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Biomedical subjects

A S Beutler

Publications and source records attributed to A S Beutler.

8 recordsLinked to original sources
PubMed1999

Tumor gene therapy made easy: allogeneic major histocompatibility complex in the C6 rat glioma model.

The C6 glioma in the immune-competent rat is a frequently used model in brain tumor gene therapy research. It displays the histologic hallmarks of the human glioblastoma and has been employed to demonstrate new mechanisms of anti-tumor immunity and therapeutic strategies. We noted that C6 tumors regressed spontaneously in three of five animals and that protective anti-tumor immunity ensued without therapeutic intervention. A review of the literature revealed that different rat strains are used as "syngeneic" host for the C6 cell glioma, namely, BDIX, BDX, Sprague-Dawley, and Wistar. Allelotyping of the RT1.A (rat MHC I homolog) by a serologic technique and of the RT1.B (rat MHC II homolog) by a newly developed molecular technique showed that C6 cells express the haplotype RT1u and are allogeneic in the preceding rat strains. Expression of the gene encoding the transactivator CIITA in C6 gliomas using an EBV-based transduction system led to induction of MHC I and II and thereby mimicked therapeutic responses that could not operate in syngeneic models. These data suggest that the C6 glioma model in the immune-competent rat should no longer be used to study gene therapy strategies, that the available data obtained in this model need to be critically reinterpreted, and that findings obtained in the C6 glioma model may not be sufficient to support a clinical trial in glioblastoma patients.

Animals↗
PubMed1996

Myogenic motor-evoked potential monitoring using partial neuromuscular blockade in surgery of the spine.

STUDY DESIGN: The authors analyzed motor-evoked potentials using transcranial electrical cortical stimulation during spinal surgery in 40 patients under conditions of partial neuromuscular blockade. OBJECTIVES: The results were used to investigate the utility of motor-evoked potential monitoring to prevent neurologic injury in spinal surgery. SUMMARY OF BACKGROUND DATA: Noninvasive transcranial electrical motor-evoked potentials are reportedly effective in predicting postoperative spinal cord deficits caused by intraoperative occurrences. However, the sensitivity and specificity of these predictions have not been assessed under conditions of partial neuromuscular blockade during a wide variety of surgical procedures. METHODS: Compound muscle action potentials were recorded at the tibialis anterior muscle while general anesthesia was maintained with nitrous oxide, etomidate, and sufentanil. Surgery was performed at all spinal levels for a variety of diagnoses. RESULTS: Reference motor-evoked potential amplitudes, measured after induction of partial neuromuscular blockade, ranged from 25 to 7562 microV (median, 600 microV). Variation in intraoperative motor-evoked potential amplitude ranged from signal loss to a 3440% increase. A decrease to less than 20% of the individual reference value for motor-evoked potential amplitude occurred at least once in nine patients (22.5%) but as not associated with postoperative motor deficits. Two patients who had loss of motor-evoked potential signal without recovery did have postoperative motor deficits. Motor-evoked potentials predicted postoperative motor improvement in six patients. An improvement threshold of 160% of the reference amplitude predicted postoperative motor improvement with 100% sensitivity and 81% specificity. CONCLUSIONS: The authors support the utility of recording transcranial electrical motor-evoked potentials in spinal surgery under partial neuromuscular blockade. Recovery of lost motor-evoked potentials was not associated with postoperative motor deficits, whereas non-recovery of lost motor-evoked potentials was.

Adolescent↗
PubMed1996

Diffuse brain invasion of glioma cells requires beta 1 integrins.

