myelin-basic-protein and Chronic-Disease

This article is focusing on the etiology of the multiple sclerosis (MS). MS is the disease of autoimmune origin, concerning mainly young people with its complicated nature. The course of the degenerative and inflammatory process is variable and unexpected, from featureless to the vast and rapidly progressing nature, leading to the considerable disability. The etiological factors are not still well known. We can distinguish different agents: genetic, immunological and environmental, among them tobacco smoking also. Authors survey the etiopathological basis of the disease.

Topics: Adjuvants, Immunologic; Adult; Autoimmunity; Causality; Chronic Disease; Comorbidity; Cytokines; Female; Humans; Male; Multiple Sclerosis; Myelin Basic Protein; Smoking; T-Lymphocytes; Tobacco Use Disorder

Early recognition of whether a product has potential as a new therapy for treating multiple sclerosis (MS) relies upon the quality of the animal models used in the preclinical trials. The promising effects of new treatments in rodent models of experimental autoimmune encephalomyelitis (EAE) have rarely been reproduced in patients suffering from MS. EAE in outbred marmoset monkeys, Callithrix jacchus, is a valid new model, and might provide an experimental link between EAE in rodent models and human MS. Using magnetic resonance imaging techniques similar to those used in patients suffering from MS pathological abnormalities in the brain, white matter of the animal can be visualized and quantified. Moreover, NMR spectroscopy, in combination with pattern recognition, offers an advanced uroscopic technique for the identification of biomarkers of inflammatory demyelination.

Topics: Animals; Animals, Outbred Strains; Antigens, CD; Biomarkers; Brain; Callithrix; CD4-Positive T-Lymphocytes; Chronic Disease; Demyelinating Diseases; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Genes, MHC Class II; Humans; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Mice; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteins; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Radiography; T-Lymphocytes, Cytotoxic; Th2 Cells

Both heterogeneity and restricted heterogeneity of the encephalitogenic myelin basic protein (MBP) peptide-specific T cell receptors (TCRs) were demonstrated in inbred animals depending on the strain-specific genetic characteristics, the stage of the disease, the compartment of the lymphocytes obtained and the methodology used. Nevertheless, the similar features of some MBP-specific TCRs demonstrated across species suggest that conservation of these autoantigen-specific molecules undoubtedly exists, even though the degree of this conservation is controversial. However, the unequivocal heterogeneity of the immune response directed at one of the most important myelin constituents, proteolipid lipoprotein (PLP), which occurs either as a primary or a secondary event during experimental allergic encephalomyelitis (EAE), indicates the complexity of the in vivo situation. Intramolecular and intermolecular spreading of antigen specificity during the course of the disease indicates that a TCR directed therapy may not be the choice of intervention in established disease even in individual strains of laboratory animals with restricted heterogeneity of the primary MBP-specific response. Studying the sequence of events, the recruited regulatory cells and cytokines, and the stromal factors controlling persistence or death of activated, memory cells in the tissue lesion, may reveal new therapeutic modalities with more universal applicabilities.

Topics: Animals; Chronic Disease; Clone Cells; Encephalomyelitis, Autoimmune, Experimental; Integrin alpha4beta1; Integrins; Leukocyte Common Antigens; Mice; Myelin Basic Protein; Myelin Proteolipid Protein; Rats; Receptors, Antigen, T-Cell, alpha-beta; Receptors, Lymphocyte Homing; T-Lymphocyte Subsets; Th1 Cells

Multiple sclerosis may ultimately be effectively treated using appropriate immunosuppressive regimens. Both cytotoxic drugs and therapeutic apheresis may constitute important therapeutic options. A number of studies are currently underway which will hopefully clarify the exact role of these treatment modalities in patients at different stages of disease.

Topics: Acute Disease; Adrenocorticotropic Hormone; Adult; Antilymphocyte Serum; Azathioprine; Cell Separation; Chronic Disease; Cyclophosphamide; Female; Humans; Immunosuppressive Agents; Lymphocyte Depletion; Male; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Plasmapheresis; Prednisone; Recurrence

Multiple sclerosis (MS) is the chronic inflammatory demyelinating disease of the CNS. Relapsing-remitting MS (RRMS) is the most common type of MS. However, the mechanisms of relapse and remission in MS have not been fully understood. While SJL mice immunized with proteolipid protein (PLP) develop relapsing-remitting experimental autoimmune encephalomyelitis (RR-EAE), we have recently observed that some of these mice were resistant to the active induction of relapsing EAE after initial clinical and histological symptoms of EAE with a severity similar to the relapsing EAE mice. To clarify the mechanism of relapsing, we examined myelin morphology during PLP

Topics: Animals; Brain; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Inflammation; Mice; Mice, Inbred Strains; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteolipid Protein; Neoplasm Recurrence, Local; Protein Isoforms; Spinal Cord

This study aims to evaluate the protective effects of progesterone on white matter injury and brain immaturity in neonatal rats with chronic hypoxia.. Three-day old Sprague-Dawley rats were randomly divided into 3 groups: (1) control (n = 48), rats were exposed to normoxia (fraction of inspired oxygen: 21% ± 0%); (2) chronic hypoxia (n = 48), rats were exposed to hypoxia (fraction of inspired oxygen: 10.5% ± 1.0%); and (3) progesterone (n = 48), rats were exposed to hypoxia and administrated with progesterone (8 mg/kg/d). Hematoxylin-eosin staining, immunohistochemistry, real-time quantitative polymerase chain reaction, and Western blot analyses were compared on postnatal day 14 in different groups. Motor skill and coordination abilities of rats were assessed via rotation experiments.. Increased brain weights (P < .05), narrowed ventricular sizes (P < .01), and rotarod experiment scores (P < .01) were better in the progesterone group than in the chronic hypoxia group. The number of mature oligodendrocytes and myelin basic protein expression increased in the progesterone group compared with the chronic hypoxia group (P < .01). The polarization of M1 microglia cells in the corpus callosum of chronic hypoxia-induced hypomyelination rats was significantly increased, whereas there were fewer M2 microglia cells. Conversely, progesterone therapy had an opposite effect and caused an increase in M2 microglia polarization versus a reduction in M1 microglia cells.. Progesterone could prevent white matter injury and improve brain maturation in a neonatal hypoxic rat model; this may be associated with inducing a switch from M1 to M2 in microglia.

Topics: Animals; Animals, Newborn; Behavior, Animal; Brain; Cell Plasticity; Chronic Disease; Disease Models, Animal; Female; Hypoxia; Leukoencephalopathies; Male; Microglia; Motor Activity; Myelin Basic Protein; Neuroprotective Agents; Oligodendroglia; Progesterone; Rats, Sprague-Dawley; White Matter

B cell-depleting therapies have recently proven to be clinically highly successful in the treatment of multiple sclerosis (MS). This study aimed to determine the effects of the novel type II anti-human CD20 (huCD20) monoclonal antibody (mAb) obinutuzumab (OBZ) on spinal cord degeneration in a B cell-dependent mouse model of MS. Double transgenic huCD20xHIGR3 (CD20dbtg) mice, which express human CD20, were immunised with the myelin fusion protein MP4 to induce experimental autoimmune encephalomyelitis (EAE). Both light and electron microscopy were used to assess myelination and axonal pathology in mice treated with OBZ during chronic EAE. Furthermore, the effects of the already established murine anti-CD20 antibody 18B12 were assessed in C57BL/6 wild-type (wt) mice. In both models (18B12/wt and OBZ/CD20dbtg) anti-CD20 treatment significantly diminished the extent of spinal cord pathology. While 18B12 treatment mainly reduced the extent of axonal pathology, a significant decrease in demyelination and increase in remyelination were additionally observed in OBZ-treated mice. Hence, the data suggest that OBZ could have neuroprotective effects on the CNS, setting the drug apart from the currently available type I anti-CD20 antibodies.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antigens, CD20; Antineoplastic Agents, Immunological; Axons; B-Lymphocytes; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Mice; Mice, Inbred C57BL; Mice, Transgenic; Microscopy, Electron; Multiple Sclerosis; Multiple Sclerosis, Chronic Progressive; Myelin Basic Protein; Myelin Proteolipid Protein; Neurofilament Proteins; Recombinant Fusion Proteins; Spinal Cord

To investigate the neuropsychological characteristics and changes in CRP, S100B, MBP, HSP-7, and NSE in serum.. Sixty-six (66) patients treated in our hospital as CCCI group were chosen for our study, and 90 patients with depression were selected as the depression group. The patients in both groups were examined with CT perfusion, depression, anxiety and cognition evaluation. Their serum CRP, S100B, MBP, HSP-70 and NSE levels were detected. Neuropsychological and serum markers characteristics were compared.. The CBF and CBV in bilateral basal ganglia, frontal lobes, greater oval center, brain stem, and left and right regions of occipital lobes of the patients in CCCI group were significantly lower than in the depression group. The HAMD and HAMA scores of CCCI group patients were significantly lower than in the depression group; CCCI group performed better regarding attention, memory, abstract terms and delayed recall. CCCI also had significantly higher total scores than the depression group. Serum CRP, S100B, MBP, HSP-70 and NSE levels in CCCI group were significantly higher than in the depression group. The differences reach statistical significance (p<0.05).. CCCI patients who are accompanied by minor depressive disorder have different degrees of cognitive impairment and experience a significant rise in serum CRP, S100B, MBP, HSP-70 and NSE.

Topics: Aged; Anxiety; Biomarkers; C-Reactive Protein; Cerebrovascular Circulation; Cerebrovascular Disorders; Chronic Disease; Depressive Disorder; Female; HSP70 Heat-Shock Proteins; Humans; Male; Middle Aged; Myelin Basic Protein; Neuropsychological Tests; Phosphopyruvate Hydratase; Polymerase Chain Reaction; Risk Factors; S100 Calcium Binding Protein beta Subunit; Tomography, X-Ray Computed

The chronic stage multiple sclerosis (MS), an inflammatory demyelinating disease of the central nervous system (CNS), remains refractory to current treatments. This refractory nature may be due to the fact that current treatments are primarily immunomodulatory, which prevent further demyelination but lack the capacity to promote remyelination. Several approaches, including transplantation of neural stem cells (NSCs) or antagonists to LINGO-1, a key part of the receptor complex for neuroregeneration inhibitors, have been effective in suppressing the acute stage of experimental autoimmune encephalomyelitis (EAE), an animal model of MS. However, their effect on the chronic stage EAE is not known. Here, we show that transplantation of NSCs had only a slight therapeutic effect when treatment started at the chronic stage of EAE (e.g., injected at day 40 postimmunization). However, NSCs engineered to produce LINGO-1-Fc, a soluble LINGO-1 antagonist, significantly promoted neurological recovery as demonstrated by amelioration of clinical signs, improvement in axonal integrity, and enhancement of oligodendrocyte maturation and neuron repopulation. Significantly enhanced NAD production and Sirt2 expression were also found in the CNS of mice treated with LINGO-1-Fc-producing NSC. Moreover, differentiation of LINGO-1-Fc-producing NSCs into oligodendrocytes in vitro was largely diminished by an NAMPT inhibitor, indicating that LINGO-1-Fc enhances the NAMPT/NAD/Sirt2 pathway. Together, our study establishes a CNS-targeted, novel LINGO-1-Fc delivery system using NSCs, which represents a novel and effective NSC-based gene therapy approach for the chronic stage of MS.

Topics: Animals; Autocrine Communication; Axons; Cell Count; Cell Differentiation; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Membrane Proteins; Mice, Inbred C57BL; Myelin Basic Protein; NAD; Neural Stem Cells; Neurons; Oligodendroglia; Paracrine Communication; Receptors, Fc; Recovery of Function; Remyelination; Sirtuin 2; Stem Cell Transplantation; Transduction, Genetic

Chronic cerebral hypoperfusion can lead to ischemic white matter injury resulting in vascular dementia. To characterize white matter injury in vascular dementia, we investigated disintegration of diverse white matter components using a rat model of chronic cerebral hypoperfusion.. Chronic cerebral hypoperfusion was modeled in Wistar rats by permanent occlusion of the bilateral common carotid arteries. We performed cognitive behavioral tests, including the water maze task, odor discrimination task, and novel object test; histological investigation of neuroinflammation, oligodendrocytes, myelin basic protein, and nodal or paranodal proteins at the nodes of Ranvier; and serial diffusion tensor imaging. Cilostazol was administered to protect against white matter injury.. Diverse cognitive impairments were induced by chronic cerebral hypoperfusion. Disintegration of white matter was characterized by neuroinflammation, loss of oligodendrocytes, attenuation of myelin density, structural derangement at the nodes of Ranvier, and disintegration of white matter tracts. Cilostazol protected against cognitive impairments and white matter disintegration.. White matter injury induced by chronic cerebral hypoperfusion can be characterized by disintegration of diverse white matter components. Cilostazol might be a therapeutic strategy against white matter disintegration in patients with vascular dementia.

