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  • 1
    Keywords: RECEPTOR ; CELLS ; EXPRESSION ; tumor ; KINASE ; TYROSINE KINASE ; ALGORITHM ; SITE ; SITES ; GENE ; GENE-EXPRESSION ; DIFFERENTIATION ; MECHANISM ; mechanisms ; MOLECULE ; LYMPHOMA ; REED-STERNBERG CELLS ; gene expression ; DESIGN ; PLASMA ; NUMBER ; B-CELLS ; RECEPTORS ; gene expression profiling ; GAINS ; chemokine ; JAK2 ; REL ; B-CELL LYMPHOMA ; FEATURES ; immunoglobulin ; secretion ; CELL LYMPHOMA ; PROFILES ; LYMPHOMAS ; EXPRESSION PROFILES ; HIGH EXPRESSION ; classical Hodgkin lymphoma ; GERMINAL-CENTER ; 9P ; mediastinal grey zone lymphoma ; primary mediastinal B cell lymphoma ; TARC
    Abstract: There are several indications that classical Hodgkin lymphoma (cHL) and at least a proportion of cases of Primary Mediastinal B cell Lymphoma (PMBL) are derived from B cells at similar stages of differentiation and share common pathogenic mechanisms. The first indication was the existence of mediastinal grey zone lymphomas as identified in the 4th International Symposium on HL, with clinical, histological and immunohistochemical features intermediate between cHL and PMBL. Second, both tumor types resemble a cell that is developmentally situated in-between the germinal center reaction and a plasma cell. Third, cHL and PMBL were found to have similar gene expression profiles, including the lack of immunoglobulin expression and low levels of B cell receptor signalling molecules, and the secretion of molecules like the chemokine TARC and the prominent expression of IL-13 receptors. Fourth, both entities were found to have common genomic aberrancies, notably in 2p15 and 9p24, the sites of the REL oncogene and the tyrosine kinase gene JAK2, respectively. Further comparison of both lymphoma types may provide further insight in the pathogenic mechanisms and allow the design of diagnostic algorithms to sort out the small number of so-called mediastinal grey zone lymphomas, that appear to be intermediate between PMBL and cHL
    Type of Publication: Journal article published
    PubMed ID: 16007868
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  • 2
    Keywords: CELLS ; EXPRESSION ; tumor ; TUMOR-CELLS ; Germany ; human ; GENE ; HYBRIDIZATION ; TISSUE ; LINES ; PATIENT ; MARKER ; TISSUES ; ANTIGEN ; SKIN ; T cells ; T-CELLS ; CELL-LINES ; DOWN-REGULATION ; SIGNAL ; IN-SITU ; DESIGN ; MUTATION ; CELL-LINE ; LINE ; ABERRATIONS ; HETEROZYGOSITY ; MELANOMA ; REGION ; MUTATIONS ; MONOCLONAL-ANTIBODIES ; MHC CLASS-I ; PHENOTYPE ; IMMUNE ESCAPE ; TUMOR ESCAPE ; in situ hybridization ; MALIGNANT-CELLS ; RE ; LEVEL ; EVENTS ; LOSSES ; CD8(+) T cell ; B2M GENE ; MEDIATED LYSIS ; PROCESSING MACHINERY
    Abstract: Purpose: Total loss of surface presentation of human leukocyte antigen (HLA) class I molecules, protecting tumor cells from the recognition by cytotoxic host CD8(+) T cells, is known to be caused by mutations in the beta 2-microglobulin (beta 2m) gene. We asked whether abnormalities of chromosome 15, harboring the beta 2m gene on 15q21, in addition to beta 2m gene mutations, are causative for the HILA class I-negative phenotype of melanoma cells. Experimental Design: To answer this, we established primary cell lines from the beta 2m-negative metastatic melanoma tissues of four different patients and analyzed them for