Vimentin expression by reed-Sternberg cells in Hodgkin's disease

Summary The expression of vimentin in Reed-Sternberg cells in 61 samples of Hodgkin's disease (HD) was examined using an avidin-biotin-peroxidase complex technique. Forty biopsies (66%) expressed vimentin, and expression was seen in all subtypes of HD. No immunophenotypic differences between vimentin-positive and vimentin-negative cases were noted. The significance of such expression is unclear, but may be related to the alterations in growth and differentiation that are typical of neoplastic cells.     

Ki-1 lymphomas in childhood: Immunohistochemical analysis and the significance of epithelial membrane antigen (EMA) as a new marker

Summary Two cases of Ki-1 lymphomas in childhood were analyzed immunohistochemically and immunoelectron microscopically. They expressed Hodgkin's disease associated antigen, Ki-1, interleukin-2 receptor (IL2R), OKT9, and HLA-DR. Histologically, the tumour cells were large in size with abundant cytoplasm and atypical nuclei. Lymph node involvement was characterized by parafollicular and marginal sinus infiltration. These features were identical to those reported in Ki-1 lymphomas. Electron microscopically tumour cells had abundant cytoplasmic organelles with pleomorphic nuclei but had no specific granules. Some tumour cells had marked interdigitation of cell membrane. Immunoelectron microscopically Ki-1 was positive on cell membrane. Tumour cells had no T-cell or B-cell antigens except for Leu-3 (T4). Unexpectedly they expressed epithelial membrane antigen (EMA) strongly. EMA was positive on cell membrane and in the cytoplasm. EMA was detected effectively in paraffin-embedded sections. Among the malignant lymphomas in childhood tested, two cases were EMA-positive. The pattern of EMA-reactivity and the histology were very similar to Ki-1 lymphomas. These results strongly suggest that Ki-1 lymphomas in childhood may arise from non-lymphoid haematopoietic cells and that EMA can be used as a new marker to distingish certain type of Ki-1 lymphomas in childhood.     

Loss of mitotic activity and the expression of vimentin in glomerular epithelial cells of developing human kidneys

Glomerular epithelial cells (GECs) have been considered as post-mitotic cells. In order to establish the stage when GECs stop dividing, the expression of proliferating cell nuclear antigen (PCNA/cyclin) in GECs during glomerulogenesis in human kidneys was studied by the streptavidin-biotin staining technique with monoclonal antibodies. Antibody specific for PCNA/cyclin reacted with almost all the cell nuclei of nephrogenic vesicles, as well as those of S-shaped bodies, the cells of which in the lower limb are progenitors of GECs. However, this reaction was markedly reduced in GECs at the capillary loop stage and entirely disappeared at the maturational stage. In contrast to the expression of PCNA/cyclin, vimentin-specific antibody did not react with nephrogenic vesicles and the lower limb of S-shaped bodies, whereas vimentin was ubiquitously expressed in the GECs at the capillary loop stage, as well as at the maturational stage. Furthermore, these two antigens were not co-expressed in the same glomerulus during glomerulogenesis, as revealed by analysis of serial sections. These results lead to the conclusion that expression of PCNA/cyclin and vimentin during glomerulogenesis is fairly stage-dependent, and GECs rapidly lose their mitotic activity at the capillary loop stage.     

Tumour-associated antigens in mammary and extramammary Paget's disease

Summary The immunoperoxidase technique was used to study the immunoreactivities of two murine monoclonal antibodies to carcinoma-associated antigens raised respectively against a human breast cancer line (MBrl) and an ovarian carcinoma (MOv2) and of a conventional anti-CEA serum in 20 cases of mammary Paget's disease of the nipple and in three cases of extramammary Paget's disease. Each of the immunoreagents stained Paget's cells in a high proportion of cases and failed to discriminate mammary from extramammary disease. The antigenic phenotypes of underlying in situ or infiltrating breast carcinomas corresponded to those of the associated Paget's disease of the nipple. The consistent immunoreactivity of eccrine and apocrine sweat glands and of normal mammary epithelia indicated an antigenic relationship between epithelia of adnexal derivation and Paget's cells.     

