RESUMO
The present study aimed to compare two different methods of extracting RNA from formalin-fixed paraffin-embedded (FFPE) specimens of diffuse large B-cell lymphoma (DLBCL). We further aimed to identify possible influences of variables--such as tissue size, duration of paraffin block storage, fixative type, primers used for cDNA synthesis, and endogenous genes tested--on the success of amplification from the samples. Both tested protocols used the same commercial kit for RNA extraction (the RecoverAll Total Nucleic Acid Isolation Optimized for FFPE Samples from Ambion). However, the second protocol included an additional step of washing with saline buffer just after sample rehydration. Following each protocol, we compared the RNA amount and purity and the amplification success as evaluated by standard PCR and real-time PCR. The results revealed that the extra washing step added to the RNA extraction process resulted in significantly improved RNA quantity and quality and improved success of amplification from paraffin-embedded specimens.
Assuntos
Fixadores , Formaldeído , Linfoma Difuso de Grandes Células B/química , Inclusão em Parafina , RNA Neoplásico , RNA , Humanos , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , RNA/química , RNA/isolamento & purificação , RNA Neoplásico/química , RNA Neoplásico/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Follicular lymphoma is clinically heterogenous, and therefore necessitates the identification of prognostic markers to stratify risk groups and optimize clinical management. It is relatively rare in patients younger than 40 years, and the clinicopathologic characteristics and biological behavior in this age group are poorly understood. In the current study, samples from a cohort of 200 patients between 19 and 40 years were evaluated retrospectively with respect to clinical, histologic, and genetic features. These were then correlated with clinical outcome. The median age at presentation was 35 years with a slight female prepoderance (56%). Most of the cases are presented with nodal disease (90%). Concomitant follicular lymphoma and diffuse large B-cell lymphoma were observed in 7 (4%) patients. Immunohistologic studies showed the expression of CD10 (91%), BCL6 (97%), BCL2 (95%), MUM1/IRF4 (12%), MDM2 (17%), and CD23 (25%). BCL2 rearrangement was present in 74%, and BCL6 in 20%. The estimated overall survival of patients was 13 years (mean). The presence of anemia, elevated lactose dehydrogenase, bone marrow involvement, and high-risk follicular lymphoma international prognostic index correlated with adverse overall survival. Our findings revealed that follicular lymphoma in young adults demonstrate similarities with that of older adults, including the frequency of presentation at various anatomic sites, grade, and adverse prognostic factors.
Assuntos
Biomarcadores Tumorais/análise , Linfoma Folicular/diagnóstico , Linfoma Difuso de Grandes Células B/diagnóstico , Adulto , Fatores Etários , Biomarcadores Tumorais/genética , Análise por Conglomerados , Feminino , Humanos , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Estimativa de Kaplan-Meier , Modelos Logísticos , Linfonodos/química , Linfonodos/patologia , Linfoma Folicular/química , Linfoma Folicular/genética , Linfoma Folicular/mortalidade , Linfoma Folicular/patologia , Linfoma Folicular/terapia , Linfoma Difuso de Grandes Células B/química , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/mortalidade , Linfoma Difuso de Grandes Células B/patologia , Masculino , Gradação de Tumores , Estadiamento de Neoplasias , Razão de Chances , Valor Preditivo dos Testes , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
Diffuse large B-cell lymphoma can be subclassified into at least two molecular subgroups by gene expression profiling: germinal center B-cell like and activated B-cell like diffuse large B-cell lymphoma. Several immunohistological algorithms have been proposed as surrogates to gene expression profiling at the level of protein expression, but their reliability has been an issue of controversy. Furthermore, the proportion of misclassified cases of germinal center B-cell subgroup by immunohistochemistry, in all reported algorithms, is higher compared with germinal center B-cell cases defined by gene expression profiling. We analyzed 424 cases of nodal diffuse large B-cell lymphoma with the panel of markers included in the three previously described algorithms: Hans, Choi, and Tally. To test whether the sensitivity of detecting germinal center B-cell cases could be improved, the germinal center B-cell marker HGAL/GCET2 was also added to all three algorithms. Our results show that the inclusion of HGAL/GCET2 significantly increased the detection of germinal center B-cell cases in all three algorithms (P<0.001). The proportions of germinal center B-cell cases in the original algorithms were 27%, 34%, and 19% for Hans, Choi, and Tally, respectively. In the modified algorithms, with the inclusion of HGAL/GCET2, the frequencies of germinal center B-cell cases were increased to 38%, 48%, and 35%, respectively. Therefore, HGAL/GCET2 protein expression may function as a marker for germinal center B-cell type diffuse large B-cell lymphoma. Consideration should be given to the inclusion of HGAL/GCET2 analysis in algorithms to better predict the cell of origin. These findings bear further validation, from comparison to gene expression profiles and from clinical/therapeutic data.
