A monoclonal antibody against nerve growth factor receptor. Immunohistochemical analysis of normal and neoplastic human tissue.

The expression of human nerve growth factor (NGF) receptor in tumors and normal tissue was investigated with the use of a monoclonal antibody recently developed against that protein. This antibody, NGFR5, reacted strongly with 100% of 25 nerve sheath tumors. Eight of nine pheochromocytomas and three of three paragangliomas also had positive results, but the immunoreactivity was restricted to the sustentacular cell population. Within cells of melanocytic lineage, there was no immunostaining of melanocytes in normal epidermis, whereas 13 of 14 benign nevi had positive results, primarily involving spindled nevocytic structures within the dermis. NGF receptor was scarcely expressed in human melanoma; 9 of 19 melanomas had positive results, but immunoreactivity was generally restricted to rare cells within the larger tumor cell population. Among nonneurogenic mesenchymal tumors, results were generally negative: 0 of 5 chondrosarcomas, 0 of 6 malignant fibrous histiocytomas, 0 of 3 meningiomas, and 1 of 8 leiomyosarcomas were immunoreactive. Carcinomas were variable in immunoreactivity: 12 of 16 squamous cell carcinomas had positive results, whereas adenocarcinomas demonstrated focal, basal epithelial immunoreactivity and neuroendocrine tumors generally had negative results. Among normal tissues, in addition to expected neural immunostaining, NGFR 5 reacted positively with several nonneural cell types, including lymphoidal follicular dendritic cells, myoepithelial cells, vascular adventitia, and basal epithelium of oral mucosa and hair follicles. Antibodies to NGF receptor may play a role in the identification of benign and malignant soft tissue lesions.

Expression of the alpha subunit of human chorionic gonadotropin in normal and neoplastic neuroendocrine cells. An immunohistochemical study.

The alpha subunit of human chorionic gonadotropin (HCG) was localized Immunohistochemically in paraffin sections of normal human tissues and neuroendocrine tumors. A small subset of dispersed neuroendocrine cells was positive in normal adult tissues, including gastric antrum, urachal remnant, anal glands and prostate. Positive cells were consistently present in perinatal lung but rare in adult lung. In contrast, the beta subunit was absent from these cells. Seventy-two of 151 extrapituitary neuroendocrine tumors (48%) were alpha subunit-positive. Thirty-three of 37 bronchial carcinoids (92%) were immunoreactive, with a high percentage of the tumors (54%) containing moderate to large numbers of positive cells. The alpha subunit was further demonstrated in 9 of 45 small cell lung carcinomas (20%), 19 of 35 extrapulmonary carcinoids (54%), 3 of 11 islet cell tumors (27%) and 8 of 13 medullary thyroid carcinomas (62%). Two of three malignant islet cell tumors were positive. Positive cells were usually few in number, except for two small cell lung carcinomas, two rectal carcinoids, one thymic carcinoid and one malignant islet cell tumor. Pheochromocytomas (n = 10) were negative. Eleven of 19 pulmonary tumorlets (58%) were alpha subunit-immunoreactive. A few beta subunit-positive cells were detected in only 6 lung lesions. The physiological significance of the imbalance of expression of HCG subunits by certain neuroendocrine cells and their tumors remains unknown.

Primitive neuroectodermal tumours of the central nervous system--an electron microscopic & immunohistochemical study.

A series of 75 poorly differentiated neoplasms of the central nervous system viz., medullo-blastoma, cerebral neuroblastoma, pineoblastoma and ependymoblastoma were studied by light microscopy (LM), electron microscopy (EM) and immunohistochemistry (IH). Although the predominant cells constituting these tumours appeared to be undifferentiated cells by LM and EM, many others showed evidence of differentiation into glial, neuronal or ependymal cell lines by EM and IH. It is therefore, concluded that all these neoplasms are best considered as primitive neuroectodermal tumours and these may be classified as such in neuro-oncology. They may be subclassified further on the basis of differentiation into one or more cell lines.

Flow cytometric study on cell kinetics of brain tumours and their cultured cells.