Expression of lymphoid-associated antigens on Hodgkin''s and Reed-Sternberg cells of Hodgkin''s disease. An immunocytochemical study on lymph node cytospins using monoclonal antibodies

The aim of this study was to elucidate the origin of Hodgkin's and Reed-Sternberg cells. Lymph node cytospins and frozen sections from 20 cases of Hodgkin's disease of different histological subtypes were immunostained by the immuno-alkaline phosphatase technique using a panel of monoclonal antibodies. As expected, the Hodgkin's and Reed-Sternberg cells of all cases were positive for the CD30 (Ki-1), CD15 (hapten X) and CD25 (Tac) antigens. In eight cases, a variable percentage of typical Hodgkin's and Reed-Sternberg cells showed a clear-cut cytoplasmic and/or surface positivity for the T-cell-associated antigens CD3, CD5, CD6 and CD4 (seven cases) or CD8 (one case), but consistently lacked B-cell and macrophage-associated markers. The best visualization of T-cell antigens was obtained in cytocentrifuge preparations and in areas of lymph node frozen sections that had been infiltrated by clusters of Hodgkin's and Reed-Sternberg cells. In two cases of Hodgkin's disease (nodular sclerosis, mixed cellularity) the neoplastic cells weakly expressed the B-cell antigens CD19 and CD22, but not T-cell or macrophage-associated markers. In 10 cases, Hodgkin's and Reed-Sternberg cells were negative for all the lymphoid- and macrophage-associated antigens. These results suggest a lymphoid (either T or B) rather than histiocytic origin for the Hodgkin's and Reed-Sternberg cells in a number of Hodgkin's disease cases.     

On the pathogenesis of sclerosis and nodularity in nodular sclerosing Hodgkin's Disease

Summary Ten cases of nodular sclerosing Hodgkin's disease involving lymph nodes were studied by electron microscopy to determine the ultrastructural composition of the nodule-stromal interphase and the collagenized regions. In addition to a few lymphocytes, rare eosinophils and neutrophils, abundant filamentous and granular electron dense material, collagen fibers and myofibroblasts were observed in all instances. Since myofibroblasts possess contractile and synthetic properties, it is likely they contribute to the retraction and sclerosis which together represent one of the morphologic hallmarks of the disease. The dense fibrosis and contractile state of such tissue may constitute a beneficial host response to contain and limit local and vascular invasion by the neoplastic cellular population, thus contributing to the relative benignity of this form of Hodgkin's disease.     

Differential expression of T cell differentiation antigens and major histocompatibility antigens on activated T cells during the cell cycle

In this report we have analyzed cell cycle-related fluctuations of both quantity and density of the T cell differentiation antigens, CD3 (T3), CD4 (T4) and CD8 (T8), as well as the major histocompatibility complex (MHC) antigens on the cell surface of activated T cells. Phytohemagglutinin-activated T cells cultured for 3 days with or without conditioned medium or for 10 days with conditioned medium and mixed lymphocyte culture-derived T cell clones were used for the analysis. Correlated measurements of the surface antigen quantity (immunofluorescence), DNA content (dye Hoechst 33342), and cell size (light scatter), not influenced by synchrony induction methods and cell fixation, were performed by dual-beam flow cytometry. Our results demonstrate that the T cell differentiation antigens, CD3, CD4 and CD8, and class I MHC antigens are increased in density in the G1 phase for all activated T cells tested. In contrast, class II MHC antigens are increased in density in the G2 phase of activated T cells maintained with conditioned medium. Since it is known that the T cell differentiation antigens and class I MHC antigens on activated T cells are necessary for proliferation of T cells, our study suggests that this effect is more significant in the G1 phase. The cell cycle changes in expression of class I and class II MHC antigens, but not of the T cell differentiation antigens, appear to be mediated by soluble factors, probably including interferon-gamma, which could produce a differential increase of class I and class II MHC antigens on G2 phase cells.     

Epstein-Barr virus-related Hodgkin's disease showing B cell lineage in an immunosuppressive patient seropositive for HTLV-I

A case of Hodgkln's disease (HD), lymphocyte depression (LD) type In an Immunosuppressive patient is described. The patient was a 48-year-old male and his parents were born In the Kyushu area, which is an endemic area for adult T cell lymphomaheukemla (ATL). He was seropositive for ATL virus (ATLV, also referred to as HTLV-I) and showed a marked Immunosuppressive condition. He developed LD-HD and Pneumocystis carinii pneumonia, and died due to respiratory failure. The Immunohistochemical and in situ hybridization analyses revealed that the Reed-Sternberglike cells In the lymph node biopsy sample were positive for Ber-H2 (CD30), Leu-M1 (CD15), L-26 (CD20), Bcl-2, p53 and EBER, the viral genome of Epstein-Barr virus (EBV).     

