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Biomedical subjects

D D Bigner

Publications and source records attributed to D D Bigner.

At least 271 records · Page 15Linked to original sources
PubMed1985

Antigenic heterogeneity of human anaplastic gliomas and glioma-derived cell lines defined by monoclonal antibodies.

The antigenic heterogeneity of human neuroectodermal tumors defined by both murine and human monoclonal antibodies (MAs) is reported; no patterns of reactivity defining degree of anaplasia, in vitro morphology, or immunogen used were apparent. We investigated the reactivity of 20 distinct murine MAs defining markers of glioma-associated or predominantly lymphoid distribution for 13 human glioma-derived (HGL) cell lines and frozen sections of 19 human glioblastoma multiforme (GBM) and six astrocytomas (AST). Methods included radioimmunoassay, immunofluorescence, immunohistochemistry, and absorption analysis. Two markers, HLA-A,B and human Thy-1, exhibited no deviation; all HGL cell lines tested bound high levels of specific MA. Individual HGL cell line reactivity with the MA panel ranged from 30 to 70%. HGL cell lines (7/13) which reacted with greater than or equal to 50% of the antiglioma MAs had the highest (30-70%) positive reactivity rates with the anti-lymphoid marker MA panel; complex antigenicity in one system correlated with multiple antigens in the other. Within the anti-lymphoid marker MA panel, subpopulations of 4/13 HGL cell lines were clearly positive for the HLA-DR (Ia) antigens; another 3/13 HGL cell lines were strongly positive for common acute lymphocytic leukemia antigen (CALLA). With the exception of Thymocyte 1 antigen (Thy-1), reactivity for early and mature T-cell markers was infrequent and sporadic. Lymphoid marker expression by HGL cell lines is highly heterogeneous, ranging from few (Thy-1 and HLA-A,B) to complex expression of Ia, T-cell, and lymphoid tumor markers. GBM and AST tissues were antigenically less complex; for each of 6/8 anti-glioma MA, 70-100% of GBM and 66-100% of AST were positive. Two MAs were highly reactive (7/10, 8/9) with GBM sections and minimally so (1/6) with AST. Antigenic expression in gliomas is complex and heterogeneous; however, clear differences in lymphoid marker expression, the identification of widely and rarely expressed glioma-associated antigens, and the potential of immunologic differentiation between GBM and AST by large panels of MAs will serve to reduce the complexity and may be of potential diagnostic or prognostic significance.

Animals↗
PubMed1985

Immunolocalization of laminin in neoplasms of the central and peripheral nervous systems.

Laminin is a basement membrane glycoprotein that is expressed in vitro by immature and neoplastic astrocytes. The expression of laminin in vivo was examined immunohistochemically in normal adult brain and 90 neoplasms of the central and peripheral nervous systems. In normal adult brain, laminin was detected in the vasculature, arachnoid, pial-glial membrane, and choroid plexus. The vasculature in all 90 tumors demonstrated intense laminin immunoreactivity. Deposits of laminin were observed at the glioma-mesenchymal junction in several neoplasms, but never between or within neuroepithelial cells. The glial basement membrane often remained intact although surrounded on both sides by invasive glioma or medulloblastoma. However, there was always fragmentation and disruption of the glial membrane in adjacent fields. Laminin expression by tumor cells was observed in 10/10 schwannomas, 9/10 fibroblastic meningiomas, 3/19 nonfibroblastic meningiomas, and 3/6 mixed glioma-sarcomas. Laminin expression in the normal nervous system and in neuroepithelial neoplasms corresponds to regions of recognized basal lamina formation, including the junction between glial and mesenchymal elements. Although invasive gliomas are able to break down the pial-glial basement membrane and gain access to the perivascular or subarachnoid space, this membrane often remains intact late in the invasive process and may represent a partial barrier to tumor invasion. Laminin may be a useful marker for schwannomas, fibroblastic meningiomas, and vascular neoplasms of the nervous system.

Basement Membrane↗
PubMed1985

Establishment and characterization of the human medulloblastoma cell line and transplantable xenograft D283 Med.