Topics: Animals; Behavior, Animal; Brain Ischemia; Carotid Stenosis; Chronic Disease; Cilostazol; Cognition; Dementia, Vascular; Diffusion Tensor Imaging; Disease Models, Animal; Hippocampus; Inflammation; Leukoencephalopathies; Myelin Basic Protein; Neuroprotective Agents; Neuropsychological Tests; Oligodendroglia; Ranvier's Nodes; Rats; Rats, Wistar; Tetrazoles; White Matter

After spinal cord (SC)-injury, a non-modulated immune response contributes to the damage of neural tissue. Protective autoimmunity (PA) is a T cell mediated, neuroprotective response induced after SC-injury. Immunization with neural-derived peptides (INDP), such as A91, has shown to promote-in vitro-the production of neurotrophic factors. However, the production of these molecules has not been studied at the site of injury.. In order to evaluate these issues, we performed four experiments in adult female Sprague-Dawley rats. In the first one, brain derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) concentrations were evaluated at the site of lesion 21 days after SC-injury. BDNF and NT-3 were significantly increased in INDP-treated animals. In the second experiment, proliferation of anti-A91 T cells was assessed at chronic stages of injury. In this case, we found a significant proliferation of these cells in animals subjected to SC-injury + INDP. In the third experiment, we explored the amount of BDNF and NT3 at the site of injury in the chronic phase of rats subjected to either SC-contusion (SCC; moderate or severe) or SC-transection (SCT; complete or incomplete). The animals were treated with INDP immediately after injury. Rats subjected to moderate contusion or incomplete SCT showed significantly higher levels of BDNF and NT-3 as compared to PBS-immunized ones. In rats with severe SCC and complete SCT, BDNF and NT-3 concentrations were barely detected. Finally, in the fourth experiment we assessed motor function recovery in INDP-treated rats with moderate SC-injury. Rats immunized with A91 showed a significantly higher motor recovery from the first week and up to 4 months after SC-injury.. The results of this study suggest that PA boosted by immunization with A91 after moderate SC-injury can exert its benefits even at chronic stages, as shown by long-term production of BDNF and NT-3 and a substantial improvement in motor recovery.

Topics: Animals; Autoimmunity; Brain-Derived Neurotrophic Factor; Chronic Disease; Disease Models, Animal; Female; Motor Activity; Myelin Basic Protein; Neurotrophin 3; Peptide Fragments; Random Allocation; Rats, Sprague-Dawley; Recovery of Function; Severity of Illness Index; Spinal Cord; Spinal Cord Injuries; Time Factors; Vaccination

Bilateral carotid artery occlusion (2-vessel occlusion, 2VO) in rats is a classic and frequently used approach to develop an animal model of chronic cerebral hypoperfusion. However, this method results in substantial mortality in rats.. We investigated whether a modified 2VO procedure, which induces bilateral carotid artery stenosis via ligation of each bilateral common carotid artery (CCA) with a 32 gauge (G) needle followed by needle removal, could produce a chronic cerebral hypoperfusion rat model with an increased survival rate. Sprague-Dawley (SD) rats were treated with the standard or modified 2VO procedure, and changes in cerebral blood flow (CBF) and survival rates were determined. On day 28, cognitive function was assessed with the Morris Water Maze (MWM) test, and neuronal survival and degeneration within the hippocampal CA1 area were measured. Damage to the white matter (WM) within the corpus striatum was assessed via Luxol fast blue (LFB) staining and analyses analyzing the levels of the myelin basic protein (MBP) protein levels.. The modified 2VO procedure induced similar cognitive impairments, hippocampal lesions and WM damage compared with the standard 2VO procedure in rats; however, it had an increased survival rate. Comparison with existing methods This novel method can be used to quickly and effectively establish a chronic cerebral hypoperfusion rat model with common materials and an improved survival rate.. Bilateral carotid artery stenosis using a 32 G needle is a useful and reliable method to develop a rat model of chronic cerebral hypoperfusion with increased survival.

Topics: Animals; Apoptosis; CA1 Region, Hippocampal; Carotid Artery, Common; Carotid Stenosis; Chronic Disease; Corpus Striatum; Disease Models, Animal; Functional Laterality; Ligation; Male; Maze Learning; Myelin Basic Protein; Needles; Neurons; Neurosurgical Procedures; Rats, Sprague-Dawley; White Matter

Chronic cerebral hypoperfusion, a sustained modest reduction in cerebral blood flow, is associated with damage to myelinated axons and cognitive decline with ageing. Oligodendrocytes (the myelin producing cells) and their precursor cells (OPCs) may be vulnerable to the effects of hypoperfusion and in some forms of injury OPCs have the potential to respond and repair damage by increased proliferation and differentiation. Using a mouse model of cerebral hypoperfusion we have characterised the acute and long term responses of oligodendrocytes and OPCs to hypoperfusion in the corpus callosum. Following 3 days of hypoperfusion, numbers of OPCs and mature oligodendrocytes were significantly decreased compared to controls. However following 1 month of hypoperfusion, the OPC pool was restored and increased numbers of oligodendrocytes were observed. Assessment of proliferation using PCNA showed no significant differences between groups at either time point but showed reduced numbers of proliferating oligodendroglia at 3 days consistent with the loss of OPCs. Cumulative BrdU labelling experiments revealed higher numbers of proliferating cells in hypoperfused animals compared to controls and showed a proportion of these newly generated cells had differentiated into oligodendrocytes in a subset of animals. Expression of GPR17, a receptor important for the regulation of OPC differentiation following injury, was decreased following short term hypoperfusion. Despite changes to oligodendrocyte numbers there were no changes to the myelin sheath as revealed by ultrastructural assessment and fluoromyelin however axon-glial integrity was disrupted after both 3 days and 1 month hypoperfusion. Taken together, our results demonstrate the initial vulnerability of oligodendroglial pools to modest reductions in blood flow and highlight the regenerative capacity of these cells.

Topics: Animals; Antigens; Axons; Blotting, Western; Brain Ischemia; Cell Count; Cell Differentiation; Cell Proliferation; Cerebrovascular Circulation; Chronic Disease; Corpus Callosum; Disease Models, Animal; Immunohistochemistry; Male; Mice; Mice, Inbred C57BL; Microscopy, Confocal; Myelin Basic Protein; Myelin Sheath; Nerve Tissue Proteins; Neural Stem Cells; Neuroglia; Oligodendroglia; Proliferating Cell Nuclear Antigen; Proteoglycans; Receptors, G-Protein-Coupled; Time Factors

Progesterone plays a protective role in the experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS). Besides spinal cord neuropathology, MS patients present a dysfunctional hippocampus. In this work we studied the therapeutic effects of the progestin Nestorone in the brain of mice with chronic EAE. Nestorone decreased clinical grade and enhanced motor behavior. In addition, it increased cell proliferation and doublecortin positive neuroblasts in the hippocampus, increased GABAergic interneurons and attenuated the number of Iba1+ microglia/macrophages, events possibly linked to enhancement of neurogenesis. Therefore, Nestorone protected against hippocampus abnormalities and improved functional outcomes of EAE mice, suggesting its potential value for MS.

Topics: Animals; Anti-Inflammatory Agents; Brain; Calcium-Binding Proteins; Chronic Disease; Doublecortin Domain Proteins; Encephalomyelitis, Autoimmune, Experimental; Female; Freund's Adjuvant; Glial Fibrillary Acidic Protein; Ki-67 Antigen; Macrophages; Mice; Mice, Inbred C57BL; Microfilament Proteins; Microglia; Microtubule-Associated Proteins; Motor Activity; Myelin Basic Protein; Myelin-Oligodendrocyte Glycoprotein; Neuropeptides; Norprogesterones; Peptide Fragments; Phosphopyruvate Hydratase

Previous studies have shown the existence of either cellular or humoral MBP-reactive elements up to 5 years after spinal cord injury (SCI), but not the presence of both after 10 years.. Twelve SCI patients, with more than 10 years of evolution, and 18 healthy blood donors were studied. Lymphocyte proliferation (colorimetric-BrdU ELISA assay) and antibody titers against MBP (ELISA Human IgG MBP-specific assay) were assessed.. SCI patients presented a significant T-cell proliferation against MBP (lymphocyte proliferation index: 3.7 ± 1.5, mean ± SD) compared to control individuals (0.7 ± 0.3; P < 0.001). Humoral response analysis yielded a significant difference (P < 0.0001) between the antibody titers of controls and SCI patients. A significant correlation between cellular and humoral responses was observed. Finally, patients with an ASIA B presented the highest immune responses.. This work demonstrates, for the first time, the existence of both cellular and humoral responses against MBP in the chronic stages (>10 years) of injury.

Topics: Adult; Antibodies; Case-Control Studies; Cell Proliferation; Chronic Disease; Female; Humans; Immunity, Humoral; Longitudinal Studies; Male; Middle Aged; Myelin Basic Protein; Paraplegia; Spinal Cord Injuries; T-Lymphocytes; Time Factors

Chronic hypoxia in rodents induces white matter (WM) injury similar to that in human preterm infants. We used diffusion tensor imaging (DTI) and immunohistochemistry to study the impact of hypoxia in the immature ferret at two developmental time points relevant to the preterm and term brain.. On ex vivo imaging, the apparent diffusion coefficient (ADC) was decreased throughout the WM after 10 days of hypoxia (hypoxia from postnatal day 10 (P10) to P20 and killed at P20 (early hypoxia P20)), corresponding to increased astrocytosis and decreased myelination. Diffusion values normalized after 10 days of normoxia (hypoxia from P10 to P20 and killed at P30 (early hypoxia P30)), but immunohistochemistry revealed significant astrocytosis and hypomyelination. In contrast, ADC and anisotropy were increased after 10 days of hypoxia at a later developmental time point (hypoxia from P20 to P30 and killed at P30 (late hypoxia P30)), with less astrocytosis and more prominent myelination.. The patterns of alteration in imaging and histology varied in relation to the developmental time at which hypoxia occurred. Normalization of diffusion measures did not correspond to the normalization of underlying histopathology.. Ferrets were subjected to 10% hypoxia and divided into three groups: early hypoxia P20, early hypoxia P30, and late hypoxia P30.

Topics: Age Factors; Animals; Astrocytes; Biomarkers; Brain; Chronic Disease; Demyelinating Diseases; Diffusion Tensor Imaging; Disease Models, Animal; Ferrets; Glial Fibrillary Acidic Protein; Hypoxia, Brain; Immunohistochemistry; Leukoencephalopathies; Myelin Basic Protein

To examine chronic changes occurring at 6 months following partial optic nerve (ON) transection, assessing optic axons, myelin, and visual function.. Dorsal ON axons were transected, leaving ventral optic axons vulnerable to secondary degeneration. At 3 and 6 months following partial transection, toluidine-blue stained sections were used to assess dimensions of the ON injury site. Transmission electron microscopy (TEM) images of ventral ON were used to quantify numbers, diameter, area, and myelin thickness of optic axons. Immunohistochemistry and fluoromyelin staining were used to assess semiquantitatively myelin protein, lipids in ventral ON, and retinal ganglion cells (RGCs) in midventral retina. Visuomotor function was assessed using optokinetic nystagmus.. Following partial ON transection, optic axons and function remained disrupted at 6 months. Although ventral ON swelling observed at 3 months (P ≤ 0.05) receded to normal by 6 months, ultrastructurally, myelinated axons remained swollen (P ≥ 0.05), and myelin thickness increased (P ≤ 0.05) due to loosening of lamellae and an increase in the number of intraperiodic lines. Axons with decompacted myelin persisted and were distinguished as having large axonal calibers and thicker myelin sheaths. Nevertheless, progressive loss of myelin lipid staining with fluoromyelin was seen at 6 months. Despite no further loss of ventral optic axons between 3 and 6 months (P ≥ 0.05), visuomotor function progressively declined at 6 months following partial transection (P ≤ 0.05).. Continued decompaction of myelin, altered myelin structure, and swelling of myelinated axons are persistent features of the chronic phases of secondary degeneration and likely contribute to progressive loss of visual function.