beta 2m gene mutations and chromosome 15 aberrations, the latter by loss of heterozygosity analysis, fluorescence in situ hybridization (FISH), and multicolor FISH. Results: Mutations at the beta 2m gene level were detected in all cell lines. The loss of heterozygosity analysis of microsatellite markers located on chromosome 15 in three of the four cell lines pointed to an extensive loss of chromosome 15 material. Subsequent molecular cytogenetic analysis revealed the coexistence of apparently normal and rearranged versions of chromosome 15 in three cell lines whereas the fourth cell line solely showed rearranged versions. Two of the four cell lines exhibited a special type of intrachromosomal rearrangement characterized by FISH signals specific for the subtelomeric region of 15q at both ends of the chromosome and one centromeric signal in between. Conclusions: Our data indicate that the complete loss of HLA class I expression in melanoma cells is due to the coincidence of the following mutational events: (a) chromosome 15 instability associated with an extensive loss of genetic material and (b) beta 2m gene mutations
    Type of Publication: Journal article published
    PubMed ID: 16740750
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  • 3
    ISSN: 1432-2307
    Keywords: DNA analysis ; Gene rearrangements ; Molecular hybridization ; Immunohistology ; Histopathology ; Malignant lymphoma ; AIDS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We investigated for rearrangements of the immunoglobulin (Ig) heavy and light chain genes and of the T cell receptorγ (TCRTγ) andβ (TCrβ) genes 45 biopsy samples from a variety of lymphoproliferative disorders. They were diagnosed histopathologically and immunophenotypically as non-Hodgkin's lymphomas (NHLs) of the B cell type (19 cases), NHLs of the T cell type (3 cases), NHLs of “undetermined“ cell type (3 cases), atypical lymphoid proliferation (1 case) and AIDS-related lymphadenopathies with florid polyclonal follicular hyperplasia (19 cases). A monoclonal proliferation of B cells was shown by DNA analysis in all 19 B cell NHLs. In two immunohistologically determined T cell NHLs (both diagnosed as mycosis fungoides) the cells had rearrangements of TCrβ gene, whereas in the third case (lymphoblastic NHL) the cells had rearrangements of Ig heavy chain and TCrγ and TCrβ genes. None of the B cell NHLs exhibited TCrγand TCrβ gene rearrangement bands. All the “undetermined” cell NHLs demonstrated rearrangements of Ig heavy chain gene associated with the germ line TCrγand TCrβ genes; in two cases light chain gene rearrangements were also found. The atypical lymphoid proliferation, in which the differential diagnosis was between a reactive or malignant process, and two out of 19 cases of florid polyclonal follicular hyperplasia showed a clonal B cell population by DNA analysis. This study indicates that there was a strong correlation between the rearrangements of specific genes and the immunophenotype of the NHL; moreover, DNA analysis of tissue biopsy specimens from phenotypically “undetermined” cell NHLs and from equivocal lymphoid proliferation using Ig and TCR gene probes yelded an answer in the cases analyzed. The significance of clonal B cell expansions found in two AIDS-related lymphadenopathies should be interpreted with caution.
