Analysis of nerve growth factor receptor expression in human neuroblastoma and neuroepithelioma cell lines.

A series of 22 neuroepithelioma and neuroblastoma cell lines were screened for expression of nerve growth factor receptor (NGFR) by flow cytometry, Western blotting, and Northern blotting. All 5 neuroepithelioma cell lines expressed cell surface NGFR, with 30-69% of cells NGFR positive, but the 17 neuroblastoma cell lines tested had a smaller percentage of cell surface NGFR-positive cells (0-21%) and 10 lines were completely lacking cell surface NGFR. SY5Y, a variant line with a neuronal phenotype derived from neuroblastoma line SKNSH, expressed much more NGFR than SHEP, a variant line with an epithelial-like phenotype also derived from SKNSH. By Western blotting, the Mr approximately 69,000 NGFR band was detected for all four neuroepithelioma cell lines tested but was visible for only 8 of 15 neuroblastoma cell lines tested. The band was most intense for neuroepithelioma cell lines SKNMC and TC32. For these two lines, a Mr approximately 56,000 and a Mr approximately 60,000 band were also detected. By Northern blotting, all three neuroepithelioma cell lines tested were positive for the 3.8 kilobase NGFR mRNA, but only 8 of 15 neuroblastoma cell lines were positive. Neuroepithelioma cell line TC32 and neuroblastoma cell line GICAN had the strongest expression of NGFR mRNA. These results demonstrate that NGFR is a biological marker for neuroepithelioma and that NGFR expression is heterogeneous for neuroblastoma cell lines. This series of neural cell lines differing in NGFR expression will be useful for future studies of regulation of NGFR expression and neuronal differentiation.

Importance of phenotypic and molecular characterization for identification of a neuroepithelioma tumor cell line, NUB-20.

A neuroblastic-like cell line (NUB-20) was derived from a case of histopathologically diagnosed metastatic neuroblastoma. The metastatic tumor and nude mouse heterotransplant resembled neuroblastoma by histological criteria, in contrast to the primary tumor, which was differentially classified as Ewing's sarcoma. However, the cell line demonstrated a unique phenotype in culture with respect to morphology, immunohistochemical markers, and sensitivity to a battery of differentiation modulators. These characteristics, together with the presence of a chromosomal translocation (11;22),(q24;q12) and amplification with enhanced expression of the c-myc protooncogene rather than N-myc, established this tumor as neuroepithelioma. Neuroepithelioma is a tumor type distinct from, but related to, neuroblastoma in its development from the neural crest lineage. These results emphasize the growing importance of cytogenetic and molecular markers in the classification and characterization of human tumors.

Immunohistochemical diagnosis of olfactory neuroblastoma.

Olfactory neuroblastoma (OLF NB) is an uncommon neurogenic tumor arising in the nasal cavity. To identify the immunohistochemical reactivity of OLF NB, we examined specimens from ten patients for the presence of neuron-specific enolase (NSE), S-100 protein (S-100) and glial fibrillary acidic protein (GFAP). Formalin-fixed paraffin sections were treated with rabbit antisera using the peroxidase-antiperoxidase (PAP) and avidin-biotin immunofluorescence methods. Results were compared with those observed in four neuroblastomas (two central and two peripheral sites), 17 medulloblastomas, four pheochromocytomas and five paragangliomas. All OLF NB were positive for NSE and S-100. NSE was uniformly distributed throughout clusters of tumor cells within tumor nodules. The S-100 reactivity was distributed in neoplastic cell nuclei and long cytoplasmic processes of cells which interconnected to form a network at the periphery of tumor cell nests. GFAP was observed only in astrocyte-like cells in one tumor. Although the S-100-positive cells in OLF NB resembled S-100-positive sustentacular cells of paraganglioma and pheochromocytoma, the network pattern was distinctive in OLF NB. The immunodiagnostic pattern of OLF NB includes the following: (a) NSE present in irregular clusters of tumor cells, (b) S-100 present in peripheral cells which form a marginal network, (c) and GFAP rarely present.

