RESUMEN
The biologic and pathologic features of B-cell malignancies bearing a translocation t(14;19)(q32;q13) leading to a fusion of IGH and BCL3 are still poorly described. Herein we report the results of a comprehensive cytogenetic, fluorescence in situ hybridization (FISH), molecular and histopathological survey of a large series of B-cell malignancies with t(14;19) or variant translocations. A total of 56 B-cell malignancies with a FISH-proven BCL3 involvement were identified with the translocation partners being IGH (n=51), IGL (n=2), IGK (n=2) and a non-IG locus (n=1). Hierarchical clustering of chromosomal changes associated with the t(14;19) indicated the presence of two different groups of IG/BCL3-positive lymphatic neoplasias. The first group included 26 B-cell malignancies of various histologic subtypes containing a relatively high number of chromosomal changes and mostly mutated IgVH genes. This cluster displayed three cytogenetic branches, one with rearrangements in 7q, another with deletions in 17p and a third one with rearrangements in 1q and deletions in 6q and 13q. The second group included 19 cases, mostly diagnosed as B-cell chronic lymphocytic leukemia (B-CLL), and characterized by few additional chromosomal changes (e.g. trisomy 12) and unmutated IgVH genes. In conclusion, our study indicates that BCL3 translocations are not restricted to B-CLL but present in a heterogeneous group of B-cell malignancies.
Asunto(s)
Leucemia de Células B/genética , Linfoma de Células B/genética , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética , Translocación Genética , Adulto , Anciano , Proteínas del Linfoma 3 de Células B , Cromosomas Humanos Par 14 , Cromosomas Humanos Par 19 , Análisis Citogenético , Femenino , Reordenamiento Génico , Genes de Inmunoglobulinas , Histocitoquímica , Humanos , Hibridación Fluorescente in Situ , Leucemia de Células B/clasificación , Leucemia de Células B/patología , Linfoma de Células B/clasificación , Linfoma de Células B/patología , Masculino , Persona de Mediana EdadRESUMEN
Indolent (low-grade) B-cell non-Hodgkin's lymphomas, such as follicular and mantle cell lymphomas, are primarily nodal lymphomas in which lymph node involvement is the dominant clinical feature. The frequency of extranodal manifestations of these primarily nodal lymphomas is often underestimated. The typical growth pattern of nodal lymphomas can be absent or not evaluable because the biopsy specimens are often small in cases with extranodal involvement. Therefore, the immunophenotype of the lymphoma cells is of great importance for the diagnosis of indolent lymphomas with extranodal localizations.
Asunto(s)
Linfoma de Células B/patología , Linfoma Folicular/patología , Linfoma de Células del Manto/patología , Biopsia , División Celular , Humanos , Inmunofenotipificación , Ganglios Linfáticos/patologíaRESUMEN
BACKGROUND AND AIMS: Gastric mucosa associated lymphoid tissue lymphoma is a well defined B cell lymphoma yet often impossible to distinguish from severe chronic gastritis on morphological grounds alone. Therefore, it was suggested to use the clonality of the immunoglobulin (Ig) heavy chain (H) genes, as detected by polymerase chain reaction (PCR), as a decisive criterion. However, there is controversy as to whether B cell clonality also exists in chronic gastritis, hence rendering this approach futile at present. METHODS: An expert panel re-examined the histology and immunohistochemistry of a total of 97 cases of gastric biopsies, including clearcut marginal zone lymphoma, chronic gastritis, and ambiguous cases, applying the Wotherspoon criteria on the basis of haematoxylin-eosin and CD20 immunostainings. In addition, a new and advanced PCR system for detection of clonal IgH gene rearrangements was independently applied in two institutions in each case. RESULTS: The overall IgH clonality assessments of both institutions were in total agreement. Overt lymphoma (Wotherspoon score 5) was clonal in 24/26 cases. Chronic gastritis (Wotherspoon scores 1 and 2) was not clonal in 52/53 cases; the clonal case being Wotherspoon score 2. Of 18 cases with ambiguous histology (Wotherspoon scores 3 and 4) four were clonal. CONCLUSIONS: Using advanced PCR technology, clonal gastritis is extremely rare, if it exists at all. Thus B cell clonality in Wotherspoon 3 and 4 cases is regarded as suitable for definitively diagnosing gastric marginal zone lymphoma.
