Evidence for the importance of extracranial venous flow in patients with idiopathic intracranial hypertension (IIH).

Idiopathic intracranial hypertension (IIH) is characterized by increased ICP without evidence for intracranial mass lesion. Although the pathogenesis remains unknown, some association was found with intracranial venous thrombosis. To our knowledge, the extracranial venous drainage was not systematically evaluated in these patients. This study compared extracranial cerebral venous outflow in eight IIH patients and eight control subjects using magnetic resonance (MR) Venography and flow measurements. In addition, the study identified extracranial factors that affect cerebral venous drainage. In six of the IIH patients, either complete or partial functional obstruction of the internal jugular veins (IJVs) coupled with increased venous outflow through secondary venous channels was documented. On average, a four-fold increase in mean venous flow rate through the epidural and/or vertebral veins was measured in IIH patients compared with the healthy subjects. In one of the healthy subjects, intracranial venous outflow was studied also during external compression of the IJVs. Over 40% of the venous outflow through the IJVs shifted to the epidural veins and intracranial pressure, measured noninvasively by MRI, increased from 7.5 to 13 mmHg. Findings from this study suggest that increased ICP in some IIH patients could be associated with increased extracranial resistance to cerebral venous outflow.

MRI study of cerebral blood flow and CSF flow dynamics in an upright posture: the effect of posture on the intracranial compliance and pressure.

Postural related changes in cerebral hemodynamics and hydrodynamics were studied using Magnetic Resonance Imaging (MRI) measurements of cerebral blood flow and cerebrospinal fluid (CSF) flow dynamics. Ten healthy volunteers (mean age 29 +/- 7) were studied in supine and upright (sitting) postures. A Cine phase-contrast MRI technique was used to image the pulsatile blood flow to the brain, the venous outflow through the internal jugular, epidural, and vertebral veins, and the bi-directional CSF flow between the cranium and the spinal canal. Previously published analyses were applied to calculate and compare total cerebral blood flow (TCBF), intracranial compliance and pressure in both postures. A lower (12%) mean TCBF was measured in the upright position compared to supine position. A considerable smaller amount of CSF flow between the cranium and the spinal canal (58%), a much larger intracranial compliance (a 2.8-fold increase), and a corresponding decrease in the MRI-derived ICP were also measured in the sitting position. These changes suggest that the increased cerebrovascular and intracranial compliances in the upright posture are primarily due to reduced amounts of blood and CSF residing in their respective intracranial compartments in the upright position. This work demonstrates the ability to quantify neurophysiologic parameters associated with regulation of cerebral hemodynamics and hydrodynamics from dynamic MR imaging of blood and CSF flows.

Relationship between total cerebral blood flow and ICP measured noninvasively with dynamic MRI technique in healthy subjects.

Cerebral blood flow and ICP are important neurophysiologic parameters known to be affected by pathology and by trauma. Limited data on the relationship between these parameters following head trauma is inconsistent with regard to whether these parameters are correlated. Data on the relationship between these parameters in the healthy state is not readily available due to a lack of noninvasive means to measure these important parameters. A recently developed noninvasive MRI-based method for simultaneous measurement of total cerebral blood flow and intracranial pressure was applied to establish the relationship between ICP and TCBF values in healthy subjects. Seventy-one simultaneous measurements of CBF and ICP were obtained from 23 healthy young adults. These results demonstrated that CBF values span over a much narrower range compared with ICP. The relationship between the inter-individual CBF and ICP measurements suggest that in the healthy state and in rest these parameters are not correlated.

Distribution of intraventricularly administered antiamyloid-beta peptide (Abeta) antibody in the mouse brain.

There is considerable interest in utilizing the intracerebroventricular (icv) route of administration of antibodies in the brain for various studies and for the therapy of malignancies, but very little is known about the anatomic extent of distribution of the antibody in brain after injection into the third ventricle. To explore the potential for icv administration of antiamyloid-beta peptide (Abeta) in reducing Abeta toxicity in brain in Alzheimer's disease, we first mapped the time course and path of transit of horseradish peroxidase (HRP)-labeled antibody. The results show that, after a single injection into the mouse third venticle, the HRP-labeled antibody is localized within the microvasculature, first that of the corticohippocampal region close to the site of injection at 3 hr. By 24 hr, the antibody is distributed throughout the hippocampus and frontoparietal cortex close to the injection site, as well as in the deep and outer cerebral cortex and cerebellar cortex remote from the injection site. The injected antibody is almost entirely removed by 4 days. Therefore, the antibody had diffused throughout all the brain by 24 hr, showing the feasibility of small quantities of anti-Abeta antibody infused into the third ventricle to reach extracellular epitopes throughout the brain parenchyma rapidly.