A new continuous cell line and transplantable xenograft, D283 Med, was derived from the peritoneal implant and ascitic fluid of a child with metastatic medulloblastoma and grew in vitro in suspension culture with spontaneous macroscopic spheroid formation. The in vitro population doubling time was 52.55 hours. Mean colony forming efficiency in an agarose medium was 1.83 +/- 0.56%. The cell line, D283 Med, grew in athymic mice as serially transplantable intracranial and subcutaneous xenografts. Intracranial tumors grew as masses of small cells with scant cytoplasm and abundant mitotic figures and prominent anuclear zones resembling neuroblastic rosettes. Subcutaneous (SQ) tumors were markedly cellular neoplasms but did not contain rosettes. They expressed glutamine synthetase, neuron-specific enolase and neurofilament protein. Glial fibrillary acidic protein and S-100 protein were not detected. The SQ tumors grew to 500 mm3 with a latency of 52.55 +/- 12.5 days and a doubling time of 9.33 +/- 2.39 days. The stemline karyotypes of the peritoneal implant and ascitic fluid cells contained an extra copy of chromosome number 11 and three marker chromosomes (8q+, 17p+, 20q+). The cultured cell line and subcutaneous and intracranial xenografts retained the three marker chromosomes and differed from the original karyotype only in that they lacked the additional copy of chromosome number 11. This cell line and transplantable xenograft may allow further analysis of the biological properties and therapeutic sensitivity of human medulloblastoma.

Animals↗
PubMed1985

Effect of glutamate analogues on brain tumor cell lines.

Glutamate analogues have been used in many different experimental approaches in neurobiology. A small number of these analogues have been classified as gliotoxic. We have examined the effect of seven glutamate analogues (five gliotoxic and two neurotoxic) on the growth and viability of four human glioma cell lines, one human medulloblastoma cell line, and one human sarcoma cell line. Aminoadipic acid and homocysteic acid predominantly affected the growth of two glioma cell lines in the presence of 4 mM glutamine. Phosphonobutyric acid predominantly affected the other two glioma cell lines and the medulloblastoma cell line in the presence of 4 mM glutamine. In medium containing no glutamine, all three analogues had marked effects on all the cell lines except the sarcoma cell line. These effects were dose dependent. We postulate that these results can in part be explained on the basis of metabolic compartmentalization.

2-Aminoadipic Acid↗
PubMed1985

Applications of monoclonal antibodies in the diagnosis and treatment of primary brain tumors.

The development of monoclonal antibodies has resulted in marked expansion in understanding the central nervous system (CNS). This has been especially true in the study of human neuroectodermal tumors where monoclonal antibodies have been used as physiological probes to define and characterize human neuroectodermal tumor-associated antigens. Utilizing monoclonal antibodies, neuroectodermal tumor-associated antigens have been described in four broad categories; biochemically defined markers, shared nervous system-lymphoid cell markers, shared neuroectodermal-oncofetal markers, and putative restricted tumor markers. Preliminary data have demonstrated the ability to localize animal and human tumors in vitro, ex vivo, and in vivo. Early application of monoclonal antibody technology to neuroimmunology and neuro-oncology has resulted in a new awareness of the complex relationships that exist within the CNS. Their specificity and reproducibility may provide the means to qualitatively and quantitatively define the phenotypic heterogeneity of human neuroectodermal tumors. Potentially, monoclonal antibodies, alone or as carriers of radionuclides, drugs, or toxins, may allow successful diagnosis and treatment of human neuroectodermal tumors.

Animals↗
PubMed1985

Aspects of immunobiology and immunotherapy and uses of monoclonal antibodies and biologic immune modifiers in human gliomas.