Assuntos
Algoritmos , Biomarcadores Tumorais/análise , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/química , Proteínas de Neoplasias/análise , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Distribuição de Qui-Quadrado , Criança , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Modelos Lineares , Linfoma Difuso de Grandes Células B/patologia , Masculino , Proteínas dos Microfilamentos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise Serial de Tecidos , Adulto JovemRESUMO
Primary cutaneous lymphomas are defined as lymphocytic neoplasias that present themselves clinically in the skin without extracutaneous disease at diagnosis and up to 6 months after it. The authors report the case of an elderly male patient, with a three- month-history of papules in the axilla which evolved into painful ulceration. Examination found deep ulcer with irregular borders ,infiltrates, in the right axilla. Physical and additional examinations did not evidence disease at distance. Histopathology revealed dense and diffuse dermic sample infiltrate of atypical lymphocytes. Imunohistochemistry shows expression of CD20 and bcl-2 antigens , with negative CD10, configuring diagnosis of cutaneous large B-cell lymphoma. In this type of cutaneous lymphoma, primary cutaneous manifestation is rare ,the incidence in men is lower and it is most commonly located in the lower limbs.
Assuntos
Linfoma Difuso de Grandes Células B/patologia , Neoplasias Cutâneas/patologia , Idoso de 80 Anos ou mais , Antígenos CD20/análise , Axila , Biomarcadores Tumorais/análise , Humanos , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/química , Masculino , Proteínas Proto-Oncogênicas c-bcl-2/análise , Neoplasias Cutâneas/químicaRESUMO
Linfomas cutâneos primários são definidos como neoplasias linfocíticas que se apresentam clinicamente na pele sem doença extracutânea no momento do diagnóstico e até por 6 meses após. Os autores relatam o caso de um paciente masculino, idoso, com história de pápulas em axila, há 3 meses, que evoluíram para ulceração. Ao exame, úlcera profunda de bordos irregulares, infiltrados, em axila direita. Exames físico e complementares não evidenciaram doença à distância. O histopatológico mostra infiltrado dérmico denso e difuso de linfócitos atípicos. A imuno-histoquímica evidencia expressão de antígenos CD20 e bcl-2, com CD10 negativo, configurando diagnóstico de linfoma cutâneo difuso de grandes células B. Neste tipo de linfoma, é rara a manifestação cutânea primária, assim como a incidência é menor em homens e a localização, mais comum em membros inferiores.
Primary cutaneous lymphomas are defined as lymphocytic neoplasias that present themselves clinically in the skin without extracutaneous disease at diagnosis and up to 6 months after it. The authors report the case of an elderly male patient, with a three- month-history of papules in the axilla which evolved into painful ulceration. Examination found deep ulcer with irregular borders ,infiltrates, in the right axilla. Physical and additional examinations did not evidence disease at distance. Histopathology revealed dense and diffuse dermic sample infiltrate of atypical lymphocytes. Imunohistochemistry shows expression of CD20 and bcl-2 antigens , with negative CD10, configuring diagnosis of cutaneous large B-cell lymphoma. In this type of cutaneous lymphoma, primary cutaneous manifestation is rare ,the incidence in men is lower and it is most commonly located in the lower limbs.
