myelin-basic-protein and Oligodendroglioma

Topics: Adolescent; Adult; Brain Neoplasms; Cell Nucleus; Child; Child, Preschool; Cytoplasm; Diagnosis, Differential; Humans; Immunohistochemistry; Male; Microscopy, Electron; Middle Aged; Myelin Basic Protein; Myelin Proteins; Myelin-Associated Glycoprotein; Oligodendroglioma; Phosphopyruvate Hydratase; S100 Proteins; Vimentin

The classification of human gliomas is currently based solely on neuropathological criteria. Prognostic and therapeutic parameters are dependent upon whether the tumors are deemed to be of astrocytic or oligodendroglial in origin. We sought to identify molecular reagents that might provide a more objective parameter to assist in the classification of these tumors. In order to identify mRNA transcripts for genes normally transcribed exclusively by oligodendrocytes. Northern blot analysis was carried out on RNA samples from 138 human gliomas. Transcripts encoding the myelin basic protein (MBP) were found in an equally high percentage of tumors that by neuropathological criteria were either astrocytic or oligodendroglial. In contrast, proteolipid protein (PLP) and cyclic nucleotide phosphodiesterase (CNP) mRNA molecules were found significantly more often in oligodendrogliomas than in astrocytomas. The strongest association with histological typing was found with the transcript for the myelin galactolipid biosynthetic enzyme UDP-galactose: ceramide galactosytransferase (CGT), which was about twice as frequently detected in tumors of oligodendroglial type. Results of glycolipid analyses were previously reported on a subset of the tumors studied herein. Statistical analyses of both molecular and biochemical data on this subset of astrocytomas, oligoastrocytomas, and oligodendrogliomas were performed to determine if a panel of markers could be used to separate astrocytic and oligodendroglial tumors. The presence of asialo GM1 (GA1) and the absence of paragloboside occurred most frequently in oligodendrogliomas. Ceramide monohexoside (CMH) levels correlated highly with the expression of mRNA for 4 myelin proteins: CGT, MBP, CNP, and PLP. The best combination of 2 markers of oligodendroglial tumors was CGT and GA1; the best combination of 3 markers was the presence of CGT, GA1, and the absence of paragloboside. We conclude that this combination of markers could be useful in distinguishing between astrocytic and oligodendroglial tumors.

Topics: 3',5'-Cyclic-AMP Phosphodiesterases; Astrocytoma; Biomarkers, Tumor; Brain Neoplasms; Cerebrosides; G(M1) Ganglioside; Galactosyltransferases; Glycolipids; Humans; Myelin Basic Protein; Myelin Proteolipid Protein; N-Acylsphingosine Galactosyltransferase; Oligodendroglioma; RNA; Survival Rate

Developments in transgenic technology have greatly enhanced our ability to understand the functions of various genes in animal models and relevant human diseases. The tetracycline (tet)-regulated transactivation system for inducing gene expression allowed us to control the expression of exogenous genes in a temporal and quantitative way. The ability to manipulate a cell-specific promoter enabled us to express one particular protein in a single type of cell. The combination of a tetracycline system and a tissue-specific promoter has led us to the development of an innovative gene expression system, which is able to express genes in a cell type-specific and time- and level-controllable fashion. An oligodendrocyte-specific myelin basic protein (MBP) gene promoter controls the reversed tet-inducible transactivator. The green fluorescent protein (GFP) gene was placed under the control of the human cytomegalovirus (CMV) basic promoter in tandem with seven tet-responsive elements (TRE), binding sites for the activated transactivator. Upon the addition of doxycycline (DOX, a tetracycline derivative), tet transactivators became activated and bound to one or more TRE, leading to the activation of the CMV promoter and the expression of GFP in oligodendrocytes. We have successfully expressed GFP and luciferase at high levels in oligodendrocytes in a time- and dose-dependent fashion. In the absence of DOX, there was almost no GFP expression in oligodendroglial cultures. Graded levels of GFP expression were observed after induction with DOX (0.5 to 12.5 microg/ml). Our data indicate that this inducible gene expression system is useful for the study of gene function in vivo and for the development of transgenic animal models relevant to human diseases such as multiple sclerosis.