Topics: Animals; Axons; Chronic Disease; Female; Fluorescent Antibody Technique, Indirect; Microscopy, Electron, Transmission; Myelin Basic Protein; Myelin Sheath; Nerve Degeneration; Nystagmus, Optokinetic; Optic Nerve; Optic Nerve Diseases; Optic Nerve Injuries; Rats; Retinal Ganglion Cells; Tubulin

Although the white matter lesions, so called leuko-araiosis, often seen in elderly people have been gaining attention due to their association with cognitive dysfunction (CD) and high risk of incident stroke, the pathological significance of these lesions still remains controversial. Therefore, in the present study, we investigated the alterations in oligodendrocytes (OLG), including oligodendrocytes progenitor cells (OPCs), myelin, and CD following chronic cerebral ischemia in rats. SD rats were subjected to bilateral common carotid artery occlusion. Immunohistochemical staining was performed at 2, 4, 6, 8, and 12weeks after the induction of ischemia with anti-NG2 (OPCs), anti-GST-π (OLG), and anti-MBP antibodies in paramedian corpus callosum (CC). CD was assessed by the Morris water maze test. There was a significant decrease in the number of GST-π positive cells at 2weeks after the start of ischemia compared with that seen in the sham group. There was a significant increase of the number of NG2 positive cells at 4weeks in the ischemia group compared with the sham group. In the ischemic group, the amount of MBP was observed to have decreased significantly at each time point compared with the sham group. CD was observed in the ischemic group than that in the sham group at all time points. Our results indicate that remyelination is strongly correlated with the recovery of cognitive dysfunction following chronic cerebral ischemia.

Topics: Analysis of Variance; Animals; Antigens; Brain Ischemia; Bromodeoxyuridine; Cell Count; Chronic Disease; Cognition Disorders; Corpus Callosum; Disease Models, Animal; Gene Expression Regulation; Maze Learning; Myelin Basic Protein; Myelin Sheath; Oligodendroglia; Proteoglycans; Rats; Rats, Sprague-Dawley; Receptor, Platelet-Derived Growth Factor alpha; Time Factors

Myelinated axons have a distinct protein architecture essential for action potential propagation, neuronal communication, and maintaining cognitive function. Damage to myelinated axons, associated with cerebral hypoperfusion, contributes to age-related cognitive decline. We sought to determine early alterations in the protein architecture of myelinated axons and potential mechanisms after hypoperfusion. Using a mouse model of hypoperfusion, we assessed changes in proteins critical to the maintenance of paranodes, nodes of Ranvier, axon-glial integrity, axons, and myelin by confocal laser scanning microscopy. As early as 3 d after hypoperfusion, the paranodal septate-like junctions were damaged. This was marked by a progressive reduction of paranodal Neurofascin signal and a loss of septate-like junctions. Concurrent with paranodal disruption, there was a significant increase in nodal length, identified by Nav1.6 staining, with hypoperfusion. Disruption of axon-glial integrity was also determined after hypoperfusion by changes in the spatial distribution of myelin-associated glycoprotein staining. These nodal/paranodal changes were more pronounced after 1 month of hypoperfusion. In contrast, the nodal anchoring proteins AnkyrinG and Neurofascin 186 were unchanged and there were no overt changes in axonal and myelin integrity with hypoperfusion. A microarray analysis of white matter samples indicated that there were significant alterations in 129 genes. Subsequent analysis indicated alterations in biological pathways, including inflammatory responses, cytokine-cytokine receptor interactions, blood vessel development, and cell proliferation processes. Our results demonstrate that hypoperfusion leads to a rapid disruption of key proteins critical to the stability of the axon-glial connection that is mediated by a diversity of molecular events.

Topics: Age Factors; Animals; Ankyrins; Axons; Cell Adhesion Molecules; Cell Adhesion Molecules, Neuronal; Chronic Disease; Corpus Callosum; Disease Models, Animal; Electron Microscope Tomography; Gene Expression Profiling; Gene Expression Regulation; Hypoxia-Ischemia, Brain; Male; Mice; Mice, Inbred C57BL; Microscopy, Confocal; Myelin Basic Protein; Myelin-Associated Glycoprotein; NAV1.6 Voltage-Gated Sodium Channel; Nerve Fibers, Myelinated; Nerve Growth Factors; Nerve Tissue Proteins; Neurofilament Proteins; Neuroglia; Neurons; Oligonucleotide Array Sequence Analysis; Optic Nerve; Ranvier's Nodes; Signal Transduction; Sodium Channels

Depressive behavior in animals is often associated with reduced levels of brain-derived neurotrophic factor (BDNF) and impaired neurogenesis in the hippocampus. Recent studies showed that T cells recognizing central nervous system (CNS)-specific antigens can regulate adult hippocampal neurogenesis and expression of BDNF. On the basis of these findings, we hypothesized that controlling CNS specific immune activity by immunization with a myelin-related peptide may have an antidepressant effect.. We investigated the impact of immunization with a CNS related peptide, on the behavioral and cellular outcomes of chronic mild stress (CMS; an animal model for depression) in rats.. Immunization with a weak agonist of a myelin-derived peptide ameliorated depressive behavior such as anhedonia (measured by sucrose preference), induced by CMS in rats. The behavioral outcome was accompanied by restoration of hippocampal BDNF levels and neurogenesis.. The results of this study introduce a novel approach of immunization with CNS-related antigens as a therapeutic means for fighting depression. Vaccination, as an antidepressant therapy, may invoke several molecular and cellular pathways that are known to be regulated by antidepressant drugs. Therefore, we suggest that immune-based therapies should be considered for treatment of depression.

Topics: Animals; Antimetabolites; Brain-Derived Neurotrophic Factor; Bromodeoxyuridine; Chronic Disease; Depressive Disorder; Enzyme-Linked Immunosorbent Assay; Exploratory Behavior; Food Preferences; Immunization; Immunohistochemistry; Motor Activity; Myelin Basic Protein; Myelin Proteins; Rats; Rats, Inbred Lew; Rats, Sprague-Dawley; Stress, Psychological; Swimming; Vaccination

CD8+ macrophages appear in the central nervous system (CNS) under various pathological conditions such as trauma and ischemia. Furthermore, macrophages expressing CD8 were found in CNS lesions of chronic, but not acute, experimental autoimmune encephalomyelitis (EAE). To further characterize cells with this phenotype, we examined CD8+ macrophages/monocytes in the CNS and peripheral organs during the course of acute and chronic EAE that had been induced by immunization of rats with myelin basic protein and myelin oligodendrocyte glycoprotein, respectively. Counting CD8+ macrophages in CNS lesions revealed that their numbers increased reaching about 60% of total infiltrating macrophages in chronic EAE, while CD8+ macrophages remained less than 5% throughout the course of acute EAE. Unexpectedly, however, higher abundance of CD8+ monocytes/macrophages in the peripheral blood was found in both acute and chronic EAE. Real-time polymerase chain reaction analysis revealed no significant difference in the levels of chemokines and chemokine receptors of blood CD8+ monocytes between acute and chronic EAE. mRNA expression of perforin, a cytotoxic substance, was up-regulated in CD8+ monocytes compared with that of CD8- monocytes in both acute and chronic EAE. These findings suggest that activated CD8+ macrophages may play a cytotoxic role in chronic EAE lesions and that cells other than CD8+ monocytes/macrophages determined the difference in CNS pathology between acute and chronic EAE. Analysis of CD8+ monocytes/macrophages may provide useful information to permit further dissect the pathomechanisms of multiple sclerosis and to develop effective immunotherapies against autoimmune diseases in the CNS.

Topics: Animals; CD8 Antigens; Chemokines; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Lymph Nodes; Macrophages; Monocytes; Myelin Basic Protein; Myelin Proteins; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Perforin; Polymerase Chain Reaction; Rats; Rats, Transgenic; Receptors, Chemokine; RNA, Messenger; Spinal Cord; Spleen; Up-Regulation

MBP-PLP fusion protein (MP4)-induced experimental autoimmune encephalomyelitis (EAE) is a model for multiple sclerosis (MS) that encompasses both a time-dependent attack on central nervous system (CNS) regions and a B cell component, mirroring important features of human multiple sclerosis. Comparing C57BL/6 with B6.129 mice immunized with MP4, we point out similarities regarding these hallmarks and thus propose that they are largely dependent on the nature of the MP4 antigen itself, while differences between the two strains suggest that additional fine-tuning is brought about by the genetic repertoire of the animal. Overall, our data imply that (i) the interplay between both the antigenic trigger and genetic variables can define the outcome of MP4-induced autoimmune encephalomyelitis in C57BL/6 and B6.129 mice and (ii) that MP4 is not only a strong neuroantigen when it comes to reproducing the dynamics in effector mechanisms as is typical of the disease but also a promising agent for studying interindividual heterogeneity derived from genetic diversity in EAE/MS.

Topics: Animals; Antigenic Variation; Antigens; B-Lymphocytes; CD8-Positive T-Lymphocytes; Central Nervous System; Chronic Disease; Cytokines; Encephalomyelitis, Autoimmune, Experimental; Genetic Predisposition to Disease; Genetic Variation; Humans; Mice; Mice, Inbred C57BL; Myelin Basic Protein; Myelin Proteolipid Protein; Recombinant Fusion Proteins

Immunohistochemistry was used to examine the expression of prostaglandin E(2) receptors EP1 and EP4 in sciatic nerves from the rat chronic constriction injury (CCI) model of neuropathic pain. At 21 days post-surgery the CCI rats had developed mechanical hyperalgesia on the operated side, and quantitative image analysis showed a highly significant doubling of the area occupied by EP1- and EP4-positive pixels in sections from CCI nerves when compared to sham-operated controls. Co-localisation studies with the marker ED1 revealed that 73% of the EP1-positive cells and 54% of the EP4-positive cells in the injured nerves represented infiltrating macrophages. Cells negative for ED1 and positive for either EP1 or EP4 were characterised as Schwann cells from their morphology and expression of myelin basic protein and S100 antigens. Similar EP1- and EP4-positive Schwann cell profiles were observed in sections of uninjured control nerves. Low levels of EP receptor expression were found in neurofilament-immunostained axons, but no consistent differences were observed in the levels of axonal EP staining between CCI and control tissue. These data provide further evidence of the importance of prostaglandins in the pathogenesis of neuropathic pain, and suggest that not only infiltrating macrophages but also Schwann cells may be involved in the modulation of these mediators in response to nerve injury.

Topics: Animals; Axons; Chemotaxis, Leukocyte; Chronic Disease; Disease Models, Animal; Immunohistochemistry; Inflammation; Ligation; Macrophages; Male; Myelin Basic Protein; Neuralgia; Peripheral Nervous System Diseases; Prostaglandins; Rats; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP1 Subtype; Receptors, Prostaglandin E, EP4 Subtype; S100 Proteins; Schwann Cells; Sciatic Neuropathy; Up-Regulation

Chronic exposure to the copper-chelating agent, cuprizone (CPZ), is an increasingly popular model for producing demyelination. More importantly, cessation of cuprizone exposure allows for full remyelination, which represents a window of opportunity for determining the influence of environmental factors on regenerative processes. In the present study, CPZ-treated animals were assessed for functional status of systemic and central cytokine responsiveness to LPS, as well as assessment for signs of body weight changes. Exposure of male C57BL/6J mice to 5 weeks of 0.2% CPZ in the diet was optimal in producing demyelination and microglial activation, as measured by myelin basic protein, CD11b, and CD45 immunohistochemistry. Acute challenge with LPS at the end of 5 weeks CPZ treatment did not alter IL-1beta, IL-6, nor TNFalpha responses in the spleen and corpus callosum. Similarly, repeated exposure to LPS during the remyelination phase (CPZ removal) did not influence these measures to LPS. Plasma corticosterone was unaffected following acute challenge of CPZ-pretreated animals, but after repeated LPS treatment, there was a significant augmentation of the corticosterone response in CPZ-pretreated mice. Interestingly, the basal concentration of IL-1beta in the corpus callosum of CPZ treated animals was significantly increased, which was in keeping with the increase in activated microglial cells. In conclusion, the cuprizone model of demyelination and remyelination does not appear to influence the systemic nor central IL-1, IL-6, and TNF responses to acute nor repeated LPS. This opens up the possibility for studying the contribution of systemic inflammatory processes on remyelination after cessation of CPZ treatment.