Assuntos
Idoso de 80 Anos ou mais , Humanos , Masculino , Linfoma Difuso de Grandes Células B/patologia , Neoplasias Cutâneas/patologia , Axila , /análise , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/química , /análise , Neoplasias Cutâneas/química , Biomarcadores Tumorais/análiseRESUMO
Diffuse large B-cell lymphoma (DLBCL) is a very infrequent neoplasm in the pediatric age group; therefore there are very few studies on the immunophenotype or genetics of these cases. We studied a series of 16 patients with nodal DLBCL occurring in patients between 10 and 18 years of age. The cases were classified according to the 2008 World Health Organization classification criteria, with application of immunohistochemistry for the detection of CD10, BCL-6, and MUM1 proteins to divide the lymphomas into germinal center and nongerminal center types. In addition, TCL1, BCL-2 expression, and the Ki-67 proliferation index were evaluated by immunohistochemistry, and c-MYC and BCL2 translocations were evaluated by fluorescence in situ hybridization. All these parameters were correlated with clinical features and outcome. Our study revealed that centroblastic morphology and the germinal center type of DLBCL are more prevalent in these young patients (63%), with 37% containing a c-MYC translocation. Only 1 case showed a BCL2 translocation, reflecting a double-hit case with features intermediate between DLBCL and Burkitt lymphoma. We found a higher frequency of BCL-2 expression than previously reported, with no direct influence on the outcome of the disease in univariate or multivariate analysis. The expression of TCL1 has not been specifically studied in nodal pediatric DLBCL before; we found a 31% incidence of TCL1 expression. MUM1 expression was observed in 44% of the cases and these positive cases showed a significant negative impact on clinical outcome. TCL1 is directly and significantly associated with the presence of c-MYC and a high proliferative index. The germinal center and nongerminal center subtypes showed significant differences for both overall survival and disease-free interval. c-MYC translocation was found in 37% of patients, and had a favorable impact on clinical outcome. We conclude that nodal pediatric and adolescent DLBCL are mainly of the germinal center type, with a generally good outcome despite the frequent expression of BCL-2 and the presence of c-MYC translocation. TCL1 expression seems to be associated with a good clinical outcome, whereas MUM1 expression predicts a poor clinical outcome.
Assuntos
Biomarcadores Tumorais/análise , Biomarcadores Tumorais/genética , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/química , Linfoma Difuso de Grandes Células B/genética , Proteínas Proto-Oncogênicas c-myc/genética , Translocação Genética , Adolescente , Criança , Proteínas de Ligação a DNA/análise , Intervalo Livre de Doença , Feminino , Humanos , Hibridização in Situ Fluorescente , Fatores Reguladores de Interferon/análise , Estimativa de Kaplan-Meier , Antígeno Ki-67/análise , Linfoma Difuso de Grandes Células B/mortalidade , Linfoma Difuso de Grandes Células B/patologia , Linfoma Difuso de Grandes Células B/terapia , Masculino , Neprilisina/análise , Valor Preditivo dos Testes , Modelos de Riscos Proporcionais , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas c-bcl-2/análise , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-6 , Estudos Retrospectivos , Medição de Risco , Fatores de Tempo , Resultado do TratamentoRESUMO
OBJECTIVE: Follicular dendritic cells (FDCs) and interdigitating dendritic cells (IDCs) are dendritic cells found in lymphoid follicles, reactive follicles and in lymphomas. The goal of this study was to evaluate the presence and distribution of FDCs and IDCs in oral lymphomas. MATERIAL AND METHODS: Immunohistochemistry reactions were applied to 50 oral lymphomas using the antibodies anti-CD21, anti-CD35 and anti-caldesmon to FDCs, and anti-S100 protein to IDCs. Caldesmon+/FDCs and S100+/IDCs were quantified in Imagelab® software. RESULTS: FDCs revealed by CD21 and CD35 were positively stained in two cases of diffuse large B-cell lymphoma, one MALT lymphoma, and in one case of mantle cell lymphoma. FDCs were immunopositive to caldesmon in all cases, as well as IDCs to S100 protein. Burkitt lymphoma presented a lower amount of caldesmon+/FDCs and S100+/IDCs than diffuse large B-cell lymphoma and plasmablastic lymphoma of the oral mucosa type. CONCLUSIONS: The microenvironment determined by neoplastic lymphoid cells in oral lymphomas is responsible by the development and expression of dendritic cells types.
