Localization of factor VIII/von willebrand factor and glial fibrillary acidic protein in the hemangioblastoma: Implications for stromal cell histogenesis

Summary The histogenesis of hemangioblastoma stromal cells is unresolved. Ultrastructural observations suggest that the stromal cells, endothelial cells, and pericytes that compose this neoplasm are all derived from angiogenic mesenchyme. The expression of factor VIII/von Willebrand factor (FVIII/vWF), a specific marker for endothelial cells, and of glial fibrillary acidic protein (GFAP), a specific marker for glial cells, was examined in 16 hemangioblastomas using the peroxidase-antiperoxidase immunohistochemical method. Endothelial cell staining for FVIII/vWF was intense in 14 tumors, weak in one, and absent in another. There was no stromal cell staining in any of the neoplasms. Process-bearing, GFAP-positive cells were observed near the tumor margin in 13 cases, and deeper in the neoplasm in 8. In two of these tumors there were also occasional GFAP-positive cells that lacked processes and had a vacuolated cytoplasm. Virtually all of the GFAP-positive cells were interpreted as trapped astrocytes rather than stromal cells. The lack of expression of FVIII/vWF by the stromal cells indicates that they are antigenically distinct from endothelial cells. Several alternatives for stromal cell histogenesis remain open. The stromal cells may be derived from endothelial cells that have undergone antigenic loss, or from angiogenic mesenchymal cells that do not express FVIII/vWF. Alternatively, the stromal cells may originate from nonangiogenic mesenchymal cells derived from the mesoderm or neuroectoderm.Supported by Grants CA-11898 and CA-22790 from the National Cancer Institute, HL-24066 from the National Heart, Lung, and Blood Institute, and IM-150 from the American Cancer Society. R.D.M. was supported by Training Grant 5T32 GM-0740304 from The National Institute of General Medical Services     

Neuronal and glial characteristics of central neurocytoma: electron microscopical analysis of two cases

We have identified two central neurocytomas which contained cells co-expressing glial fibrillary acidic protein and synaptophysin defined by double-label immunostaining. Dual-positive cells were mostly polygonal in shape and with a morphological appearence similar to that of reactive astrocytes. This distinct morphology could be used to distinguish cells expressing glial fibrillary acidic protein from cells with round and clear cytoplasm which did not express glial fibrillary acidic protein and which composed the majority of the tumor. Samples containing polygonal cells were selected for electron microscopy from toluidine blue-stained semithin sections. Ultrastructural findings were similar in both neurocytomas, with both being composed predominantly of round cells with clear cytoplasm corresponding to the clear cells identified by light microscopy. Dense-core vesicles and clear vesicles were frequently observed in the cell processes. Apart from these clear cells, polygonal cells with electron-dense cytoplasm were noted. Paralleling the results of double immunostaining, these polygonal cells contained both dense-core vesicles and intermediate, presumably glial filaments. Microtubules and lipofuscin granules were also observed. These results suggest that cells expressing glial fibrillary acidic protein in central neurocytoma include tumor cells with both neuronal and glial characteristics. Received: 25 January 1995 / Revised: 9 June 1995 / Revised, accepted: 1 December 1995     

Immunohistochemical study of fibronectin in hemangioblastomas and hemangiopericytomas

Summary Eight hemangioblastomas and two hemangiopericytomas were studied using indirect immunoperoxidase stains for fibronectin (FN) and glial fibrillary acidic protein (GFAP) in formalin-fixed, paraffinembedded surgical specimens. Stromal cells in hemangioblastomas were GFAP-negative and showed variable FN expression, while GFAP-positive cells were FN-negative, thus suggesting that the stromal cells are not derived from astrocytes. Hemangiopericytoma cells were poorly to intermediately FN-positive. The origin of stromal cells is discussed in the light of their fine structure and the immunohistochemical stains with other cell markers.     

Proliferating potential of folliculo-stellate cells in human pituitary adenomas

Summary Folliculo-stellate cells (FS cells) in 40 pituitary adenomas and portions of anterior pituitary adjacent to the tumor in 26 cases were investigated immunohistochemically, using polyclonal antisera to S-100 protein (S-100) and glial fibrillary acidic protein (GFAP). The objective was to clarify the histological behavior of the FS cells.In most pituitary adenomas there were few or no S-100-or GFAP-positive cell, in comparison with numerous positive cells in the parts of the adenohypophyses compressed by adenomas. However, positive FS cells were observed in some types of pituitary adenomas. Growth hormone and prolactin producing adenomas frequently contained significant amounts of FS cells. In non-functioning adenomas, an unique case of FS cell adenoma was present. The adenoma was composed mainly of FS cells and immature glandular cells. The FS cells were sometimes located around follicles containing Periodic acid Schiff-positive material. Therefore, the FS cell adenoma is characterized by S-100- and GFAP-positive FS cells and PAS-positive follicles. In this type of adenoma, FS cells seemed to be the main proliferating component.In parts of the adenohypophyses adjacent to the adenomas, GFAP0-positive FS cells were numerous. In the pathological conditions FS cells may possess the potential of reactive proliferation.     