Asunto(s)
Gastritis/diagnóstico , Linfoma de Células B de la Zona Marginal/diagnóstico , Neoplasias Gástricas/diagnóstico , Algoritmos , Antígenos CD20/metabolismo , Biomarcadores de Tumor/metabolismo , Enfermedad Crónica , Células Clonales/patología , Diagnóstico Diferencial , Gastritis/patología , Reordenamiento Génico de Cadena Pesada de Linfocito B , Genes de Inmunoglobulinas , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Linfoma de Células B de la Zona Marginal/patología , Células Madre Neoplásicas/patología , Proyectos Piloto , Reacción en Cadena de la Polimerasa/métodos , Reproducibilidad de los Resultados , Neoplasias Gástricas/patologíaRESUMEN
Control of mycobacterial infection by the cellular immune system relies both on antigen-presenting cells and on T lymphocytes. The quality of an effective cellular immune response is dependent on functional signal transduction residing in the cytoplasmic tails of the T-cell receptor CD3 components. In order to investigate potential effects of mycobacteria on T-cell receptor signalling, we examined the protein expression of T-cell signal transduction molecules (CD3zeta, ZAP-70, p59fyn, p56lck). In Western blots of peripheral blood mononuclear cells of Mycobacterium tuberculosis infected patients, only the CD3zeta-chain showed a marked reduction in protein expression. To investigate the situation in situ, immunoenzymatic and immunofluorescence stainings for CD3epsilon and CD3zeta expression were performed on sections of normal lymphoid tissue, M. leprae infected and sarcoid tissue. CD3epsilon and CD3zeta expression were similar with respect to intensity, localization and the number of cells stained in normal lymphoid tissue and in sarcoid granulomas. In contrast, the granulomas of M. leprae infected tissues showed a significantly reduced expression of CD3zeta compared to CD3epsilon. Using double immunofluorescence analysis, virtually no CD3zeta expression could be detected in comparison to the CD3epsilon expression in the lesions. Apparently, mycobacteria are capable of significantly reducing CD3zeta-chain expression, which may be restored by cytokines. IL-2-enhanced zeta-chain expression and T-cell effector functions, defined by interferon-gamma release, in M. tuberculosis-specific and human leucocyte antigen-DR restricted CD4+ T cells isolated from granuloma lesions from patients with pulmonary tuberculosis. Because CD3zeta is essential for CD3 signalling and for eliciting T-cell effector functions, reduced CD3zeta protein expression could result in altered signal transduction and inefficient T-cell effector functions. Alternatively, reduced CD3zeta-chain expression may protect T cells from repetitive TCR stimulation associated with anergy or apoptosis.
Asunto(s)
Complejo CD3 , Proteínas de la Membrana/metabolismo , Infecciones por Mycobacterium/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Técnica del Anticuerpo Fluorescente , Granuloma/inmunología , Humanos , Técnicas para Inmunoenzimas , Interleucina-2/inmunología , Lepra Lepromatosa/inmunología , Proteína Tirosina Quinasa p56(lck) Específica de Linfocito/sangre , Tonsila Palatina/inmunología , Proteínas Tirosina Quinasas/sangre , Proteínas Proto-Oncogénicas/sangre , Proteínas Proto-Oncogénicas c-fyn , Sarcoidosis Pulmonar/inmunología , Transducción de Señal/inmunología , Tuberculosis Pulmonar/inmunología , Proteína Tirosina Quinasa ZAP-70RESUMEN
Angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) is a peculiar T cell lymphoma, as expanding B cell clones are often present besides the malignant T cell clones. In addition, large numbers of Epstein-Barr virus (EBV)-infected B cells are frequently observed. To analyze the differentiation status and clonal composition of EBV-harboring B cells in AILD, single EBV-infected cells were micromanipulated from lymph nodes of six patients with frequent EBV(+) cells and their rearranged immunoglobulin (Ig) genes analyzed. Most EBV-infected B cells carried mutated Ig genes, indicating that in AILD, EBV preferentially resides in memory and/or germinal center B cells. EBV(+) B cell clones observed in all six cases ranged from small polyclonal to large monoclonal expansions and often showed ongoing somatic hypermutation while EBV(-) B cells showed little tendency for clonal expansion. Surprisingly, many members of expanding B cell clones had acquired destructive mutations in originally functional V gene rearrangements and showed an unfavorable high load of replacement mutations in the framework regions, indicating that they accumulated mutations over repeated rounds of mutation and division while not being selected through their antigen receptor. This sustained selection-free accumulation of somatic mutations is unique to AILD. Moreover, the survival and clonal expansion of "forbidden" (i.e., Ig-deficient) B cells has not been observed before in vivo and thus represents a novel type of viral latency in the B cell compartment. It is likely the interplay between the microenvironment in AILD lymph nodes and the viral transformation that leads to the survival and clonal expansion of Ig-less B cells.