Serum amylase and lipase elevation is associated with intracranial events.

Serum amylase and lipase elevation has been observed in trauma patients and patients with traumatic intracranial bleeding. However, the causes of this elevation have not been clearly elucidated. A further question remains as to whether other intracranial events are associated with such enzyme elevation as well. We retrospectively reviewed 75 patients consecutively admitted to Cook County Hospital Neurosurgical Intensive Care Unit over a 3-month period for trauma, infection, tumor, or other space-occupying lesions with an unstable condition or neurological deficit. Eleven patients (15%) had elevated amylase and lipase levels. The patients were divided into two groups: Group I (n = 64) had normal and Group II (n = 11) had raised amylase and lipase levels [amylase 402 +/- 444 U/L with normal < or = 125 U/L and lipase 474 +/- 313 U/L with normal < or = 55 U/L]. All Group II patients suffered an intracranial event. Twenty-four Group I (38%) and 10 Group II (91%) patients required craniotomy (P < 0.01). No patients had clinical or radiographic evidence of pancreatitis. In summary, intracranial events are associated with serum amylase and lipase elevation probably through centrally activated pathways. Because of the lack of diagnostic value, routine pancreatic enzyme monitoring should not be performed in this patient population.

Immunogene therapy with interleukin-2-secreting fibroblasts for intracerebrally metastasizing breast cancer in mice.

OBJECT: Breast cancer is the second leading cause of cancer-related death in American women. Brain metastases occur in 15 to 30% of patients with breast cancer, and this usually results in death. Despite the availability of surgery, radiotherapy, and chemotherapy, the prognosis for patients with breast cancer that has metastasized to the intracerebral region remains poor. This study was designed to determine if an intracerebrally metastasizing breast tumor could be treated successfully with a cellular vaccine consisting of allogeneic fibroblasts (H-2K) modified to secrete interleukin (IL)-2. METHODS: The authors used EO771 breast cancer cells, derived from a spontaneously arising breast-cancer tumor in C57BL/6 mice. The authors first determined the length of survival of C57BL/6 mice that had been injected with varying numbers of EO771 cells into the right frontal lobes and found that 100% of those animals that received a dose of 10(4) cells died within 41 days, whereas 100% of the group that received 10(3) cells died within 23 days. Based on these results, 5 x 10(4) EO771 cells were injected into the right frontal lobe of C57BL/6 mice and the animals were treated intracerebrally with a single intratumoral injection of 10(6) allogeneic fibroblasts genetically engineered to secrete IL-2. The allogeneic fibroblasts transfected with the IL-2 gene formed large quantities of IL-2 as measured by an enzyme-linked immunosorbent assay. Control groups of animals were treated with either allogeneic fibroblasts transfected with the retroviral vector, but not the IL-2 gene, or with media. The effects of this treatment on the animal's survival and the tumor's histopathological characteristics were investigated. The results indicate a significant prolongation (p < 0.005) of survival in animals with intracerebrally metastasizing breast cancer treated only with IL-2-secreting allogeneic fibroblasts. Tumor did not develop in four of 10 animals in the IL-2-treated group, and these animals were rechallenged at 90 days by intracerebral injection of tumor, but no treatment cells, and compared with four naive animals receiving intracerebral tumor. Again, animals that previously had been treated with IL-2-secreting fibroblasts had a markedly prolonged survival (p < 0.05) compared with control animals following a second challenge with tumor cells. Histopathological examination revealed smaller tumors associated with lymphocytic infiltrations in the treated animals. CONCLUSIONS: This work represents a new treatment for breast cancer that has metastasized to the brain in which a cellular vaccine consisting of allogeneic fibroblasts genetically engineered to secrete cytokines is used as a novel means for delivery of immunogene therapy and demonstrates the induction of long-term immunity against tumor.