With the aid of flow cytometry (FCM), distribution of DNA content in 40 cases of brain tumour, primary culture cell, and secondary culture cell can be determined and chronological change after subculture is studied from the analysis of their cell cycle. In most primary cultures, proliferating index (PI) is likely to decrease, which suggests that environmental change might affect the growth activity. In comparison with that of the original sample, DNA-histogram of the secondary culture can be divided into the following 3 types: the type recovering to the original pattern ("adapting type"), in which astrocytoma, ependymoma, glioblastoma and medulloblastoma are included, 2) the type increasing more at G2 + M phase than the original ("proliferating type"), in which meningioma and some of glioblastoma are included, and 3) the type decreasing so far as to induce degeneration or death ("degenerating type"), in which pituitary adenoma and neurinoma are included. FCM is of great usefulness for the study of cell kinetics of a tumour cell undergoing culture and the present method will be available for the respective study of biological characteristics of the cultured cell, established cell line or sensitivity test for antineoplastic agents.

Immature neural elements in immature teratomas. An immunohistochemical and ultrastructural study.

Immature neural tissue pathobiology in teratomas may have important implications for clinical prognosis, nervous system embryology, and neurological oncology. However, immunohistochemical and ultrastructural examinations of these neoplasms have been scarce. The authors examined immunohistochemically the immature neural elements in nine immature teratomas. Using modified peroxidase-antiperoxidase (PAP) immunoperoxidase (IP) techniques, they evaluated the immunoreactivity to glial fibrillary acidic protein (GFAP), neuron-specific enolase (NSE), neurofilament (NF), chromogranin (CG), and vimentin (VM). All nine teratomas were immunoreactive for both GFAP and NSE, one was reactive for NF, and five (56%) were immunoreactive for vimentin. All cases were nonreactive for chromogranin. In addition, ultrastructural examination (electron microscopic [EM]) was performed on eight of these tumors. By EM examination, both astrocytes and oligodendroglia were identified in varying stages of development. Astrocytes often displayed abundant intermediate filaments. However, primitive undifferentiated cells were also found. Neuronal differentiation included long cell processes with tubules and filaments, vesicles, rare dense-core granules, and synapses. Ependymal differentiation (cilia, microvilli, prominent junctions) was observed in two cases. Pigmented retinal epithelium was seen in one tumor. No ambiguous (hybrid) cells were identified. Cellular interactions usually resembled the relationships found in normal adult brain tissue.

The expression of subunits of human chorionic gonadotropin (hCG) by nontrophoblastic, nonendocrine, and endocrine tumors.

The value of the subunits of human chorionic gonadotropin (hCG) as tumor markers is controversial. The production of hCG-alpha and hCG-beta by 214 nontrophoblastic exocrine and by 416 endocrine tumors was analyzed by using immunocytochemical technics. hCG-alpha immunoreactivity was found in 131 of 416 endocrine and in 8 of 214 nonendocrine tumors. hCG-beta could not be visualized specifically with the use of two polyclonal antisera and one monoclonal antibody. The authors conclude that (1) hCG-alpha is neither a tissue nor a tumor-specific marker; (2) hCG-alpha is produced by a variable proportion (21-55%) of endocrine tumors arising in various organs but not by ileal carcinoids, paragangliomas, pheochromocytomas, nor Merkel cell tumors; (3) hCG-alpha is a marker of malignancy only in pancreatic endocrine tumors; and (4) hCG-beta is rarely, if ever, produced by tumors.

Gastric submucosal tumours of neurogenic origin with neuroaxonal and Schwann cell elements.