Assuntos
Humanos , Células Dendríticas Foliculares/química , Células Dendríticas/química , Linfoma não Hodgkin/química , Neoplasias Bucais/química , Proteínas de Ligação a Calmodulina/análise , Imuno-Histoquímica , Linfoma de Zona Marginal Tipo Células B/química , Linfoma de Zona Marginal Tipo Células B/patologia , Linfoma Difuso de Grandes Células B/química , Linfoma Difuso de Grandes Células B/patologia , Linfoma de Célula do Manto/química , Linfoma de Célula do Manto/patologia , Linfoma não Hodgkin/patologia , Neoplasias Bucais/patologia , /análise , /análise , /análiseRESUMO
OBJECTIVE: Follicular dendritic cells (FDCs) and interdigitating dendritic cells (IDCs) are dendritic cells found in lymphoid follicles, reactive follicles and in lymphomas. The goal of this study was to evaluate the presence and distribution of FDCs and IDCs in oral lymphomas. MATERIAL AND METHODS: Immunohistochemistry reactions were applied to 50 oral lymphomas using the antibodies anti-CD21, anti-CD35 and anti-caldesmon to FDCs, and anti-S100 protein to IDCs. Caldesmon+/FDCs and S100+/IDCs were quantified in Imagelab software. RESULTS: FDCs revealed by CD21 and CD35 were positively stained in two cases of diffuse large B-cell lymphoma, one MALT lymphoma, and in one case of mantle cell lymphoma. FDCs were immunopositive to caldesmon in all cases, as well as IDCs to S100 protein. Burkitt lymphoma presented a lower amount of caldesmon+/FDCs and S100+/IDCs than diffuse large B-cell lymphoma and plasmablastic lymphoma of the oral mucosa type. CONCLUSIONS: The microenvironment determined by neoplastic lymphoid cells in oral lymphomas is responsible by the development and expression of dendritic cells types.
Assuntos
Células Dendríticas Foliculares/química , Células Dendríticas/química , Linfoma não Hodgkin/química , Neoplasias Bucais/química , Proteínas de Ligação a Calmodulina/análise , Humanos , Imuno-Histoquímica , Linfoma de Zona Marginal Tipo Células B/química , Linfoma de Zona Marginal Tipo Células B/patologia , Linfoma Difuso de Grandes Células B/química , Linfoma Difuso de Grandes Células B/patologia , Linfoma de Célula do Manto/química , Linfoma de Célula do Manto/patologia , Linfoma não Hodgkin/patologia , Neoplasias Bucais/patologia , Receptores de Complemento 3b/análise , Receptores de Complemento 3d/análise , Proteínas S100/análiseRESUMO
Ki-1 anaplastic large cell non-Hodgkin lymphoma is a well recognized clinical entity. The main clinical feature includes lymphadenopathy with mediastinal sparing. In the extranodal disease, which occurs in approximately half of the cases, the skin is the most common site; bone marrow, lung and central nervous system are generally not involved. Anaplastic large cell lymphoma is more common among young people, in whom the prognosis is more favorable. Primary anaplastic large cell lymphoma of the lung is a rare clinical entity. Its clinical expression is similar to a high grade malignant lymphoma, and in most cases the diagnosis is made in advanced stages. We present a pediatric case with ALK-1 positive anaplastic large cell lymphoma of the lung.