Recent progress in brain tumor biology research has helped us to understand the causes of our failure to improve patient survival with current therapeutic approaches. Present combination regimens of surgery, radiotherapy, and chemotherapy fail to address adequately two inherent biologic properties of brain tumors: tumor-induced host immunosuppression and tumor cell heterogeneity. Future success in immunotherapy will depend on our ability to dissect the mechanisms involved in tumor-host interactions. Also, therapies may have to be individualized. The potential for MAs in research and clinical application is great owing to their unlimited discriminative power for molecular and functional characterization. Differential diagnosis of difficult cases, such as parenchymatous tumors or neoplastic cells in CSF, with panels of MAs will be available in the near future. The use of MAs to direct localization and therapy of brain tumors is also promising, as evidenced by the preliminary data from our laboratory and from those of other workers. Other applications of MAs, such as prevention of invasion and metastasis by targeting cell surface molecules involved in cell-matrix attachment function, are also under study. Future refinement in MA-specificity and in enhancement of MA-delivery to tumor sites will significantly complement new therapeutic approaches. As brain tumors evade immunosurveillance through active participation in inducing tumor-specific immunosuppression, successful immunotherapy, either passive serotherapy or active immunization, will be best achieved in patients with a slight or moderate immunosuppressive state. Alteration of immune status with various biologic response modifiers to boost host reactivity against tumors will be an important adjunct in our arsenal against brain tumors. IFNs, with their direct tumoricidal activity, and lymphokines (such as interleukin-2) may be such reagents with a promising future.

Antibodies, Monoclonal↗
PubMed1985

Influence of glutamine on the growth of human glioma and medulloblastoma in culture.

Cellular supply of glutamine, an essential substrate for growth, is derived from extracellular fluid and de novo synthesis. We investigated the relative importance of these sources to the growth of six human anaplastic glioma- and one human medulloblastoma-derived permanent cell lines. Exogenous glutamine was limiting for the proliferation of glioma-derived lines D-54 MG, U-118 MG, and U-251 MG. In contrast, medulloblastoma-derived line TE-671 and glioma-derived lines U-373 MG, D-245 MG, and D-259 MG grew in the absence of supplemental glutamine. Two cell lines with contrasting glutamine requirements, D-54 MG and TE-671, were used to explore the pharmacological interference with glutamine metabolism. DL-alpha-Aminoadipic acid, a reported glutamic acid analogue with gliotoxic properties, significantly inhibited the growth of both lines. These effects were reversed by increasing glutamine, suggesting that the major action of DL-alpha-aminoadipic acid is as a glutamine antagonist. In contrast, the glutamine synthetase inhibitor delta-hydroxylysine demonstrated activity only against TE-671. Acivicin and 6-diazo-5-oxo-L-norleucine, glutamine analogues available for clinical use, reduced the proliferation of both cell lines at pharmacological concentrations. Methionine sulfoximine, a glutamine synthetase inhibitor previously used clinically, produced marked growth inhibition only against TE-671. These findings indicate that the synthesis and utilization of glutamine are potentially exploitable targets for the chemotherapy of some human gliomas and medulloblastomas.

2-Aminoadipic Acid↗
PubMed1985

Combination chemotherapy in vitro exploiting glutamine metabolism of human glioma and medulloblastoma.

The human glioma-derived cell line D-54 MG and the human medulloblastoma-derived cell line TE-671 have been shown to be sensitive in culture to the pharmacological interference with glutamine metabolism by acivicin, 6-diazo-5-oxo-L-norleucine, and methionine sulfoximine. Using as a guide the multiple contributions of glutamine to the biosynthesis of proteins, purines, and pyrimidines, we now have identified six additional antimetabolites active against these lines in vitro at clinically relevant concentrations. The 50% growth-inhibitory levels of the drugs against D-54 MG in 6-day continuous exposure experiments were: L-asparaginase, 0.057 IU/ml; 5-fluorouracil, 0.5 micrograms/ml; 6-mercaptopurine, 0.8 micrograms/ml; actinomycin D, 0.0007 micrograms/ml; N-phosphonacetyl-L-aspartic acid, 2.3 micrograms/ml; and 5-azacytidine, 0.2 micrograms/ml (3-day exposure. The corresponding 50% growth-inhibitory values in TE-671 were: L-asparaginase, 0.54 IU/ml; 5-fluorouracil, 1.5 micrograms/ml; 6-mercaptopurine, 4.7 micrograms/ml; actinomycin D, 0.00044 micrograms/ml; N-phosphonacetyl-L-aspartic acid, 4.5 micrograms/ml; and 5-azacytidine, 0.49 micrograms/ml. Dipyridamole up to 10 micrograms/ml was inactive against both lines. The isobologram method was used to evaluate the effectiveness of several two-drug combinations which were biochemically designed. The sums of the optimal fractional inhibitory concentrations for the pairs were: acivicin plus L-asparaginase, 0.14; acivicin plus methionine sulfoximine, 0.40; 6-diazo-5-oxo-L-norleucine plus methionine sulfoximine, 0.60; acivicin plus 6-mercaptopurine, 1.0, all in TE-671; and acivicin plus 5-fluorouracil, 0.79, in D-54 MG. Our findings suggest that an antimetabolite regimen exploiting glutamine sensitivity might improve the chemotherapy of some human gliomas and medulloblastomas.