Asunto(s)
Linfocitos B/virología , Reordenamiento Génico de Linfocito B , Herpesvirus Humano 4/aislamiento & purificación , Linfadenopatía Inmunoblástica/inmunología , Linfoma de Células T/inmunología , Mutación , Adulto , Anciano , Anciano de 80 o más Años , Antígenos CD20 , Diferenciación Celular , Células Clonales , Femenino , Genes de Inmunoglobulinas , Genes Codificadores de la Cadena gamma de los Receptores de Linfocito T , Humanos , Linfadenopatía Inmunoblástica/virología , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/virología , Activación de Linfocitos , Linfoma de Células T/virología , Masculino , Micromanipulación , Persona de Mediana Edad , ARN Viral/aislamiento & purificación , Latencia del VirusAsunto(s)
Anticuerpos Monoclonales/inmunología , Proteínas de Ciclo Celular/inmunología , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Inmunohistoquímica/métodos , Antígeno Ki-67/análisis , Ciclo Celular , Proteínas de Ciclo Celular/análisis , Células Cultivadas , Humanos , Antígeno Ki-67/inmunología , Leucocitos Mononucleares/química , Linfoma Folicular/química , Linfoma Folicular/diagnóstico , Componente 6 del Complejo de Mantenimiento de Minicromosoma , Neoplasias/diagnóstico , Oocitos/químicaRESUMEN
Angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) is defined in the current lymphoma classifications as a T-cell non-Hodgkin's lymphoma. However, in approximately one third of the cases of this lymphoproliferative disease rearrangements of T-cell receptor (TCR) genes indicating clonal expansion of T cells are not detectable. It is currently believed that these cases may represent early stages of a lymphoma with a minor oligoclonal T-cell population. In the present study, 18 lymph nodes with the characteristic histology of AILD were investigated for clonal T-cell receptor gene rearrangements by analysis of DNA extracted from whole tissue sections. Dominant T-cell clones were detected in 12 of these cases. Single CD4(+) and CD8(+) T cells and proliferating Ki67(+) cells of seven cases were micromanipulated from frozen tissue sections. TCRbeta gene rearrangements were amplified from these cells by polymerase chain reaction and sequenced. In all informative cases, the clonal gene rearrangements were only detected among CD4(+), and not among CD8(+) T cells, indicating that the tumor clones in AILD usually derive from CD4(+) T cells. Minor clonal T-cell populations in those cases in which no clone was found by whole-tissue DNA analysis were not detectable even at single cell resolution. T-cell clones in 4 of 10 cases were found to express similar TCRbeta chains, indicating a potential role of (super) antigen triggering in at least some cases of AILD.
