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A J Suckling

Publications and source records attributed to A J Suckling.

At least 19 recordsLinked to original sources
PubMed1989

Identification of a common idiotype on myelin oligodendrocyte glycoprotein-specific autoantibodies in chronic relapsing experimental allergic encephalomyelitis.

The myelin/oligodendrocyte glycoprotein (MOG) is a target antigen for autoantibody-mediated demyelination in chronic relapsing experimental allergic encephalomyelitis (CREAE) in the Strain 13 guinea pig. Anti-idiotypic antibodies directed against a demyelinating MOG-specific monoclonal antibody (8-18C5) have identified a cross-reactive idiotype in 10/10 CREAE sera. In nine of these sera inhibition studies demonstrated that this idiotype was at or in close proximity to the combination site of guinea pig anti-MOG autoantibodies. This cross-reactive anti-MOG idiotype may represent an important target for the anti-idiotypic regulation of demyelinating antibody responses in CREAE.

Animals↗
PubMed1989

The origin and specificity of intrathecal IgG in chronic relapsing experimental allergic encephalomyelitis.

The source of IgG in the cerebrospinal fluid (CSF) in guinea pigs with chronic relapsing experimental allergic encephalomyelitis (CR-EAE) was investigated using quotient analysis of total IgG and albumin concentrations and by computing CSF-plasma ratios of specific IgG concentrations. Increased blood-CSF barrier (B-CSFB) permeability was shown by elevated albumin quotients in both relapse and remission phases of CR-EAE and intrathecal production of IgG was indicated by raised ratios of IgG to albumin in the CSF. Intrathecal IgG synthesis was greatest in guinea pigs which had little B-CSFB damage. When enzyme-linked immunosorbent assays (ELISA) for whole cord, myelin basic protein (MBP) or Mycobacterium tuberculosis were performed with CSF and plasma adjusted to the concentration of total IgG, the CSF/plasma ratios of ELISA results for specific antibodies were less then unity and ratios for whole cord and MBP were lower than those for M. tuberculosis. There was thus no evidence for a selective increase in the CSF of antibody specific either for the neuroantigens tested or for adjuvant components. The CSF-plasma ratios for each specific antibody were inversely correlated with the extent of total IgG intrathecal synthesis, suggesting that much of the antibody production within the CNS is the result of polyclonal B cell activation.

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PubMed1988

Stimulation of protein phosphorylation in mixed glial cell primary cultures and subcultures by the phorbol ester 12-O-tetradecanoylphorbol-13-acetate.

Glial cell primary cultures consisting of protoplasmic and fibrous astrocytes, oligodendrocytes and progenitor glial cells incubated in medium containing 0.5% foetal calf serum and treated with 25 nM 12-o-tetradecanoylphorbol-13-acetate (TPA) for periods between 15 and 60 min showed a stimulation of protein phosphorylation which was most prominent in a polypeptide with a molecular weight of about 80,000 Da. Glial subcultures consisting mainly of Type 2 astrocytes, oligodendrocytes and progenitor glia showed a similar TPA stimulation of 80,000 Da protein phosphorylation detectable within 1 min of phorbol ester addition. TPA treatment of primary glial cultures led to an enhancement of phospholipid turnover but exposure of primary glial cultures to concentrations of TPA up to 250 nM caused no morphological change in protoplasmic astrocytes. 4-Phorbol (4-PH) or dimethylsulfoxide (DMSO) was without effect on protein phosphorylation or lipid turnover in glial cultures.

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PubMed1988

Protein kinase C activity in soluble fractions from glial cells in primary culture and subcultures.

Protein kinase C (calcium + phospholipid-dependent kinase) activity has been measured in soluble 100,000 g fractions from mixed glial cells in primary culture; in 12 day cultures the specific activity (mean +/- S.D.) was 184 +/- 10 pmol 32P incorporated/10 min/mg protein. In glial cell subcultures lacking protoplasmic astrocytes protein kinase C specific activity was lower. An inhibitor of protein kinase C in 100,000 g supernatants was removed by chromatography through DE-52 anion exchange resin increasing the specific activity of the calcium + phospholipid-dependent kinase about 20 times. Protein kinase C was also associated with membrane fractions from glial cells; the membrane-associated enzyme had a higher specific activity than in the cytoplasm.

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PubMed1988

Plasma immunoglobulin responses and disease severity in chronic relapsing experimental allergic encephalomyelitis.