Topics: Acute Disease; Animals; Body Weight; CD11b Antigen; Chelating Agents; Chronic Disease; Corpus Callosum; Corticosterone; Cuprizone; Cytokines; Demyelinating Diseases; Disease Models, Animal; Leukocyte Common Antigens; Lipopolysaccharides; Male; Mice; Mice, Inbred C57BL; Microglia; Myelin Basic Protein; Myelin Sheath; Nerve Regeneration; Spleen

Local inflammation may be a precipitating event in autoimmune processes. In this study, we demonstrate that regulated influx of monocytes and dendritic cells (DC) into the CNS causes an acute neurological syndrome that results in a demyelinating encephalomyelitis. Expansion of monocytes and DC by conditional expression of Flt3 ligand in animals expressing CCL2 in the CNS promoted parenchymal cell infiltration and ascending paralysis in 100% of the mice within 9 days of Flt3 ligand induction. Depletion of circulating monocytes and DC reduced disease incidence and severity. Unlike the classical models of experimental autoimmune encephalomyelitis, depletion of CD4+ and CD8+ T cells did not affect disease induction. T cells and demyelinating lesions were observed in the CNS at a later stage as a result of organ-specific inflammation. We propose that alterations in the numbers or function of monocytes and DC coupled to dysregulated expression of chemokines in the neural tissues, favors development of CNS autoimmune disease.

Topics: Acute Disease; Animals; Cell Movement; Chemokine CCL2; Chronic Disease; Dendritic Cells; Doxycycline; Encephalomyelitis, Autoimmune, Experimental; Leukocyte Count; Ligands; Membrane Proteins; Meningoencephalitis; Mice; Mice, Transgenic; Monocytes; Myelin Basic Protein; Spinal Cord

In humans, studies of autoreactive T cells that mediate multiple sclerosis have been largely confined to testing peripheral blood lymphocytes. Little is known how such measurements reflect the disease-mediating autoreactive T cells in the CNS. This information is also not available for murine experimental allergic encephalomyelitis (EAE); the low number of T cells that can be obtained from the blood or the brain of mice prevented such comparisons. We used single-cell resolution IFN-gamma ELISPOT assays to measure the frequencies and functional avidities of myelin basic protein (MBP:87-99)-specific CD4 cells in SJL mice immunized with this peptide. Functional MBP:87-99-specific IFN-gamma-producing cells were present in the CNS during clinical signs of EAE, but not during phases of recovery. In contrast, MBP:87-99-specific T cells persisted in the blood during all stages of the disease, and were also present in mice that did not develop EAE. Therefore, the increased frequency of MBP:87-99-reactive T cells in the blood reliably reflected the primed state, but not the inflammatory activity of these cells in the brain. The functional avidity of the MBP:87-99-reactive T cells was identical in the brain and blood and did not change over 2 mo as the mice progressed from acute to chronic EAE. Therefore, high-affinity T cells did not become selectively enriched in the target organ, and avidity maturation of the MBP:87-99-specific T cell repertoire did not occur in the observation period. The data may help the interpretation of measurements made with peripheral blood lymphocytes of multiple sclerosis patients.

Topics: Acute Disease; Animals; Autoantigens; Autoimmunity; Central Nervous System; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Humans; Interferon-gamma; Lymph Nodes; Male; Mice; Myelin Basic Protein; Peptide Fragments; Spleen; T-Lymphocyte Subsets; Time Factors

To elucidate the pathomechanisms of relapses of autoimmune disorders and to develop immunotherapy against relapses, we induced acute monophasic and chronic relapsing (CR) experimental autoimmune encephalomyelitis (EAE) in DA rats. Immunopathological and cytokine-chemokine analyses demonstrated that the number of infiltrating macrophages was significantly elevated in the CR-EAE than in acute EAE lesions and that IFN-gamma and IP-10 in the spinal cord were significantly upregulated during the first attack and relapse of CR-EAE, respectively, than at the peak of acute EAE. In vivo administration of decoy chemokine receptor plasmid DNAs encoding the binding sites of CXCR3 and CCR2 suppressed the development of relapse of CR-EAE. Importantly, multiple injections of DNAs did not elicit the antibody production against chemokine receptors. Taken together, these findings demonstrated that neutralization therapy with decoy chemokine receptor DNAs is effective to control autoimmune diseases.

Topics: Acute Disease; Animals; Antigens; Cell Movement; Cell Proliferation; Central Nervous System; Chemokines; Chronic Disease; Cytokines; DNA; Down-Regulation; Encephalomyelitis, Autoimmune, Experimental; Genetic Therapy; Immunization; Immunotherapy; Macrophages; Myelin Basic Protein; Myelin Proteins; Myelin Sheath; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Rats; Rats, Inbred Strains; Receptors, Chemokine; Recurrence; RNA, Messenger; T-Lymphocytes

Findings are that progression of chronic manganese intoxication comes along with development of humoral immune response to antibodies of myelinic coating of nerve fibers. That is expressed by increased occurrence of reliable titers of antibodies to basic myeloprotein G and increased serum IL-6 level. The authors suggest using determination of serum antibodies to myeloprotein G and IL-1, IL-6 to specify severity of chronic manganese intoxication.

Topics: Adult; Antibodies; Biomarkers; Chronic Disease; Cytokines; Disease Progression; Humans; Interleukin-1; Interleukin-6; Male; Manganese Poisoning; Middle Aged; Myelin Basic Protein; Myelin Sheath; Occupational Diseases; Welding

Experimental allergic encephalomyelitis (EAE) is the animal model for multiple sclerosis. The present study underlines the importance of sodium phenylacetate (NaPA), a drug approved for urea cycle disorders, in inhibiting the disease process of adoptively transferred EAE in female SJL/J mice at multiple steps. Myelin basic protein (MBP)-primed T cells alone induced the expression of NO synthase (iNOS) and the activation of NF-kappaB in mouse microglial cells through cell-cell contact. However, pretreatment of MBP-primed T cells with NaPA markedly inhibited its ability to induce microglial expression of iNOS and activation of NF-kappaB. Consistently, adoptive transfer of MBP-primed T cells, but not that of NaPA-pretreated MBP-primed T cells, induced the clinical symptoms of EAE in female SJL/J mice. Furthermore, MBP-primed T cells isolated from NaPA-treated donor mice were also less efficient than MBP-primed T cells isolated from normal donor mice in inducing iNOS in microglial cells and transferring EAE to recipient mice. Interestingly, clinical symptoms of EAE were much less in mice receiving NaPA through drinking water than those without NaPA. Similar to NaPA, sodium phenylbutyrate, a chemically synthesized precursor of NaPA, also inhibited the disease process of EAE. Histological and immunocytochemical analysis showed that NaPA inhibited EAE-induced spinal cord mononuclear cell invasion and normalized iNOS, nitrotyrosine, and p65 (the RelA subunit of NF-kappaB) expression within the spinal cord. Taken together, our results raise the possibility that NaPA or sodium phenylbutyrate taken through drinking water or milk may reduce the observed neuroinflammation and disease process in multiple sclerosis patients.

Topics: Acute Disease; Administration, Oral; Adoptive Transfer; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Differentiation; Cell Line; Cell Movement; Chronic Disease; Disease Progression; Encephalomyelitis, Autoimmune, Experimental; Enzyme Inhibitors; Female; Growth Inhibitors; Immunosuppressive Agents; Injections, Intraperitoneal; Lymphocyte Activation; Mice; Mice, Inbred Strains; Microglia; Myelin Basic Protein; NF-kappa B; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Phenylacetates; Phenylbutyrates; Severity of Illness Index; Spinal Cord; T-Lymphocyte Subsets; Transcription Factor RelA; Tyrosine

Since the basic mechanisms behind the beneficial effects of IFN-beta in multiple sclerosis (MS) patients are still obscure, here we have investigated the effects of IFN-beta gene disruption on the commonly used animal model for MS, experimental autoimmune encephalomyelitis (EAE). We show that IFN-beta knockout (KO) mice are more susceptible to EAE than their wild-type (wt) littermates; they develop more severe and chronic neurological symptoms with more extensive CNS inflammation and demyelination. However, there was no discrepancy observed between wt and KO mice regarding the capacity of T cells to proliferate or produce IFN-gamma in response to recall Ag. Consequently, we addressed the effect of IFN-beta on encephalitogenic T cell development and the disease initiation phase by passive transfer of autoreactive T cells from KO or wt littermates to both groups of mice. Interestingly, IFN-beta KO mice acquired a higher incidence and augmented EAE regardless of the source of T cells. This shows that the anti-inflammatory effect of endogenous IFN-beta is predominantly exerted on the effector phase of the disease. Histopathological investigations of CNS in the effector phase revealed an extensive microglia activation and TNF-alpha production in IFN-beta KO mice; this was virtually absent in wt littermates. This coincided with an increase in effector functions of T cells in IFN-beta KO mice, as measured by IFN-gamma and IL-4 production. We suggest that lack of endogenous IFN-beta in CNS leads to augmented microglia activation, resulting in a sustained inflammation, cytokine production, and tissue damage with consequent chronic neurological deficits.

Topics: Adjuvants, Immunologic; Adoptive Transfer; Animals; Autoantibodies; Autoantigens; Cells, Cultured; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Gene Deletion; Genetic Predisposition to Disease; Immunophenotyping; Incidence; Inflammation; Interferon-beta; Macrophage Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Myelin Basic Protein; Myelin Sheath; Peptide Fragments; Severity of Illness Index; T-Lymphocyte Subsets; Th1 Cells; Th2 Cells

The relationship between autoreactivity to myelin antigens and disease progression in multiple sclerosis (MS) is not fully understood. We addressed this relationship by cross-sectionally comparing an objective measure of MS disability with immune cytokine responses to myelin proteins. The ELISPOT assay was used to determine the ex vivo interferon gamma (IFNgamma) and interleukin-10 (IL-10) production by peripheral blood mononuclear cells (PBMCs) in response to peptides spanning the entire proteolipid protein (PLP) and myelin basic protein (MBP) molecules in 20 patients with relapsing-remitting (RR) MS and 27 age- and sex-matched healthy controls. MS patients showed significantly higher MBP-induced IFNgamma responses and PLP-induced IL-10 responses compared with healthy controls. Using the Multiple Sclerosis Functional Composite (MSFC), a new multifactorial measure of disability, MS patients showed a significant correlation between the IFNgamma response to PLP peptides and MBP peptides, and disability. In contrast, in MS patients, there was no correlation between the MSFC and the response to unrelated control antigens or mitogens. These data show that myelin-specific T lymphocytes secreting the inflammatory cytokine IFNgamma correlate with functional impairment in MS, supporting an antigen-specific link between the immune response to myelin and disability in MS.

Topics: Adolescent; Adult; Cells, Cultured; Chronic Disease; Cross-Sectional Studies; Cytokines; Disability Evaluation; Disease Progression; Female; Humans; Interferon-gamma; Interleukin-10; Lymphocyte Activation; Magnetic Resonance Imaging; Male; Middle Aged; Multiple Sclerosis, Relapsing-Remitting; Myelin Basic Protein; Myelin Proteolipid Protein; Peptide Fragments; T-Lymphocytes; Up-Regulation

Functional loss after spinal cord injury (SCI) is caused, in part, by demyelination of axons surviving the trauma. Administration of guanosine (8 mg/kg/day, i.p.) for 7 consecutive days, starting 5 weeks after moderate SCI in rats, improved locomotor function and spinal cord remyelination. Myelinogenesis was associated with an increase in the number of mature oligodendrocytes detected in guanosine-treated spinal cord sections in comparison with controls. These data indicate that guanosine-induced remyelination resulted, at least in part, from activation of endogenous oligodendrocyte lineage cells. These findings may have significant implications for chronic demyelinating diseases.

Topics: Animals; Chronic Disease; Female; Guanosine; Motor Activity; Myelin Basic Protein; Myelin Sheath; Nerve Fibers, Myelinated; Rats; Rats, Wistar; Recovery of Function; Spinal Cord Injuries

Cerebral white matter (WM) lesions are observed frequently in human ischemic cerebrovascular disease and have been thought to contribute to cognitive impairment. This type of lesion can be experimentally induced in rat brains under chronic cerebral hypoperfusion by the permanent occlusion of both common carotid arteries. However, it remains uncertain whether chronic ischemia can damage both the gray and white matter, and whether it can induce demyelination with or without axonal damage. Therefore, we examined axonal damage using immunohistochemistry for the amyloid beta/A4 precursor protein (APP), chromogranin A (CgA) and demyelination using immunohistochemistry for the encephalitogenic peptide (EP) in this model. Severe WM lesions such as vacuolation and the loss of nerve fibers appeared in the optic nerve and optic tract after 3 days of ligation, and less intense changes were observed in the corpus callosum, internal capsule, and fiber bundles of the caudoputamen after 7 days with Klüver-Barrera and Bielschowsky staining. These WM lesions persisted even after 30 days. The APP, CgA, and EP-immunopositive fibers increased in number from 1 to 30 days after the ligation in the following WM regions: the optic nerve, optic tract, corpus callosum, internal capsule, and fiber bundles of the caudoputamen. In contrast, only a few APP, CgA, or EP-immunopositive fibers were detected in the gray matter regions, including the cerebral cortex and hippocampus. These results indicate that the WM is more susceptible to chronic cerebral hypoperfusion than the gray matter, with an involvement of both axonal and myelin components. Furthermore, immunohistochemistry for APP, CgA, and EP is far superior to routine histological staining in sensitivity and may become a useful tool to investigate WM lesions caused by various pathoetiologies.