Assuntos
Neoplasias Pulmonares/química , Linfoma Difuso de Grandes Células B/química , Proteínas de Membrana/análise , Receptores de Activinas Tipo II , Criança , Humanos , Neoplasias Pulmonares/diagnóstico , Linfoma Difuso de Grandes Células B/diagnóstico , MasculinoRESUMO
El linfoma no Hodgkin anaplásico de células grandes, positivo al antígeno Ki-1, es una entidad bien reconocida. La característica clínica usual es la linfadenopatía con extensión mediastinal y en la enfermedad extranodal, que ocurre en la mitad de los casos, es la piel el sitio más común, con afección poco frecuente a médula ósea, pulmón y sistema nervioso. La mayor frecuencia de presentación del linfoma anaplásico de células grandes es en la población joven, con pronóstico más favora ble que en la población adulta. El linfoma anaplásico de células grandes primario de pulmón es una entidad clínica rara. Su comportamiento clínico corresponde a un linfoma de alto grado y en la mayoría de los casos se presentan al diagnóstico como una enfermedad en estadio avanzado. Se informa un caso pediátrico de linfoma anaplásico de células grandes primario de pulmón, ALK 1 positivo, que es un marcador pronóstico y específico de esta entidad.
Ki 1 anaplastic large cell non Hodgkin lymphoma is a well recognized clinical entity. The main clinical feature includes lymphadenopathy with mediastinal sparing. In the extranodal disease, which occurs in approximately half of the cases, the skin is the most common site; bone marrow, lung and central nervous system are generally not involved. Anaplastic large cell lymphoma is more common among young people, in whom the prognosis is more favorable. Primary anaplastic large cell lymphoma of the lung is a rare clinical entity. Its clinical expression is similar to a high grade malignant lymphoma, and in most cases the diagnosis is made in advanced stages. We present a pediatric case with ALK1 positive anaplastic large cell lymphoma of the lung.
Assuntos
Criança , Humanos , Masculino , Neoplasias Pulmonares/química , Linfoma Difuso de Grandes Células B/química , Proteínas de Membrana/análise , Receptores de Activinas Tipo II , Neoplasias Pulmonares/diagnóstico , Linfoma Difuso de Grandes Células B/diagnósticoRESUMO
We evaluated the expression of apoptosis-regulating proteins (p53, Bcl-2, Bax, Bak and Mcl-1) in paraffin-embedded tissues of 33 patients with diffuse large B cell non-Hodgkin's lymphoma, and assessed the relationship of these proteins to clinical outcome and response to chemotherapy. Our results showed that p53 expression was an independent immunohistochemical parameter related to a poor prognosis in these lymphomas. Bcl-2, Bax, Bak and Mcl-1 proteins, though highly expressed in almost all cases were not associated with prognosis or response to treatment.
Assuntos
Apoptose/genética , Linfoma Difuso de Grandes Células B/mortalidade , Proteínas de Neoplasias/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/análise , Feminino , Humanos , Imuno-Histoquímica , Linfoma de Células B/química , Linfoma de Células B/mortalidade , Linfoma de Células B/patologia , Linfoma Difuso de Grandes Células B/química , Linfoma Difuso de Grandes Células B/patologia , Masculino , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Proteína de Sequência 1 de Leucemia de Células Mieloides , Prognóstico , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Estudos Retrospectivos , Análise de Sobrevida , Resultado do Tratamento , Proteína Supressora de Tumor p53/metabolismo , Proteína Killer-Antagonista Homóloga a bcl-2 , Proteína X Associada a bcl-2Assuntos
AMP Cíclico/biossíntese , Histamina/fisiologia , Linfoma Difuso de Grandes Células B/química , Receptores Histamínicos H1/análise , Receptores Histamínicos H2/análise , Humanos , Linfoma Difuso de Grandes Células B/metabolismo , Receptores Histamínicos H1/efeitos dos fármacos , Receptores Histamínicos H2/efeitos dos fármacos , Sistemas do Segundo Mensageiro/fisiologia , Células Tumorais CultivadasRESUMO
An 8-year-old boy was seen with a cutaneous Ki-1 anaplastic, large cell lymphoma with multiple lesions. Some of the lesions showed spontaneous regression. During more than seven years of disease no systemic involvement was observed, but recurrent, self-healing lesions did appear. Histopathologic examination of five lesions revealed a variety of findings, from an inflammatory infiltrate to a highly anaplastic pattern. The neoplastic cells expressed Ki-1 and leukocyte common antigens. Ultrastructurally, those cells showed ruffled indentations. The differential diagnosis includes microvillous malignant lymphoma. The patient has had a four-year follow-up without relapses.