Cerebral medulloepithelioma with bone, cartilage, and striated muscle. Light microscopic and immunohistochemical study.

A two-and-a-half-year-old girl had a cerebral medulloepithelioma with histologic evidence of ependymal, astroglial, oligodendroglial, and neuroblastic differentiation, as well as islands of cartilage and bone, and a microscopic focus of striated rhabdomyoblasts. Immunohistochemical staining for glial fibrillary acidic protein and S-100 protein with the peroxidase-antiperoxidase technique revealed positive staining in the various differentiating neural elements of the tumor. Mesectodermal differentiation, or the production by neuroectoderm of tissues usually considered to be mesodermal, is being increasingly recognized in the fields of embryology and tissue culture. Although this is the likely explanation for the composition of this tumor, other possibilities are considered as well.

Evidence for neural origin and PAS-positive variants of the malignant small cell tumor of thoracopulmonary region ("Askin tumor").

The differential diagnoses of childhood and adolescent tumors composed of small round cells include a distinctive clinicopathological entity called malignant small cell tumor (MSCT) of the thoracopulmonary region in childhood. In the present study, 15 such tumors that fulfilled the criteria by Askin et al. were examined for features of possible neural differentiation by light and electron microscopy (EM). With hematoxylin-eosin stain (H&E) the tumors were made up of small undifferentiated cells; rosette formation was noticed in four cases. By immunohistochemistry all 15 tumors were positive for neuron/specific enolase (NSE), which is a specific marker for neural elements and their tumors including neuroblastomas. Ten of 15 MSCT had positive PAS staining. Ultrastructurally dense core (neurosecretory) granules and cell processes indicative of neuronal differentiation could be recognized in 10 of 14 tumors. The dense core granules were often atypical. Filamentous cytoskeleton, never observed in Ewing's sarcoma, was often present. Based on the current results, MSCT of the thoracopulmonary region can be considered a peripheral neuroectodermal tumor with the possible origin in intercostal nerves. MSCTs are generally misdiagnosed as Ewing's sarcoma due to their primitive appearance in H&E sections and their periodic acid-Schiff positivity. NSE immunostaining, preferably augmented by electron microscopy, is necessary for their correct diagnosis.

Some differences in steroid receptors between meningeal and neuroepithelial tumors.

In our study we evaluated the presence of steroid hormone receptors in a large series of endocranial tumors and investigated the possible correlations between receptor presence and levels and patient's age and sex. Our data indicate that steroid receptor behavior in the neoplasias considered is quite different from that observed in endocrine-related tumors, such as breast and endometrial cancer. Moreover, in neuroepithelial tumors an interesting difference between male and female subjects has been found regarding the receptor distribution in poor differentiated neoplasias.

Peripheral neuroepithelioma in childhood. Immunohistochemical demonstration of epithelial differentiation.

Peripheral neuroepithelioma arising from the chest wall of a 4-year-old girl is described. She died of local recurrence 15 months after surgery. Light- and electron-microscopic as well as immunohistochemical findings confirmed the neuroectodermal nature of the tumor. A distinctive histologic feature was the presence of clustered epitheliallike cells, which immunohistochemically stained positive for both keratin and carcinoembryonic antigen. The epithelial nature of these peculiar cells is presented.

Olfactory neuroblastoma. Cytodiagnostic features in a case with ultrastructural and immunohistochemical correlation.

