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Biomedical subjects

C S Raine

Publications and source records attributed to C S Raine.

At least 19 recordsLinked to original sources

Effects of anti-white matter serum on myelin and lipid synthesis in brain prisms.

Tissue prisms prepared by choping whole mouse brain maintained respiratory capacity and ultrastructural integrity of 3 h in vitro. Normal rabbit serum (ca. 25%) caused no morphological change but inhibited the synthesis of galactolipids by the prisms. Heating the serum abolished the inhibition. Complement containing anti-white matter rabbit serum destroyed myelin and inhibited galactolipid synthesis to a greater degree than did normal serum. Structures other than myelin were unaffected by the antiserum. Incubation in the presence of heated anti-white matter serum eliminated the myelin destruction but resulted in specific morphological changes characterized by the doubling of the myelin lamellae at the intraperiod line. Immunoperoxidase studies suggest specific binding of immunoglobulin to components of myelin located at the intraperiod lone. These changes were similar to those found in organotypic cultures. Heated antiserum did not inhibit galactolipid synthesis but addition of complement (normal guinea pig serum) to the heated antiserum restored only that portion of the inhibition which exceeded that caused by normal serum. Heat labile factors in normal rabbit serum which inhibit myelin lipid synthesis in the prisms must be corrected for in studies in which the heating of serum is used to demonstrate that the effect is complement dependent. The prism system is simpler than that of organotypic cultures and may be useful in the study of myelinotoxic factors.

Animals

Late-onset malignant astrocytoma in a case of multiple sclerosis. Clinical, neuropathological, virological, and tissue culture studies.

An unusual case of concurrent MS and anaplastic astrocytoma is presented. MS was diagnosed in a female patient at the age of 22 years. A left side thalamotomy was performed for relief of severe intention tremor at age 28 and at age 32 she received immunosuppressive therapy for 1 year. At the age of 36 after a severe exacerbation of her symptoms a left side fronto-temporal tumor was diagnosed and a subtotal neurosurgical extirpation was performed. Histopathologically, the tumor was an anaplastic astrocytoma, which was further substantiated by electron microscopy and establishment of a permanent cell line in vitro. The cultured tumor cells were negative for measles virus by immunofluorescence. The relationship between the reactive astrocytes in MS plaques and astrocytic neoplasia is discussed.

Adult

Oligodendrocyte staining by multiple sclerosis serum is nonspecific.

The immunofluorescent staining properties of 65 serum samples from 54 patients with multiple sclerosis (MS), 63 samples from 55 patients with other neurological diseases (OND), and sera from 14 healthy normal individuals were examined on frozen sections of bovine and human brain. When tested on bovine brain sections, positive oligodendrocyte staining was present in 63% of MS sera, 43% of OND sera, and 29% of normal sera. The percentages were lower with human brain tissue. Astrocyte and myelin staining was common. F(ab')2 fragments purified from selected positive and negative MS and control sera gave no staining, though IgG fractions from the same sera prior to pepsin digestion gave positive staining. When tested against antihuman IgM and IgA conjugates, the same positive sera and their IgG-depleted globulin fractions gave minimal or no staining. These results indicate that oligodendrocyte staining is not specific for MS, is not due to specific antibody, and is probably the result of nonspecific binding to Fc receptors.

Animals

Multiple sclerosis: circulating antigen-reactive lymphocytes.

Circulating lymphocyte populations were examined in 85 patients with multiple sclerosis (MS), 26 of whom showed exacerbations; 48 patients with other neurological diseases (OND); 14 patients suffering from psychiatric disorders; and 2 normal subjects. The study involved the assay of early (active, high-affinity rosetting) T-cells, myelin basic protein (MBP)-reactive early T-cells, late (total, 24-hour rosetting) T-cell levels were significantly lower in MS (p less than 0.01) than in OND subjects. Exacerbations in MS were usually accompanied by further decreases in early T-cells. The lower levels of early T-cells in MS and their fluctuations are believed to reflect disease activity. MBP-reactive early T-cells were more frequently increased in MS (75% of cases) than OND (50%), and while this might be indicative of increased sensitization against myelin antigens, it was found not to be an MS-specific phenomenon.

Adult

Experimental allergic encephalomyelitis. Frequent sampling of blood alters disease course.