Plasma IgG, IgA and IgM responses in various stages of chronic relapsing experimental allergic encephalomyelitis (CR-EAE) were investigated by ELISA and rocket immunoelectrophoresis. Autoantibody levels were elevated in acute EAE but immunoglobulin responses were maximal in chronic disease. Plasma IgG and IgA specific for the whole cord, myelin and MBP correlated closely with the clinical signs of post-acute disease; in age-matched groups, levels were lower in animals in remission or with no further disease than in those in relapse or with a stable chronic disease course. Sequential sampling revealed a significant increase in neuroantigen-specific IgG (with MBP as the dominant autoantigen) during the onset of a relapse. Lipid-specific antibody levels were raised throughout CR-EAE but constituted only a small proportion of the total response against neural antigens. Determination of total immunoglobulin concentrations suggested a general suppression of IgG responses in guinea pigs in remission. The strong correlations found between antibody levels and the severity of chronic disease provide further evidence that antibody-mediated mechanisms can play a major role in the pathogenesis of CR-EAE.

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PubMed1988

Autoantibody responses in the cerebrospinal fluid of guinea pigs with chronic relapsing experimental allergic encephalomyelitis.

We report the use of the ELISA technique to measure IgG specific for whole cord, myelin, myelin basic protein and Mycobacterium tuberculosis in the cerebrospinal fluid (CSF) of Strain 13 guinea pigs in different stages of chronic relapsing experimental allergic encephalomyelitis (CR-EAE). Specific antibody levels to all 4 antigen preparations were related to the severity of clinical signs, with the highest levels of IgG in the CSF of guinea pigs in relapse or in stable chronic disease. Total IgG levels in the CSF, though elevated throughout the course of CR-EAE, did not show any association with the category of disease. Control animals inoculated with complete Freund's adjuvant (CFA) alone showed CSF IgG levels specific for M. tuberculosis that were not significantly different from those in animals with chronic EAE, indicating that CFA may itself induce a late-acting increase in blood-brain barrier permeability.

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PubMed1987

Activated T-cells and macrophages in the cerebrospinal fluid and the spinal meningeal exudate in chronic relapsing experimental allergic encephalomyelitis.

An analysis has been made of the cell types which mark with monoclonal antibodies against T cells, macrophages and the IL-2 receptor (anti-Tac) in the blood, cerebrospinal fluid (CSF) and spinal meningeal exudates taken from guinea pigs in the relapse and remission stages of chronic relapsing experimental allergic encephalomyelitis (CR-EAE). Whilst the T-cell and macrophage content of blood remained unchanged throughout the course of CR-EAE, T cells accounted for the majority of the CSF pleocytosis associated with relapsing disease but both T cells and macrophages populated the meningeal exudate in substantial numbers. Activated T cells (Tac+) rose in number in blood only after the onset of relapse but formed a far higher proportion of the CSF pleocytosis or meningeal exudate than in paired blood samples. Meningeal exudate cells from Freund's adjuvant-inoculated, but not uninoculated animals, also showed an increase in Tac+ cell levels. In addition, the meningeal exudate contained a substantial number of cells which did not label with anti-T or anti-macrophage antibodies and which did not vary in absolute numbers throughout the course of disease.

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PubMed1987

Lymphocytic activation in peripheral blood and cerebrospinal fluid during the course of chronic relapsing experimental allergic encephalomyelitis.

A monoclonal antibody against the human interleukin-2 receptor (anti-Tac) has been found to cross-react with an antigen on the surface of guinea pig leucocytes. Cells marking with anti-Tac and with an anti-pan T cell monoclonal antibody have been quantitated in the peripheral blood and cerebrospinal fluid (CSF) of guinea pigs with chronic relapsing experimental allergic encephalomyelitis (CR-EAE). T cells account for about 90% of peripheral blood leucocytes in all animals whilst in the CSF, T cells are the major contributor only when there is a pleocytosis. The proportion of T cells marking with anti-Tac, a measure of T cell activation, in blood and CSF of control animals is 12%, rising to 23% in blood in the post-acute phase of the disease. However, a fall in the blood Tac/T ratio to 13% occurs during the first 10 days of relapse with a subsequent rise to 30-35%. This change is related to the time after onset of relapse irrespective of the subsequent course of the disease. From first relapse onwards CSF lymphocytes show a greater level of activation than lymphocytes from paired peripheral blood samples but the proportion of Tac+ cells in CSF does not increase with increasing CSF pleocytosis. The data is consistent with migration of activated T cells from blood to CSF at the onset of relapse.

