, 95°C, 90°C) with 4 IHC stains that are commonly used in routine practice: CD3, Ki67, CK20, and Melan A (n=10 for each antibody/epitope retrieval temperature combination). A scoring system was devised, the slides were scored in a blinded manner, and statistical analysis was performed. For comparison, the same study was performed in a laboratory near sea level (Atlanta, GA). RESULTS: At high altitude, lower HIER temperatures resulted in improved staining patterns, as quantified by stronger staining intensity and greater area of the slides stained. The scores obtained with HIER temperatures of 95°C and 90°C were higher than those obtained with HIER of 100°C, and the difference was found to be statistically significantly for some antibody/epitope retrieval temperature combinations (P<0.05). This effect was not seen in the laboratory near sea level. CONCLUSIONS: We show that alternate epitope retrieval recommendations are warranted for laboratories at high altitude. Furthermore, we suggest that manufactures should consider how their instruments will perform at high altitude as they further automate the process of IHC.
Assuntos
Altitude , Complexo CD3 , Temperatura Alta , Imuno-Histoquímica/métodos , Complexo CD3/química , Humanos , Imuno-Histoquímica/normas , Queratina-20/química , Antígeno Ki-67/química , Antígeno MART-1/química , Inclusão em Parafina , Controle de QualidadeRESUMO
BACKGROUND: The in situ detection of parasite antigens in tissue sections by immunohistochemistry (IHC) is a diagnostic alternative for human American tegumentary leishmaniasis (ATL), but has not been used for the diagnosis of cutaneous lesions in dogs with ATL. This study describes the results of IHC for the detection of amastigote forms and other Leishmania sp. antigen-positive cells and compares the results of IHC, histopathology and cytopathology for the diagnosis of canine ATL. In addition, possible cross-reactivity with sporotrichosis is analyzed. METHODS: Forty paraffin-embedded biopsies and 40 smears of cutaneous lesions from dogs with ATL, confirmed by isolation and characterization of Leishmania (Viannia) braziliensis, and 40 paraffin-embedded biopsies of cutaneous lesions from dogs with sporotrichosis, confirmed by isolation of Sporothrix schenckii in culture (control group), were studied. RESULTS: Immunohistochemistry was more sensitive in detecting amastigote forms than cytopathology and histopathology, with a positivity rate of 70% (n=28) versus 37.5% and 22.5% for histopathology and cytopathology, respectively. Cytoplasmic staining of mononuclear and endothelial cells was detected by IHC, which was highly specific since no cytoplasmic staining of these cells or staining of fungal structures was observed in sporotrichosis fragments. CONCLUSIONS: In view of the higher sensitivity of IHC in detecting Leishmania sp. antigen and patterns of positivity for Leishmania sp. antigen compared to histopathology or cytopathology and the absence of cross-reactions with sporotrichosis, we recommend this technique for the diagnosis of canine tegumentary leishmaniasis.
Assuntos
Doenças do Cão/diagnóstico , Imuno-Histoquímica/veterinária , Leishmania braziliensis/isolamento & purificação , Leishmaniose Cutânea/veterinária , Animais , Antígenos de Protozoários/imunologia , Doenças do Cão/imunologia , Cães , Imuno-Histoquímica/métodos , Imuno-Histoquímica/normas , Leishmania braziliensis/imunologia , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/imunologia , Sensibilidade e EspecificidadeRESUMO
There is a lack of standardization of a best practice protocol for Phosphatase and Tensin Homolog (PTEN) assessment by immunohistochemistry in anatomic pathology routine practice. We performed immunohistochemistry for 19 antibodies against PTEN, eleven of which were excluded during the standardization step. Immunohistochemistry of the remaining eight antibodies was performed on a Tissue Microarray containing 55 prostate and 40 renal carcinoma samples. Fluorescent in situ hybridization (FISH) was used as reference standard for immunohistochemistry specificity evaluation. Concerning nuclear staining, polyclonal (Cat#22034-1-AP); 6H2.1 mMAb (Cat#ABM-2052), Y184 RabMAb (Cat#NB110-57441) and 217702 mMAb antibodies presented the highest agreement with fluorescent in situ hybridization (p<0.001 for all) and with regard to cytoplasmic staining, Y184 RabMAb (Cat#NB110-57441); polyclonal (Cat#22034-1-AP) and 217702 mMAb presented the highest agreement (p<0.001 for all). Our results indicate that several commercially available antibodies do not show reliability of sensitivity and specificity for PTEN evaluation and we propose 6H2.1 mMAb (Cat#ABM-2052) as the antibody of choice for laboratory standardization and best practice in clinical routine, which demonstrated excellent sensitivity for both nuclear and cytoplasmic staining, specificity for PTEN by Western blot and good correlation with PTEN status by FISH with regard to nuclear staining.