Topics: Animals; Anti-Bacterial Agents; Brain Neoplasms; Dose-Response Relationship, Drug; Doxycycline; Gene Expression Regulation, Neoplastic; Green Fluorescent Proteins; Humans; Kinetics; Luminescent Proteins; Myelin Basic Protein; Oligodendroglia; Oligodendroglioma; Promoter Regions, Genetic; Rats; Recombinant Proteins; Time Factors; Transcription, Genetic; Transfection

We examined the expression of glial- and neuronal-specific mRNAs within human gliomas using in situ hybridization. We found that low-grade astrocytomas contained a high number of proteolipid protein (PLP) mRNA-positive cells and that the number of PLP-stained cells decreased markedly with increasing tumor grade. Interestingly, the ratio of PLP mRNA-stained cells:myelin basic protein (MBP) mRNA-stained cells in normal white matter and low-grade astrocytoma was about 2:1 but approached 1:1 with increasing tumor grade. This parameter appeared to be a good indicator of tumor infiltration in astrocytomas, so we tested this in the analysis of other gliomas. Unlike astrocytomas, oligodendrogliomas were found consistently to contain few PLP mRNA- or MBP mRNA-expressing cells. In contrast, gemistocytic astrocytomas, typically highly invasive tumors, contained high numbers of PLP-positive cells and a ratio of PLP mRNA:MBP mRNA-stained cells of about 1.5:1, similar to low-grade astrocytomas. Nonradioactive in situ hybridization also enabled the morphological identification of specific cells. For example, gemistocytic astrocytes, which were found to be strongly vimentin mRNA positive, contained little glial fibrillary acidic protein mRNA and did not stain for PLP or MBP mRNAs. Neuronal mRNAs, such as neurofilament 68, were observed in small numbers of entrapped neurons within gliomas but were uninformative with respect to predicting tumor grade. Our results suggest that oligodendrocytes survive low-grade tumor infiltration and that glial tumor cells, unlike cell lines derived from them, do not express oligodendrocyte or neuronal mRNAs. In addition, the expression of mRNAs for the two major myelin protein genes, PLP and MBP, could be used to predict the grade and extent of tumor infiltration in astrocytomas.

Topics: Astrocytoma; Brain Neoplasms; Gene Expression Regulation, Neoplastic; Glial Fibrillary Acidic Protein; Glioma; Humans; In Situ Hybridization; Keratins; Myelin Basic Protein; Myelin Proteolipid Protein; Neurofilament Proteins; Neuroglia; Neurons; Oligodendroglia; Oligodendroglioma; RNA, Messenger; RNA, Neoplasm; Vimentin

Pathological differentiation of oligodendroglioma and mixed oligoastrocytoma from astrocytoma is difficult, relying on morphological characteristics due to the lack of reliable immunohistochemical stains. Oligodendrocytes, the presumed cell of origin of oligodendrogliomas, highly express the genes encoding myelin basic protein (MBP) and proteolipid protein (PLP). We analyzed the expression of these genes to determine whether they might be useful molecular markers of oligodendrocytic tumors. MBP and PLP were highly expressed in all oligodendrogliomas and minimally expressed in glioblastomas multiforme. MBP was highly expressed in mixed oligoastrocytomas, whereas PLP expression was minimal. The association between tumor classification and expression of the MBP and PLP genes was statistically significant. Expression of these genes may serve as a useful molecular marker for some subtypes of human gliomas.

Topics: Adult; Aged; Biomarkers, Tumor; Brain Neoplasms; Female; Gene Expression Regulation, Neoplastic; Glioblastoma; Glioma; Humans; Male; Middle Aged; Myelin Basic Protein; Myelin Proteolipid Protein; Oligodendroglia; Oligodendroglioma; Reverse Transcriptase Polymerase Chain Reaction; Survival Analysis; Time Factors

Two human Golli (for gene expressed in the oligodendrocyte lineage)-MBP (for myelin basic protein) cDNAs have been isolated from a human oligodendroglioma cell line. Analysis of these cDNAs has enabled us to determine the entire structure of the human Golli-MBP gene. The Golli-MBP gene, which encompasses the MBP transcription unit, is approximately 179 kb in length and consists of 10 exons, seven of which constitute the MBP gene. The human Golli-MBP gene contains two transcription start sites, each of which gives rise to a family of alternatively spliced transcripts. At least two Golli-MBP transcripts, containing the first three exons of the gene and one or more MBP exons, are produced from the first transcription start site. The second family of transcripts contains only MBP exons and produces the well-known MBPs. In humans, RNA blot analysis revealed that Golli-MBP transcripts were expressed in fetal thymus, spleen, and human B-cell and macrophage cell lines, as well as in fetal spinal cord. These findings clearly link the expression of exons encoding the autoimmunogen/encephalitogen MBP in the central nervous system to cells and tissues of the immune system through normal expression of the Golli-MBP gene. They also establish that this genetic locus, which includes the MBP gene, is conserved among species, providing further evidence that the MBP transcription unit is an integral part of the Golli transcription unit and suggest that this structural arrangement is important for the genetic function and/or regulation of these genes.

