Murray B. Bornstein’s research while affiliated with Albert Einstein College of Medicine and other places

Longterm infection of cultured hamster dorsal root ganglia with Hallé SSPE virus The morphogenesis of Hallé SSPE virus in cultures of hamster dorsal root ganglia (DRG) over a period of 60 days was followed by light and electron microscopy and by immunoperoxidase techniques. The Hallé virus was originally isolated from the lymph node of a patient with SSPE, and had been passaged only in non-neural cell lines. In organotypic cultures of hamster DRG, all cells except the neurons became infected. Large intracytoplasmic and intranuclear inclusions were commonly found in fibroblasts, but never in Schwann cells. The latter cells rounded up and were lost from the culture by 20 days post-infection (PI). Cell-free virus was detected by 7 days PI, reached a maximum at 30–40 days PI, and even at 60 days PI, substantial amounts of virus were still recorded. Budding virus particles were seen throughout the course of the infection. The formation of syncytia was rare. The present results are compared with previous data on the growth of measles virus in hamster DRG, and the growth of measles and SSPE viruses in cultures of hamster cerebellum. It was concluded that among the available strains of SSPE and measles virus, the Hallé strain is unique in its failure to infect nerve cells.

Multiple sclerosis and experimental allergic encephalomyelitis: specific demyelination of CNS in culture Cultures of mouse spinal cord tissue coupled to dorsal root ganglia, in which both CNS and PNS myelin develop, have been exposed for various periods of time to sera from patients with multiple sclerosis (MS) and animals with experimental allergic encephalomyelitis (EAE) induced by inoculation with whole CNS tissue in complete Freund's adjuvant. In both instances (MS and EAE), there was a selective depletion of CNS myelin with no adverse effect visible on PNS myelin in the same cultures. Prolonged exposure to demyelinating sera led to a sclerotic appearance in the CNS portion of cultures while PNS myelination progressed unimpeded. The results lend weight to the hypothesis that MS results from an immunological mechanism directed specifically against the CNS.

The implication of insulin-like growth factor I (IGF-I) in the myelination and the repair of myelin that occur after a demyelinating process was evaluated in organotypic cultures of embryonic nerve tissue. The amount of myelin of mouse spinal cord explants exposed to media supplemented with IGF-I beginning on the first day of explantation was recorded by light-microscopic examination and quantitation of the 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity. After 9 days in vitro (DIV), the cultures treated with medium supplemented with 0.1-1 microgram/ml IGF-I showed a greater amount of myelin and an increase over the controls in CNPase activity between 50 and 80% at 16 DIV and 100% at 21 DIV. Total demyelination with a concomitant reduction of about 80% in the CNPase activity resulted when anti-white matter antiserum and complement were added to the nutrient medium of fully myelinated cultures. This effect was partially reverted when IGF-I was included in the demyelinating medium. The higher inhibition, about 50%, was obtained with concentrations of IGF-I between 0.1 and 0.5 micrograms/ml. To study the effect of IGF-I on remyelination, well-myelinated cultures were completely demyelinated, maintained in that state for 2 or 15 DIV and after that allowed to remyelinate for 14 days. Cultures exposed to medium supplemented with 0.01-0.1 microgram/ml IGF-I showed a degree of remyelination similar to that of the normal nutrient medium-fed cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Astrocytes produce factors that control the growth and differentiation of many cell types within the CNS as well as play a role in the generation of the immune response. The extent to which these two functions interact has received less attention. We now report that astrocyte cultures established from rat brain endogenously express mRNA and low levels of secreted biologically active protein for the monocyte growth and differentiation factor colony stimulating factor-1 (CSF-1). Exposure of astrocytes to interleukin-1 (IL-1) and/or tumor necrosis factor (TNF) upregulated the expression of CSF-1 mRNA and protein. Following treatment with 100 U/ml of TNF, IL-1, or TNF+IL-1, maximum CSF-1 mRNA expression was observed at 3 hr. In the presence of IL-1 an increase in biologically active CSF-1 was detected in the astrocyte conditioned medium at 6 hr. These data indicate that the expression of CSF-1 by astrocytes can be modulated by exposure to the cytokines IL-1 and TNF. To determine whether CSF-1 provides a mitogenic signal for microglia during development, mouse spinal cord organotypic cultures were exposed to recombinant mouse CSF-1 (rmCSF-1), resulting in proliferation of microglia by 7 days and an increase in the number of ramified microglia over ameboid microglia by 14 days.