Asunto(s)
Reordenamiento Génico de la Cadena beta de los Receptores de Antígenos de los Linfocitos T/genética , Genes Codificadores de la Cadena beta de los Receptores de Linfocito T/genética , Linfoma de Células T/genética , Subgrupos de Linfocitos T/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Regiones Determinantes de Complementariedad/genética , ADN de Neoplasias/genética , Amplificación de Genes , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la PolimerasaRESUMEN
Progressively transformed germinal centers (PTGCs) are histologic structures mainly composed of small resting B cells and intermingled proliferating centroblast-like cells. The B-cell differentiation processes within PTGCs and their relation to classical germinal centers (GC) and to lymphocyte-predominant Hodgkin disease (LPHD), with which PTGCs are often associated, are largely unknown. To address these issues, single small resting (Ki67-) and proliferating (Ki67+) centroblast-like cells were isolated from 7 PTGCs of 5 lymph nodes, and rearranged immunoglobulin genes were amplified and sequenced. Most small resting B cells were clonally unrelated, and most carried unmutated immunoglobulin gene rearrangements resembling mantle zone B cells. Small resting B cells with mutated immunoglobulin gene rearrangements may represent centrocytes, memory B cells, or both. Among the centroblast-like Ki67+ cells, expanded B-cell clones were observed in 6 of 7 PTGCs analyzed. Clonally related V region genes showed extensive intraclonal diversity, and the mutation pattern indicated stringent selection of the cells for the expression of functional antigen receptors. Thus, somatic hypermutation, clonal expansion, and selection occur also in the disorganized PTGC microenvironment, as in classical GCs. In lymph nodes affected by PTGCs, no clonal expansion across the borders of individual PTGCs was observed, distinguishing PTGCs from LPHD.
Asunto(s)
Linfocitos B/inmunología , Centro Germinal/patología , Enfermedad de Hodgkin/inmunología , Adolescente , Adulto , Linfocitos B/química , Linfocitos B/citología , Diferenciación Celular , Niño , Femenino , Reordenamiento Génico de Linfocito B , Genes de Inmunoglobulinas , Centro Germinal/citología , Centro Germinal/inmunología , Enfermedad de Hodgkin/genética , Enfermedad de Hodgkin/patología , Humanos , Inmunohistoquímica , Antígeno Ki-67/análisis , Antígeno Ki-67/inmunología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/patología , Masculino , Modelos InmunológicosRESUMEN
Three cases of adenosquamous cell carcinoma and squamous cell carcinoma of the upper urinary tract are presented. The fact that the urothelium normally has no glandular or squamous structures renders the pathogenesis of these tumours interesting. The process is assumed to begin with an urothelial metaplasia resulting from a reaction to chronic irritation, leading to dedifferentiation, dysplasias and, in the end, to a squamous cell carcinoma or adenocarcinoma. The relevant medical histories include chronic episodes of pyelonephritis or nephrolithiasis. Diagnosis, therapeutic approaches and prognosis of these rare tumours are discussed.
Asunto(s)
Carcinoma Adenoescamoso/patología , Carcinoma de Células Escamosas/patología , Neoplasias Renales/patología , Neoplasias Ureterales/patología , Anciano , Carcinoma Adenoescamoso/cirugía , Carcinoma de Células Escamosas/cirugía , Femenino , Humanos , Neoplasias Renales/cirugía , Masculino , Persona de Mediana Edad , Neoplasias Ureterales/cirugíaAsunto(s)
Adenocarcinoma de Células Claras/genética , Carcinoma de Células Renales/genética , Deleción Cromosómica , Proteínas de Unión al ADN/genética , Neoplasias Renales/genética , Monoéster Fosfórico Hidrolasas/genética , Factores de Transcripción/genética , Proteínas Supresoras de Tumor , Adenocarcinoma de Células Claras/patología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Carcinoma de Células Renales/patología , Cromosomas Humanos Par 10/genética , Análisis Mutacional de ADN , Genes Supresores de Tumor/genética , Humanos , Hibridación Fluorescente in Situ , Interfase , Neoplasias Renales/patología , Fosfohidrolasa PTENRESUMEN
In rare cases of B-cell chronic lymphocytic leukemia (B-CLL), large cells morphologically similar to or indistinguishable from Hodgkin/Reed-Sternberg (HRS) cells of Hodgkin's disease (HD) can be found in a background of otherwise typical B-CLL. To test these HRS-like cells for a potential clonal relationship to the B-CLL cells, single cells were micromanipulated from immunostained tissue sections, and rearranged immunoglobulin genes were amplified from HRS-like cells and B-CLL cells and sequenced. The same variable (V) gene rearrangements with shared and distinct somatic mutations were found in HRS-like and B-CLL cells from 1 patient, which indicates derivation of these cells from 2 distinct members of a germinal-center B-cell clone. Separate clonal V gene rearrangements were amplified from HRS-like and B-CLL cells from 2 other patients, showing concomitant presence of 2 distinct expanded B-cell clones. Epstein-Barr virus (EBV) was detected in the HRS-like cells of these 2 latter cases, indicating clonal expansion of an EBV-harboring B cell in the setting of B-CLL. There is evidence that HRS-like cells in B-CLL, like HRS cells in HD, derive from germinal-center B cells. In all cases, somatic mutations have been detected in the rearranged V genes of the HRS-like cells, and in 1 of the EBV-positive HRS-like cell clones, somatic mutations rendered an originally functional V gene rearrangement nonfunctional. We speculate that the HRS-like cells in B-CLL represent potential precursors for HRS cells causing HD.