Topics: Alternative Splicing; Amino Acid Sequence; Animals; Base Sequence; Chromosome Mapping; Exons; Genes; Humans; Mice; Molecular Sequence Data; Myelin Basic Protein; Oligodendroglioma; RNA, Neoplasm; Sequence Alignment; Sequence Homology, Amino Acid; Tissue Distribution; Transcription, Genetic

A 30-year-old man suffered for a year of a typical syndrome of cerebellar tumor. At suboccipital craniectomy a soft tumor infiltrating both hemispheres and vermis, filling up part of the IV ventricle was found. After subtotal removal of the neoplasm the postoperative course was poor and the patient died 5 weeks later. Biopsy material consisted of three types of tissue: 1. large nests of carrot-shaped, hyperchromatic cells, 2. fields of "halo" cells presenting myelin basic protein (MBP) immunoreactivity and 3. fields and scattered strongly GFAP-positive cells. The histological and immunocytochemical pattern of the neoplasm indicates differentiation of the tumor into oligodendrogliomatous and astrocytomatous line being an uncommon example of dual glial differentiation capability in medulloblastoma.

Topics: Adult; Astrocytoma; Cell Differentiation; Cerebellar Neoplasms; Fatal Outcome; Glial Fibrillary Acidic Protein; Humans; Male; Medulloblastoma; Myelin Basic Protein; Neoplasm Invasiveness; Oligodendroglioma

Four mixed oligodendrogliomas and astrocytomas were investigated by electron microscopy and immunohistochemistry (GFAP, NSE and MBP). GFAP-positive oligodendroglioma cells and their transitional cells to GFAP-negative oligodendroglioma cells were present, suggesting successive morphological changes of astrocytic tumor cells. NSE-positive cells, suggestive of residual neurons, also exhibited round nuclei and perinuclear halos. On electron microscopy, oligodendroglioma cells that showed glial filaments, vascular end-feet and zonulae adherentes were occasionally present. The tumor cells with or without astrocytic characteristics showed common features of cytoplasmic organelles. These findings suggest that most oligodendroglioma cells in mixed gliomas are of an astrocytic nature and that characteristic microscopic features of oligodendroglioma are of a common cellular form that can be taken by various types of cells under certain circumstances.

Topics: Astrocytoma; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; Microscopy, Electron; Myelin Basic Protein; Oligodendroglioma; Phosphopyruvate Hydratase

The present study was undertaken to evaluate the utility of pathologic features and specific immunohistochemical studies in estimating the prognosis of oligodendroglioma. The pathological diagnosis of an oligodendroglioma was made on HE stained-sections according to WHO classification. Sixteen oligodendrogliomas, twelve mixed oligoastrocytomas and ten anaplastic oligodendrogliomas were immunotested by the peroxidase-antiperoxidase (PAP) method with anti-GFAP serum, anti-S-100 serum and anti-MBP (Myelin basic protein) serum and by the avidin biotin peroxidase-complex (ABC) method with anti-vimentin serum and ant-Leu 7 monoclonal antibody. GFAP positive cells were interpreted as reactive astrocytes, neoplastic astrocytes and neoplastic oligodendrocytes, S-100 positive cells were interpreted as reactive astrocytes and neoplastic astrocytes. Leu 7 positive cells were found in only one case of anaplastic oligodendroglioma. Anti-Leu 7 could not be considered as a specific marker for oligodendroglioma. Of the anaplastic oligodendroglioma 60% displayed MBP positively and 70% displayed vimentin positively. NSE positive cells were found in a few anaplastic oligodendrogliomas. The present study has not so far uncovered any marker that is restricted to oligodendrogliomas. However GFAP may be useful to assess the extent of reactive astrocytes and neoplastic astrocytes in the oligodendroglioma or mixed oligoastrocytoma. MBP and vimentin will help to determine the malignancy of oligodendroglioma.

Topics: Adult; Aged; Biomarkers, Tumor; Brain Neoplasms; Child; Female; Glial Fibrillary Acidic Protein; Glioma; Humans; Immunohistochemistry; Male; Middle Aged; Myelin Basic Protein; Oligodendroglioma; Phosphopyruvate Hydratase; Prognosis; S100 Proteins; Vimentin

To search for a marker for oligodendroglioma, immunohistochemical analysis of myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) were performed on tissues from 38 oligodendrogliomas. MBP was detected in 14 such tumors, positive cells being characterized by having rather rich cytoplasm containing a large nucleus and a prominent nucleolus. They seemed to be relatively poorly differentiated cells corresponding to oligodendroglioma grade II in the WHO classification, and appeared to have some cytological resemblance to immature oligodendrocytes in the newborn brain. Mature grade I tumor cells with a small dark nucleus and a perinuclear halo, and grade III anaplastic cells were MBP-negative. GFAP-positive cells were also found in tumors containing MBP-positive cells. A double immunostain for MBP and GFAP demonstrated that some tumor cells were simultaneously stained with both. Presumably the oligodendroglioma cells that showed such dual immunostaining would intrinsically express a myelin-forming glial phenotype. MBP may be applicable as a marker for oligodendrogliomas.