Diffuse invasion of brain tissue by single tumor cells is a characteristic feature of gliomas and a major reason why these tumors cannot be completely resected. The molecular basis of brain invasion is poorly understood. We regulated the expression of beta 1 integrins, the major group of extracellular matrix receptors, in astrocytic tumor cells by using a tetracycline-dependent transcription control system. Rat C6 glioma cells were stably transfected with (a) the tetracycline-controlled transactivator (tTA) gene, (b) antisense beta 1 cDNA under the control of a tTA/tetracycline-responsive promoter, and (c) the beta-galactosidase (lacZ) gene for histochemical identification. In one clone, C6TL beta, beta 1 protein levels were unaffected in the presence of tetracycline, but they were reduced by 60% in the absence of tetracycline because of production of antisense mRNA. C6TL beta cells were transplanted into the striatum of nude mice. After 14 days in the presence of tetracycline in the drinking water, tumors showed diffuse brain invasion, mainly along vascular basement membranes. In the absence of tetracycline, however, tumor cells were compact and generally well delineated from the surrounding brain tissue. These data, ie, blocking of brain invasion by antisense beta 1 mRNA, either because of disturbed interaction of beta 1 with brain extracellular matrix components or interference with beta 1-dependent signaling pathways, strongly suggest that beta 1 integrins are required for diffuse brain invasion of gliomas.

Animals↗
PubMed1995

Retrovirus-mediated expression of an artificial beta-endorphin precursor in primary fibroblasts.

Peptides are of potential interest in the field of gene therapy but require modification by genetic engineering to facilitate their secretion. Amino terminal addition of a signal peptide is not always sufficient to achieve this goal, as found in this study for beta-endorphin. To overcome this problem, addition of the pre-pro-sequence of mouse nerve growth factor to beta-endorphin was tested. Retrovirus-mediated expression of a hybrid construct of the pre-pro-sequence of nerve growth factor and human beta-endorphin in primary fibroblasts resulted in the secretion of beta-endorphin immunoreactivity at a rate of 620 pg/h/10(6) cells. Analysis of the secreted beta-endorphin immunoreactivity with reverse-phase HPLC, immunoassays using three different antibodies, and an assay for the specific displacement of [3H][D-Ala2,N-MePhe4,Gly-ol5]enkephalin from mu-opioid receptors suggests that the pre-pro-sequence is cleaved off from the pre-pro-sequence/beta-endorphin construct prior to secretion, resulting in bona fide beta-endorphin. Transplantation of beta-endorphin-secreting cells into brain or spinal cord may provide a gene therapy approach for the treatment of chronic, opioid-sensitive pain states.

3T3 Cells↗
PubMed1995

Expression of biologically active basic fibroblast growth factor by genetically modified rat primary skin fibroblasts.

Basic fibroblast growth factor (FGF-2) is normally expressed as a cell-associated protein, and accordingly it is not clear how it exerts its action on target cells in vivo. It has been proposed that cells release, by death or other mechanisms, small amounts of FGF-2 that then acts in an autocrine manner. To address the question of whether it is necessary that FGF-2 remain cell associated or needs to be secreted from cells to have biological activity, we expressed the 18-kDa form of FGF-2 in primary fibroblasts as a cell-associated (FGF-2-B) or as a secreted (FGF-2-S) protein. FGF-2 protein is detected in cell lysates and membrane fractions of both cell types, whereas it is present in significant amounts only in the conditioned medium of FGF-2-S cells. No FGF-2 is detected in control (untransfected) cells. FGF-2-S cells also grow faster than the control of FGF-2-B cells. Yet, when evaluated for their ability to promote the survival of embryonic hippocampal neurons in vitro, both the cell types are active, establishing the activity of the transgene product. We conclude that FGF-2 is active when engineered to be expressed as a cell-associated form or secreted from cells.

Animals↗
PubMed1995

Pilocytic astrocytoma presenting as an extra-axial tumor in the cerebellopontine angle: case report.

Glial tumors in the cerebellopontine angle (CPA) are rare. Four histologically distinct types of glial tumors of the CPA have been described in the literature as ependymoma, medulloblastoma, mixed glial tumor, and fibrillary astrocytoma. This case report describes a pilocytic astrocytoma of the CPA. A 58-year-old man with a hearing loss had an extra-axial tumor in the left CPA that extended into the internal auditory canal. The characteristics of the tumor on magnetic resonance imaging scans differed from those of typical CPA tumors. It adhered avidly to the cochlear and vestibular nerves, which had to be sacrificed for gross total resection. Microscopic examination showed the typical features of an adult-type pilocytic astrocytoma.

Astrocytoma↗