Twelve cases of gastric submucosal tumours, originally diagnosed as leiomyoma and leiomyosarcoma, were investigated by staining with the neurogenic markers S-100 and neuron specific enolase (NSE). Three cases were, in addition, studied by electron microscopy. The tumours stained negatively for desmin, indicating that they were not of smooth muscle origin. All stained positively for S-100 and NSE. The ultrastructural features in the cases examined by electron microscopy were reminiscent of autonomic nerve structures normally present in the gastric wall. These included fine cytoplasmic processes of Schwann cells, wrapped with complete or incomplete basement membranes, and structures analogous to post-ganglionic neuroaxonal components. These tumours appear to represent a distinct type of gastric neoplasm originating from autonomic nerve elements in the stomach wall. They are different from previously described schwannomas or neurofibromas. In some of the tumours, identifiably neural elements are relatively a minor component, and the majority of the tumour cells are of undetermined origin. It is suggested that these cells may be poorly differentiated, lacking their antigenic determinants specific to neural differentiation.

The immunohistochemistry of gastrointestinal stromal tumors. Evidence supporting an origin from smooth muscle.

To study the histogenesis of gastrointestinal stromal tumors, 79 cases were evaluated for desmin (DES), vimentin (VIM), and S-100 protein immunoreactivity by the avidin-biotin immunoperoxidase procedure on paraffin-embedded, Bouin's-fixed tissue sections. All tumors showed weak vimentin positivity. Trapped non-neoplastic smooth muscle and nerve twigs were often noted, particularly at the tumor periphery. Significant tumor S-100 positivity was not identified in our series. Similarly, glial fibrillary acidic protein (GFAP) immunoreactivity (performed in 11 desmin-negative tumors) was not detected within either gastrointestinal stromal tumors or enteric glial cells. Fifty-three percent (53%) of all gastrointestinal stromal tumors (GIST) displayed positive tumor cell desmin immunoreactivity. All 10 esophageal and all four colorectal tumors were diffusely desmin positive and unequivocal smooth muscle lesions. In contrast, only 17 of 37 (46%) benign and six of nine (67%) malignant gastric tumors were desmin positive. Similarly, four of 10 (40%) benign and one of nine (11%) malignant small-bowel tumors expressed desmin. Several gastric neoplasms with prominent nuclear palisading resembling schwannian Antoni A regions were nonetheless desmin positive. Epithelioid gastric tumors were more frequently desmin positive than nonepithelioid tumors; however, this positivity did not attain statistical significance. Two gastrointestinal stromal tumors that were desmin negative in paraffin-embedded material had detectable antigen in frozen sections. Gastric and small-bowel tumors measuring less than 3 cm were significantly more often desmin positive than those 3 cm or greater. We conclude that the method of fixation, tissue preparation, and immunostaining may significantly affect the expression of desmin. Although the histogenesis of gastrointestinal stromal tumors remains controversial, most of these tumors show evidence of smooth muscle differentiation.

Immunohistopathologic spectrum of the glial fibrillary acidic protein in neurooncology.

To conventional histologic and special staining techniques as well as electron microscopy, immunohistological methods have recently been added to characterize neuroectodermal and non-neuroectodermal tumors of both the central and the peripheral nervous system. An ever increasing panel of these morphologic techniques enables us today, to characterize precisely such tumors in or around the central and peripheral nervous system, sometimes only hampered by preservation of tissue inadequate for the respective methods.

[Immunohistochemical localization of S-100 protein in granular cell myoblastoma]. / Localización inmunohistoquímica de la proteína S-100 en el mioblastoma de células granulares.

Three cases of granular cell myoblastoma have been studied in order to determine the presence and distribution of the S-100 specific protein in the neoplastic cells, using immunocytochemical staining techniques, through the modified avidin-biotin method. Positive immunostaining was observed in the three cases studied. The comparative study of various control cases histogenetically originating from neuroectoderm (melanoma) and specifically from Schwann cells, as also the presence of strongly positive staining in Schwann cells of peripheral nerve fibres situated inside and outside the tumor, support the concept of the neurogenic origin of this interesting tumor.

Immunohistochemistry in neuro-oncology.

The technical modalities of the immunohistochemical demonstration of antigens of neuroectodermal differentiation and of mesenchymal nature in neural tissue are discussed. On the main antigens (S-100 protein, GFAP, Vimentin, Neuronal specific Enolase, neurofilaments, myelin basic protein, carboanhydrase C, FVIII/RAg, laminin, fibronectin) the most important findings are described and critically considered. They are interpreted in the light of the present knowledge on the problems of differentiation of the oncotypes and on particular aspects of bio-pathology of brain tumors.