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  • 4
    ISSN: 1432-2307
    Keywords: Laminin ; Basement membrane ; Follicular dendritic reticulum cell ; B-cell lymphoma ; Immunohistology ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary We performed a comparative immunohisto-cytochemical study of the distribution patterns of laminin and follicular dendritic reticulum cells (DRCs) within their follicular microenvironment in both nodular or diffuse B-cell non Hodgkin's lymphomas (NHLs). Twenty nine cases of immunophenotypically diagnosed B-cell NHLs (19 of follicular center cell origin-FCCL- and 10 of the diffuse well differentiated lymphocytic type-WDLL-) and five reactive lymph nodes with follicular hyperplasia were analyzed by immunoperoxidase and immunofluorescence techniques. Serial frozen sections and cytospin preparations were tested either with single antibodies anti laminin and DRC-1, or paired reagents in double labeling immunofluorescence. Our results indicated consistently that within both the reactive germinal centers and the neoplastic nodules of FCCL laminin immunostaining visualized a punctate-granular pattern apart from the linear vascular basement membrane positivity. Double immunofluorescence assay demonstrated that there was a close parallelism between this laminin staining pattern and DRC-1 distribution showing a well developed DRCs meshwork; in the diffuse tumour areas of both FCCL and WDLL, laminin immunoreactivity was found only in those cases in which nests of DRCs were observed. Double immunofluorescence studies performed on cytospin preparations demonstrated that the groups of cells containing DRC-1 positive cells, contained a positivity for laminin, although within the cell the staining for DRC-1 was intense and diffuse, while that for laminin was granular and more sparse. Our results suggested that these laminin and DRC-1 positive reactive sites may be present on the same cells. Since the reduction in number or loss of both DRCs and their related immunostaining for laminin within the microenvironment was consistently associated with a loss of nodularity by lymphoma cells, whereas nodularity in reactive and neoplastic conditions was associated with a rich DRCs meshwork and the related laminin immunostaining, a trapping function of DRCs exercised in the presence of laminin should be considered.
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  • 5
    ISSN: 1432-2307
    Keywords: Reed-Sternberg cells ; Vimentin ; Lymph node pathology ; Immunohistology ; DNA analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary There are few data on the reactivity of Reed-Sternberg (RS) cells with antibodies against vimentin. In a preliminary survey of biopsy specimens from 16 cases of Hodgkin's disease (HD), we found that the antivimentin (V9) monoclonal antibody stained RS cells in 6 cases. We therefore examined vimentin expression on RS cells immunohistologically in 38 Bouin-fixed and paraffin-embedded lymph nodes with HD [lymphocyte predominance (LP) 4; nodular sclerosis (NS) 23; mixed cellularity (MC) 7; lymphocyte depletion (LD) 4]. The results were correlated with the histopathological features, the immunohistological phenotype of the RS cells, and the findings obtained from molecular genetics studies (available in 13 cases). RS cells were found to express strong and diffuse cytoplasmic staining for vimentin in 13 cases, all of the NS subtype. No differences in antigenic expression on RS cells were found between the vimentin-positive and negative cases within the NS subtype. DNA analysis revealed no B- or T-cell clonal populations in the tested samples. The results indicated that RS cells were immunostained by anti-vimentin (V9) antibody with a relatively high frequency, but only in the NS subtype of HD. This subtype, however, was heterogeneous according to vimentin immunostaining on RS cells. The significance of this finding concerning the RS cell origin in this subset is discussed.
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  • 6
    ISSN: 1432-2307
    Keywords: CD 68 ; Monoclonal antibody ; Immunohistochemistry ; Monocyte/macrophage-related cells ; Fibrous histiocytoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary A monoclonal antibody (Ki-M6) against the CD 68 antigen, which labels cells of the monocyte/macrophage system, was tested on Bouin-fixed, paraffin-embedded samples of normal, reactive and neoplastic tissues by an avidin-biotin-peroxidase complex method, with the aim of establishing its value in diagnostic pathology. In normal human tissues, Ki-M6 reactivity was confined to the so-called resident macrophages populating normal organs under physiological conditions. Moreover, restricted reactivity against cells of macrophage lineage was observed in reactive and inflammatory lesions. Granulocytes, monocyte/macrophage-related immune accessory cells, and other analysed normal tissue structures did not reveal any reactivity. Ki-M6 was strongly reactive with the cases of benign (4/4) and malignant (15/15) fibrous histiocytomas, in addition to the true histiocytic lymphomas (3/3). Cases of granular cell tumour (2/3) showed strong reactivity with Ki-M6, whereas only few immunoreactive cells, with weak staining, were seen in the other Ki-M6-positive neoplasms [neurofibroma (3/3), benign schwannoma (1/2), ganglioneuroma (1/1), malignant schwannoma (5/9), melanoma (9/28), dermatofibrosarcoma protuberans (1/1), myelomonocytic leukaemia (3/3)]. Among the epithelial malignancies tested (47 cases), Ki-M6 was positive only in renal cell carcinoma (11/14). Malignant lymphomas of the Hodgkin (56 cases) and non-Hodgkin type (67 cases) were uniformly non-reactive. From these data, Ki-M6 appears to be an excellent marker of monocyte/macrophage-related cells and appears to be a reliable indicator for fibrous histiocytomas and true histiocytic malignancies. The availability of this additional antibody capable of staining routinely processed tissue is of practical interest.