Assuntos
Adenocarcinoma/metabolismo , Neoplasias Renais/metabolismo , PTEN Fosfo-Hidrolase/genética , Neoplasias da Próstata/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Humanos , Imuno-Histoquímica/normas , Hibridização in Situ Fluorescente , Rim/metabolismo , Rim/patologia , Neoplasias Renais/genética , Neoplasias Renais/patologia , Masculino , PTEN Fosfo-Hidrolase/metabolismo , Guias de Prática Clínica como Assunto , Próstata/metabolismo , Próstata/patologia , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Padrões de Referência , Análise Serial de TecidosRESUMO
Toxoplasma gondii is a zoonotic agent of great importance in veterinary and public health. The aim of this study was to identify T. gondii by IHC (immunohistochemistry) in different sheep tissues and to determine if an association exists between the results obtained by this method and those obtained by the Modified Agglutination Test (MAT). Tissue specimens of twenty-six sheep seroreactive for T. gondii were selected for histopathological evaluation. The presence of T. gondii was investigated in brain, liver and heart samples by IHC and a possible anti-T. gondii antibody cross reactions with other parasites. McNemar's, Chi-square and Fisher's Exact Tests were applied for the statistical analysis of the results. The analysed tissues showed at least one of the following histopathological changes: mild-to-moderate congestion, focal polymorphonuclear inflammatory infiltrate and multifocal or focal mononuclear inflammatory infiltrate. Sarcocystis spp. were identified in the histological sections from both the heart and diaphragm tissues of 88.5% (23/26) of the animals. A total of 46.2% (12/26) of the T. gondii seroreactive sheep was also positive for T. gondii by IHC in at least one organ (brain, liver or heart). The liver IHC-positivity for T. gondii was statistically equivalent to the global individual IHC-positivity, according to McNemar's test. In addition, IHC allowed the detection of T. gondii in infected animals regardless of the titration observed in the MAT. The statistical difference observed between the three organs when comparing the low titration group, suggested that the heart might be the most suitable organ to detect T. gondii infection by IHC. The IHC results in this study revealed that almost half of MAT positive animals could serve as potential sources of infection for humans because bradyzoites were identified in different tissues, regardless of the MAT titration.
Assuntos
Testes de Aglutinação/veterinária , Imuno-Histoquímica/veterinária , Doenças dos Ovinos/diagnóstico , Toxoplasmose Animal/diagnóstico , Testes de Aglutinação/normas , Animais , Anticorpos Antiprotozoários/metabolismo , Encéfalo/parasitologia , Diafragma/parasitologia , Coração/parasitologia , Imuno-Histoquímica/normas , Fígado/parasitologia , Ovinos , ToxoplasmaRESUMO
Gastrointestinal stromal tumor is the most common clinically significant mesenchymal neoplasm of the gastrointestinal tract. The expression of the intermediate filament cytokeratin in gastrointestinal stromal tumor is not frequently reported in the literature. The aim of this study was to investigate the immunohistochemical expression of several types of cytokeratin in a large number of cases (n=687), including a pan-cytokeratin marker (AE1/AE3 cocktail antibodies), high-molecular weight cytokeratins (34ßE12 antibody), and individual cytokeratins 8 (35ßH11 and CAM5.2 antibodies), 7, 14, and 20. Ki-67 antigen was used for the determination of cell proliferation index, and the correlation between Ki-67 and cytokeratin expression was evaluated. Cytokeratin expression was also correlated with several clinicopathologic parameters. The expression of pan-cytokeratin was observed in 24 (3.5%) cases, with variable intensity. Only 1 of 687 (0.1%) cases showed cytokeratin 14 expression. All 687 cases revealed no expression of high-molecular weight cytokeratins, cytokeratins 7, 8, and 20. No significant statistical association was found between AE1/AE3 immunoreactivity and several clinicopathologic parameters, including sex, tumor location and size, cell morphology, mitotic count, risk of aggressive behavior, and Ki-67 antigen cell proliferation index. However, statistical correlation between AE1/AE3 immunoreactivity and a higher age at diagnosis was detected. These results show that cytokeratin expression is not frequent in gastrointestinal stromal tumor, but caution is necessary to avoid erroneous diagnoses.