Asunto(s)
Linfocitos B/patología , Centro Germinal/patología , Enfermedad de Hodgkin/patología , Leucemia Linfocítica Crónica de Células B/patología , Células Madre Neoplásicas/patología , Células de Reed-Sternberg/patología , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Transformación Celular Neoplásica , Transformación Celular Viral , Células Clonales/patología , ADN de Neoplasias/genética , Células Madre de Carcinoma Embrionario , Infecciones por Virus de Epstein-Barr/patología , Resultado Fatal , Femenino , Reordenamiento Génico de Cadena Pesada de Linfocito B , Genes de Inmunoglobulinas , Herpesvirus Humano 4/aislamiento & purificación , Humanos , Región Variable de Inmunoglobulina/genética , Inmunofenotipificación , Leucemia Linfocítica Crónica de Células B/genética , Leucemia Linfocítica Crónica de Células B/virología , Persona de Mediana Edad , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Infecciones Tumorales por Virus/patologíaRESUMEN
Among the four subtypes of Hodgkin disease (HD), lymphocyte-predominant (LP) HD is now generally considered as a separate entity. The B cell nature of the typical Hodgkin and Reed-Sternberg (HRS) cells and their variants (L and H, lymphocytic and histiocytic cells) in LP HD has long been suspected, but the question of whether these cells represent a true tumor clone is unclear. We previously demonstrated clonal Ig gene rearrangements in one case of LP HD. In the present study, five cases of LP HD were analyzed by micromanipulation of single HRS cells from frozen tissue sections and DNA amplification of rearranged Ig heavy chain genes from those cells. Clonal V gene rearrangements harboring somatic mutations were detected in each case. In three cases ongoing somatic mutation was evident. This shows that HRS cells in LP HD are a clonal tumor population derived from germinal center B cells. The pattern of somatic mutation indicates that HRS cells in LP HD are selected for antibody expression. This, and the presence of ongoing mutation discriminates LP from classical HD.
Asunto(s)
Linfocitos B/inmunología , Enfermedad de Hodgkin/patología , Subgrupos Linfocitarios/inmunología , Adolescente , Adulto , Linfocitos B/patología , Niño , Células Clonales , Reordenamiento Génico de Linfocito B , Enfermedad de Hodgkin/inmunología , Humanos , Región Variable de Inmunoglobulina/genética , Subgrupos Linfocitarios/patología , Persona de Mediana Edad , Datos de Secuencia MolecularRESUMEN
In the immunobiological characterization of lymph node cells, sinus-lining cells (SLCs) have been given little attention mainly due to the difficulties in their recognition. Ki-M9 is a new monoclonal antibody (MAb) selected for its unique capability to visualize SLCs in human lymph nodes. The details were established by light and electron microscopy and immunoprecipitation of the corresponding biosynthetically labeled antigen. Ki-M9 recognizes a 70-kd protein localized on the surface membrane of SLCs. In the lymphoid tissue, a mild reactivity was exclusively encountered on follicular dendritic reticulum cells in the germinal centers of secondary lymphoid follicles. In other organs, some squamous epithelial and myoepithelial cells were recognized by this antibody. Immunomonitoring of SLCs on light and electron microscopic levels revealed their dendritic morphology, lack of phagosomes, and their close association with type IV collagen fibers. Considering the occurrence of typical dendritic SLCs on the front line of antigen flood, we propose that SLCs be investigated for a possible antigen-binding property.