Previous studies have shown that Strain 13 guinea pigs sensitised for experimental allergic encephalomyelitis (EAE) as adults usually develop an acute, fatal form of disease while animals inoculated as juveniles usually display a chronic relapsing form. The present study reports that following repeated short-interval blood sampling by cardiac puncture for the estimation of lymphocyte populations, some adult Strain 13 guinea pigs sensitised for acute EAE unexpectedly survived and developed chronic EAE, while a group of juveniles sensitised for chronic EAE and bled under the same conditions, developed a more severe, acute form of EAE. It is suggested that this reversal of disease course was related to the depletion of circulating factors.

Age Factors

Chronic relapsing experimental allergic encephalomyelitis. Correlation of circulating lymphocyte fluctuations with disease activity in suppressed and unsuppressed animals.

Groups of juvenile Strain 13 guinea pigs sensitized for chronic relapsing experimental allergic encephalomyelitis (EAE) with isogeneic central nervous system (CNS) tissue in complete Freund's adjuvant (CFA) were either left to develop late-onset chronic EAE (unsuppressed), or given a series of injections of bovine myelin basic protein (MBP) in incomplete Freund's adjuvant (IFA) to suppress the disease. All unsuppressed animals developed disease and all suppressed animals remained healthy over a 27-month period of study. some unsuppressed and suppressed animals were rechallenged with CNS tissue in CFA 12 or 26 months post-inoculation (PI). Unsuppressed animals all became sick 2-4 weeks after rechallenge, while rechallenged, suppressed animals were protected, indicating that the suppression was permanent. Pathologic findings in the CNS complemented the clinical changes. Circulating lymphocyte studies were performed on animals from all groups. Early (active, high-affinity rosetting) T cell levels in unsuppressed animals showed significant decreases during exacerbations (P less than 0.01) and normal values during remissions. After rechallenge, circulating early T cells decreased in unsuppressed animals with the development of signs. In suppressed animals, early T cells showed significant elevations during, and for a short time after, the period of suppressive injections, and normal values afterwards. These levels did not change significantly after rechallenge. Late (total, 24 hour rosetting) T cell and B cell values showed minor fluctuations only which did not correlate with disease activity. These results indicate that chronic relapsing EAE can be successfully suppressed with MBP in IFA, that this suppression is permanent and that the immunologic findings presented correlate well with the clinical and pathologic facets of the disease. the findings are presented in terms of their relevance to multiple sclerosis.

Animals

Acute experimental allergic encephalomyelitis. Myelin basic protein-reactive T cells in the circulation and in meningeal infiltrates.

MBP-reactive early T cells were determined simultaneously in the circulation and in inflammatory cells from the meninges of guinea pigs with acute EAE. In addition, early and late T cells as well as B cells were estimated in both compartments. Within the circulation, early T cells were found to decrease at the onset of clinical signs, and MBP-reactive early T cells were significantly higher than in normals. Different changes were found in inflammatory cells from the meninges. In this cell population, early T cells were increased but did not show significant, further elevation after incubation with MBP. This may indicate that the majority of early T cells in meningeal infiltrates had been previously activated by MBP in vivo.

Acute Disease

Heterogeneity of virus particles in measles virus.

A heterogeneous population of virions is generated by measles virus-infected cells. These particles are partially separable by sucrose density centrifugation into three peaks. Each population is stable and contains infectious particles. The particles of all three populations contain at least six polypeptide species that differ between particle populations only in quantity. All three populations contain a 50S RNA species, and the heaviest density peak also contains an additional species of 43S RNA. The difference between these results and previous studies with measles virions will be discussed.

Animals

Experimental allergic neuritis. Ultrastructure of serum-induced myelin aberrations in peripheral nervous system cultures.

Mature myelinated cultures of mouse dorsal root ganglia (sometimes grown in combination with spinal cord tissue) have been exposed to unheated and heated (complement-inactivated) serum from rabbits with experimental allergic neuritis (EAN). Experimental cultures were examined for periods ranging from several hours to approximately 2 weeks of exposure. Some cultures were exposed to EAN serum for approximately 1 week then returned to normal medium for examination of the reversibility of the lesions. Unheated EAN serum induced demyelination of peripheral nervous system fibers within 96 hours of exposure. Following removal of the EAN serum, affected fibers remyelinated. Heated EAN serum produced a type of myelin swelling identical with that described previously in spinal cord explants exposed to serum from rabbits with experimental allergic encephalomyelitis. The lamellar spacing of the peripheral nervous system meylin was increased to approximately 23 nm. and the normal bifilar intraperiod line was increased to four leaflets. Some hypermyelination was seen. The swelling was incompletely reversible following removal of the heated serum. These findings are discussed in terms of their relevance to immunemediated demyelination and peripheral neuropathy associated with the hypergammaglobulinemic states.