Analysis of magnetic resonance imaging-based blood and cerebrospinal fluid flow measurements in patients with Chiari I malformation: a system approach.

OBJECT: A pilot study was performed to assess noninvasively the change in intracranial compliance (ICC) and intracranial pressure (ICP) in patients with Chiari I malformation who undergo foramen magnum decompression. The working hypothesis was that the main effect of the decompressive surgery is a change in ICP. Noninvasive cine phasecontrast magnetic resonance (MR) imaging is a motion-sensitive dynamic MR imaging technique that allows for visualization and quantitation of tissue motion and flow. The authors' group has used dynamic phase-contrast MR imaging to visualize and quantify pulsatile blood and cerebrospinal fluid (CSF) flow in the craniospinal system. METHODS: A system approach has been used to characterize the hemodynamic-hydrodynamic coupling in the craniospinal system and to derive measures for ICC and ICP. Magnetic resonance imaging-based ICC and ICP values are derived from the ratio of the volume and pressure changes that occur naturally during each cardiac cycle. The authors conducted a prospective study of four patients, three of whom were studied before and after decompressive surgery; significant change in MR imaging-derived ICC and ICP values was documented in only one of the three surgically treated patients. A significant change in the dynamics of the intracranial volume change (ICVC) during the cardiac cycle, however, was observed in all three patients. In healthy individuals the ICVC waveform usually consists of the following sequence: monotonic increase in intracranial volume (ICV) during the systolic phase due to increased blood inflow, monotonic decrease in ICV caused by the onset of CSF outflow into the spinal canal, and increase in the venous outflow. A nonmonotonic decline in the ICVC waveform has been observed in all patients with headaches, and a relatively normal waveform was found in those without headaches or whose headaches were resolved or alleviated by the surgery. A "partial-valve" mechanism is proposed as an explanation for the abnormal ICVC dynamics. The monotonic decline in ICVC is interrupted by a "premature" reduction in the CSF outflow. This may be caused by a displacement of the hindbrain into the cervical spinal canal during the systolic phase. This obstructs the CSF flow at the later part of the systolic phase such that the ICV does not continue its gradual decline. Postsurgery, the ICVC waveforms presented a more normal-appearing ICVC dynamics profile. CONCLUSIONS: Magnetic resonance imaging measurement of transcranial CSF and blood flow may lead to a better understanding of the pathophysiology of Chiari malformations and may prove to be an important diagnostic tool for guiding for the treatment of patients with Chiari I malformation.

Treatment of the T98G glioblastoma cell line with antisense oligonucleotides directed toward mRNA encoding transforming growth factor-alpha and the epidermal growth factor receptor.

Antisense oligonucleotides (oligos) complementary to mRNA encoding transforming growth factor-alpha (TGF-alpha) and its target, the epidermal growth factor receptor (EGFR), are efficacious against human prostate and breast cancers carried in athymic nude mice. Glioblastomas, also regulated by EGFR expression, would appear to be similarly susceptible, and we now employ them against the T98G tumor model. T98G cells were distributed into wells and allowed to adhere prior to addition of oligos (12.5 microM) directed against TGF-alpha and/or EGFR for 6 d of treatment before thymidine radiolabeling. Supplemental media and oligos (25 microM final concentration) were added after d 3. Statistically significant inhibition by oligos directed against TGF-alpha, EGFR, and their combination was 13.8%, 26.3%, and 18.1%, respectively. In a subsequent experiment cells were incubated with increasing amounts of each oligo and their combination for 3 d prior to radiolabeling. Statistically significant inhibition of growth for either oligo at every concentration was found. Cells incubated with 6.25, 12.5, 25, and 50 microM antisense directed against TGF-alpha had a mean inhibition of 29.3%, 33.3%, 21.7%, and 46.6%, respectively. Cells similarly treated with oligos against EGFR had a mean inhibition of 77.9%, 80.3%, 82.0%, and 83.7%, respectively, and cells incubated with 6.25, 12.5, 25 and 50 microM of each oligo had a mean inhibition of 74.7%, 70.6%, 70.8%, and 76.3%, respectively. Lastly, in a paired experiment, cells treated with 0, 0.39, 0.78, 1.56, 3.125, and 6.25 microM of oligos, either specifically directed against EGFR or a random control, for 3 d were evaluated for both thymidine incorporation and EGFR expression. Statistically significant inhibition of 3H-thymidine incorporation was seen in cells with the oligo specifically directed against EGFR at 3.125 microM and 6.25 microM when compared to non-oligo containing controls. This was accompanied by a comparable significantly decreased expression of a low-MW reactive derivative of EGFR at 3.125 microM and 6.25 microM in Western blots, and of a high-MW reactive EGFR at 6.25 microM. The significant effect against high-MW EGFR was observed vs both the non-oligo containing control and the random sequence. Oligo concentrations between 0.78 and 1.5 microM also resulted in decreased expression of the low-MW form, but not significant differences in thymidine radiolabeling. In recovery experiments, cells treated initially with greater oligo concentrations required significantly increased time to recover, particularly in cells treated with EGFR directed oligos. Intracellular uptake and nuclear localization was demonstrated with FITC tagged oligos. In summary, even at relatively low oligo concentrations and short exposure, oligos against TGF-alpha, and particularly EGFR, significantly inhibit in vitro growth of the T98G glioblastoma, possibly mediated by decreased EGFR expression.