GFAP expression of human Schwann cells in tissue culture

We have studied the expression of the intermediate filament (IF) proteins, vimentin and glial fibrillary acidic protein (GFAP), in cultured human Schwann cells (SC) from patients with different neuropathies and normal control cases. SC cultures from sural nerve biopsies of 8 subjects with axonal neuropathies, 8 with demyelinating neuropathies and 3 normal controls were included in this study and processed with double immunofluorescence technique, using anti-vimentin and anti-GFAP antibodies, during the 2nd, 4th and 6th week of culture. Five cultures incubated with anti-GFAP antibodies were also processed for immunoelectron microscopy. Specificity tests of the used antibodies were performed. We have found that: (1) cultured human SC constantly express vimentin; (2) SC from normal controls are GFAP-negative in the first period of culture; (3) SC from pathologic nerves can contain GFAP-immunoreactive IF and the percentage of GFAP-positive SC is higher in axonal than in demyelinating neuropathies; (4) during the permanence in culture human SC from both normal and pathologic cases acquire the ability to synthesize GFAP. The obtained data suggest that the removal from axonal contact and the resulting loss of myelinating function induce a cytoskeletal cellular response in human SC characterized by the cytoplasmic accumulation of GFAP-immunoreactive IF.     

Cerebral granular cell tumors: report of a case and a note on their nature and expected behavior

Summary A case of cerebral granular cell tumor (GCT) is reported. Histologically, the growth was composed of benign astrocytes, granular cells and transitional forms between both elements. Glial fibrillary acidic protein was detected in the glial component and, to a lesser extent, in the granular cells. Alpha-1-antichymotrypsin was demonstrated in the latter component only. Ultrastructural study also supported the evidence that neoplastic astrocytes became granular cells. The survey of the literature and our own results suggest that GCTs in this particular location, even when histologically benign, seem to have a worse prognosis than the low-grade supratentorial astrocytomas.     

Glial fibrillary acidic protein and intermediate filaments in human glioma cells

Summary Cultured human glioma cells were studied by double indirect immunofluorescence technique using antisera against intermediate filaments and glial fibrillary acidic protein. With both antisera cytoplasmic fibrillar fluorescence was seen. Perinuclear bundles of intermediate-sized filaments, induced by vinblastine treatment, were strongly stained with both antisera. The degree of codistribution of the two types of antigenic determinants varied considerably from cell to cell. These results suggest that two types of filament-related antigenic determinants can be present in the same cell, and also that glial fibrillary acidic protein-related filaments may possess functional similarities to the intermediate filaments found in other cells. Glial fibrillary acidic protein remains as a useful and specific antigenic marker for the study of glial cells in vitro.     

Medulloblastomas in childhood: histological factors influencing patients' outcome

Sixty-five medulloblastomas in infancy and childhood treated from 1965 through 1981 were reviewed, and the correlation between histological findings of medulloblastomas and clinical course of the patients was studied. Thirty-five patients died but the remaining 30 are alive and without clinical evidence of recurrence 5 years or more after surgery. Certain histological features on light microscopic examinations (e.g., pleomorphism of tumor cells, nuclear-cytoplasmic ratio, mitotic index, degree of vascularity and endothelial proliferation) do influence patient outcome with statistical significance (P<0.05). Thirty out of 65 medulloblastomas were examined further, using immunohistochemical methods with glial fibrillary acidic protein (GFAP), neuron specific enolase (NSE), and factor VIII/vW factor (F VIII/vWF). GFAP stain was negative in 20%, NSE stain in 13.3% and F VIII/vWF stain in 16.7% of the medulloblastomas studied. Desmoplastic medulloblastomas showed a strong tendency toward positive NSE and GFAP staining in the glomerular portion. There was no correlation between patient outcome and the results of applied immunohistochemical studies. Our data indicate that certain histological features may influence patient outcome, but the degree and pattern of cellular differentiation do not predict outcome.     