Assuntos
Biomarcadores Tumorais/biossíntese , Neoplasias Gastrointestinais , Regulação Neoplásica da Expressão Gênica , Queratina-14/biossíntese , Queratina-20/biossíntese , Queratina-7/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antineoplásicos/química , Proliferação de Células , Feminino , Neoplasias Gastrointestinais/metabolismo , Neoplasias Gastrointestinais/patologia , Humanos , Imuno-Histoquímica/métodos , Imuno-Histoquímica/normas , Antígeno Ki-67/biossíntese , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
O6-Methyguanine-DNA methyltransferase (MGMT) repairs DNA damage and acts as a tumor suppressor in normal cells by preventing DNA mutations. Several antibodies against MGMT are used for immunohistochemical assessment of this marker and no universal standard is adopted. We evaluated the immunohistochemical expression of MGMT with 5 commercially available primary antibodies in 59 invasive breast carcinomas. A tissue microarray was constructed using 59 invasive breast carcinoma samples. Five primary antibodies against MGMT were used for immunohistochemistry, including clones MT3.1, SPM287, and MT23.2. Heat-induced antigen retrieval and polymer-based immunohistochemistry were performed. Stains were analyzed by microscopy and automated digital slide technology. qRT-PCR was performed for all tumors. Clone SPM287 had the highest sensitivity (p<0.001), and clone MT3.1 had the lowest sensitivity (p<0.001). Clone MT23.2 generated higher levels of cytoplasmic staining, which was not observed with the other antibodies (p<0.001). Fifty-six samples (94.9%) showed hypoexpression of MGMT compared with normal breast, as measured by qRT-PCR (p<0.001). Only clone SPM287 correlated significantly with the qRT-PCR results (p=0.027). Antibody clone SPM287 is the most sensitive and specific antibody for the immunohistochemical evaluation of MGMT, rendering it a reliable and effective reagent for research and clinical practice in breast cancer.
Assuntos
Anticorpos Monoclonais , Biomarcadores Tumorais/análise , Neoplasias da Mama/enzimologia , Carcinoma Ductal de Mama/enzimologia , Metilases de Modificação do DNA/análise , Enzimas Reparadoras do DNA/análise , Imuno-Histoquímica/normas , Proteínas Supressoras de Tumor/análise , Especificidade de Anticorpos , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/patologia , Feminino , Humanos , Guias de Prática Clínica como Assunto , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Análise Serial de TecidosRESUMO
Breast cancer accounts for approximately one quarter of all cancers in females. HER2 gene amplification or HER2 protein overexpression, detected in about 20% of breast carcinomas, predicts a more aggressive clinical course and determines eligibility for targeted therapy with trastuzumab. HER2 testing has become an essential part of the clinical evaluation of all breast carcinoma patients, and accurate HER2 results are critical in identifying patients who may be benefited from targeted therapy. This study investigated the concordance in the results of HER2 immunohistochemistry assays performed in 500 invasive breast carcinomas between a reference laboratory and 149 local laboratories from all geographic regions of Brazil. Our results showed an overall poor concordance (171 of 500 cases, 34.2%) regarding HER2 results between local and reference laboratories, which may be related to the low-volume load of HER2 assays, inexperience with HER2 scoring system, and/or technical issues related to immunohistochemistry in local laboratories. Standardization of HER2 testing with rigorous quality control measures by local laboratories is highly recommended to avoid erroneous treatment of breast cancer patients.
Assuntos
Biomarcadores Tumorais/análise , Neoplasias da Mama/diagnóstico , Imuno-Histoquímica/normas , Hibridização in Situ Fluorescente/normas , Receptor ErbB-2/análise , Anticorpos Monoclonais/administração & dosagem , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Brasil , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Feminino , Humanos , Imuno-Histoquímica/métodos , Hibridização in Situ Fluorescente/métodos , Variações Dependentes do Observador , Valor Preditivo dos Testes , Controle de Qualidade , Receptor ErbB-2/genética , Padrões de Referência , TrastuzumabRESUMO
Identifying breast cancers with HER2 overexpression or amplification is critical as these usually imply the use of HER2-targeted therapies. DNA (amplification) and protein (overexpression) HER2 abnormalities usually occur simultaneously and both in situ hybridisation and immunohistochemistry may be accurate methods for the evaluation of these abnormalities. However, recent studies, including those conducted by the Association for Quality Assurance of the Spanish Society of Pathology, as well as the experience of a number of HER2 testing National Reference Centres have suggested the existence of serious reproducibility issues with both techniques. To address this issue, a joint committee from the Spanish Society of Pathology (SEAP) and the Spanish Society of Medical Oncology (SEOM) was established to review the HER2 testing guidelines. Consensus recommendations are based not only on the panellists' experience, but also on previous consensus guidelines from several countries, including the USA, the UK and Canada. These guidelines include the minimal requirements that pathology departments should fulfil in order to guarantee proper HER2 testing in breast cancer. Pathology laboratories not fulfilling these standards should make an effort to meet them and, until then, are highly encouraged to submit to reference laboratories breast cancer samples for which HER2 determination has clinical implications for the patients.