Astrocytes produce factors that control the growth and differentiation of many cell types within the CNS as well as play a role in the generation of the immune response. The extent to which these two functions interact has received less attention. We now report that astrocyte cultures established from rat brain endogenously express mRNA and low levels of secreted biologically active protein for the monocyte growth and differentiation factor colony stimulating factor‐1 (CSF‐1). Exposure of astrocytes to interleukin‐1 (IL‐1) and/or tumor necrosis factor (TNF) upregulated the expression of CSF‐1 mRNA and protein. Following treatment with 100 U/ml of TNF, IL‐1, or TNF+IL‐1, maximum CSF‐1 mRNA expression was observed at 3 hr. In the presence of IL‐1 an increase in biologically active CSF‐1 was detected in the astrocyte conditioned medium at 6 hr. These data indicate that the expression of CSF‐1 by astrocytes can be modulated by exposure to the cytokines IL‐1 and TNF. To determine whether CSF‐1 provides a mitogenic signal for microglia during development, mouse spinal cord organotypic cultures were exposed to recombinant mouse CSF‐1 (rmCSF‐1), resulting in proliferation of microglia by 7 days and an increase in the number of ramified microglia over ameboid microglia by 14 days.

Since multiple sclerosis (MS) is believed to be an immune-mediated disease, it follows that its therapies should be directed towards modulating the immune system. Current MS treatments, which include the use of exogenous steroids that are immunosuppressive, do not meet therapeutic objectives. delta 9-Tetrahydrocannabinol (THC), an active component of marijuana, has been shown to be immunosuppressive. To test THC's ability to suppress an immune-mediated disease, experimental autoimmune encephalomyelitis (EAE), the laboratory model of MS, was used. Lewis rats and strain 13 guinea pigs were administered THC either before inoculation for EAE or treated with THC after injection. Control animals received placebo. The effect of dose, in addition to the timing of treatment, was also investigated. All animals treated with placebo developed severe clinical EAE 10-12 days post-injection (d.p.i.) and more than 98% died by 15 d.p.i. THC-treated animals had either no clinical signs or mild signs with delayed onset (13-15 d.p.i.) with survival greater than 95%. Examination of central nervous system tissue revealed a marked reduction of inflammation in the THC-treated animals. Therefore, as THC has been shown to inhibit both clinical and histologic EAE, it may prove to be a new and relatively innocuous agent for the treatment of immune-mediated diseases.

This study explores the longitudinal assessment of visual evoked potentials (VEPs) in the rabbit as a method for defining factors underlying functional and structural changes associated with optic neuritis and the inflammatory demyelinating diseases. In rabbits with experimental autoimmune encephalomyelitis (EAE) induced by sensitization with guinea pig spinal cord myelin, injection of lymphokines into the posterior chamber of one eye (monocular challenge) produces an early inflammatory response in the retina and optic nerve, and an alteration in the VEP, all limited to the injected eye and its projections. The earliest changes in the timing and distribution of the cortical VEP occur within hours of ocular challenge and precede histopathological evidence of structural demyelination at the light microscope level. Prechallenge assessment allows the induced monocular prechiasmal effects to be distinguished from the more diffuse electrophysiological findings associated with EAE (i.e. those due to sensitization alone). In sensitized/challenged animals there is a clear correspondence between electrophysiological and morphological measures of dysfunction at the time points sampled. These results suggest that this model system affords an excellent opportunity to examine the precise structural correlates of the early functional changes associated with the onset of inflammatory demyelination within the CNS. Furthermore, the stability of the system provides the capacity to monitor alterations over the complete course of inflammation, demyelination and remyelination, induced by experimental manipulations.