Gliosis in the spinal cords of rats with experimental allergic encephalomyelitis: immunostaining of carbonic anhydrase and vimentin in reactive astrocytes

Spinal cord sections from rats sensitized to develop experimental allergic encephalomyelitis (EAE) were immunostained with antibodies against glial fibrillary acidic protein (GFAP), carbonic anhydrase, and vimentin, to see whether the latter two antigens could be detected in GFAP-positive reactive astrocytes. Sixteen days after sensitization (16 dpi) there was intense carbonic anhydrase immunostaining in GFAP-positive cells in the spinal cords of EAE rats, particularly in the white matter. At 13 and 20 dpi carbonic anhydrase immunostaining in astrocytes was less intense, and in the spinal cord white matter of control animals carbonic anhydrase was not detected in the few GFAP-positive cells. In the spinal cords of EAE rats vimentin immunostaining was observed in inflammatory cells and astrocytes. In the latter, GFAP and carbonic anhydrase were colocalized with vimentin. The data suggest that carbonic anhydrase expression in astrocytes is an acute response to injury and that vimentin can be detected in astrocytes, as well as inflammatory cells, as early as 16 dpi.     

Peculiar pattern of glial fibrillary acidic protein production in extracranial metastatic tumor cells of malignant astrocytoma

Summary An 83-year-old woman suffered from malignant astrocytoma originating in the temporal lobe. Autopsy revealed its extracranial metastasis to the liver, lung and bone marrow. The tumor tissue at the primary site was composed of plump, process-forming cells and small cells with scanty cytoplasm, and showed dural invasion. In the metastatic areas, most of the tumor cells were small cells, although proliferation of the plump cells in contact with perivascular connective tissue was marked, particularly in the liver. These plump cells were positively stained with antiserum to glial fibrillary acidic protein (GFAP), showing that the collagenous tissue was able to induce increased production of GFAP by the glial tumor cells.     

Immunohistochemical observations on rat radial glia: relationship with the origin of ethylnitrosourea-induced tumors

Summary Gliomas induced in the rat by transplacental administration of ethylnitrosourea (ENU) are intensely immunoreactive for vimentin and scarcely for glial fibrillary acidic protein (GFAP). Since tumoral transformation takes place during the late fetal and early postnatal period, the sequential expression of the two glial antigens has been investigated in this age period in ENU-treated and control rats. Immunohistochemical and immunoelectron microscopical methods have been employed. Vimentin was widely expressed starting from embryonal day 14 (E 14) in the processes of radial glia; as long as radial glia was present, vimentin decorated it. GFAP was, at earliest, observed at E 20 and expressed by glial cells with a stellate, i.e., mature shape. No GFAP-positive radial process was observed. No difference was found between ENU-treated and control rats. Since ENU is most effective in producing tumors when administered at the 16–17th day of fetal life, vimentin-positive radial glia is a candidate target of ENU. The similarity of intermediate filament pattern between radial glia in the late fetal life and tumors induced by transplacental ENU suggests that radial glia might be the cell of origin.Supported in part by M. P. I. 60% grant, Rome     

Gliomatosis cerebri: report of a case presenting as a focal cerebral mass

Summary Gliomatosis cerebri (syn. astrocytomatosis cerebri) is a rare diffuse neoplastic condition affecting all parts of the brain. The clinical and pathological features of an unusually diffuse and histologically uniform case of gliomatosis cerebri are presented. Many tumour cells stained positively for glial fibrillary acid protein, confirming the astrocytic derivation of this neoplasm. Differing views on the nature of gliomatosis cerebri are briefly discussed.     

Axonal regeneration in old multiple sclerosis plaques

Summary Cryostat sections of two old plaques removed at autopsy from the spinal cord of a 62-year-old man with multiple sclerosis of 24-year duration were studied by indirect immunofluorescence with antibodies to neurofilament proteins, glial fibrillary acidic protein (GFAP), glial hyaluronate-binding protein (GHAP), vimentin and laminin. The neurofilament monoclonal antibodies used in this study reacted with phosphorylated epitopes of the two large polypeptides of the neurofilament triplet (NF 150K, NF 200K). As previously reported [Dahl D, Labkovsky B, Bignami A (1989) Brain Res Bull 22:225–232], the neurofilament antibodies either stained axons in the distal stump of transected sciatic nerve in the early stages of regeneration or late in the process, i.e., after regenerating axons had reached the distal stump of the transected sciatic nerve. Both multiple sclerosis plaques were positive for GFAP and vimentin, but negative for GHAP, while astrocytes in myelinated spinal cord white matter stained with both GFAP and GHAP antibodies. Laminin immunoreactivity in the plaques and normal spinal cord was confined to blood vessels. One plaque was almost devoid of axons as evidenced by indirect immunofluorescence with neurofilament antibodies. Another plaque was packed with bundles of thin axons running an irregular course in the densely gliosed tissue. Axons in the plaque only stained with neurofilament antibodies reacting with sciatic nerve in the early stages of regeneration while axons in the surrounding myelinated white matter were decorated by all neurofilament antibodies, regardless of the time of appearance of immunoreactivity in crushed sciatic nerve. It is concluded that reactive astrocytes forming glial scars do not constitute a non-permissible substrate for axonal growth.Supported by NIH grant NS 13034 and by the Veterans Administration     