Topics: Amyloid beta-Protein Precursor; Animals; Axons; Brain Ischemia; Cerebral Infarction; Chromogranin A; Chromogranins; Chronic Disease; Demyelinating Diseases; Immunohistochemistry; Male; Myelin Basic Protein; Nerve Fibers, Myelinated; Peptide Fragments; Prosencephalon; Rats; Rats, Wistar; Wallerian Degeneration

It has been suggested that Golli proteins, structurally related to myelin basic proteins (MBPs), have a role in autoimmune processes. We studied the expression of these proteins in multiple sclerosis (MS) and determined that the number of Golli-immunoreactive (ir) cells was significantly higher around lesions of chronic MS than in control white matter. Golli proteins were expressed in the adult oligodendrocyte precursor cells (OPCs), activated microglia/macrophages, and some demyelinated axons around MS lesions. Their expression in adult OPCs indicates remyelination attempts, whereas the expression in the subpopulation of microglia/macrophages suggests roles in the immune processes of MS. In addition, Golli proteins may be markers of axonal transection, which is characteristic for MS.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Brain; Brain Chemistry; Chronic Disease; Female; Humans; Immunohistochemistry; Macrophages; Male; Microglia; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Nerve Tissue Proteins; Oligodendroglia; Stem Cells; Transcription Factors

Chronic ischemic brain injuries were studied in 7- and 14-day-old rat pups, which were subjected to bilateral carotid artery occlusion (BCAO) on postnatal day 1. BCAO preferentially injured white matter in the corpus callosum, subcortex and internal capsule areas while largely spared cortical neurons. White matter rarefaction in the corpus callosum was observed in 12 out of the 17 BCAO rat brains and significantly enlarged lateral ventricles were found in five out of seven P14 BCAO rat brains. These white matter changes were similar to injuries found in newborn infants with periventricular leukomalacia (PVL). White matter injuries in the 7-day-old BCAO rat brain were accompanied with increased activation of microglia/macrophages, as indicated by ED1 and OX42 positive immunostaining. Immature oligodendrocytes in the 7-day-old BCAO rat brain, as indicated by O4+/O1+ staining, were much fewer than in the sham-operated rat brain. Immunostaining for myelin basic protein (MBP) at the fimbria hippocampus and the internal capsule areas in the 7-day-old BACO rat brain was also much less than in the control rat brain. Consistent with the immunostaining data, MBP mRNA expression in the 7-day-old, but not in the 14-day-old, BCAO rat brain was significantly less than in the control rat brain. The overall results suggest that pre-oligodendrocytes and immature oligodendrocytes might be major targets for chronic ischemic insults and activated microglia/macrophages are possibly involved in the process of white matter injury.

Topics: Animals; Animals, Newborn; Biomarkers; Brain; Brain Ischemia; Chronic Disease; Immunohistochemistry; Macrophages; Microglia; Myelin Basic Protein; Oligodendroglia; Rats; Rats, Sprague-Dawley; RNA, Messenger; Staining and Labeling

Theiler's murine encephalomyelitis virus (TMEV) infection produces a chronic inflammatory disease of the spinal cord white matter, with striking similarities to both experimental allergic encephalomyelitis (EAE) and human multiple sclerosis (MS). The first phase of demyelination in this model appears to be dependent on a delayed-type hypersensitivity (DTH) response to viral antigens, driven by CD4+, Th1 lymphocytes. Macrophages, recruited in the infected CNS, would be responsible for most of the myelin damage. Recently, new populations of CD4+ lymphocytes were demonstrated in infected mice, this time with specificity for myelin antigens, particularly PLP. This suggests that, in the chronic phase of the disease, an autoimmune mechanism of demyelination, similar to EAE, may participate in the process of myelin destruction. The present study represents a first step in exploring the functional activity of these anti-myelin lymphocytes that emerge during the chronic phase of the disease. Lymphocytes were removed from chronically infected animals, they were stimulated with the major PLP encephalitogenic epitope for SJL/J mice, and they were added to organotypic myelinated spinal cord cultures for different lengths of time. Results show that lymphocytes stimulated with the major PLP epitope have a powerful capacity for demyelinating these cultures, while MBP stimulated lymphocytes and lymphocytes from control animals do not. This study, suggests that the anti-myelin response that emerges during the chronic phase of the infection is functionally active. A similar phenomenon of epitope spreading from virus to organ specific antigens may take place in humans and be involved in a number of immune-mediated diseases, including MS.

Topics: Animals; Cardiovirus Infections; Cells, Cultured; Chronic Disease; Demyelinating Diseases; Encephalitis; Epitopes; Immunization; Lymphocytes; Mice; Mice, Inbred Strains; Myelin Basic Protein; Myelin Proteolipid Protein; Myelin Sheath; Organ Culture Techniques; Ovalbumin; Theilovirus

This study was undertaken to better understand the role of cytokines in the pathogenesis, especially in the mechanisms of relapse, of experimental autoimmune encephalomyelitis (EAE). For this purpose, we induced acute and chronic relapsing (CR) EAE in DA rats and determined several immunological parameters in rats at various stages of two types of EAE. Histopathological analysis revealed that there was no significant difference in the severity of inflammation in the spinal cord lesions between the two groups. However, demyelination was observed only in rats with CR EAE. Cytokine analysis by competitive PCR demonstrated that levels of TNF-alpha, IL-6 and IL-12 p40 mRNA in the spinal cord at the first attack of CR EAE were significantly higher than those at the peak stage of acute EAE. The mRNA expression of anti-inflammatory cytokines, IL-10 and TGF-beta1, was generally low in both acute EAE and the first attack of CR EAE and upregulated at later stages of CR EAE. These findings suggest that persistent high-level expression of pro-inflammatory cytokines is closely associated with demyelination and relapse of EAE. In contrast, anti-inflammatory cytokines play only a minor role in the relapse.

Topics: Acute Disease; Amino Acid Sequence; Animals; Chronic Disease; Cytokines; Encephalomyelitis, Autoimmune, Experimental; Inflammation; Interferon-gamma; Interleukins; Molecular Sequence Data; Myelin Basic Protein; Peptide Fragments; Rats; Rats, Inbred Strains; Recurrence; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spinal Cord; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Topics: Animals; Antigenic Variation; Antigens, Viral; Autoantigens; Autoimmune Diseases; Autoimmunity; Chronic Disease; Disease Models, Animal; Disease Progression; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Humans; Immunity, Cellular; Immunization; Lymphocyte Subsets; Mice; Models, Immunological; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Self Tolerance

Experimental autoimmune encephalomyelitis (EAE) is a widely used animal model for multiple sclerosis (MS). EAE is typically initiated by CD4(+) T helper cell type 1 (Th1) autoreactivity directed against a single priming immunodominant myelin peptide determinant. Recent studies have shown that clinical progression of EAE involves the accumulation of neo-autoreactivity, commonly referred to as epitope spreading, directed against peptide determinants not involved in the priming process. This study directly addresses the relative roles of primary autoreactivity and secondary epitope spreading in the progression of both EAE and MS. To this end we serially evaluated the development of several epitope-spreading cascades in SWXJ mice primed with distinctly different encephalitogenic determinants of myelin proteolipid protein. In a series of analogous experiments, we examined the development of epitope spreading in patients with isolated monosymptomatic demyelinating syndrome as their disease progressed to clinically definite MS. Our results indicate that in both EAE and MS, primary proliferative autoreactivity associated with onset of clinical disease invariably regresses with time and is often undetectable during periods of disease progression. In contrast, the emergence of sustained secondary autoreactivity to spreading determinants is consistently associated with disease progression in both EAE and MS. Our results indicate that chronic progression of EAE and MS involves a shifting of autoreactivity from primary initiating self-determinants to defined cascades of secondary determinants that sustain the self-recognition process during disease progression.

Topics: Acute Disease; Adult; Animals; Antigenic Variation; Autoantigens; Autoimmune Diseases; Autoimmunity; Brain; Chronic Disease; Disease Models, Animal; Disease Progression; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Female; Humans; Immunity, Cellular; Immunization; Immunodominant Epitopes; Male; Mice; Models, Immunological; Multiple Sclerosis; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Myelin-Associated Glycoprotein; Myelin-Oligodendrocyte Glycoprotein; Peptide Fragments; Self Tolerance; Spinal Cord; Spleen; Th1 Cells

Myelin basic protein (BP)-specific T lymphocyte cell lines were selected from the lymph nodes (LN) of BP-immunized, H-2d, CXJ-1 mice prior to the onset of clinical disease. These CD4+ T cells induced severe acute experimental autoimmune encephalomyelitis (EAE) in MHC-compatible (H-2d), lymphocyte-deficient (SCID) mice (C.B-17scid/scid). The incidence of disease was much higher in immunodeficient SCID mice (71%) than in syngeneic immunocompetent CXJ-1 mice (5%). SCID mice with EAE had an acute progressive paralytic disease with inflammation and myelin loss detected in the spinal cord. Eighty-six percent (12/14) of mice followed for more than 2 weeks had 1 or more relapses of EAE. These results demonstrate that clinical remission and relapse of EAE can be induced by the single adoptive transfer of a LN-derived BP-specific T cell line in the absence of host-derived effector and regulatory lymphocytes. Furthermore, the data demonstrate that the pathogenic potential of BP-specific T cells is greater in lymphocyte-deficient SCID mice compared with immunocompetent mice, suggesting that autoreactive T cells are controlled by potent inhibitory mechanisms associated with regulatory lymphocytes. These results are relevant to mechanisms of disease remission and relapse mediated by lymphocytes involved in paralytic inflammatory diseases such as multiple sclerosis (MS).

Topics: Animals; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line; Cell Transplantation; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Female; Immunophenotyping; Leukocyte Common Antigens; Macrophage-1 Antigen; Mice; Mice, SCID; Myelin Basic Protein; Recurrence; Spinal Cord; Vaccination

An animal model of chronic cerebral hypoperfusion was developed with coiled clips applied to both carotid arteries of adult Mongolian gerbils for between 1 week and 2 months. In the brain of this animal model, rarefaction of white matter with dilatation of the ventricles was frequently observed. To better understand the mechanism of white matter alteration under cerebral hypoperfusion, the chronological sequence of molecular changes in the cerebral white matter of the animal model was determined.. Specially designed coiled clips were placed around both carotid arteries of Mongolian gerbils to create stenosis without occlusion. Changes in levels of myelin basic protein (MBP) as a marker of myelin, neurofilament H (NFH) as a marker of axonal proteins, and glial fibrillary acidic protein (GFAP) in astroglia after 2 months of cerebral hypoperfusion were analyzed with Western blotting and enzyme-linked immunosorbent assay.. Western blotting of the white matter after 2 months of hypoperfusion showed that the levels of MBP and NFH decreased, whereas that of GFAP increased. The time course of MBP and NFH changes determined with enzyme-linked immunosorbent assay revealed that the change of MBP preceded that of NFH.. In the present study it was shown that the damage to myelin precedes that to the axon in the white matter in a chronic cerebral hypoperfusion animal model, suggesting that the change in myelin is the primary pathological event in the cerebral white matter under chronic hypoperfusion. The present study may help in understanding the mechanisms of white matter pathology in leukoaraiosis.

Topics: Animals; Blotting, Western; Brain; Brain Chemistry; Brain Ischemia; Chronic Disease; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Gerbillinae; Glial Fibrillary Acidic Protein; Myelin Basic Protein; Neurofilament Proteins; Time Factors

Chronic relapsing experimental autoimmune encephalomyelitis (crEAE), a model for multiple sclerosis, was used to test 2 regimens of insulin-like growth factor-I (IGF-I) treatment. We induced crEAE by injecting 3x10(7) myelin basic protein-(MBP) sensitized lymph node cells into adult female SJL/J mice. Fifty-one mice, divided randomly into 4 groups, were used in the first trial. Two groups received IGF-I (a gift of Cephalon, Inc.) 0.6 mg/kg/d subcutaneously from day 7 to day 16 and the other two groups received placebo injections. IGF-I treatment reduced clinical deficits during the first attack and during 2 subsequent relapses. Image analysis of immunostained and histological sections showed that IGF-I treatment reduced BBB defects and both the numbers and sizes of inflammatory, demyelinating, and demyelinated lesions. Twelve mice that had recovered from their first attack were used in our second trial to evaluate possible adverse effects of prolonged treatment with a higher dose of IGF-I. Six received 1.2 mg/kg/d for 6 weeks (days 19-63). No adverse effects of IGF-I treatment were identified. The eyes, hearts, livers, and kidneys of IGF-I-treated mice were normal histologically and their spleens also appeared normal except for mild to moderate microscopic increases in lymphopoesis. Our results suggest that prolonged IGF-I treatment is well tolerated and that the anti-inflammatory effects of IGF-I have a major role in reducing clinical deficits and lesion severity in crEAE. These effects, if present in multiple sclerosis, may benefit patients with this disease.