Disruption of Sertoli cell vimentin filaments in prepubertal rats: An acute effect of butylparaben in vivo and in vitro

Parabens are p-hydroxybenzoic acid ester compounds widely used as preservatives in foods, cosmetics, toiletries and pharmaceuticals. We have recently shown that butylparaben induces spermatogenic cell apoptosis in prepubertal rats. We have conducted the present study for further information. Three-week-old male Sprague–Dawley rats (n = 8) were given a single oral dose of 1000 mg/kg butylparaben. The rats were sacrificed under anesthesia at 3, 6 and 24 h after administration and their testes were collected for histopathological and immunohistochemical examination. Results showed a gradual collapse of Sertoli cell vimentin filaments and decreased actin staining intensity without accompanying changes in the pattern of tubulin expression, while spermatogenic cells became separated from the basement membrane and sloughed into the lumen in the butylparaben-treated rats, compared to the controls. To determine the direct effects of butylparaben on Sertoli cells, primary Sertoli cell cultures with and without butylparaben treatment were examined. Toluidine blue staining in butylparaben treated-cultured Sertoli cells showed an increased number and size of vacuoles in their cytoplasm. In agreement with the in vivo experiment, the in vitro study also clearly demonstrated disruption of vimentin filaments in Sertoli cells after butylparaben treatment. Considering both our present and previous reports, we can speculate that butylparaben-induced disruption of Sertoli cell vimentin filaments may lead to precocious release of spermatogenic cells from underlying Sertoli cells, and the released cells may undergo apoptosis owing to loss of support provided by the Sertoli cells.     

Glial cell differentiation in neuron-free and neuron-rich regions

Summary The development of the human fetal hippocampus and dentate gyros has been studied immunocytochemically. The first glial cells to appear are vimentin-positive radial glial cells. A gradual transition from vimentin to glial fibrillary acidic protein (GFAP) reactivity in the radial glial cells occurs at week 8. The GFAP-positive radial glial cells transform into astrocytes from week 14. A population of small S-100-positive somata which morphologically and spatially are distinct from GFAP-positive radial glial cells and their transformed progeny, are found as early as week 9.5 in the hippocampus during the period of peak neurogenesis. The well-defined immunoreactivity of the morphologically homogenous cell subpopulation for S-100 protein, which has been used as an astrocytic marker in the adult hippocampus, indicates that astrocytes may differentiate at very early gestational ages in human fetuses. The S-100-positive astrocytes are thought to be derived from ventricular zone cells, which at the time of their appearance do not express any of the applied astrocytic markers (S-100, GFAP, vimentin). It is suggested that the S-100-positive astrocytic cell population interacts with the first incoming projection fibers, so modulating the pattern of connectivity.     

锰对大鼠生精细胞Caspase-3 mRNA调控及支持细胞波形蛋白表达的影响

目的 探讨锰对大鼠生精细胞Caspase-3 mRNA调控及支持细胞波形蛋白（vimentin）表达的影响。 方法 将正常雄性SD大鼠48只,随机分为1个对照组和2个染锰组,给予各组大鼠腹腔注射生理盐水和15mg/kg、30mg/kg氯化锰。分别染锰4周和6周,随机处死各组大鼠8只。应用末端脱氧核糖核酸转移酶介导的dUTP缺口末端标记技术（TUNEL） 、原位杂交法和免疫组织化学链酶亲和素生物素过氧化物酶复合物（SABC）法进行检测研究。 结果 1. 与空白对照组比较,15mg/kg、30mg/kg染锰组生精细胞凋亡指数（AI）均升高（ P<0.05, P<0.01）,Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞波形蛋白（vimentin）阳性细胞率均显著降低（ P<0.01）。2. 染锰剂量相同,染锰6周组与染锰4周组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞vimentin阳性细胞率均显著降低（ P<0.01）。3. 染锰时间相同,30mg/kg MnCl2组与15mg/kg MnCl2组比较,生精细胞AI与Caspase-3 mRNA阳性细胞率均显著升高（ P<0.01）,支持细胞vimentin阳性细胞率均显著降低（ P<0.01）。4. 各组大鼠生精细胞AI和Caspase-3 mRNA阳性细胞率呈正相关（ r =0.842, P<0.01）,与支持细胞vimentin阳性细胞率呈负相关（ r =-0.859, P<0.01）,各组大鼠生精细胞Caspase-3 mRNA阳性细胞率和支持细胞vimentin阳性细胞率呈负相关（ r =-0.975, P<0.01）。 结论 染锰大鼠生精细胞Caspase-3 mRNA表达增加导致生精细胞凋亡增加;支持细胞vimentin表达下降;这可能是锰生殖毒性的重要分子机制之一。     

Immunocytochemical localization of vimentin in stellate cells (Folliculo-stellate cells) of the rat, cat and rabbit pituitary pars distalis

Summary The occurrence of vimentin, a specific intermediate filament protein, has been studied by an indirect immunoperoxidase method in the anterior pituitary gland of adult rats, cats and rabbits of both sexes. In the three species studied, the immunoreaction product was detected in the cytoplasm of stellate-shaped cells scattered throughout the pars distalis. These stellate cells showed long cytoplasmic processes which could be seen between the secretory cells, and occasionally, encircling them. These processes sometimes reach the vasculo-connective septa. The marginal cells lining the anterior layer of the hypophyseal cleft of the rat and the cat pituitary glands also showed a positive immunoreaction. Finally, cyst or follicle-like structures lined by immunostained cells with basal processes could be observed in the anterior lobe of the rat, but not in that of the rabbit or the cat.These findings support the previously held view that folliculo-stellate cells and marginal cells of the anterior pituitary gland have a common nature and suggest that these cell types might be derived from glial neuroectodermic cells.     