In a case of olfactory neuroblastoma, originally misdiagnosed as an undifferentiated carcinoma, cytologic examination of material scraped from the superior nasal vault revealed tumor cells suggestive of neuroblastoma. The most significant cytodiagnostic feature was the presence of a fibrillary cytoplasm with ill-defined borders. Also noteworthy were the smudged hyperchromatic nuclei and structures resembling rosettes or pseudorosettes. The diagnosis was confirmed by electron microscopy, which revealed the presence of dense-core neurosecretory granules, clear vesicles, neurotubules and neurofilaments, and by immunohistochemistry, which showed positive staining for neuron-specific enolase but negative staining for keratin and glial fibrillary acidic protein. Since olfactory neuroblastoma has a relatively good prognosis and aggressive surgical resection may be curative, it is important that this tumor be distinguished from other small cell malignancies arising in the nasal cavity. The present case shows that the diagnosis can be made by the cytologic examination of scrapings from the tumor.

[Immunohistochemical identification of olfactory esthesioneuromas. Apropos of 16 cases]. / Identification immunohistochimique des esthésioneuromes olfactifs. A propos de 16 observations.

We report anatomoclinical and immunohistochemical analysis of sixteen cases of esthesioneuroblastomas. Microscopic study confirm difficulty of diagnostic for this tumors. Results of S 100 protein reaction for Schwann cells identification, NSE and HNK1 reaction for nervous cells and KL1 reaction for epithelial cells drawn from olfactory mucosa, allow definition of immunologic ENO profile. Pattern immunologic criteria are defined by S 100, NSE or/and HNK1, and eventually KL1 positive reactions permit differential diagnosis with other nervous tumors or undifferentiated carcinomas of nasal fossa. Histo-prognostic patterns are defined by S 100 reactivity distributed in neoplastic cells and cytoplasmic process of cells, to form a continuous network in well differentiated ENO and discontinuous network in undifferentiated forms of ENBO. These results confirm histogenesis of this tumor derived from olfactory mucosa and emphasized only two distinct types: neuro epithelial tumors corresponding to ENEO and cases of ENBO and nervous tumors grouping ENCO and any cases of ENBO.

[Neuroendocrine tumor of the nasal cavity (esthesioneuroblastoma). Apropos of a case with paraneoplastic Cushing's syndrome]. / Tumeur neuroendocrine de la cavité nasale (esthésioneuroblastome). A propos d'un cas avec syndrome de Cushing paranéoplasique.

Presentation of a 48-y old woman who developed a neuroendocrine tumor of the nasal cavities. This lesion progressed rapidly despite an extensive resection and repeated chemotherapy. The patient refused radiotherapy. Before her death, 28 months later, she exhibited a paraneoplastic Cushing-like syndrome. At autopsy, restricted to the brain, there was a 5 cm diameter tumor invading the frontal area without alteration of the hypothalamus or the pituitary gland. Routine histology and electron microscopy confirmed the neuroendocrine nature of the tumor. Immunohistochemistry revealed the tumor to be positive only for neurone specific enolase, negative for S-100 protein, neurofilament and ACTH. The pituitary gland was positive for most usual hormones (GH, PRL, TSH, LH, FSH) but only few cells were slightly positive for ACTH. Many Crooke cells were observed. These findings suggest that the tumor secreted an ACTH-like substance (not detected actually by immunochemistry) that stimulated the activity of the adrenal cortex but inhibited normal production of ACTH at the pituitary gland level.

An immunohistochemical study of the primitive and maturing elements of human cerebral medulloepitheliomas.

Four examples of human cerebral medulloepithelioma were studied immunohistochemically with a panel of antibodies and antisera to neuronal and glial proteins. The tumors, in addition to primitive medullary epithelium, contained areas of neuroblastic, ganglionic, astrocytic, ependymoblastic and ependymal differentiation, and in one tumor, areas resembling polar spongioblastoma. Tumor cells throughout the primitive medullary epithelium displayed focal immunoreactivity for vimentin, glial fibrillary acidic (GFA) protein and for the neuron-associated class III beta-tubulin isotype. Neuroblasts showed immunoreactivity for the class III beta-tubulin isotype, microtubule-associated protein 2 and neuron-specific enolase. Immunoreactivity for neurofilament epitopes and synaptophysin was detected in areas of ganglionic differentiation and coincided with the demonstration of neurofibrils in Bielschowsky's silver impregnations. Vimentin was the only marker detected in ependymoblastic and ependymal rosettes or in areas of polar spongioblastoma, as well as in mesenchymal cells. The results indicate that, even in very primitive neoplastic neuroepithelium, immunocytochemical evidence of early commitment of some of the cells to a neuronal or glial lineage can be demonstrated. The neuron-associated class III beta-tubulin isotype appears to be one of the earliest markers indicative of neuronal differentiation in normal and neoplastic primitive neuroepithelium.