Topics: Animals; Blood-Brain Barrier; Body Weight; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunohistochemistry; In Situ Hybridization; Insulin-Like Growth Factor I; Macrophages; Magnetic Resonance Imaging; Mice; Mice, Inbred Strains; Myelin Basic Protein; Myelin Sheath; Nerve Regeneration; Neuroglia; Recurrence; RNA Probes

Experimental autoimmune encephalomyelitis (EAE), a model for human multiple sclerosis, is an inflammatory disease of the CNS mediated by autoreactive T lymphocytes directed against the neuroantigen, myelin basic protein (MBP). EAE is inducible in the Lewis rat, which exhibits an acute monophasic disease, and in selected mouse strains, which show a remitting-relapsing or chronic course of paralysis. We examined the effects of neuroendocrine modulation by restraint stress on these models of EAE. In Lewis rats, daily cycles of restraint resulted in significant suppression of both clinical and histopathologic changes of EAE. Suppression of EAE was more pronounced in the female than in the male rat, which follows from the higher endogenous corticosterone levels in the female. Mechanistic studies suggested that stress affected the processing of MBP or the T-cell idiotype. In the relapsing murine model of EAE, B10.PL mice were restrained beginning either before MBP challenge or after the establishment of relapsing disease. We observed a striking inhibition of EAE clinical signs in mice stressed before challenge relative to nonstressed controls. Interestingly, approximately 10 days after termination of the stress period, clinical signs returned and were as severe or more severe than in control nonstressed animals. Stress administered after relapsing EAE was established had no protective effect. In vitro parameters revealed that only stress initiated before disease induction significantly reduced the frequency of MBP-specific lymphocytes in the spleen and lymph nodes. Both Th1 and Th2 cytokine responses were suppressed in stressed mice. T-cell receptor transgenic mice exposed to restraint showed a marked decreased in the number and functional activity of transgene-positive lymphocytes. In summary, elevated levels of endogenous neuroendocrine hormones exert a profoundly suppressive effect on both acute and chronic models of autoimmune CNS injury.

Topics: Acute Disease; Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Male; Mice; Mice, Inbred Strains; Mice, Transgenic; Myelin Basic Protein; Neurosecretory Systems; Rats; Rats, Inbred Lew; Receptors, Antigen, T-Cell; Recurrence

Stress disturbs homeostasis by altering the equilibrium of various hormones which have a significant impact on immune responses. Few studies have examined the influence of stressors on autoimmune disease in animal models. In our work, we studied the effects of long-term exposure (14 days) to chronic varied stress (CVS) in a model of experimental autoimmune encephalomyelitis (EAE) in Wistar rats. We studied whether the exposure to CVS before or after the immune challenge would correlate with differences in the clinical course of the disease. We also examined whether the CVS would modulate the magnitude of the cellular or the humoral immune response. We observed opposite effects on the clinical signs in animals stressed before or after the immune challenge. The clinical signs of the disease were attenuated in animals stressed before but not after the immune challenge. Relationships were found in the modulation of the clinical severity related to the time of exposure to the CVS, the histological alterations and the proliferative results. Stressed animals with milder clinical signs presented an exacerbated humoral response against myelin antigens while stressed animals with more severe clinical symptoms exhibited a significantly diminished one. Besides, we detected the presence of specific IgG1 associated with the exposure to CVS before the induction of EAE. Our results show that, depending on the timing of the exposure of Wistar rats to the CVS, the neuroendocrine disbalance favors a more pronounced humoral or cellular profile of the response.

Topics: Animals; Antibodies; Antibody Formation; Antigens; Cattle; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Enzyme-Linked Immunosorbent Assay; Female; Immunoglobulin G; Myelin Basic Protein; Myelin Sheath; Rats; Rats, Wistar; Stress, Physiological

Some autoimmune diseases are thought to arise after an infection. Infectious agents can initiate a chronic inflammatory response associated with autoimmune reactions. Chagas disease, caused by the intracellular parasite Trypanosoma cruzi, is an excellent model for autoimmune disease induced by an infection. The chronic disease is characterized by rich inflammatory infiltrate in myocardial and nervous tissues, with virtually no demonstrable parasites. We were able to demonstrate the presence of antibody to myelin basic protein (MBP) in the serum from T. cruzi chronically infected mice. Lymphocytes from mice immunized with T. cruzi-derived soluble extract antigen (TCSE) proliferate in response to MBP in vitro. Lymphocytes from animals immunized with MBP also were activated by TCSE in vitro. By studying the overlapping peptides from the MBP molecule, we were able to identify two regions responsible for the cross-reactivity.

Topics: Amino Acid Sequence; Animals; Antibodies, Protozoan; Antigens, Protozoan; Autoimmune Diseases; Chagas Disease; Chronic Disease; Cross Reactions; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Immunization; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred CBA; Molecular Sequence Data; Myelin Basic Protein; Peptide Fragments; Trypanosoma cruzi

During chronic relapsing experimental autoimmune encephalomyelitis (EAE), T lymphocytes specific for myelin protein epitopes are stimulated in vivo. When epitopes are unique from the disease-initiating myelin protein epitope, this phenomenon has been termed "epitope spreading". These T-lymphocyte responses have been detected primarily in lymph node and spleen during the relapsing phase of disease. If myelin proteins are sequestered behind the blood brain barrier, a fundamental question arises: where does the in vivo stimulation of T lymphocytes occur during relapsing EAE? While it has been thought that epitope spreading may occur within the central nervous system (CNS), here we present data supporting a novel hypothesis. Epitope spreading during EAE may not occur within the CNS, but rather within lymphoid tissues. Both myelin basic protein (MBP) and proteolipid protein (PLP) are expressed at the RNA and protein level in lymph node, thymus and spleen of SJL mice with relapsing EAE. This myelin protein expression occurs within T lymphocytes, B lymphocytes and macrophages. Further, T-lymphocyte lines from SJL mice specific for the immunodominant and subdominant epitopes of MBP and PLP can recognize endogenous protein within cells derived from lymphoid tissues. Thus, immunologically relevant myelin proteins are endogenously produced and presented within lymphoid tissues. The hypothesis that epitope spreading occurs within lymphoid tissues would explain how myelin protein-specific T lymphocytes become activated outside the CNS to allow their passage through the blood brain barrier to form new CNS lesions during relapses.

Topics: Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Immune Tolerance; Lymphoid Tissue; Mice; Models, Immunological; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Recurrence; T-Lymphocytes

Monitoring the TCR repertoire is indispensable for the assessment of T cell-associated autoimmune diseases and subsequent TCR-based immunotherapy. In the present study, we examined the TCR repertoire of spinal cord T cells of Lewis rats by CDR3 spectratyping during chronic relapsing experimental autoimmune encephalomyelitis (EAE) induced by immunization with spinal cord homogenate. It was found that Vbeta8.2 spectratype with the shortest CDR3 expanded oligoclonally throughout the course of the disease. In addition, Vbeta12 spectratype expansion was observed at the first and second attacks of EAE. Sequence analysis revealed that clones with the DSSYEQYF sequence, which is a representative sequence of myelin basic protein (MBP)-reactive T cell clones, constituted the predominant population in the Vbeta8.2 family. Surprisingly, Vbeta12 also used the identical amino acid sequence in the CDR3 region. These findings indicate that although infiltrating T cells in the central nervous system are activated polyclonally, the TCR repertoire remains unchanged throughout the course. Moreover, the finding that the predominant CDR3 amino acid sequence of Vbeta8.2 and Vbeta12 spectratypes is identical with that of MBP-induced EAE suggests that a single Ag in spinal cord homogenate, possibly MBP, is involved in disease development.

Topics: Animals; Autoimmune Diseases; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; Guinea Pigs; Myelin Basic Protein; Peptide Fragments; Polymerase Chain Reaction; Rats; Rats, Inbred Lew; Receptors, Antigen, T-Cell, alpha-beta; Recurrence; Sequence Alignment; Sequence Analysis, DNA; Spinal Cord

Central nervous system tissue from multiple sclerosis and non-multiple sclerosis subjects was studied for the expression of exon 2 myelin basic protein gene products at the protein and message levels by immunocytochemistry and in situ hybridization, respectively. The exon 2-encoded protein sequence is normally expressed during development (myelination) within the 21.5- and 20.2-kd isoforms of myelin basic protein and is downregulated in the adult central nervous system where the 18.5- and 17.2-kd isoforms predominate, the latter devoid of exon 2 owing to alternative splicing. Exon 2 myelin basic protein gene products were readily demonstrable in multiple sclerosis samples, the highest levels correlating with remyelination in chronic lesions while normal adult central nervous system and non-multiple sclerosis material showed very low levels and fetal human central nervous system tissue (a positive control) showed high levels. We conclude that recapitulation of ontogenetic events during myelin repair accounts for the increased expression of the exon 2-encoded protein sequence in the adult central nervous system during multiple sclerosis, an event that might underly the previously observed T-cell activation to this protein sequence during relapses.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Central Nervous System; Chronic Disease; Exons; Female; Fetus; Genes; Genes, Developmental; Humans; Immunohistochemistry; In Situ Hybridization; Male; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Myelin Sheath; Nervous System Diseases

We have studied a case of acute, fulminating multiple sclerosis (MS) (Marburg type) at the pathological and biochemical levels. Postmortem examination of the brain revealed extensive areas of gross rarefaction in the hemispheric white matter. Histologically, well-demarcated areas of demyelination with a large influx of macrophages and a subtle perivascular infiltration of lymphocytes were seen with relative preservation of the axis cylinders. Myelin basic protein (MBP) was isolated and purified [correction of purifed] from noninvolved white matter. It was slightly larger in molecular weight than MBP from normal brain or from chronic MS brain. The increase in mass was accounted for, in part, by the deimination of 18 of 19 arginyl residues to citrulline, making the patient's MBP much less cationic than MBP from normal white matter. When expressed as the ratio of least cationic form of MBP to the most cationic (C-8/C-1), the normal ratio was 0.82, chronic MS 2.5, and the patient in this study 6.7. Because the ratio of 6.7 was similar to 7.5 found for a 15-month-old infant, MBP was considered to be of the immature form. The data are consistent with a genetic factor influencing the charge microheterogeneity of MBP. The resulting less cationic MBP cannot carry out its normal function of compacting multilayers.

Topics: Adult; Antibodies, Monoclonal; Arginine; Blotting, Western; Brain; Brain Chemistry; Chronic Disease; Citrulline; Demyelinating Diseases; Fatal Outcome; Female; Humans; Lymphocytes; Macrophages; Magnetic Resonance Imaging; Multiple Sclerosis; Myelin Basic Protein

Murine chronic relapsing experimental autoimmune encephalomyelitis (CR-EAE) is a model of inflammatory demyelinating disease of the central nervous system (CNS) with similarity to multiple sclerosis (MS) in humans. Mice with confirmed neurologic deficits from CR-EAE were treated by oral administration of whole bovine myelin to investigate the effect of long-term oral delivery of myelin antigens on clinical disease and on the inflammatory response in the CNS. EAE-positive mice were fed doses of 1 mg, 10 mg, or 20 mg of bovine myelin every other day for 6 months. We found that prolonged oral delivery of neuroantigen suppressed inflammatory and demyelination foci in the CNS of myelin-treated mice with no exacerbation of clinical disease status compared with the control group. Analysis of histologic sections of brain and spinal cords with hematoxylin-eosin (H&E) and Luxol fast blue (LFB) staining showed a decrease in the inflammatory cell infiltration and active centers of demyelination, respectively. Furthermore, after 6 months of treatment, there was no increased sensitization to myelin antigens seen, as measured by antimyelin basic protein (MBP) or anti-proteolipid apoprotein (PLP) antibodies. These results demonstrate that prolonged oral administration of myelin antigens in diseased animals has an ameliorating effect on the pathologic process and supports its potential long-term use in humans with MS.