Effect of thyroid deficiency on the development of glia in the hippocampal formation of the rat: an immunocytochemical study

The development of glia in the hippocampal formation of normal and hypothyroid rats was studied using immunocytochemical staining for either glial fibrillary acidic protein (GFAP) or vimentin. Light microscopy showed lower GFAP immunoreactivity in the radial glial processes of young hypothyroid rats compared to normal animals. These processes followed the known path of neuroblast migration toward the proliferative zone of the dentate gyrus until the end of the 1st postnatal week. Vimentin immunoreactivity showed that the glial processes were present and therefore immature at least with respect to their cytoskeletal composition. We propose that this early defect in the maturation of the radial glial fibers accounts for the final deficit in the granule cells of the dentate gyrus. Later in development, thyroid deficiency also reduced the density and number of GFAP-labeled astrocytes and the growth of their processes. This observation is in complete disagreement with the glial hypertrophy induced by thyroid deficiency in the cerebellum. The considerably increased histogenetic cell death observed in the cerebellum of young hypothyroid rats could in turn induce glial hypertrophy, whereas the hippocampal formation, where a normal low number of cell deaths is observed, is only subjected to the general depressive effect of thyroid deficiency on cell maturation.     

Hemangioblastomas: Immunohistochemical and ultrastructural study of the stromal cells

An immunohistochemical study of 36 hemangioblastomas (Hmbl) from 32 patients was performed to clarify the cytogenesis of stromal cells (SC). In 19 of 29 Hmbl, SC revealed glial fibrillary acidic protein-immunolabeling with an antigen retrieval by trypsinization, and were also positive for S-100 protein, αB crystallin, neuron-specific enolase and vimentin. Ultrastructurally, SC contained lipid droplets, unevenly distributed intermediate filaments with occasional myelin figures, primitive desmosomal junctions and, although rare, Rosenthal fibers or a cilium. It is concluded that SC associated with Hmbl exhibit markers consistent with an astrocytic origin.     

Denervated skeletal muscle stimulates migration of Schwann cells from the distal stump of transected peripheral nerve: An in vivo study

We have tested the stimulation of Schwann cell migration from the distal stump of a 1 week transected sciatic nerve of adult rats by denervated skeletal muscle. Migrating Schwann cells were distinguished by the presence of non-specific cholinesterase (nChE) activity and glial fibrillary acidic protein (GFAP) at a distance of about 6 mm among denervated muscle fibres 4 weeks after insertion of the distal stump. In addition, the distal stump was introduced into the open end of a silicone chamber packed with artificial fibrin sponge (Gelaspon®) soaked in homogenate from intact or denervated muscles. A larger amount of migrated Schwann cells was observed in the chambers filled with homogenate from denervated muscles. An alteration in the amounts of Schwann cells migrating into the silicone chambers observed after histochemical staining (nChE or GFAP) was supported by biochemical measurements of the nChE activity. The biochemical assessment of the nChE activity revealed the increased amounts of migrated Schwann cells in proportion to the protein contents of homogenates from the denervated muscles. In addition, heating of homogenate from the denervated muscles resulted in a diminution of Schwann cell migration. Bromodeoxyuridine incorporation did not show an increased proliferation of Schwann cells inside the chambers following application of homogenate from the denervated muscles in comparison with the homogenate from the innervated muscles. Our results suggest a stimulation of Schwann cell migration from the distal stump of the transected sciatic nerve by soluble factor(s) produced by denervated skeletal muscles.     