Glial fibrillary acidic protein in medulloblastomas and other embryonic CNS tumours of children.

Investigation of GFAP in 50 medulloblastomas showed a few GFAP-positive tumour cells in 5 cases only; 17 tumours were negative, and 28 showed a "pseudopositivity", i.e. GFAP-bearing cells were identified as reactive or degenerating astrocytes, intermingled with tumour elements. A high GFAP content was seen in 2 small-cell gliomas of the cerebellum, whereas 3 pineoblastomas, 2 neuroblastomas of CNS, and one medulloepithelioma were negative. GFAP is a very good method for identificating astrocytes, but does not seem to be reliable for identifying the origin of undifferentiated tumours such as medulloblastomas. In these neoplasms glial differentiation is lacking or extremely rare, GFAP-positivity being mostly an artifact. The investigation of small tumour samples or the positivity of a single cell are inadequate data for a correct evaluation of the findings, especially taking in mind that GFAP of degenerated astrocytes can be phagocytised by cells other than glial (e.g., macrophages, epithelial and meningioma cells). The importance of carefully checking the whole structure of the tumour is stressed, GFAP positivity or negativity being not a sufficient criterion for its nosological classification.

Primitive neuroectodermal (neuroepithelial) tumour of soft tissue of the neck in a child: demonstration of neuronal and neuroglial differentiation.

A 4-year-old girl had a pathologically proven primitive neuroectodermal (neuroepithelial) tumour of soft tissue in the left posterolateral aspect of the neck. The neoplasm consisted of primitive neuroepithelial cells forming Homer Wright rosettes, mature ganglion cells and astrocytes. Astroglia were identified by localization of cytoplasmic glial fibrillary acidic protein (GFAP). Striking similarity is noted between the current tumour and those found in the central nervous system, including cerebellar medulloblastomas. The diverse cellular elements of the present primitive neuroectodermal neoplasm suggest an origin of the tumour from the neuroectodermal component of an ectomesenchymal remnant of the neural crest. Differentiation of the neuroectodermal component of the neural crest into primitive neuroepithelial cells could result in the occurrence of a primitive neuroectodermal neoplasm which may further differentiate into neurons and neuroglia.

Immunohistochemical demonstration of keratin in canine neuroepithelioma.

Morphological features and immunoreactivity for cytokeratin (CK), glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE) of three canine neuroepitheliomas and three canine ependymomas were investigated. Neuroepitheliomas were in three German shepherds as intradural-extramedullary solitary masses, with spinal cord displacement between T10 and L2. Histologically, they contained tubules and acini, lined by epithelial cells with focal squamous metaplasia, rosette-like structures, and polygonal to spindle-shaped cells between tubules. Acini were empty or filled with a homogeneous, eosinophilic periodic acid-Schiff (PAS)-positive material. Mitotic indices varied from low to moderate. Ependymomas occurred in the third (two cases) and fourth ventricle in adult boxers. Histologically, they were composed of cells with an ill-defined, scant amphophilic cytoplasm, with a central round euchromatic nucleus; cells formed pseudorosettes, with a central fibro-vascular stroma. Neuroepitheliomas stained for CK, but ependymomas did not. Both failed to stain for GFAP, NSE, or phosphotungstic acid hematoxylin (PTAH). Thus, antibodies to cytokeratin are useful to distinguish neuroepitheliomas from ependymomas.