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  • 7
    ISSN: 1432-2307
    Keywords: Hodgkin's disease ; Epstein-Barr virus ; Latent membrane protein ; Vimentin ; Messenger RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The presence of Epstein-Barr virus (EBV) genome in Hodgkin's and Reed-Sternberg (HRS) cells, as detected using in situ hybridization (ISH) with biotinylatedBamHI “V” probes, along with the expression of EBV-encoded latent membrane protein (LMP) and vimentin was examined in paraffin-embedded sections of 39 immunomorphologically characterized cases of Hodgkin's disease (HD). ISH demonstrated EBV in HRS cells in 15 of 39 cases, whereas LMP expression was detected in 11 of 39 cases, only in the presence of EBV genome detection. With the exception of 1 case, in which HRS cells expressed B-cell-associated antigens, the LMP-positive cases included specimens in which HRS cells were of non-B, non-T phenotype. LMP expression showed a stronger association with lymphocyte depletion (LD) (3/3) and mixed cellularity (MC) (6/11) than with lymphocyte predominance (0/5) or nodular sclerosis (2/20) subtypes. Vimentin expression on HRS cells was found in all the LMP-expressing cases and only in a fraction (13/28) of LMP-negative cases. This study supports the view that HD represents a heterogeneous group of diseases also in terms of EBV association, LMP expression being strongly related to the “aggressive” LD and MC histological subtypes. In light of the supposed interactions between vimentin and LMP, their coexpression on HRS cells, as detected in this study, provides further evidence for a significant role of EBV in the development of a proportion of HD cases.
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  • 8
    ISSN: 1432-2307
    Keywords: Hodgkin's disease ; Anaplastic large cell lymphoma ; Immunohistochemistry ; In situ immunophenotyping ; Monoclonal antibodies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Morphological and immunohistological studies were carried out on a series of 137 lymphomas including CD30+ anaplastic large cell (ALC) lymphomas (48 cases) and non-lymphocyte predominant Hodgkin's disease (HD) (89 cases), with the aim of assessing in situ expression of a combination of antibodies including anti-CD30/BerH2, epithelial membrane antigen (EMA), CD15 and CD45, in addition to other monoclonal antibodies suitable for paraffin tissues. A greater proportion of cases of ALC lymphomas than of HD exhibited positivity for CD45 (91.7% vs 17.6%), EMA (56.2% vs 4.5%), CD43 (53.6% vs 13.1%) and CD45RO (39.5% vs 3.5%), whereas Reed-Sternberg (RS) cells in HD most frequently expressed CD15 (93.2% vs 20.8%) antigen. Moreover, in 35 of 48 (72.9%) ALC lymphomas tumour cells expressed the CD30+, CD45+, CD15−, EMA− or+ phenotypic profile, while in the same percentage (62/ 85) of HD cases RS cells were found to express the CD30+, CD45−, CD15+, EMA− profile. This study suggests that the differential expression of CD45, EMA, and CD15 may be used in the separation of ALC lymphomas and HD. However, co-expression of CD30, CD45 and CD15 antigens by RS cells in HD (14/85 cases, 16.5% in this series) and by tumour cells in ALC lymphomas (9/48 cases, 18.7% in this series) may be encountered in a non-negligible fraction of cases.
