Naked DNA vaccination differentially modulates autoimmune responses in experimental autoimmune encephalomyelitis

Vaccination with naked DNA represents a therapeutic strategy currently under consideration in multiple sclerosis (MS). In this study, we tested the potential therapeutic effect of vaccination with a naked DNA construct encoding proteolipid protein (pRc/CMV-PLP) upon the outcome of subsequent sensitization for experimental autoimmune encephalomyelitis (EAE) actively-induced in SJL mice with PLP139-151 peptide in adjuvant. Intramuscular vaccination with the naked DNA pRc/CMV-PLP construct led to PLP expression in local muscle tissue that persisted for about 8 weeks. Early sensitization for EAE (4 weeks after DNA vaccination) caused recipient mice to develop a severe, exacerbated form of disease (in comparison to control mice), while late sensitization (>10 weeks) resulted in a milder, ameliorated form. In the groups sensitized <10 weeks post-DNA vaccination with pRc/CMV-PLP induction of a Th1-type cytokine response was noted. In contrast, sensitization >10 weeks post-DNA vaccination led to peripheral tolerance as evidenced by a decrease in T cell proliferation and cytotoxic T cell response, no Th2 response, and no increase in apoptosis. These data are novel in that they demonstrate a differential effect of DNA vaccination and have important implications for its use as a mechanism to enhance or modulate immune reactivity.     

Tumor necrosis factor mediates myelin and oligodendrocyte damage in vitro

Recombinant human tumor necrosis factor (rhTNF) has been tested for its effect on myelinated cultures of mouse spinal cord tissue. As controls, recombinant human interferon gamma (rhIFN) and interleukin-2 (rhIL-2) were tested, as well as T-cell supernatants, antigalactocerebroside serum, and normal culture medium. It was found that rhTNF induced delayed-onset (18-24 hr) oligodendrocyte necrosis and a type of myelin dilatation peculiar to this system. Some nerve fibers progressed to demyelination by 72 hours. The myelin dilatation was not reversible by return to normal feeding solution for 3 days. In contrast, rhIFN, rhIL-2, T-cell supernatants, and normal medium had little or no effect on cultures. This mechanism differs from other immune-mediated mechanisms in that it appears that a physiological (not structural) demyelination occurs initially without overt destruction of the myelin sheath. These observations are relevant to the evolution of the multiple sclerosis plaque: dysfunction of ionic channels might contribute to the eventual demise of oligodendrocytes and axons in the longstanding lesion.     

On the development of CNS lesions in natural canine distemper encephalomyelitis

Acute lesions within spinal cord white matter have been studied by light and electron microscopy in 3 dogs suffering from the acute form of canine distemper encephalomyelitis (CDE). Prominent features of these lesions were viral inclusions, giant cell formation, cellular degeneration, myelin breakdown and phagocytic activity by cells believed to be derived from local glia. The viral inclusions occurred in giant cells, many astrocytes, macrophages and occasional oligodendroglia. Only suggestions of active viral replication from cell membranes were present. On the basis of the above features, these CDE lesions were classed as being acute. Perivascular inflammation and parenchymal invasion by haematogenous cells were lacking. However, older, gliotic, demyelinated lesions were always associated with inflammation. The pattern of demyelination in acute CDE lesions differed from those seen in other conditions, in particular the autoimmune demyelinating diseases. In acute CDE lesions, individual fibres became separated from others by rings of cells, the processes of which systematically stripped the myelin from the outer layers of the sheath inwards until a naked segment of axon remained. Some of the macrophages were recognisable as astroglia. Elsewhere, unequivocal astrocytes containing myelin debris were common. The results suggest that inflammation in acute CDE lesions is not a primary event, and that viral invasion causes breakdown of tissue which is accompanied pari passu by myelin destruction. The latter might be related to the non-specific release of host factors (viz. hydrolytic enzymes) or humoral factors during the cellular degeneration. Local cells appeared to participate in the process of myelin phagocytosis. Overt inflammation and damage by haematogenous cells were features only of chronic lesions and have been described previously in studies on chronic CDE lesions. The results are interpreted in terms of their relevance to the study of human subacute sclerosing panencephalitis, of which CDE is considered the animal analogue, and multiple sclerosis, the paradigm of the human demyelinating diseases.     