Immunohistochemical demonstration of vimentin in human cerebral tumors.

The distribution of vimentin (VIM) has been histochemically investigated in 53 cerebral tumors and compared in gliomas to that of glial fibrillary acidic protein (GFAP). In gliomas VIM is less positive than GFAP, but shows the same distribution. It cannot be considered as indicating immaturity of glial tumor cells. VIM is also positive in glial processes of cerebellar pilocytic astrocytomas, in Schwann cells of neurinomas and in endothelial cells of all oncotypes. In medulloblastomas, VIM decorates reactive glia cells. A diffuse positive reaction has been observed in meningiomas. In hemangioblastomas, besides intervascular and endothelial cells, groups of polygonal cells are intensely positive for both VIM and GFAP. The interpretation of VIM in cerebral tumors is largely based on the distribution patterns of this intermediate filament in the developing CNS of rodents.

An immunohistochemical study of glial and neuronal markers in primary neoplasms of the central nervous system.

Paraffin-embedded tissues from 56 primary neoplasms of the central nervous system and seven cases of non-neoplastic reactive astrocytosis were examined by immunoperoxidase techniques on serial sections using monoclonal antibodies to glial fibrillary acidic protein (GFAP) and the 68 kDa neurofilament subunit and monospecific polyclonal antibodies to alpha- and gamma-enolase. gamma-Enolase was present in all neoplasms of neuronal origin, but was also present in anaplastic gliomas (particularly in giant cells), in some well-differentiated astrocytomas and reactive astrocytes. The cells containing gamma-enolase in these cases appeared morphologically identical to those containing alpha-enolase and GFAP in adjacent serial sections. No relationship was found between gamma-enolase immunoreactivity and cellular anaplasia in the gliomas studied. Subependymal neoplasms from patients with tuberose sclerosis exhibited evidence of both astrocytic and neuronal differentiation, sometimes in morphologically distinct cell populations, consistent with their suggested origin from a primitive cell line.

S-100 protein as a marker for melanocytic and other tumours.

The majority of melanocytic tumours are easily diagnosed but they become a problem when they are amelanotic and the tumour cells resemble those of other tumours. This applies particularly to secondary melanoma. Detection of S100 protein is a useful identifying marker. S100 protein, so named for its solubility in saturated ammonium sulphate, is derived from brain tissue. It is a dimer and belongs to a calcium binding group of proteins. The protein was first thought to be in neural or neural crest derived tissues but has been found in chondrocytes, adipocytes, myoepithelial cells, dendritic cells of lymphoid tissue, Langerhans cells and T lymphocytes. The protein is present in a high proportion of malignant melanomas and nevocytic nevi of skin, but is less positive in eye melanomas. It is present in gliomas, Schwannomas and neurofibromas but not in neurone derived tumours such as neuroblastomas. Chondromas, chondrosarcomas, liposarcomas, some osteogenic sarcomas and some histiocytic tumours are positive. The tumours that do not contain S100 protein are listed. Pending development of melanoma-directed monoclonal antibodies, the use of anti-serum to S100 protein plus anti-keratin and anti-leukocyte reagents is useful in the identification of tumours of doubtful histogenesis.

Bombesin in human neuroendocrine (NE) neoplasms.