Assuntos
Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , DNA de Neoplasias/análise , Genes erbB-2 , Imuno-Histoquímica/métodos , Hibridização In Situ/métodos , Serviço Hospitalar de Patologia/normas , Manejo de Espécimes/métodos , Algoritmos , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/tratamento farmacológico , Carcinoma Ductal de Mama/patologia , Ensaios Clínicos Fase III como Assunto/estatística & dados numéricos , Feminino , Controle de Formulários e Registros/normas , Humanos , Imuno-Histoquímica/normas , Hibridização In Situ/normas , Estudos Multicêntricos como Assunto , Serviço Hospitalar de Patologia/organização & administração , Serviço Hospitalar de Patologia/estatística & dados numéricos , Garantia da Qualidade dos Cuidados de Saúde/organização & administração , Kit de Reagentes para Diagnóstico , Reprodutibilidade dos Testes , Espanha , Manejo de Espécimes/normas , TrastuzumabRESUMO
Some mycobacterial infections, such as tuberculosis, are characterized by apoptosis in infected or by-stander mononuclear immune cells. For localized (paucibacillary, PB) and diseminated (multibacillary, MB) leprosy, characterized by polarized Thl-like vs, Th2-like immune responses, respectivelly little is known about lesional apoptosis. We analyzed sections of paraffin-embedded, untreated leprosy lesions from 21 patients by an indirect immunofluorescent terminal deoxynucleotide-transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay. Some TUNEL (+) PB sections were then reacted with phycoerythein-conjugated (red)antibodies against T cells, monocytes, or antigen-presenting (Langerhans) cells. TUNEL (+) bodies were detected in 9 of 16 PB lesions (56%) and in 1 of 5 MB lesions (20%). Some TUNELL (+) bodies in PB disease were CD3+ (T cell), as well as CD4+ (T-helper) or cd8+ (T-cytotoxic) Apoptosis characterizes PB and MB leprosy lesions and may be more freqeunt in PB disease. In PB disease, some TUNEL (+) bodies may derive from T cells
Assuntos
Humanos , Hanseníase Dimorfa/imunologia , Hanseníase Dimorfa/patologia , Hanseníase Tuberculoide/imunologia , Hanseníase Tuberculoide/patologia , Hanseníase Virchowiana/imunologia , Hanseníase Virchowiana/patologia , Hanseníase/imunologia , Hanseníase/patologia , Imuno-Histoquímica , Mycobacterium leprae/patogenicidade , Apoptose/fisiologia , Apoptose/imunologia , Biópsia , Imuno-Histoquímica/normasRESUMO
OBJECTIVE: To assess the contribution of immunocytochemistry (ICC) to aspiration biopsy cytology (ABC), in a diagnostic context, on routine, previously stained cytologic specimens. STUDY DESIGN: Among 5,221 consecutive cases of ABC, 5.3% were subjected to ICC in the clinical-morphologic context. One hundred of these cases, with a final clinical and histopathologic diagnosis, were studied to determine the contribution of this ancillary technique to the final cytologic diagnosis. All cases had histopathologic study and prospective ICC, performed on usual smears, alcohol fixed and previously stained by the Papanicolaou technique, and were subjected to an avidinbiotin-peroxidase complex method. RESULTS: ICC was contributory in 82% of cases. The contribution of ICC to ABC of lymphoid tissue, thyroid and related organs, soft tissue and miscellaneous cases was, respectively, 84% (39 cases), 88% (26), 72% (18) and 76% (17). ICC was noncontributory in 18 cases, due mainly to misleading interpretation (6%), uncharacteristic profile (5%) and inconclusive immunostain (7%). CONCLUSION: ICC could be successfully applied in routine ABC specimens since the usually investigated antigenic determinants are preserved, allowing previous morphologic study and screening of the smears. The principal contribution of ICC applied to lymph nodes, thyroid and soft tissue aspirates was, respectively, confirmation of metastatic neoplasms, differential of follicular versus C-cell proliferation and assessment of mesenchymal lineages.