Animals

Disorganization of myelinogenesis in tissue culture by anti-CNS antiserum.

The presence of decomplemented anti-CNS antiserum profoundly affects myelinogenesis in cultured mouse embryo spinal cord. Light and electron microscope study has shown that oligodendroglia differentiate and produce an abundance of cell processes which surround the oligodendrocytes in a chaotic, disorganized array. Where the cell processes chance to meet, they form a kind of aberrant swollen myelin. Rarely, the oligodendroglial processes ensheath axons. For the most part, the available axons remain unmyelinated. On removal of the decomplemented antiserum, oligodendroglia differentiate and form normal myelin around the available axons. Myelination of peripheral nervous system (dorsal root ganglion) axons in the same preparations is unaffected by the presence of the antiserum. Thus, under these circumstances, the message from the neuron to the oligodendrocyte to make myelin is apparently intact, yet there is interference with the ability of the oligodendroglial cell process to find, attach to and encircle CNS axons with a normal myelin sheath.

Animals

Chronic relapsing experimental allergic encephalomyelitis: CNS plaque development in unsuppressed and suppressed animals.

Central nervous system (CNS) lesion morphology has been studied in inbred Strain 13 guinea pigs sensitized for chronic relapsing EAE in which the disease was either left to develop (unsuppressed) or was suppressed with injections containing myelin basic protein (MBP). Pathologic changes correlated well with clinical activity. In unsuppressed chronic EAE animals, active clinical disease was invariably matched by acute inflammation in the CNS. In more chronic states, the CNS displayed fibrosis and remyelination while relapses showed the CNS to contain recent changes superimposed upon old lesions. In animals in which the disease was suppressed by injections of MBP, clinical signs did not develop. However, some early subclinical changes were seen morphologically. These lesions were able to remyelinate early on and there was no progression in lesion formation. Apparently, therefore, MBP had a beneficial effect upon the course of the disease and had promoted structural repair. It thus appears that MBP therapy might be one effective approach for the prevention of chronic relapsing EAE. The findings should prove relevant to future MBP trials in multiple sclerosis.

Animals

Suppression of chronic allergic encephalomyelitis: relevance to multiple sclerosis.

The expression of chronic relapsing experimental allergic encephalomyelitis in strain 13 guinea pigs was suppressed with a single series of injections of myelin basic protein in incomplete Freund's adjuvant. The suppression appeared permanent, and subsequent rechallenge with central nervous system antigen failed to elicit exacerbations.

Animals

Encephalitogenic properties of purified preparations of bovine oligodendrocytes tested in guinea pigs.

The present study has investigated central nervous system disease in guinea pigs inoculated with emulsions containing purified preparations of bovine oligodendroglia and their fractions isolated with or without trypsinization, whole bovine white matter or myelin basic protein (MBP). The MBP content of the oligodendroglial fractions was determined by radioimmunoassay. It was found that oligodendroglia prepared from trypsinized fresh brain contained minute amounts of MBP and did not induce disease. The corresponding cell fraction from non-trypsinized frozen brain was rich in MBP and induced disease. Bovine white matter and MBP induced typical experimental allergic encephalomyelitis (EAE). The structural preservation of the non-encephalitogenic trypsinized MBP-poor cells was very good and that of the encephalitogenic MBP-rich non-trypsinized cells very poor. It has been concluded that the encephalitogenicity observed was due to MBP, rather than to a specific oligodendroglial antigen.

Animals

Characterization of antioligodendrocyte serum.

An antioligodendrocyte serum (AOS) has been raised in rabbits against preparations of isolated bovine oligodendrocytes. The antibody was assayed by two techniques. By complement fixation with isolated oligodendrocytes, the titer of the antibody was 1:64 to 1:128. By indirect immunofluorescence testing of oligodendrocyte suspensions and frozen brain sections, the titer of the AOS was 1:256 to 1:512. When unabsorbed AOS was used, immunofluorescent staining proved it specific for bovine oligodendrocytes in suspension and in sections, and for human oligodendrocytes in sections. In cell suspensions, the staining was membrane related and in sections, cytoplasmic. The oligodendrocyte staining could be totally removed by absorption of AOS against oligodendrocyte suspensions, whereas absorption against bovine myelin, bovine myelin basic protein, and bovine neurons did not affect the staining reaction. AOS also stsined Schwann cells, a property possible related to antigens shared with oligodendrocytes. It is concluded that AOS is specific for oligodendrocytes and can now be applied to fundamental and disease-rel

Animals