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  • 9
    ISSN: 1569-8041
    Keywords: AIDS ; bcl-6 ; genetic lesion ; lymphoma ; oncogene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Non-Hodgkin's lymphoma (NHL) represents a major complicationof AIDS. Systemic AIDS-related NHLs (AIDS-NHLs) derive from B cells and areclassified into four distinct groups, including small noncleaved-celllymphoma (SNCCL), diffuse large-cell lymphoma (DLCL), anaplastic large-celllymphoma (ALCL), and body-cavity-based lymphoma (BCBL). The molecularpathogenesis of AIDS-NHL is characterized by the association of specificgenetic lesions with distinct AIDS-NHL categories. Genetic lesions ofAIDS-NHL involve proto-oncogenes (c-myc, Ras), tumor suppressor loci (p53,6q), and viral infection (Epstein–Barr virus, human herpesvirus type8). Design: The aim of this work was to define the involvement of the bcl-6gene in AIDS-related lymphomagenesis by investigating the distribution ofbcl-6 structural alterations throughout the pathologic spectrum of AIDS-NHL.Both gross rearrangements and mutations in the 5′ noncoding regions ofthe gene were investigated. Results: Gross rearrangements of bcl-6 are confined to a fraction ofAIDS-DLCL cases among AIDS-NHLs. Conversely, mutations of the 5′noncoding regions of bcl-6 are detected in a large proportion ofAIDS-SNCCLs, AIDS-DLCLs and AIDS-ALCLs independent of the concomitantpresence of bcl-6 rearrangements. Conclusions: Mutations of the 5′ noncoding regions of bcl-6 represent the most frequent genetic lesion presently detectable amongsystemic AIDS-NHLs. The frequency of these mutations and their location in theproximity of bcl-6 regulatory regions suggest that they may play a rolein AIDS-related lymphomagenesis.
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  • 10
    ISSN: 1569-8041
    Keywords: eosinophils ; Hodgkin's disease ; Reed–;Sternberg cells ; TNF-like ligands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Even though the presence of a prominent tissue eosinophiliarepresents a common histopathologic feature of Hodgkin's disease (HD),eosinophils have been mainly regarded as ‘innocent’ bystanders recruited andactivated during the cellular reaction typical of HD. To evaluate theputative role of eosinophils or eosinophil-derived cytokines on tumor-cellregulation in HD, we have analyzed these cells for the functional expressionof surface ligands (L) of the tumor necrosis factor (TNF) superfamily, whosespecific receptors are known to transduce proliferation signals at thesurface of Hodgkin (H) and Reed–;Sternberg (RS) cells. Materials and methods: Eosinophils from peripheral blood of healthydonors and patients with HD, primary hypereosinophilic syndrome (HES), orsecondary hypereosinophilia (HE), were purified by density gradientcentrifugation and immunomagnetic depletion of residual granulocytes. Results: By immunostaining and mRNA analysis, we were able to show thateosinophils from normal donors and patients with HD, HES, and HE express anumber of receptors and ligands of the TNF superfamily, including CD40,CD40L, CD30L, CD95/Fas, CD95/FasL and 4-1BB. In addition, we provideevidence that cytokines regulating eosinophil proliferation and activation,i.e., interleukin (IL)-5, IL-3, and granulocyte-macrophagecolony-stimulating factor, are able to enhance the cellular density ofseveral TNF superfamily ligands and/or receptors at the surface of culturedeosinophils. Finally, we have shown that native CD40L and CD30L at thesurface of purified eosinophils are functionally active and able totransduce proliferative signals on CD40+ and CD30+ target cells, includingcultured H-RS cells. Conclusions: Our data suggest that eosinophils may act as importantelements in the pathology of HD by providing cellular ligands forTNF-superfamily receptors (CD40, CD30, CD95/Fas) able to transduceproliferation and antiapoptotic signals at the surface of H-RS cells. Thepresence on eosinophils of receptors for TNF ligands expressed by activatedT cells (i.e., OX40L, FasL, CD40L, 4-1BBL), also suggest that eosinophils maycontribute to the deregulated network of interactive signals between H-RScells, T cells, and other surrounding reactive cells.
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