Assuntos
Biópsia por Agulha/métodos , Carcinoma de Células Grandes/patologia , Imuno-Histoquímica/métodos , Metástase Linfática/patologia , Neoplasias da Glândula Tireoide/patologia , Adenocarcinoma/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Biópsia por Agulha/normas , Carcinoma Medular/patologia , Criança , Pré-Escolar , Feminino , Humanos , Imuno-Histoquímica/normas , Tecido Linfoide/patologia , Linfoma/patologia , Masculino , Melanoma/patologia , Mesoderma/patologia , Pessoa de Meia-Idade , Estudos Prospectivos , Neoplasias da Próstata/patologia , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Aplication of flow cytofluorometry to blood transfusion: flow citofluorometry has been applied to transfusion medicine for nearly twenty years now. Many of these applications have been for purposes, but several have been suggested for more routine uses. In this summary it is expected to reach of aplication the description of more intervention in the transfusion medicine: detection and quantitation of protein bound to blood cells; detection and quantitation of minor cell population; detection and quantitation of cellular antigens; detection of platelet and WBC antibodies; demostration of platelet activation; demos-tration and quantitation of mocyte interaction with IgG-sensitized RBCs
Assuntos
Humanos , Citometria de Fluxo/métodos , Técnicas In Vitro , Imuno-Histoquímica/normas , Antígenos de Plaquetas Humanas/análise , Sobrevivência Celular , Células Precursoras Eritroides/classificação , Quimera , Citometria de Fluxo , Eritrócitos/imunologia , Corantes Fluorescentes , Imunoglobulina G/sangue , MosaicismoRESUMO
Aplication of flow cytofluorometry to blood transfusion: flow citofluorometry has been applied to transfusion medicine for nearly twenty years now. Many of these applications have been for purposes, but several have been suggested for more routine uses. In this summary it is expected to reach of aplication the description of more intervention in the transfusion medicine: detection and quantitation of protein bound to blood cells; detection and quantitation of minor cell population; detection and quantitation of cellular antigens; detection of platelet and WBC antibodies; demostration of platelet activation; demos-tration and quantitation of mocyte interaction with IgG-sensitized RBCs (AU)
Assuntos
Humanos , Técnicas In Vitro , Citometria de Fluxo/métodos , Imuno-Histoquímica/normas , Citometria de Fluxo/estatística & dados numéricos , Eritrócitos/imunologia , Imunoglobulina G/sangue , Quimera , Mosaicismo , Sobrevivência Celular , Corantes Fluorescentes/diagnóstico , Antígenos de Plaquetas Humanas/análise , Células Precursoras Eritroides/classificaçãoRESUMO
El diagnóstico de los linfomas cutáneos tiene importantes implicancias para los pacientes. Tradicionalmente el diagnóstico de cualquier lesión cutánea está basado en criterios clínicos e histopatológicos. Ninguno de estos criterios es absoluto y en los últimos años se ha destacado la trascendencia de la inmunohistoquímica y la biología molecular en relación con el diagnóstico y pronóstico. La inmunohistoquímica permite definir subpoblaciones de linfocitos, estableciendo el fenotipo, categorizando algunos cuadros y destacando en algunos casos el pronóstico a partir del CD30 (+) o (-). La biología molecular define el genotipo, aportando el concepto de monoclonalidad que sugiere malignidad. Por otra parte, proponemos una clasificación de los linfomas cutáneos primarios que ha sido recientemente introducida
Assuntos
Humanos , Imuno-Histoquímica/normas , Linfoma de Células B/diagnóstico , Linfoma Cutâneo de Células T/diagnóstico , Diagnóstico Diferencial , Rearranjo Gênico da Cadeia delta dos Receptores de Antígenos dos Linfócitos T/imunologia , Linfoma de Células B/classificação , Linfoma de Células B/genética , Linfoma Cutâneo de Células T/classificação , Linfoma Cutâneo de Células T/genética , Biologia Molecular/tendências , Reação em Cadeia da Polimerase/tendênciasRESUMO
El diagnóstico de los linfomas cutáneos tiene importantes implicancias para los pacientes. Tradicionalmente el diagnóstico de cualquier lesión cutánea está basado en criterios clínicos e histopatológicos. Ninguno de estos criterios es absoluto y en los últimos años se ha destacado la trascendencia de la inmunohistoquímica y la biología molecular en relación con el diagnóstico y pronóstico. La inmunohistoquímica permite definir subpoblaciones de linfocitos, estableciendo el fenotipo, categorizando algunos cuadros y destacando en algunos casos el pronóstico a partir del CD30 (+) o (-). La biología molecular define el genotipo, aportando el concepto de monoclonalidad que sugiere malignidad. Por otra parte, proponemos una clasificación de los linfomas cutáneos primarios que ha sido recientemente introducida (AU)