Topics: Biomarkers, Tumor; Brain Neoplasms; Glial Fibrillary Acidic Protein; Humans; Immune Sera; Immunohistochemistry; Myelin Basic Protein; Oligodendroglioma

The histologic feature of oligodendroglioma is constituted by the monotonously arranged sheets of tumor cells intersected with delicate vascular stroma. The shape of the tumor cells is usually uniform and occasionally rather pleomorphic. In order to clarify biological characteristics of the tumor cells at level of cell differentiation, immunohistochemical stains for myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) with PAP method were performed on 13 oligodendrogliomas obtained by surgical removal. The cells stained positive with MBP immunostain were encountered in 5 tumors and their incidence was ranging from 1% to more than 30% of all tumor cells. The MBP-positive cells were characterized by having a large clear nucleus with a prominent nucleolus and relatively abundant cytoplasm, that seemed to mimic myelin-forming glia appearing in normal developing brain. The cells with poor cytoplasm or surrounded by a perinuclear halo were MBP-negative, and the pleomorphic cells in majority of anaplastic oligodendrogliomas were also negative. On the other hand, with GFAP stain the positive cells were observed in 8 tumors. It is still controversial to interpret the occurrence of GFAP-positive cells in oligodendrogliomas. In this study the GFAP-positive cells were morphologically identical to the MBP-positive cells, so that both MBP and GFAP are supposed to exist in the same tumor cells. It is assumed that some of oligodendroglioma cells which showed such immunostainabilities could possess an intrinsic character to express a phenotype of myelin-forming glia. Possibly, MBP substance is applicable as one of immunohistochemical markers for oligodendroglioma cells to indicate a certain cell differentiation.

Topics: Adult; Aged; Brain Neoplasms; Female; Glial Fibrillary Acidic Protein; Humans; Immunologic Techniques; In Vitro Techniques; Male; Middle Aged; Myelin Basic Protein; Neoplasm Proteins; Oligodendroglioma; Staining and Labeling

Twenty-eight oligodendrogliomas and seven oligoastrocytomas were immunotested by the peroxidase-antiperoxidase (PAP) method with antiglial fibrillary acidic protein (GFAP) serum, anti-Leu 7 monoclonal antibody (Mab), anti-myelin-associated glycoprotein (MAG) Mab, anti-myelin basic protein (MBP) serum, anti-carbonic anhydrase C (CA C) serum and anti-neuron-specific enolase (NSE) serum. The immunoreactivity of their vascular pattern was studied with Ulex europaeus type I lectin (UEA I). According to their morphology and distribution GFAP-positive cells were respectively interpreted as reactive astrocytes, neoplastic astrocytes and neoplastic oligodendrocytes. Reactive astrocytes were found in the tumor, around the tumor and surrounding the supporting blood vessels. Neoplastic astrocytes were mainly found in the oligoastrocytomas and usually closely intermingled with neoplastic oligodendrocytes. GFAP-positive neoplastic oligodendrocytes were found in the typical oligodendrogliomatous areas. They had central nuclei and GFA positivity was mainly found in the perinuclear cytoplasm. They correspond to the "gliofibrillary oligodendrocytes" described by Herpers and Budka. Of the oligodendrogliomas 91% displayed Leu 7 positivity, but anti-Leu 7 cannot be considered as a specific marker for oligodendrogliomas since other neuroepithelial tumors have been reported to react with this antibody. MAG-, CA C- and NSE-positivities were found in a number of tumor cells in a few oligodendrogliomas. All the tumor cells were MBP-negative, but myelin sheaths and fragments of myelin in the infiltrated white matter were clearly demonstrated by this antiserum. UEA I strikingly demonstrated the vascular pattern of the tumors, and its usefulness as a discriminating marker for the supportive endothelial cells was confirmed.

Topics: Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Carbonic Anhydrases; Epitopes; Glial Fibrillary Acidic Protein; Histocytochemistry; Humans; Immunochemistry; Lectins; Myelin Basic Protein; Myelin Proteins; Myelin-Associated Glycoprotein; Oligodendroglioma; Phosphopyruvate Hydratase; Plant Lectins