Antimetabolites, Antineoplastic↗
PubMed1985

Comparison of intravenous versus intracarotid therapy with 1,3-bis(2-chloroethyl)-1-nitrosourea in a rat brain tumor model.

Currently numerous clinical trials are in progress utilizing intracarotid (i.c.) 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) for the treatment of malignant gliomas based upon the proposed focal nature of these tumors and the assumption that the i.c. route delivers higher levels of drug to the tumor. To date, however, increased efficacy in an animal model has not been clearly demonstrated for the i.c. delivery of BCNU. We have evaluated the dose-response curve for the i.v. and i.c. administration of BCNU in a commonly utilized experimental brain tumor model, the 9L rat gliosarcoma. An initial toxicity trial utilizing the i.p. 10% lethal dose (LD10) of BCNU by the i.v. and i.c. routes failed to demonstrate any significance in toxicity between the two routes. Tumor-bearing animals were then treated on Day 15-16 after tumor inoculations with 1, 10, 25, 50, 75, and 100% of the LD10 dose by either the i.v. or the i.c. route. Both i.v. and i.c. BCNU gave maximum survival increases at 75-100% LD10 doses, and there was no therapeutic advantage seen from i.c. delivery. However, at 50% of the LD10 dose (6.65 mg/kg), triplicate experiments demonstrated that the i.c. but not the i.v. dose maintained maximum efficacy equivalent to 100% of the LD10 given either i.v. or i.c. When the dose was reduced to 25% of the LD10 dose (3.33 mg/kg), two of three experiments showed efficacy of the i.c. delivery of this lower drug dosage to be equivalent to 100% of the LD10 given i.v. or i.c. The i.v. dosage resulted in a significant reduction in survival in all three trials. At 10% of the LD10 dose (1.30 mg/kg), neither the i.v. nor the i.c. administration retained equivalent efficacy to 100% of the LD10. However, in one of two trials, the i.c. groups had statistically better survival than controls, while in neither experiment was any advantage over controls seen in the i.v. treated groups. At 1% of the LD10 dose, neither the i.v. nor the i.c. route demonstrated any therapeutic efficacy. From our data, the advantage of the i.c. delivery of BCNU in the intracranial 9L rat gliosarcoma appears to be in the fact that significantly lower dosages than those given i.v. may be utilized to achieve equivalent survival with potentially less systemic toxicity.

Animals↗
PubMed1984

In vivo measurement of regional brain tissue pH using positron emission tomography.

Carbon-11-labeled dimethyloxazolidinedione ([11C]DMO) was injected intravenously into human subjects, and serial positron emission tomographic (PET) scans were obtained until brain-blood equilibration was achieved or could be accurately predicted from dynamic PET and 11C blood data. Knowledge of regional brain-blood partition coefficients for DMO, together with measurements of arterial blood hematocrit and pH, permitted the calculation of regional brain tissue and tumor pH (rpH). [11C]DMO PET rpH values were similar to rpH values derived from quantitative autoradiographic measurements of [14C]DMO concentrations in rat brain slices.

Brain↗
PubMed1984

Characterization of three restricted specificity monoclonal antibodies raised against the human glioma cell line D-54 MG.