Topics: Administration, Oral; Animals; Antibody Formation; Cattle; Chronic Disease; Demyelinating Diseases; Disease Progression; Encephalomyelitis, Autoimmune, Experimental; Female; Mice; Mice, Inbred Strains; Myelin Basic Protein; Myelin Proteolipid Protein; Recurrence; Time Factors

Chronic relapsing experimental autoimmune encephalomyelitis (EAE), induced in mice by the injection of myelin basic protein (MBP), is a T cell-mediated autoimmune disease characterized by periods of paralysis and remission. We have shown previously that the oral administration of MBP or MBP peptides renders Lewis rats refractory to EAE. This study was undertaken to examine the conditions necessary to produce oral tolerance in a chronic relapsing model of EAE in B10.PL mice. The optimal tolerizing regimen for the mouse was found to be a single feeding of 20 mg of MBP suspended in PBS. To determine the ability to suppress chronic disease, a range of doses (0.4-100 mg) was administered orally in a single dose before challenge. Larger oral doses (20 or 100 mg) of MBP provided the best protection from EAE, while 0.4 mg exacerbated the clinical course of disease. Secretion of the proinflammatory cytokines, IL-2 and IFN-gamma, were lowest in the group fed 20 mg. A single feeding of MBP before challenge or as late as the first day of clinical signs showed significant protection over the relapsing disease course. Once relapsing EAE was established, multiple oral doses of MBP were required to achieve suppression of clinical signs of disease. These findings suggest that vehicle, dosage, and timing are important considerations in the successful application of oral tolerance strategies for suppression of chronic disease processes.

Topics: Administration, Oral; Animals; Autoimmune Diseases; Chronic Disease; Cytokines; Desensitization, Immunologic; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Immune Tolerance; Male; Mice; Myelin Basic Protein; Recurrence

T cell receptor (TCR)-recognizing regulatory cells, induced after vaccination with self-reactive T cells or TCR peptides, have been shown to prevent autoimmunity. We have asked whether this regulation is involved in the maintenance of peripheral tolerance to myelin basic protein (MBP) in an autoimmune disease model, experimental autoimmune encephalomyelitis (EAE). Antigen-induced EAE in (SJL x B10.PL)F1 mice is transient in that most animals recover permanently from the disease. Most of the initial encephalitogenic T cells recognize MBP Ac1-9 and predominantly use the TCR V beta 8.2 gene segment. In mice recovering from MBP-induced EAE, regulatory CD4+ T cells (Treg) specific for a single immunodominant TCR peptide B5 (76-101) from framework region 3 of the V beta 8.2 chain, become primed. We have earlier shown that cloned B5-reactive Treg can specifically downregulate responses to Ac1-9 and also protect mice from EAE. These CD4 Treg clones predominantly use the TCR V beta 14 or V beta 3 gene segments. Here we have directly tested whether deletion/blocking of the Treg from the peripheral repertoire affects the spontaneous recovery from EAE. Treatment of F1 mice with appropriate V beta-specific monoclonal antibodies resulted in an increase in the severity and duration of the disease; even relapses were seen in one-third to one-half of the Treg-deleted mice. Interestingly, chronic disease in treated mice appears to be due to the presence of Ac1-9-specific T cells. Thus, once self-tolerance to MBP is broken by immunization with the antigen in strong adjuvant, TCR peptide-specific CD4 Treg cells participate in reestablishing peripheral tolerance. Thus, a failure to generate Treg may be implicated in chronic autoimmune conditions.

Topics: Adoptive Transfer; Animals; CD4-Positive T-Lymphocytes; Chronic Disease; Crosses, Genetic; Down-Regulation; Encephalomyelitis, Autoimmune, Experimental; Female; Immune Tolerance; Immunodominant Epitopes; Lymphocyte Activation; Lymphocyte Depletion; Mice; Mice, Inbred Strains; Myelin Basic Protein; Peptide Fragments; Receptors, Antigen, T-Cell, alpha-beta; Recurrence

Because of its proximity to the central nervous system, the cerebrospinal fluid (CSF) represents an important source of T cells that potentially could mediate putative autoimmune diseases such as multiple sclerosis (MS). To overcome the low CSF cellularity, we evaluated culture conditions that could expand CSF T cells, with a focus on the expression of T-cell receptor V beta genes utilized by T cells specific for the potentially encephalitogenic autoantigen myelin basic protein (BP). Expansion of "activated" CSF cells with IL-2/IL-4 plus accessory cells optimally retained BP-responsive T cells that over-expressed V beta 1, V beta 2, V beta 5, or V beta 18, compared to expansion using supernatants from PHA-stimulated blood cells, or anti-CD3 antibody that led to different V gene bias and rare reactivity to BP. Sequential evaluation of paired CSF and blood samples from a relapsing remitting MS patient indicated that BP-reactive T cells were present in CSF during the period of clinical activity, and the pattern of BP recognition in CSF was partially reflected in blood, even after CSF reactivity had dissipated during remission. Over-expressed V beta genes were not always constant, however, since in three sequential evaluations of a chronic progressive MS patient, V beta genes over-expressed in the first BP-reactive CSF switched to a different V beta gene bias that was present in the second and third CSF samples. Blood samples reflected each pattern of CSF V beta gene bias, but retained the initial bias for at least 4 months after its disappearance from CSF. These data indicate that selective expansion of IL-2/IL-4-responsive CSF cells favors growth of the BP-reactive subpopulation, and, in a limited number of patients studied, reflected clinical disease activity. In comparison, blood T cells provided a partial but longer lasting reflection of the CSF BP reactivity and V beta gene bias.

Topics: Adult; Aged; Base Sequence; Cell Division; Chronic Disease; Female; Gene Expression Regulation; Humans; Male; Middle Aged; Molecular Sequence Data; Multiple Sclerosis; Myelin Basic Protein; Phenotype; Polymerase Chain Reaction; Receptors, Antigen, T-Cell, alpha-beta; Recurrence

Acute and monophasic experimental autoimmune encephalomyelitis (EAE) was induced in rats by immunization with myelin basic protein (MBP). Proliferative responses of lymph node cells to major and minor encephalitogenic and nonencephalitogenic determinants of the MBP molecules were measured at various time intervals after the immunization. Results of these experiments revealed that additional responses to minor determinants which had been observed at the late stage of mouse EAE (Lehmann et al., Nature 356, 155, 1992) were very weak and short-lived in the rat. Furthermore, the response to the minor encephalitogenic determinant was not recognized throughout the course of EAE. Coimmunization with synthetic peptides, corresponding to the major and minor determinants, induced T-cell response only to the major determinant. These findings suggest that poor generation of T cells reactive with the minor encephalitogenic epitope is attributable to peptide competition between these two determinants. The present results, together with those reported in mice, strongly suggest that differences in the clinical course of EAE, i.e., acute monophasic or chronic relapsing, is closely related to the presence or absence of "determinant spread" to minor encephalitogenic epitope(s).

Topics: Acute Disease; Amino Acid Sequence; Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Lymph Nodes; Molecular Sequence Data; Myelin Basic Protein; Peptide Fragments; Rats; T-Lymphocytes

In the present study a tetranucleotide (TGGA)n repeat polymorphism 5' to the myelin basic protein (MBP) gene was evaluated in a group of HLA-class II-typed, chronic progressive multiple sclerosis (MS) patients. This polymorphism has been reported by others to be associated with MS. Contrary to these reports we observed similar allele frequencies in patients and controls. Our results indicate that there is no association between MS and a polymorphism 5' to the MBP gene.

Topics: Alleles; Base Sequence; Chronic Disease; HLA-DR Antigens; Humans; Molecular Sequence Data; Multiple Sclerosis; Myelin Basic Protein; Polymorphism, Genetic

Cells from 25 mice at different stages of experimental autoimmune encephalomyelitis (EAE) adoptively transferred using lymph node cells sensitized to a synthetic encephalitogenic peptide of myelin basic protein (p87-99) were examined for reactivity to a different encephalitogenic myelin antigen, proteolipid protein (PLP). Cellular reactivity to a synthetic encephalitogenic peptide of PLP (pPLP) was found in 3/5 mice with acute EAE, 4/9 with chronic EAE, 1/6 mice with EAE in remission, and 0/5 during relapse. No proliferation to pPLP was seen in naive mice or in healthy mice immunized with p87-99. Two of 13 mice developed typical EAE after the serial transfer of cells from animals with p87-99-induced EAE had been activated with pPLP in vitro. Controls consisted of recipients of (a) pPLP-activated cells from naive donors or donors immunized with a nonencephalitogenic MBP peptide, (b) cells from mice immunized with MBP activated in vitro with irrelevant antigen, or (c) ovalbumin-activated cells serially transferred from mice with adoptively transferred EAE. No control mice developed signs. These results suggest that during the course of myelin breakdown caused by an immune response to one myelin component (MBP), autoimmune reactivity to at least one additional myelin antigen (PLP) can arise. Furthermore, the acquired cellular reactivity to additional myelin components may be sufficient in some cases to induce disease itself, perhaps lending insight into mechanisms involved in disease progression.

Topics: Amino Acid Sequence; Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunotherapy, Adoptive; Lymphocyte Activation; Mice; Molecular Sequence Data; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Peptide Fragments; Recurrence

Topics: Animals; Autoimmune Diseases; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Genes; Mice; Myelin Basic Protein; Receptors, Antigen, T-Cell, alpha-beta; Spinal Cord; T-Lymphocytes

Chronic relapsing experimental allergic encephalomyelitis (EAE) was induced in Thy-1.1 congenic SJL/J mice by the adoptive transfer of myelin basic protein (MBP)-responsive lymph node cells from Thy-1.2 SJL/J mice. The Thy-1 congenic mouse strain was constructed on the SJL (Thy-1.2) background by the initial cross with the AKR (Thy-1.1) strain and does not reject Thy-1.2+ T cells. Quantitative immunocytochemical analysis of the central nervous system (CNS) of Thy-1.1 recipients showed preferential trafficking of Thy-1.2+ T cells to the meninges and white matter, beginning prior to onset of clinical signs. At 7 days post-transfer (dpt), Thy-1.2+ donor cells constituted 2.5% of the infiltrating cells and reached peak values (ca. 10%) during the first attack. At later stages (up to ten relapses), Thy-1.2+ T cells constituted 2-5% of the infiltrate. In control mice injected with irrelevant antigen-stimulated Thy-1.2+ T cells, only the occasional Thy-1.2+ T cell could be demonstrated up to 14 dpt. This is the first study showing unequivocally the presence of MBP-stimulated, adoptively transferred T cells within the CNS of recipients throughout the course of EAE, particularly during later relapsing stages. These results indicate that the persistent presence of antigen-specific T cells may be required for the recruitment of non-CNS antigen-responsive immune cells.

Topics: Animals; Antigens, Surface; Cell Movement; Central Nervous System; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Immunization, Passive; Lymph Nodes; Membrane Glycoproteins; Mice; Mice, Inbred Strains; Myelin Basic Protein; T-Lymphocytes; Thy-1 Antigens; Time Factors

It had been demonstrated previously that the administration of transforming growth factor-beta 1 (TGF-beta 1) reduced the clinical severity of experimental allergic encephalomyelitis (EAE). Treatment with the related immunosuppressive molecule, TGF-beta 2, resulted in similar inhibition of T cell activation and proliferation in vitro. Long-term treatment was effective in reducing clinical severity of EAE and the number of relapses in mice receiving either myelin basic protein- or peptide-91-103-specific T cell lines. When examined histologically, mice that had received TGF-beta 2 demonstrated significantly less inflammation and demyelination in the central nervous system. Examination of other organs demonstrated no pathology or deleterious side effects from long-term TGF-beta 2 therapy. These findings have relevance for the use of TGF-beta 2 as a therapeutic agent for the human demyelinating disease, multiple sclerosis.

Topics: Amino Acid Sequence; Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Lymphocyte Activation; Mice; Mice, Inbred Strains; Molecular Sequence Data; Myelin Basic Protein; Time Factors; Transforming Growth Factor beta

Chronic relapsing experimental allergic neuritis (crEAN) was induced by repeated transfers of P2-protein reactive T lymphocyte lines. Clinically, each intravenous transfer of P2-reactive T cells induced a relapse of the disease with weight loss and flaccid paresis of the hindlimbs followed by recovery. After multiple transfers, recovery from disease was incomplete, leading to increasing neurological deficit during the remissions. The pathology of the lesions during exacerbations was characterized by massive inflammation in the peripheral nervous system, associated with extensive endoneurial oedema, nerve fibre destruction and wallerian degeneration. Selective primary demyelination and remyelination was found in the minority of affected nerve fibres. No onion bulbs were present in chronic lesions. In the central nervous system partial degeneration of the posterior columns reflected the extent of wallerian degeneration in the peripheral nerves and spinal roots. In addition, during stages of active disease some T lymphocytes and upregulation of Ia antigen expression were found in the spinal cord.