Asunto(s)
Células Dendríticas/citología , Ganglios Linfáticos/citología , Animales , Anticuerpos Monoclonales/inmunología , Antígenos de Superficie/inmunología , Células Cultivadas , Células Dendríticas/inmunología , Humanos , Inmunoglobulina M/inmunología , Inmunohistoquímica , Inmunofenotipificación , Ganglios Linfáticos/inmunología , Ratones , Ratones Endogámicos BALB C , Microscopía ElectrónicaRESUMEN
By immunization with nuclear lysates of L428 cells, we raised a monoclonal mouse antibody, Ki-S2 (IgG1). In Western blots, this antibody recognizes a nuclear antigen with an apparent molecular mass of 100 kD, termed p100. Protein sequencing of p100 showed that this is a hitherto unknown protein. Immunohistochemical examination of cryostat and paraffin sections of nearly all human tissue types and neoplasms showed that p100 was exclusively expressed in the nuclei of a fraction of proliferating cells. Cell sorting and fluorescence-activated cell sorting analysis of stimulated peripheral blood mononuclear cells showed that p100 was exclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis. During mitosis, this protein is strictly associated with the spindle pole and with the mitotic spindle, whereas during S and G2, p100 is diffusely distributed throughout the cell nucleus. Immediately after completion of cytokinesis, p100 was rapidly degraded. In L428 cells, p100 is phosphorylated at least during mitosis. It has a turnover time of about 1 hour. Studies on routinely processed paraffin sections of specimens of malignant lymphoma, benign and malignant nevocellular tumors, and breast cancer showed that in all cases less than 40% of the Ki-67-positive growth fraction expressed p100. Thus, p100 might prove to be a more reliable measure of cellular proliferation and one that is more closely correlated to cancer prognosis, beyond its general biologic relevance as a cell cycle protein.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Fase G2 , Mitosis , Proteínas Nucleares/metabolismo , Fase S , Animales , Anticuerpos Monoclonales , Proteínas de Ciclo Celular/aislamiento & purificación , Endonucleasas , Citometría de Flujo , Humanos , Ratones , Peso Molecular , Proteínas Nucleares/aislamiento & purificación , Especificidad de ÓrganosRESUMEN
Various studies have reported the utility of anti-CD34 staining in the differential diagnosis of dermal spindle-cell tumors. To investigate whether monoclonal antibody Ki-M1p might add practical diagnostic information, we examined a total of 120 cutaneous spindle cell neoplasms using a panel of markers. Anti-CD34 antibody QBEnd/10 consistently stained dermatofibrosarcomas, Kaposi's sarcomas, neurofibromas, and, to a lesser extent, hemangiopericytomas. A positive reaction was also found in > 18% of the dermatofibromas. Ki-M1p staining showed an intense immunoreaction in all dermatofibromas, whereas no reactivity was observed in dermatofibrosarcomas. In addition, a subset of cells was labeled in atypical fibroxanthomas and Kaposi's sarcomas. Neurofibromas, spindle-cell hemangioendotheliomas, and hemangiopericytomas were negative. Dermatofibrosarcomas and atypical fibroxanthomas also moderately expressed smooth muscle-specific actin. Immunohistochemically, a discrimination between dermatofibrosarcomas and neurofibromas was possible only by means of an antibody against the nerve growth factor receptor. We conclude that the combination of several antibodies, in particular anti-CD34 and Ki-M1p, may improve the accuracy of diagnostic immunohistochemistry in the field of cutaneous spindle cell tumors. We speculate that dermatofibroma is primarily a macrophage-rich inflammatory lesion in which cytokine secretion induces a secondary proliferation of fibroblasts, whereas dermatofibrosarcoma is likely to issue from primitive dermal cells of uncertain origin.