Monoclonal antibodies ( MCAs ) have been derived from a fusion of P3-NS1/1-Ag 4-1 (NS1) myeloma cells and splenocytes immunized to human glioma cell line D-54 MG. MCAs 2F3 , 4C7 , and 5B7 were analyzed by cell surface radioimmunoassay (CS-RIA), quantitative absorption, indirect immunofluorescence, and peroxidase-anti-peroxidase (PAP) immunohistology of unfixed tissue samples. MCA 2F3 exhibits the most highly restricted pattern of reactivity we have observed, reacting only with 5/12 glioblastoma cell lines and 1/4 fetal skin lines by CS-RIA, and to 9/11 glioblastoma tissue samples by PAP and absorption analysis; this MCA is totally nonreactive with melanomas, neuroblastomas, meningiomas, and control non-central nervous system tumors, and to adult and fetal tissues including brain, thymus, spleen, liver, lung, heart, gut, skin, and muscle by PAP analysis. MCAs 4C7 and 5B7 demonstrate neuroectodermal tumor cross-reactivity profiles, reacting with either melanomas ( 5B7 ) or melanomas and neuroblastomas ( 4C7 ); both are reactive with fetal skin, brain, and thymus of less than or equal to 16 weeks of gestational age. Other than this latter fetal antigen reactivity, these MCAs share the same negative reactivity profile described above for MCA 2F3 . Data from experiments using control or 0.02% EDTA-treated confluent cell monolayers of D-54 MG as antibody absorbents showed that the antigens detected are present in the extracellular matrix material remaining following cell removal. The data presented here establish that these highly restrictive anti-human glioma cell line MCAs are expressed in primary human gliomas; that the markers defined are developmental in nature, in that they are expressed by human fetal tissue, but not by adult tissue; and that in conjunction with previously characterized specificities, these markers of antigenic heterogeneity will be valuable in model system studies of therapeutic response heterogeneity.

Absorption↗
PubMed1984

Iodine-125 interstitial irradiation of virally induced dog brain tumours.

Brain tumours were induced by intracerebral inoculation of beagle puppies with Avian sarcoma virus (ASV) suspension. The inoculation produced solitary or multiple sarcomas in three dogs and intraventricular anaplastic gliomas in four dogs. At the time of the first tumour positive CT-control Iodine-125 seeds (activity 8.5-10.5 mCi) were placed into the lesion. The radioactive Iodine-125 implants produced sharply delineated calcifying necroses with vital unaffected tumour outside the necrosis. The necrotizing and calcifying effect was apparent after 18 days and complete after 97 days when a transitional zone was no longer detectable. The response of neoplastic tissue to low dose rate interstitial irradiation was distinctly different in terms of the volume of necrosis when compared with Iodine-125 radionecroses in healthy brain tissue.

Animals↗
PubMed1984

Carotid artery injections in 40- to 99-g Fischer rats: technical note and evaluation of blood flow by various injection techniques.

Despite extensive clinical use of brain tumor chemotherapy via the internal carotid artery (ICA), demonstration of efficacy and toxicity screening of ICA chemotherapy in brain tumor models has been limited. A method for performing ICA injections in 40- to 99-g Fischer rats is described, with documentation of its effect upon cerebral blood flow. A 33 gauge cannula was secured into a PE-10 catheter and, after ligation of the external carotid (ECA) and the pterygopalatine arteries, injections were made into the common carotid artery (CCA). The reliability of this method compared to CCA injection with and without ligation of the ECA was evaluated utilizing 15-micrometers 103Ru microspheres. With this technique, 68.3 +/- 11.19% of the microspheres lodged in the brain, compared to 28.9 +/- 21.3% to 32.7 +/- 19.8% for the other techniques (P less than 0.01). With this technique, regional perfusion of brain tumors can be done in the avian sarcoma virus rat glioma model, the D-54 MG human glioma-immunosuppressed rat transplantation model, and any of the large rat brain tumor models. The relevance of this method of experimental regional perfusion for the preclinical assessment of the efficacy and of the toxicity of chemotherapy and immunotherapy via the ICA is discussed.

Animals↗
PubMed1984

Intracerebral transplantation of a human glioma line in immunosuppressed rats.