Bombesin is a 14 amino acid peptide isolated from amphibian skin which was found to have stimulatory effects upon gastric and pancreatic secretions, release of gastrointestinal hormones, gallbladder contraction and bronchoconstriction. It is present in amphibian gastric endocrine cells, avian proventriculus endocrine cells and avian brain. In mammals it is present mainly in nerve cells and fibers. The only mammalian endocrine cell shown to date to have bombesin is the P-cell in fetal lung. Bombesin is also found in mammalian brain, with its highest concentration in the hypothalamus. We examined several groups of human neuroendocrine neoplasms for the presence of bombesin by immunohistochemistry. Our findings indicate that bombesin is present 68% of bronchial carcinoids, 65% of pulmonary neuroendocrine carcinomas, 62% of neuroendocrine carcinomas of the skin, 5-10% of pheochromocytomas and extraadrenal paragangliomas and 35% of gastrointestinal carcinoids and neuroendocrine carcinomas. Parallel studies in a wide variety of non neuroendocrine neoplasms failed to reveal the presence of bombesin. We conclude that bombesin is a highly specific marker of neuroendocrine differentiation and thus a valuable tumor marker. Furthermore, its specificity compares favorably with another neuroendocrine marker, neuron specific enolase, an enzyme thought to be present only in neural tissues and neuroendocrine cells but recently found in non neural human tissues and non neuroendocrine neoplasms.

The contribution of immunohistochemistry to the diagnosis of neuroendocrine tumors.

The use of specific secretory products, hormones, and neuroregulators as diagnostic tools for neuroendocrine tumors is illustrated. Results of extensive immunohistochemical and cytologic investigations are discussed in the light of other pathologic and clinical findings to serve as a basis for tumor classification and as a help in prognostic evaluation. Endocrine tumors of the pancreas, gut, lung and urogenital tract are dealt in some detail.

Value of S-100 protein in the diagnosis of soft tissue tumors with particular reference to benign and malignant Schwann cell tumors.

Two hundred and two benign and malignant soft tissue lesions were studied for the presence of S-100 protein by means of the peroxidase-antiperoxidase technique on formalin-fixed, paraffin-embedded tissue. Virtually all benign nerve sheath tumors (neurofibroma, neurilemoma, and granular cell tumor) contained numerous immunoreactive S-100-positive cells. Only one-half (18 of 36) of malignant schwannomas contained the protein, suggesting that its presence is an expression of differentiation in Schwann cell tumors. S-100 protein was not identified within pure neuroblastic tumors (neuroblastoma, neuroepithelioma) but could be identified within rare cells of the ganglioneuroblastoma and within the Schwann cell component of ganglioneuroma. It was also identified within most melanocytic tumors (cellular blue nevus, clear cell sarcoma, and melanoma). In fact, its constant presence in melanoma indicates that it may prove to be an independently reliable method for diagnosing amelanotic forms. It is also sporadically present within a variety of mesenchymal lesions including lipoma, liposarcoma, synovial chondromatosis, chondrosarcoma, fibromatosis, histiocytosis X, and chordoma. Although S-100 protein is highly characteristic of neural crest-derived tumors, it is not restricted to them and, consequently, must be interpreted cautiously. It may prove helpful in select situations such as the distinction of (a) benign nerve sheath tumors from other benign mesenchymal tumors such as fibrous histiocytomas, (b) cellular neurilemomas from malignant schwannomas, (c) malignant schwannomas from conventional fibrosarcoma (d) malignant melanomas from many carcinomas, and, possibly (e) juvenile xanthogranulomas from histiocytosis X.

An immunoperoxidase study of S-100 protein distribution in normal and neoplastic tissues.

The presence of S-100 protein was immunohistochemically studied in many types of formalin-fixed and paraffin-embedded tumors (260 cases). Peripheral nerve tumors, i.e., schwannomas, neurofibromas, granular cell tumors, and neurogenic sarcomas were demonstrated to contain variable amounts of S-100 protein in the tumor cell cytoplasm and nuclei. In ganglioneuromas and ganglioneuroblastomas, neoplastic Schwann cells or satellite cells were positive for S-100 protein. About one-half of the cases of carcinoid tumors stained weakly for S-100 protein. In addition to these nervous tissue and carcinoid tumors, chondrosarcoma, chordomas, pleomorphic adenomas of the salivary gland, and Langerhans cell granulomatosis were also shown to produce S-100 protein. In many types of breast tumors and other lesions, S-100 protein positive cells were likely to correspond to the distribution of myoepithelial cells. These results indicate that S-100 protein is not strictly specific to nervous tissue and its tumors; however, the immunohistochemical demonstration of S-100 protein can be a useful diagnostic tool in tumor diagnosis.