Asunto(s)
Anticuerpos Monoclonales , Antígenos CD34/análisis , Antígenos de Superficie/análisis , Biomarcadores de Tumor/análisis , Proteínas Nucleares/análisis , Neoplasias Cutáneas/patología , Autoantígenos , Citoplasma/patología , Dermatofibrosarcoma/patología , Diagnóstico Diferencial , Endotelio Vascular/patología , Hemangiopericitoma/patología , Humanos , Inmunofenotipificación , Neurofibroma/patología , Complejo de la Endopetidasa Proteasomal , Sarcoma de Kaposi/patología , Sensibilidad y EspecificidadRESUMEN
We report five novel monoclonal antibodies (Ki-S1, Ki-S4, Ki-S6, Ki-S7, and Ki-S8) reactive with a proliferation-related nuclear antigen. In immunoprecipitation and Western blot experiments using crude nuclear extracts, they recognized a protein of 170 kD that, after proteolytic digestion of the immunoprecipitate and sequencing of the resulting peptides, was identified as the alpha-isoform of human topoisomerase II. This was confirmed by testing the antibodies on a highly purified enzyme preparation. Crossreactivity with topoisomerase II beta was ruled out by testing the antibodies on crude extracts from yeast cells expressing the beta-isoform exclusively. The antibodies bind the antigen with different affinities and at different epitopes, apparently located within the carboxyl third of the enzyme. All five antibodies are suitable for archival material after adequate antigen retrieval, thereby enabling retrospective studies. This report illustrates the tissue and subcellular distribution of the antigen through the cell cycle by immunohistochemistry and confocal fluorescence microscopy. The antibodies will be useful tools in further analysis of morphological and functional aspects of topoisomerase II and may serve diagnostic purposes, as well as providing prognostic information in tumor pathology.
Asunto(s)
Anticuerpos Monoclonales , Antígenos de Neoplasias/química , ADN-Topoisomerasas de Tipo II/química , Isoenzimas/química , Especificidad de Anticuerpos , Sitios de Unión , Western Blotting , División Celular , Línea Celular , Proteínas de Unión al ADN , Humanos , Microscopía ConfocalRESUMEN
To study the distribution pattern of interleukin-6 (IL-6)-producing cells in normal human lymph nodes, we applied the in situ reverse transcription-polymerase chain reaction technique. We describe a new modification of this technique for monitoring small amounts of specific nucleotide sequences in conventional paraffin sections. This technique differs in at least two respects from those described earlier. The two decisive steps are: 1) the reverse transcription of mRNA and the subsequent amplification of cDNA by polymerase chain reaction are performed by a new single enzyme capable of both reaction types in one and the same medium without buffer exchange; and 2) for the specific detection of the amplified cDNA, a modified version of the primed in situ labeling technique was used. The technique, carried out on normal human lymph nodes, traces a low load of IL-6 mRNA in fibroblasts, endothelial cells, and a minor population of T lymphocytes in the pulp region. High levels of expression were encountered in about 20% of perisinusoidal pulp macrophages. In addition, moderate activity was detectable in sinus lining cells. Because no major activity was found in the germinal centers of the lymphoid B follicles and in the T zone, it is suggested that the plasma cell differentiation ensuing from primary and secondary B-cell immunization is mainly effected by the sinus lining cells as well as perifollicular and perisinusoidal pulp macrophages capable of producing high amounts of IL-6.