MR-Intracranial pressure (ICP): a method to measure intracranial elastance and pressure noninvasively by means of MR imaging: baboon and human study.

PURPOSE: To develop a noninvasive method for intracranial elastance and intracranial pressure (ICP) measurement. MATERIALS AND METHODS: Intracranial volume and pressure changes were calculated from magnetic resonance (MR) imaging measurements of cerebrospinal fluid (CSF) and blood flow. The volume change was calculated from the net transcranial CSF and blood volumetric flow rates. The change in pressure was derived from the change in the CSF pressure gradient calculated from CSF velocity. An elastance index was derived from the ratio of pressure to volume change. The reproducibility of the elastance index measurement was established from four to five measurements in five healthy volunteers. The elastance index was measured and compared with invasive ICP measurements in five patients with an intraventricular catheter at MR imaging. False-positive and false-negative rates were established by using 25 measurements in eight healthy volunteers and six in four patients with chronically elevated ICP. RESULTS: The mean of the fractional SD of the elastance index in humans was 19.6%. The elastance index in the five patients with intraventricular catheters correlated well with the invasively measured ICP (R:(2) = 0.965; P: <.005). MR imaging-derived ICPs in the eight healthy volunteers were 4.2-12.4 mm Hg, all within normal range. Measurements in three of the four patients with chronically elevated ICP were 20.5-34.0 mm Hg, substantially higher than the normal limit. CONCLUSION: MR imaging-derived elastance index correlates with ICP over a wide range of ICP values. The sensitivity of the technique allows differentiation between normal and elevated ICP.

Cytokine immunogene therapy.

The prognosis for patients with either a primary or metastatic brain tumor is poor. Clearly new forms of therapy to improve the long-term survival of patients with malignant brain tumors are urgently needed. The authors are in the process of developing a new and novel form of treatment for primary and metastatic brain tumors in which they use genes involved in growth repression. In particular most tumors fail to induce an antitumor immune response strong enough to kill the tumor. Under appropriate circumstances, however, immunity can be produced in unique structures on the tumor cells known as antigens. To prepare the vaccine, genes are transferred into a fibroblast cell line that causes the cell to produce cytokines, the potent proteins known to stimulate the immune system. These cells are subsequently injected into the tumor bed, resulting in the development of an antitumor immune response. In experiments described in this manuscript, the authors have investigated a number of ways of augmenting the immune response by administering this type of cellular vaccine. They found that mice with a primary intracerebral glioma, melanoma, or breast cancer treated with this allogeneic cytokine-secreting vaccine survived significantly longer than untreated mice. Additionally the vaccine was found to stimulate a systemic antitumor immune response, as shown by immunocytotoxic studies, histopathological examination, and delayed immune memory responses. In summary, these results indicate that immunogene therapy is a promising new targeted therapy for the treatment of intracerebral malignant tumors.

Prolongation of survival of mice with glioma treated with semiallogeneic fibroblasts secreting interleukin-2.