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PubMed1987

IgG subclass responses and immediate skin sensitivity in guinea-pigs with chronic relapsing experimental allergic encephalomyelitis.

In strain-13 guinea-pigs inoculated for chronic relapsing experimental allergic encephalomyelitis (CR-EAE), IgG1 and IgG2 subclass antibody responses were investigated using single radial immunodiffusion and enzyme-linked immunosorbent assays (ELISA) for IgG1 and IgG2 specific for whole cord, myelin, myelin basic protein and Mycobacterium tuberculosis. The early acute stage revealed no increase in IgG1 but was associated with increased levels of IgG2 specific for neural and adjuvant components. Throughout the chronic phase of the disease, there were increased levels of IgG of both subclasses specific for the antigens tested but a preferential synthesis of IgG1. Levels of both IgG1 and IgG2 specific for neuroantigens were lowest in those guinea-pigs which did not develop signs of chronic disease. Immediate skin sensitivity against a wide range of neural antigens was not demonstrated though positive results may have been masked by the ability of myelin basic protein to induce non-specific mast cell degranulation and by altered histamine responsiveness in disease. Guinea-pigs with chronic paralysis had a lower skin sensitivity to histamine, compound 48/80 and M. tuberculosis than those in remission.

Animals↗
PubMed1987

Cerebrospinal fluid interleukin 1 like activity during chronic relapsing experimental allergic encephalomyelitis.

Cerebrospinal fluid (CSF) and plasma samples were taken from strain 13 guinea pigs in various stages of chronic relapsing experimental allergic encephalomyelitis (CREAE). The samples were assayed for interleukin 1 (IL-1) in the C3H/Hej mouse thymocyte assay. After removal of inhibitors, IL-1 was detectable in low amounts in plasma (5 U/ml) throughout the course of the disease but was raised in the acute phase (12 U/ml). CSF IL-1 was, however, only present in low amounts (6 U/ml) during the acute phase but was elevated (18 U/ml) during the chronic stages of CREAE. During the relapse phase levels of IL-1 correlated with the total leucocyte count in the CSF. On gel filtration of CSF, IL-1 activity eluted at approximately 15 kD and could not be attributed to leakage of plasma IL-1 during CSF puncture or IL-2 activity.

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PubMed1986

Cyclosporin-A treatment of experimental allergic encephalomyelitis: changes in immunological regulation and blood-CSF barrier function.

Strain 13 guinea pigs at 18-21 days of age were sensitised with spinal cord and Freund's complete adjuvant to induce experimental allergic encephalomyelitis (EAE). Treatment with cyclosporin-A (CS-A) from one day before sensitisation until from 12 to 39 days after sensitisation resulted in a suppression of the disease. Suppression was indicated by an absence or reduction in severity of clinical signs together with a lack of increase in blood-cerebrospinal fluid (CSF) barrier permeability to proteins and a lower white cell count in the CSF from treated animals. When CS-A treatment was withdrawn, clinical disease reappeared but changes in IgG concentration in the CSF indicated that intrathecal synthesis of IgG had occurred, which was not the case in the untreated disease. Modification by CS-A of the immunoregulatory processes involved in the development of EAE, provides a model to study the special conditions which operate in neuroimmunological disorders.

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PubMed1986

Chronic relapsing experimental allergic encephalomyelitis. Inhibition of lymphocyte mitogenesis by disease-related serum factors.

Peripheral blood mononuclear cells (PBMC) from strain 13 guinea pigs at various stages of chronic relapsing experimental allergic encephalomyelitis (CREAE) showed significantly reduced reactivity to the polyclonal T cell mitogens, phytohaemagglutinin and concanavalin A. Serum taken at the same time revealed an increase in inhibitors of lymphocyte mitogenesis. The factors identified appeared to inhibit the initial stages of lymphocyte proliferation: they were not cytotoxic for lymphocytes nor were they complement-dependent.

Animals↗
PubMed1986

Quantitative analysis of the cellular constituents of the cerebrospinal fluid in chronic relapsing experimental allergic encephalomyelitis.