The effects of prolonged beta-adrenoceptor blockade on heart weight and cardiac intracellular potentials in rabbits.

Estimates were made in vivo in rabbits of the relative beta-receptor blocking potency and duration of action of propranolol and practolol. In further experiments groups of litter mates were injected twice daily with approximately equi-active amounts of propranolol or practolol, or with saline, for several weeks. The heart weights of the treated animals were significantly lower than those of the controls, the water contents were higher, and the dry weight differences were highly significant; -16.8% after 2 mg/kg bd propranolol for six weeks and -33.8% after 10 mg/kg practolol. (The treated animals grew less rapidly than the controls; when corrected for body weight these figures were -11.9% and -20.4%, respectively.) In the practolol group, but not the propranolol group, the duration of the atrial intracellular potentials was prolonged. There was no evidence that the prolonged treatment with either drug had a negative inotropic effect, or reduced positive inotropic responses to isoprenaline.     

Myelin basic protein and myelin-associated glycoprotein in chronic, relapsing experimental allergic encephalomyelitis

One-micron plastic sections of spinal cords from SJL/J mice with chronic relapsing experimental allergic encephalomyelitis (EAE) were reacted immunocytochemically with antiserum to myelin basic protein and myelin-associated glycoprotein. The distribution of myelin basic protein and myelin-associated glycoprotein in myelin sheaths was compared in acute and chronic areas of demyelination. No difference in the size of the lesion was seen with the two antisera. Myelin-associated glycoprotein was seen periaxonally in both normal myelin sheaths and sheaths which showed extensive splitting and ballooning as seen with toluidine blue stain and myelin basic protein antiserum. At least at the level of the light microscope, myelin basic protein antiserum gave intense staining of myelin while antiserum to myelin-associated glycoprotein showed little or no affinity to stain the myelin sheath itself, in contrast to other recent electron microscope observations. A few myelin basic protein or myelin-associated glycoprotein-containing oligodendrocytes were seen in lesion areas and remyelination by oligodendrocytes was rare. These observations are in agreement with findings from other models of EAE and multiple sclerosis where a primary loss of myelin has been implicated.     

A new concept of indexing tibiofibular torsion: a pilot study using dry bones

A newly developed clinical method of indexing tibial torsion uses the medial surface of the tibia as the proximal reference; however, the selection of a specific landmark on the medial surface has not been justified. Three different surfaces relating to the tibial tuberosity were tested using 24 dry tibial bones to determine which provides the most accurate and reliable landmark for use as the proximal reference. The medial surface of the tibia at the inferior point of the tibial tuberosity was the most reliable proximal reference that yielded the highest level of association between the newly developed clinical method and true tibial torsion (r = 0.77). The new method has the potential to describe the anatomy of the leg and to improve the clinical measurement of tibiofibular torsion.     

Scrapie in the rat: an electron-microscope study

Summary Both well formed amyloid bodies showing clear concentric lamination and smaller masses of amyloid material have been found in the brains of rats suffering from scrapie. Amyloid appears to form within astrocyte processes and glial cell bodies, as well as shrunken nerve cells. Rarely, it was present in degenerate axis cylinders. The individual fibrils which make up amyloid deposits appeared to have a spiral constitution, and deposits large enough to be seen in the light microscope were commonly strikingly symmetrically located.

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Zusammenfassung In den Gehirnen von Scrapie-kranken Ratten wurden wohlgeformte Amyloidkörperchen mit deutlicher konzentrischer Schichtung und kleinere amyloide Massen gefunden. Amyloid scheint sich in Astrocytenfortsätzen und Gliazellkörpern sowie in geschrumpften Nervenzellen zu bilden. Selten ist es in degenerierten Achsenzylindern vorhanden. Die Einzelfibrillen, welche die Amyloidablagerungen aufbauen, scheinen spiralige Form zu haben. Die großen im Lichtmikroskop sichtbaren Ablagerungen sind gewöhnlich auffallend symmetrisch im Gehirn lokalisiert.