OBJECT: The current prognosis for patients with malignant brain tumors remains poor, and new therapeutic options are urgently needed. We previously have shown that prolongation of survival can be achieved in C57BL/6 mice (H-2b) with a syngeneic intracerebral or subcutaneous glioma when treated with allogeneic mouse fibroblasts (H-2k) genetically engineered to secrete interleukin-2 (IL-2). Like other antigens, tumor-associated antigens are recognized by cytotoxic T lymphocytes in the context of determinants specified by the major histocompatibility complex class I locus. Because the rejection of allogeneic major histocompatibility complex determinants has the property of an immune adjuvant, immunotherapy of glioma with a cellular immunogen that combines the expression of both syngeneic class I determinants and allogeneic antigens could have advantages over an immunogen that expresses syngeneic or allogeneic determinants alone. METHODS: To investigate this question in a mouse glioma model, we further modified allogeneic mouse fibroblasts (H-2k), not only for IL-2 secretion, but also for the expression of H-2Kb class I determinants. We tested the immunotherapeutic properties of these semiallogeneic cells (LM-IL-2/H-2Kb) in C57BL/6 mice with Gl261 glioma in both subcutaneous and intracerebral glioma models. RESULTS: C57BL/6 mice with either a subcutaneous or intracerebral glioma treated solely by injection of these IL-2-secreting semiallogeneic cells had significantly prolonged survival rates compared with untreated mice or mice treated with cells secreting only IL-2 or cells lacking the H-2Kb determinants. In some instances, the mice treated with the semiallogeneic cells survived indefinitely, suggesting total eradication of the glioma. When a 51Cr release assay was used, the specific immunocytotoxicity measured by release of isotopes from labeled Gl261 glioma cells coincubated with spleen cells from mice immunized with the semiallogeneic IL-2-secreting cells was significantly higher than that of spleen cells from nonimmunized mice or mice immunized with allogeneic cells lacking syngeneic major histocompatibility complex determinants. In addition, antibody depletion studies using monoclonal antibodies against CD8+ and natural killer/lymphokine-activated killer cells demonstrated a specific CD8+ immunocytotoxic response in animals immunized with the semiallogeneic IL-2-secreting cells compared with only a natural killer/lymphokine-activated killer response in mice immunized with the allogeneic IL-2-secreting cells. CONCLUSION: The augmented immune response against glioma in mice treated with the semiallogeneic IL-2-secreting cells points toward a new form of immunotherapy, "immuno-gene therapy," for patients with malignant intracerebral glioma.

In vivo establishment of T98G human glioblastoma.

Human derived T98G glioblastoma has long been utilized as an in vitro model for epidermal growth factor receptor (EGFR)-mediated growth regulation. Recently, T98G has been employed to develop new types of therapy directed at limiting EGFR expression such as by administration of antisense oligonucleotides directed against EGFR encoding mRNA. A major limitation to extending this model for in vivo application is that T98G implanted s.c. or intracerebrally has been reported not to grow in nude mice. In an effort to extend this model to permit in vivo studies, we evaluated the use of Matrigel and orthotopic (intracranial) implantation techniques. When equal volumes of Matrigel were mixed with T98G cell suspensions, tumors developed at both flank and orthotopic locations. Four groups of nude mice were inoculated into the flanks with either 10(5), 10(6), 4 x 10(6) or 10(7) T98G cells in a 150 microliters total volume with Matrigel. In 1/5, 3/5, 1/5 and 1/3 mice receiving 10(5), 10(6), 4 x 10(6) and 10(7) cells, respectively, tumors developed 11, 15, 15 and 15 weeks, respectively, following inoculation. Out of 4 mice inoculated orthotopically (intracranially into the frontal lobe) with only 4 x 10(4) cells and Matrigel, 2 developed tumors. However, all mice (4/4) inoculated orthotopically with 4 x 10(5) cells in a 10 microliters total volume with Matrigel developed tumors. Two were identified histologically following a scheduled sacrifice at 36 and 60 days and two more at 103 and 118 days after sacrifice following abnormal behavior. The best tumor establishment efficacy combined orthotopic implantation of 4 x 10(5) T98G cells with Matrigel. These techniques permit the use of T98G glioblastoma as an in vivo model for new forms of therapy.

Development of a new mouse brain tumor model using implantable micro-cannulas.