Cerebrospinal fluid (CSF) was taken from guinea pigs in various stages of chronic relapsing experimental allergic encephalomyelitis (CR-EAE). The leucocytes in CSF samples were counted and subjected to immunocytochemical analysis using monoclonal antibodies selectively recognising guinea pig T cells, macrophages or Ia antigens. The results showed that total leucocyte numbers and the proportion of macrophages in CSF were elevated in the acute phase of CR-EAE but samples of CSF from animals in early relapse did not show a significant elevation in leucocyte count or macrophage content. In addition the level of T cell activation was higher in CSF than in peripheral blood during disease and was highest in the acute and immediately post-acute phases of the CR-EAE.

Animals↗
PubMed1986

Chronic relapsing experimental allergic encephalomyelitis: cyclosporin-A treatment of relapsing and remitting disease.

Adult strain 13 guinea pigs in the relapsing-remitting phase of chronic relapsing experimental allergic encephalomyelitis were treated with cyclosporin-A (CS-A) at an oral dose of 35 mg kg-1 body wt. for periods of 17-32 days. Of the animals treated in relapse or remission the majority (6/11) showed a deterioration in clinical condition during treatment with the remaining 5 showing no change and 6/8 developed more severe clinical signs after treatment had ceased. In contrast, a similar group of untreated or vehicle-only treated relapsing-remitting animals developed a worse or better clinical condition in similar numbers when assessed over a similar time period with the majority (22/33) showing no change in clinical signs. Blood white cell counts and plasma IgG values remained constant throughout treatment and no clinical signs of drug toxicity were evident. Cerebrospinal fluid (CSF) mononuclear cell counts rose initially only in CS-A treated animals but fell to levels below those of controls by the end of treatment. These results are discussed in the context of the effectiveness of CS-A therapy in autoimmune diseases of the central nervous system.

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PubMed1984

The effect of Freund's adjuvants on blood-cerebrospinal fluid barrier permeability.

In guinea pigs with experimental allergic encephalomyelitis induced by spinal cord homogenate--complete Freund's adjuvant (CFA) emulsions an increase in the albumin permeability of the blood--cerebrospinal fluid barrier occurred from day 10 post-inoculation (p.i.) onward. In animals inoculated with CFA alone an increased albumin permeability was also demonstrated but only between days 5 and 10 after inoculation; by day 14-16 p.i. the barrier permeability had returned to control values. A similar change was seen in animals inoculated with incomplete Freund's adjuvant (IFA) only. However, both CFA and CFA-cord induced a strong humoral immune response which was not seen in animals inoculated with IFA alone. These results may have important consequences for the understanding of the development of inflammatory diseases of the central nervous system.

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PubMed1984

Infection of rat brain primary cell cultures with an avirulent A7 strain of Semliki Forest virus.

The ability of A7 Semliki Forest Virus (SFV) to infect primary brain cell cultures has been examined using cultures prepared from 1-2-day neonatal rat cerebral hemispheres. These cultures, characterised immunocytochemically using cell-specified markers, contain mainly GFAP+ protoplasmic astrocytes and smaller multiprocessed A2B5+ cells, probably fibrous astrocytes. 10% of the cells are GC+ oligodendrocytes and some neurones are also present. These cultures support virus growth and a cytopathic effect was observed. Using double labelling techniques with the cell-specific markers and anti-SFV antibody A7 has been shown to readily infect cells which carry either the A2B5+ antigen or galactocerebroside marker. Protoplasmic astrocytes (GFAP+/A2B5-) are not readily infected under the conditions used. The protein labelling studies using [35S]methionine show that host cell protein synthesis is not completely shut off and continues in the astrocyte protein region. These results suggest that cells derived from a common A2B5+, GFAP-, GC- progenitor glial cell, i.e. GC+ oligodendrocytes and A2B5+/GFAP+ fibrous astrocytes, are more readily infected than other brain cell types including the protoplasmic astrocytes.

Animals↗
PubMed1984

Cerebrospinal fluid sampling from guineapigs: sample volume--related changes in protein concentration in control animals and animals in the relapsing phase of chronic relapsing experimental allergic encephalomyelitis.

Cerebrospinal fluid (CSF) was removed from guineapigs by puncture of the cisterna magna and the total sample volume of 200-360 microl divided into 40 microl aliquots. After determination of albumin and IgG in these CSF aliquots it was found that successive samples gave different results. In general, up to 100 microl CSF could be removed before the protein concentration began to increase. In animals with chronic relapsing experimental allergic encephalomyelitis (CR-EAE) the rise in albumin concentration was accompanied by a corresponding fall in the number of white cells in later samples.

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