Asunto(s)
Interleucina-6/genética , Ganglios Linfáticos/química , Reacción en Cadena de la Polimerasa , ARN Mensajero/análisis , Transcripción Genética , Humanos , Inmunohistoquímica , Ganglios Linfáticos/patología , Adhesión en ParafinaRESUMEN
Besides specific cellular-mediated T cell responses, B cell related humoral responses have been demonstrated during the course of graft-versus-host disease after semiallogeneic transplantation of cellular antigen. Following semiallogeneic small bowel transplantation, there are, besides others, two specific forms of antigen-presenting cells, namely sinus lining cells (SLCs) and follicular dendritic cells (FDCs) which mediate primary and secondary humoral immune responses, respectively. This study was aimed to clarify the role of these dendritic cell entities after transplantation of small bowel grafts in a one-sided graft-versus-host (GvH) model for untreated and immunosuppressed (15-deoxyspergualin) recipient animals. As graft-versus-host disease progressed, SLCs and FDCs were eliminated in donor and recipient graft-versus-host associated target tissues (spleen and mesenteric lymph nodes) of untreated animals, whereas these dendritic cells prevailed in immunosuppressed recipients. 15-deoxyspergualin successfully prevented GvHD and significantly prolonged the mean survival time of untreated rats (16.0 +/- 4.5 d) for at least 21 d. Based on the immunosuppressive efficacy of 15-deoxyspergualin on the survival and function of SLCs and FDCs, an unaltered development of germinal centers and B cell proliferation within mesenteric lymph nodes and spleen was maintained
Asunto(s)
Células Dendríticas Foliculares/fisiología , Modelos Animales de Enfermedad , Enfermedad Injerto contra Huésped/patología , Intestino Delgado/trasplante , Animales , Células Presentadoras de Antígenos/fisiología , Movimiento Celular/fisiología , Femenino , Citometría de Flujo/métodos , Supervivencia de Injerto/efectos de los fármacos , Guanidinas/farmacología , Inmunosupresores/farmacología , Intestino Delgado/efectos de los fármacos , Intestino Delgado/patología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/patología , Ganglios Linfáticos/trasplante , Vasos Linfáticos/efectos de los fármacos , Vasos Linfáticos/patología , Vasos Linfáticos/trasplante , Masculino , Ratas , Ratas Endogámicas Dahl , Ratas Endogámicas Lew , Linfocitos T/fisiología , Trasplante Heterotópico/métodos , Trasplante Isogénico/métodosRESUMEN
OBJECTIVE: The course of metastatic renal cell carcinoma shows a broad range of interindividual variation that cannot be sufficiently predicted by tumor stage and grade. The aim of this study was to establish the prognostic value of DNA ploidy and the proliferation marker Ki-67 in renal cell carcinoma. METHODS: Both parameters were measured simultaneously in 100 tumors and then correlated with the classic prognostic criteria pathologic stage and tumor differentiation grade as well as clinical course (early tumor progression). RESULTS: DNA ploidy correlated well with staging, grading and early progression. The proliferation index (Ki-67) correlated well with tumor stage and histopathological grade but not with the clinical course (early progression). CONCLUSION: Due to the diverse biological potential of renal cell carcinoma observation of further clinical course, including late tumor progression, will be necessary to determine whether one of these two indicators can provide additional information beyond what differentiation grade and tumor stage already tell us.
Asunto(s)
Biomarcadores de Tumor/análisis , Carcinoma de Células Renales/diagnóstico , ADN de Neoplasias/análisis , Antígeno Ki-67/análisis , Neoplasias Renales/diagnóstico , Aneuploidia , Carcinoma de Células Renales/patología , Diploidia , Progresión de la Enfermedad , Femenino , Citometría de Flujo , Humanos , Riñón/patología , Neoplasias Renales/patología , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , Estudios ProspectivosRESUMEN
A patient with acute monoblastic leukemia (AML, M5A) was treated successfully in December 1987. In 1993 after 6 years in complete remission, she presented with an intracutaneous nodular mass on her right upper arm which was resected in toto and shown to be undifferentiated monoblastic leukemia. Two further chloroma lesions were excised in July 1994 and March 1995 respectively. Bone marrow cytology and histology always showed a continuing complete remission with no evidence of leukemia relapse. In July 1995 she presented with a disseminated skin infiltrate and a relapse with 80% monoblasts in the bone marrow. After one course of chemotherapy (Idarubicin/Ara-C), a second complete remission was achieved and her leukemic skin infiltrate disappeared completely. This case illustrates that chloromas of the skin can occur as late as 6 years after treatment for AML and also emphasizes that the occurrence of a chloroma does not necessarily mean immediate leukemia relapse. It also stresses that a second complete remission can be achieved with standard AML-induction therapy despite widespread leukemic skin infiltrates in such patients.