锂-匹罗卡品致痫大鼠海马胶质纤维酸性蛋白的表达及意义

目的观察其海马经HE染色后组织病理学、胶质纤维酸性蛋白免疫反应阳性表达细胞在LPS中各观察时间点海马CA1、CA3、齿状回的表达,探讨其致机制。方法锂-匹罗卡品急性诱导SD癫痫持续状态模型鼠形成后,采用免疫组化和图像分析方法观察海马HE染色组织病理学、胶质纤维酸性蛋白免疫反应阳性表达细胞。结果模型组各时间点海马细胞形态出现病理性改变,部分细胞脱失,胞浆浓缩,胞核固缩深染;胶质纤维酸性蛋白免疫反应阳性表达细胞亦显著上调（P〈0.05）。结论Pilo诱导SD大鼠癫痫发作后存在显著的海马神经元结构和胶质细胞的损伤,以胶质细胞损伤更显著,胶质纤维酸性蛋白持续高表达可能是这种功能异常的胶质细胞增生的重要原因,也可能是锂-匹罗卡品致癫痫发作的重要因素之一。     

Distribution of metallothionein in the human central nervous system

The distribution of metallothionein (MT), a metal-binding protein, was examined immunohistochemically in the normal human brain and spinal cord. Paraffinembedded brain tissue from three patients who had died from a non-neurological disease and were free of histopathological central nervous system alterations were processed. The results of the present study demonstrate that MT is readily detectable in a subgroup of astrocytes in the normal human brain. MT staining is most intense on grey matter astrocytes that bear short stout processes and which probably represent protoplasmic astrocytes. Using anti-MT and anti-glial fibrillary acidic protein immunostaining, we could demonstrate two subpopulations of astrocytes that were mutually exclusive. The functional significance of MT-expression in protoplasmic astrocytes is not entirely clear. Metal detoxification is only one of the many postulated functions of MT. The finding that staining for MT permits subtyping of astrocytes may be of great importance in glia research and surgical pathology of the human brain. © 1993 Wiley-Liss, Inc.     

Argyrophilic tau-positive twisted and non-twisted tubules in astrocytic processes in brains of Alzheimer-type dementia: an electron microscopical study

This report concerns pathological astrocytic tubular structures (astrocytic tubules, As-Tbs) that coexist with glial filaments in astrocytic processes in brains with presenile-onset Alzheimer-type dementia. The formation of As-Tbs appears to be related to the duration of disease and the intensity of Alzheimer histopathology. In three cases in which the disease was of extremely long duration, As-Tbs were found in the frontal and temporal neocortices, the temporal pole and the hippocampus using electron microscopy, whereas they were not found in two cases with a long, but not extremely long, illness duration. As-Tbs were almost exclusively found in the highly devastated neuropil, and we could not find them in regions of moderate neuronal degeneration despite intensive inspection. As reported previously, some As-Tbs was seen adjacent to extracellular neurofibrillary tangles (NFTs) and in perivascular astrocytes. Our novel finding is that they can exist independently from these, in the highly devastated neuropil. Two types of As-Tbs were observed, twisted tubules with periodic constrictions at 50- to 80-nm intervals and non-twisted tubules where no constrictions were seen but which had a 15-nm fuzzy outer contour. They were positively stained by anti-human tau antibody, an antibody that does not recognize extracellular NFTs. Thus, it is most likely that As-Tbs are not the sequestration of extracellular NFTs, and that they are of astrocytic origin. Moreover, As-Tbs showed argyrophilia. As-Tbs appear indistinguishable from dystrophic neurites under the light microscope. The present data suggest that they may be more widely distributed in the damaged cerebral neuropil than previously thought. Received: 20 June 1996 / Revised: 20 July 1997 / Accepted: 31 July 1997     

Development of stroma in malignant lymphomas of the brain compared with epidural lymphomas

Summary The relation of lymphoma cells to gliomesenchymal stroma within nervous tissue was studied by peroxidase-antiperoxidase immunostaining of formalin-fixed and paraffin-embedded surgical specimens for fibronectin (FN), factor VIII-related antigen and glial fibrillary acidic protein in 17 malignant non-Hodgkin lymphomas of the brain. For comparison, 9 non-Hodgkin lymphomas, 6 Hodgkin lymphomas, and 19 plasmacytomas of the spinal or cranial epidural spaces were studied with the same methods. Lymphoma cells were consistently negative for all markers. All lymphomas of the brain showed conspicuous concentric perivascular circles of immunoreactivity for FN in parts infiltrating brain tissue. Such structures are considered to derive from splitting of basal laminae of preexisting brain vessels; they were not seen in tumors of the epidural space. Cells with conspicuous FN content were found in brain as well as in epidural lymphomas. A monohistiocytic origin of those cells was confirmed by presence of monohistiocytic markers lysozyme and -1-antichymotrypsin. Thus, additional immunostaining for FN seems to be useful for detecting monohistiocytes/macrophages in brain tumors.