Regulated expression of cell surface antigens during B cell development

The progression of lymphocytes along the B cell developmental pathway is marked by the regulated appearance and disappearance of a variety of cell surface markers including immunoglobulin. In this review, several of these antigens are discussed in the context of their possible functions during normal B cell differentiation.     

Adenomatoid odontogenic tumour: co-expression of keratin and vimentin

Summary Immunohistochemical observations of intermediate sized proteins in five cases of adenomatoid odontogenic tumour (AOT) are described. The immunohistochemical detections of keratins were made with polyclonal antiserum (TK, 41–65 kDa) and three monoclonal keratin antibodies (KL1: 55–57 kDa; PKK1: 40, 45, and 52.5 kDa and nos. 19, 18, 8; K8.12: nos. 16, 13) and vimentin and desmin monoclonal antibodies. Histologically, the tumour epithelia could be divided into two types: type A cells were a spindle or columnar shape and formed solid, ductal, tubular or whorled structures. Type B cells were small and compact cells at the periphery of the A cell-containing focus. Immunohistochemically, the type A cells showed very slight reaction with all antibodies to keratins, whereas the type B cells indicated slight-to-moderate expression of keratin and vimentin, and showed coexpression. Both types of cell showed a negative reaction for desmin. Only one case was associated with cystic lesions, and the cyst-lining was composed of thin squamous epithelium. Keratin expression in this epithelium was strong. In the histogenesis of AOT it was postulated that the tumour cells may have originated from undifferentiated odontogenic epithelium or stratum intermedium cells.     

The expression of the CD3 antigen in Hodgkin''s disease

Tissue from 86 cases of Hodgkin's disease, fixed in formalin and embedded in paraffin, was immunostained for the T-cell marker CD3. Of these cases, 20 were selected on the basis of previous reactivity of Reed-Sternberg cells for T-cell associated antigens in frozen sections whilst the remaining 66 were retrieved from the routine pathology files. Five of the 20 selected cases and 22 of the retrieved cases showed predominantly cytoplasmic positivity in a subpopulation of Hodgkin and Reed-Sternberg cells. CD3 positive cells were present in all subtypes of Hodgkin's disease including three of nine lymphocyte predominance cases. It therefore appears that some Hodgkin and Reed-Sternberg cells can express the major T-cell antigen CD3. Although these findings are open to other interpretations, they are consistent with the hypothesis that at least some cases of Hodgkin's disease arise from activated T-cells.     

Expressions of cyclin E, A, and B1 in Hodgkin and Reed–Sternberg cells: Not suppressed by cyclin-dependent kinase inhibitor p21 expression

p21 Is involved in the control of the mammalian cell cycle through the binding and inhibition of cyclin-dependent kinases. The cyclins are dependent on the phases of the cell cycle, and divided into two classes: mitotic cyclins (A, B1, B2) and G1 cyclins (C, D1, D2, D3, E). The product of the p21 gene is a potent downstream effector of the p53 tumor-suppressor gene function. The Hodgkin and Reed- Sternberg (H & RS) cells in Hodgkin's disease are reported to frequently express p53, p21, and nuclear proliferative activity (Ki-67). To clarify the relationship of p21, p53 and cyclins, we performed the immunohistochemistry of p53, p21, Ki-67, cyclin D1, cyclin E, cyclin A and cyclin B1, using 11 cases with Hodgkin's disease. In addition, we performed p53 gene sequencing of exon 5-8, and in situ hybridization of Epstein-Barr virus (EBV) EBER-1 region, whose products have reported to induce the expression of cyclin D. In this study, in all cases, Ki-67 was expressed in almost all H & RS cells, and p53 and p21 were expressed in H & RS cells. No p53 gene mutations were detected in any case, and p53 protein overexpression did not correlate with p53 gene mutations. The number of p21-positive H & RS cells was significantly related with that of the p53-positive cells. The cyclins E, A, B1 and D1 were also expressed in H & RS cells. Unexpectedly, the expression of the cyclins was not suppressed by p21 and p53 expression. In addition, the existence of EBV was not related to the expression of cyclins. It is considered that H & RS cells are, indeed, in cell cycle and commonly express the cell cyclins, and that the cell cycle of H & RS cells may not be specifically fixed in the G1, S, G2 or M phases.