A model was developed for in vivo study of the human glioma-derived D-54 MG cell line in the brains of immunosuppressed Fischer 344 rats. The rats were injected with horse anti-rat thymocyte serum before and after intracerebral inoculation with 5 or 10 microliters of a D-54 MG tumor cell suspension. Reproducible mortality distributions were obtained, with deaths occurring 18 to 34 days after intracerebral inoculation. Tumors grew as well circumscribed intracerebral masses with sheets of anaplastic cells, areas of necrosis bordered by concentrated nuclei, and minimal lymphocytic infiltration. Cytogenetic analysis revealed the same general chromosome distribution and markers in the heterotransplanted glioma cells as in the cultured line. Blood-brain barrier disruption was demonstrated by intracerebral tumor staining after intravenous injection of Evans blue dye. The in vivo growth of D-54 MG in immunosuppressed rats provides a reliable experimental model for the study of chemotherapy, immunodiagnosis, and immunotherapy of a human glioma-derived tumor in an animal sufficiently large to evaluate intracarotid or intratumoral injection of therapeutic agents.

Animals↗
PubMed1984

Blood-brain barrier disruption in immature Fischer 344 rats.

Methods for transiently disrupting the blood-brain barrier (BBB) which are consistent with survival are described for immature Fischer 344 rats weighing 40 to 99 gm. A catheter was retrogradely inserted into the external carotid artery to the level of the bifurcation. Perfusion of 1.4 M mannitol or 1.6 M arabinose, at a rate of 0.01 to 0.1 ml/sec for 30 seconds, resulted in transient BBB disruption as measured by Evans blue dye (EBD) staining. Higher flow rates or perfusion with 10% to 30% dimethyl sulfoxide were associated with a mortality rate ranging from 0% to 44%. Perfusion with 0.9% sodium chloride or intrafemoral artery perfusion with 1.4 M mannitol did not disrupt the BBB. Optimum BBB disruption as measured by EBD staining was achieved with 1.6 M arabinose at 0.026 ml/sec for 30 seconds, at which time all of the 42 experimental animals had BBB disruption; all of the animals so treated survived 2 weeks following perfusion. This technique will allow the efficacy of delivering chemotherapeutic agents following BBB disruption to be tested in several of the more commonly used small-animal models for brain-tumor research.

Animals↗
PubMed1984

Comparison of various methods for delivering radiolabeled monoclonal antibody to normal rat brain.

Different methods were evaluated for delivering iodine-125 monoclonal antibodies (Mab's) to the central nervous system in 40- to 99-gm Fischer rats. By evaluating interhemispheric, interregional, and brain:blood ratios of Mab's, the efficacy of intracarotid (IC) or intravenous (IV) administration of Mab's with and without prior IC perfusion with 0.9% NaCl (normal saline, NS), 1.4 M mannitol, or 1.6 M arabinose, or of femoral artery perfusion with 1.4 M mannitol was evaluated. No difference was seen between IC and IV administration of Mab's with or without prior perfusion. Intracarotid perfusion with hyperosmolar agents was required to disrupt the blood-brain barrier (BBB) and to significantly elevate brain levels of Mab's. The brain and blood levels of Mab's were elevated in all regions of the brain following hyperosmolar BBB disruption. However, the levels were significantly higher in the ipsilateral hemisphere, with cross-over occurring primarily in the vascular distribution of the contralateral anterior cerebral artery. Intracarotid hyperosmolar perfusion produced 450% to 500% increases in ipsilateral and 240% to 280% increases in contralateral hemispheric brain:blood Mab ratio levels compared to those achieved with NS perfusion. For IC perfusion of mannitol or arabinose, flow rates ranging from 0.017 to 0.052 ml/sec were equally effective in disrupting the BBB. Insignificant morbidity and mortality rates were noted up to 2 weeks following BBB disruption. Additional ligation of major extracranial branches of the external and internal carotid arteries prior to IC perfusion did not result in a selective increase in hemispheric Mab levels. Temporally, following hyperosmolar BBB disruption, brain:blood Mab ratios remained elevated bilaterally at 7 days after Mab delivery, with the ipsilateral hemispheric levels remaining significantly elevated compared with the contralateral hemispheric levels until Day 5, when the ratio returned to the nonperfused range. Catheterization was required in the small animals and was performed under magnification in 10 to 20 minutes, with less than an 8% overall morbidity and mortality. The methodology developed should prove helpful in delivery of Mab's or other agents in rat tumor models and experimental models for other disease entities.

Animals↗