The objective of this study was to develop a brain tumor model in a mouse where gene therapy could be delivered either directly into a pre-established tumor bed or prophylacticly prior to tumor delivery (protective treatment). Micro-cannulas were constructed from metal tubing, implanted into the right frontal lobe of mice, and then secured in place in the skull with cement. Experiments evaluating the usefulness, reproducibility and morbidity of the system were performed. It was found that tumor cells could reproducibly be delivered into the right frontal lobe of the mice. Two tumors could be precisely delivered into the same area following injections at different times. Repeat injections were performed without a stereotaxic frame and without the need for repeat intracerebral needle tracts. There were no noticeable side effects of maintaining the cannulas in place for long periods of time. In summary, this system is useful for studying the effects of various treatment strategies on established brain tumors in a mouse model which more closely simulates the clinical situation. It obviates the need for time consuming stereotaxic procedures or repeat invasive intracerebral injections.

Survival and toxicity of an allogeneic cytokine-secreting fibroblast vaccine in the central nervous system.

OBJECTIVE: In previous studies, we determined that C57BL/6 mice (H-2b) with intracerebral (i.c.) malignant glioma or melanoma treated with allogeneic fibroblasts genetically engineered to secrete interleukin (IL)-2 results in prolonged survival and a cellular antitumor response when the treatment is administered intratumorally (via i.c. injection). For this study, we evaluated the survival and toxicity of the cytokine-secreting cellular vaccine administered directly into the central nervous system in both syngeneic (C3H H-2k) and allogeneic (C57BL/6 H-2b) mice using bioassay and polymerase chain reaction techniques. METHODS: First, brain sections were prepared at varying time intervals (2-60 d) after i.c. injection of the IL-2-secreting fibroblasts (H-2k) into allogeneic and syngeneic mice, and the presence of the cells was detected by polymerase chain reaction for the neomycin gene, which was used as a selection marker for the gene transfer. As a second means of assessing survival of the IL-2-secreting cells, an in vitro bioassay technique was used. Brain sections were prepared at varying time intervals (2-60 d) after i.c. injection of the IL-2-secreting cells into allogeneic and syngeneic mice. Cells were disassociated and grown in media containing neomycin. RESULTS: A positive signal by polymerase chain reaction was no longer detectable 14 days after injection into allogeneic C57BL/6 (H-2b) mice. In contrast, under similar circumstances, the IL-2-secreting cellular vaccine could be detected for more than 55 days in histocompatible (C3H) syngeneic mice (H-2k). Allogeneic cells could be recovered from tissue culture for only 2 to 5 days after i.c. injection of the modified cells. In contrast, syngeneic cells were recovered for up to 28 days after i.c. injection. At no time did the mice exhibit signs of neurological deficit or adverse affects on their survival (>60 d). CONCLUSION: An allogeneic cytokine-secreting cellular vaccine has a limited lifespan in the central nervous system, surviving only 14 days or less, and has no apparent adverse effects. These results indicate the advantage of using a modified allogeneic cell line as the delivery vehicle for gene therapy in the treatment of an i.c. neoplasm.

Intracerebral versus subcutaneous immunization with allogeneic fibroblasts genetically engineered to secrete interleukin-2 in the treatment of central nervous system glioma and melanoma.

OBJECTIVE: The purpose of this study was to determine the optimal route of delivery of gene therapy for an intracerebral (IC) tumor. In previous studies, treatment of an IC tumor with the IC administration of a cellular vaccine consisting of allogeneic fibroblasts genetically engineered to secrete cytokines prolonged survival. Systemic delivery of gene therapy is of significant clinical interest. METHODS: In this study, allogeneic fibroblasts engineered to secrete interleukin (IL)-2 (LM-IL-2 cells) were administered either subcutaneously or intracerebrally to C57BL/6 mice with IC glioma. In addition, fibroblasts genetically engineered to express (antibody-defined) melanoma-associated antigens and to secrete IL-2 (RLBA-IL-2) were injected either intracerebrally or subcutaneously into mice bearing IC melanoma. RESULTS: The results indicate a significant prolongation of survival in mice with IC glioma treated intracerebrally with LM-IL-2 cells, relative to the survival of mice with IC glioma treated subcutaneously with LM-IL-2 cells or untreated mice with glioma. The specific release of isotope from 51Cr-labeled glioma cells coincubated with spleen cells from animals treated either subcutaneously or intracerebrally with LM-IL-2 cells was significantly greater than the release of isotope from glioma cells coincubated with spleen cells from nonimmunized mice. In a similar fashion, the survival of mice with IC B16 melanoma immunized intracerebrally with RLBA-IL-2 cells was significantly longer than nonimmunized mice injected with B16 cells alone. In contrast, the survival of mice with IC melanoma treated by subcutaneous injection with RLBA-IL-2 cells was not significantly different than that of untreated mice. Using a 51Cr-release assay, the specific release of isotope from labeled B16 cells coincubated with spleen cells from mice immunized either intracerebrally or subcutaneously with RLBA-IL-2 cells was significantly higher than that of B16 cells coincubated with cells from nonimmunized mice. CONCLUSIONS: Direct IC administration of fibroblasts genetically engineered to secrete IL-2 was more effective in prolonging survival than peripheral subcutaneous administration in the treatment of mice with IC glioma or melanoma.

Insulin-like growth factors in central nervous system tumors.

Insulin-like growth factors (IGFs) appear to play a role in the development of tumors in general and brain tumors in particular. Specific receptors for IGFs have been identified in normal human and rat brain, and evidence suggests that components of the IGF signal transduction system may play a role in the transformation process. Secretion of IGFs by a variety of human brain tumors has been confirmed, and these growth factors appear to have an autocrine stimulatory effect on these tumors. IGFs circulate in the blood stream bound to at least six distinct binding proteins which may modulate the effects of these growth factors on target tissues. Sex steroids may also regulate the behavior of certain brain tumors such as meningiomas at least in part through their effects on the expression of IGFs and their binding proteins. Recently, antisense gene technology against certain IGFs or their receptors have resulted in potent antitumor effects in the case of several gliomas, although the mechanism for this remains unclear.

Civilian cases of tangential gunshot wounds to the head.

A series of 168 civilian cases of tangential gunshot wounds to the head is presented. Neurologic deficits on presentation were generally minimal. Computed tomographic (CT) scans were performed in 51% of patients, and abnormal CT findings were noted in 35% (18% of all patients). Major operative procedures were required in 9% of the patients. Serious sequelae of tangential injuries are described even with patients who initially have no neurologic abnormality. We suggest that a CT scan is warranted in all cases of tangential gunshot wounds to the head.

Gunshot wounds to the head in civilian practice.

An aggressive surgical strategy was applied to cranial gunshot wound victims at Cook County Hospital in Chicago from 1983 to 1992. A series of 480 patients with an overall mortality rate of 34% is presented. A total of 150 patients underwent craniotomy with an operative mortality rate of 21%. Criteria for operation were Glasgow Coma Scale scores of 3 through 7 without hypotension or fixed and dilated pupils or Glasgow Coma Scale scores of 8 through 15 with intracranial bone fragments or significant clot. This study supports previous reports that even patients with severe neurological deficits and massive cerebral damage can benefit from aggressive treatment and make satisfactory recoveries.

Prolonged survival of mice with glioma injected intracerebrally with double cytokine-secreting cells.

A novel approach toward the treatment of glioma was developed in a murine model. The genes for both interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were first transfected into a mouse fibroblast cell line that expresses defined major histocompatibility complex (MHC) determinants (H-2k). The double cytokine-secreting cells were then cotransplanted intracerebrally with the Gl261 murine glioma cell line into syngeneic C57BL/6 mice (H-2b) whose cells differed at the MHC from the cellular immunogen. The results indicate that the survival of mice with glioma injected with the cytokine-secreting allogeneic cells was significantly prolonged, relative to the survival of mice receiving equivalent numbers of glioma cells alone. Using a standard 51Cr-release assay, the specific release of isotope from labeled Gl261 cells coincubated with spleen cells from mice injected intracerebrally with the glioma cells and the cytokine-secreting fibroblasts was significantly higher than the release of isotope from glioma cells coincubated with spleen cells from nonimmunized mice. The cellular antiglioma response was mediated by natural killer/lymphokine-activated killer and Lyt-2.2+ (CD8+) cells. The increased survival of mice with glioma and the specific immunocytotoxic responses after immunization with fibroblasts modified to secrete both IL-2 and IFN-gamma indicate the potential of an immunotherapeutic approach to gliomas with cytokine-secreting cells.