Topics: Animals; Cell Line; Chronic Disease; Myelin Basic Protein; Myelin P2 Protein; Neuritis, Autoimmune, Experimental; Peripheral Nerves; Rats; Rats, Inbred Strains; Recurrence; T-Lymphocytes

Development of experimental allergic encephalomyelitis (EAE) in the SJL (H-2s) mice is associated with a T cell-dependent autoimmune response to the C-terminal part of the myelin basic protein (MBP). In this study the influence of both H-2 and non-H-2 genetic background on EAE induced with the MBP89-101 peptide is described. Analysis of different H-2q haplotype strains, B10G, B10Q, SWR and NFR/N, showed that the B10 background is relatively resistant to disease induction. Both SWR and NFR/N were susceptible to EAE showing that the H-2q haplotype is permissive for EAE development induced with MBP89-101 and that the T cell receptor (TcR) haplotype or complement C5 deficiency exert no significant influence on disease susceptibility. In a series of H-2-congenic strains on the B10 background only B10RIII (H-2r) mice were susceptible to EAE. The B10RIII mice developed a severe EAE with early onset and chronic progressive or relapsing course of disease. In addition, B10RIII mice treated with Freund's complete adjuvant and pertussis toxin alone showed an early monophasic disease. The clinical observations were confirmed by immunohistopathologic analysis of the central nervous system. In these studies, we also applied antibodies to different TcR V beta elements which showed no specific limitation of the used TcR among infiltrating T cells in the target tissue in any of the strains. It is concluded that an MBP peptide-specific disease can be induced in three different haplotypes and it is possible that shared structures between the As, Aq and Ar molecules are of importance for the trigger of encephalitogenic T cells with different TcR V elements. The presently described chronic EAE model induced in the B10RIII mice will be of value as a model for multiple sclerosis.

Topics: Adjuvants, Immunologic; Amino Acid Sequence; Animals; Cerebellum; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Haplotypes; Major Histocompatibility Complex; Mice; Mice, Inbred Strains; Molecular Sequence Data; Myelin Basic Protein; Peptide Fragments; Pertussis Toxin; Spinal Cord; Time Factors; Virulence Factors, Bordetella

Chronic relapsing experimental allergic encephalomyelitis (CR-EAE) is an autoimmune demyelinating disease of the central nervous system and serves as an experimental model of human multiple sclerosis. Amino acid residues p91-103 of myelin basic protein are encephalitogenic in SJL mice and transfer of T cell lines that recognize this epitope results in CR-EAE. We show here that coculture of T cells in the presence of p91-103 that has been chemically cross-linked to the antigen presenting cells renders the T cell lines tolerant to the antigen. Injection of p91-103 coupled splenocytes into animals that had received encephalitogenic p91-103 reactive T cells significantly reduced the incidence and severity of EAE. Furthermore, treatment of mice with a single injection of antigen coupled splenocytes after they had recovered from their initial paralytic attack prevented the development of subsequent clinical relapses in all animals. These studies indicate that this effect is long lasting and can be successfully accomplished in an established autoimmune disease. Hence this form of immunotherapy may be considered as a therapeutic modality in the treatment of autoimmune diseases when the autoantigens are known.

Topics: Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunotherapy, Adoptive; Lymphocyte Activation; Mice; Myelin Basic Protein; Peptide Fragments; T-Lymphocytes

Thymidine incorporation proliferation assays to whole bovine P2 protein and its 58-81 and 14-25 synthetic peptides were performed on blood mononuclear cells from ten patients with Guillain-Barré syndrome (GBS), six patients with chronic idiopathic demyelinating polyradiculoneuropathy (CIDP), and age and sex matched normal subjects. The only patients whose cells showed any response were two out of four with very early GBS. One responded to P2 and both synthetic peptides. One responded to P2 but to neither peptide. The results support a role for cell mediated immunity to P2 protein in some patients with Guillain-Barré syndrome.

Topics: Adult; Aged; Child; Chronic Disease; Demyelinating Diseases; Female; Humans; Lymphocyte Activation; Male; Middle Aged; Myelin Basic Protein; Myelin P2 Protein; Peptides; Polyradiculoneuropathy

Chronic relapsing experimental allergic encephalomyelitis was induced by the passive transfer of [14C]thymidine-labeled myelin basic protein (MBP)-sensitized lymphocytes from MBP-immunized mice to naive syngeneic recipients. Labeled lymphocytes were localized and quantitated in the central nervous system during acute and chronic disease and clinical relapses. The results have shown that MBP-immune T cells home to the central nervous system endothelium 24 hours prior to and during initial clinical disease (5 to 7 days posttransfer). Unexpectedly, labeled MBP-immune cells never migrated far from blood vessels and, despite the presence of massive parenchymal inflammatory cell infiltration, almost invariably remained within the perivascular area. Quantitation revealed that labeled cells represented a minority (usually 1% to 4%) of the inflammatory cells during acute and early chronic disease. Furthermore, labeled cells could not be demonstrated in the central nervous system at the time of clinical relapse. We conclude that in this model, MBP-immune lymphocytes act exclusively from a perivascular location to orchestrate the influx of inflammatory cells that are predominantly of recipient derivation.

Topics: Acute Disease; Animals; Cell Movement; Central Nervous System; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Endothelium; Female; Immunization, Passive; Mice; Mice, Inbred Strains; Myelin Basic Protein; Spinal Cord; T-Lymphocytes

Topics: Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Lymph Nodes; Lymphocyte Activation; Myelin Basic Protein; Rats; Rats, Inbred Lew; Rats, Inbred Strains

Juvenile strain-13 guinea pigs challenged with whole central nervous system (CNS) tissue in complete Freund's adjuvant (CFA) developed chronic-relapsing (CR) experimental allergic encephalomyelitis (EAE). The animals that recovered from the first clinical episode were divided into three groups. One group was left untreated, one group was treated with three intracardiac injections of 100 micrograms glutaraldehyde-fixed myelin basic protein (MBP)-liposomes (MBP-L-GA) given once a week, and one group was treated with cytochrome c-liposomes (CYC-L-GA). The animals treated with MBP-liposomes were very well protected against further relapses. In vitro proliferative responses of peripheral blood lymphocytes (PBL) were performed repeatedly on most animals. The lymphocytes exhibited excellent proliferative responses to MBP, proteolipid apoprotein (PLP) and whole myelin, as well as to purified protein derivative (PPD) and concanavalin-A (ConA). High proliferative responses were recorded over the entire period of observation which lasted 12-22 months, each time the animals were tested in remission or in full relapse. However, a sharp decrease in proliferative responses was observed in most animals when the assay was performed 24-48 h before to 24 h after entering a relapse. The results demonstrate the presence of long-term and sustained cell-mediated responses to two distinct neuroantigens, and show fluctuations of both neuroantigen-specific and nonspecific responses concordant with a well-defined phase of the disease. Isoelectric focusing and immunofixation was performed on sera and cerebrospinal fluids obtained at the time of sacrifice. The pattern showed clear oligoclonal IgG bands (OB) in the samples obtained from untreated, CYC-L-GA-treated as well as in the MBP-L-GA-treated animals.

Topics: Animals; Cell Division; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunity, Cellular; Immunoglobulins; Isoelectric Focusing; Liposomes; Lymphocytes; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Oligoclonal Bands; Recurrence

Chronic relapsing experimental allergic encephalomyelitis (CREAE) was induced in SJL mice following the adoptive transfer of a T cell line derived from mice immunized with a synthetic peptide corresponding to residues 89-100 of the guinea pig myelin basic protein (MBP) molecule. This cell line proliferated to both the peptide and MBP and induced CREAE characterized by a series of relapses with eventual stabilization. Central nervous system (CNS) inflammation and demyelination were prominent neuropathologic features of both the acute and relapsing phase of the disease. The chronic phase was characterized by CNS lesions containing chronically demyelinated fibers with remyelination and some fiber drop-out, but little inflammation. The induction of CREAE in SJL mice by a cell line specific for residues 89-100 demonstrates that T cell recognition of this epitope is required for successful disease induction in this strain of mouse.

Topics: Animals; Brain; Cell Division; Cell Line; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Immunization, Passive; Lymph Nodes; Mice; Mice, Inbred Strains; Myelin Basic Protein; Peptide Fragments; Recurrence; Spinal Cord; T-Lymphocytes

We have examined the possibility that Ag-specific CTL responses may play a role in the pathogenesis of CREAE by using an effector T cell line (LN400) specifically reactive to the SJL encephalitogenic epitope defined by myelin basic protein MBP residues(90-101). The LN400 cell line was capable of adoptively transferring CREAE to naive SJL mice and proliferated specifically to synthetic peptides corresponding to MBP residues(90-101) and an N-acetylated analogue of this epitope, as well as MBP. Moreover, the cell line generated Ag-specific CTL responses only against syngeneic targets that had been pulsed with these Ag. Targets pulsed with irrelevant Ag were not lysed. These CTL responses were MHC restricted to H-2s and were inhibited if targets were preincubated with mAb specific for relevant class II Ag. No inhibition was seen if targets were preincubated with mAb specific for class I Ag, indicating that the CTL responses generated by this L3T4+ Lyt-2.2- cell lines were class II restricted. Studies designed to detect nonspecific CTL through a bystander mechanism failed to demonstrate significant lysis of bystander targets by this Ag-specific cell line. These findings have relevance in defining potential mechanisms of disease induction in this model autoimmune disease.

Topics: Animals; Cell Line; Chronic Disease; Cytotoxicity, Immunologic; Encephalomyelitis, Autoimmune, Experimental; Epitopes; Guinea Pigs; Histocompatibility Antigens Class II; Mice; Mice, Inbred BALB C; Myelin Basic Protein; Peptide Fragments; Phenotype; Recurrence; T-Lymphocytes, Cytotoxic

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.

Topics: Albumins; Animals; Antibody Specificity; Autoantibodies; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunoglobulin G; Multiple Sclerosis; Myelin Basic Protein; Spinal Cord

Neurologic complications of brain tissue-derived rabies vaccine may be chronic or progressive. Lymphocytes and serum from three patients with this form of encephalitis were studied. Two patients had positive lymphoproliferation to purified myelin, two had antibody to white matter, and one had antibody to myelin basic protein. No patient had antibody to proteolipid protein, myelin-associated glycoprotein, or cerebroside.

Topics: Antibodies; Brain; Chronic Disease; Encephalitis; Enzyme-Linked Immunosorbent Assay; Humans; Myelin Basic Protein; Rabies Vaccines; Recurrence

To investigate the role of anti-myelin antibodies in chronic relapsing experimental allergic encephalomyelitis (CR-EAE), sera from SJL/J mice with CR-EAE actively induced by inoculation with spinal cord homogenate in complete Freund's adjuvant (CFA) were compared with sera from mice to whom CR-EAE was passively transferred by lymph node cells (LNC) stimulated with myelin basic protein (BP). Sera were obtained serially from mice during both remissions and relapses of disease and were evaluated for the presence of anti-myelin antibodies using an avidin-biotin-immunoperoxidase technique. Four of six mice with CR-EAE induced with cord-CFA were positive for anti-myelin antibodies 15-124 days after inoculation, with 16 of 18 sera positive in these four mice. Two mice inoculated with cord-CFA did not have detectable serum anti-myelin antibodies, despite a clinical and histopathological picture indistinguishable from the antibody-positive mice. None of seven mice with CR-EAE passively transferred by BP-stimulated LNC had detectable anti-myelin antibodies in 30 sera obtained 7-141 days after cell transfer. We conclude that serum anti-myelin antibodies probably do not play a significant role in the pathogenesis of CR-EAE in SJL/J mice.

Topics: Animals; Autoantibodies; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunization, Passive; Immunoenzyme Techniques; Mice; Mice, Inbred Strains; Myelin Basic Protein; Recurrence; Spinal Cord

In vivo administration of anti-CD4 mAb (GK1.5) has been shown to be effective in preventing acute and relapsing experimental allergic encephalomyelitis (EAE). In the present report we have studied the depletion of CD4+ cells by a single dose of GK1.5 on the immune response to myelin basic protein and in the development of EAE. Our studies show that depletion of CD4 cells in mice that had received encephalitogenic CD4+ T cells altered the kinetics of acute and relapsing EAE, but did not prevent disease altogether. The in vitro T cell proliferative response to myelin basic protein in lymph node cells was maintained in the presence of significant depletion of CD4+ cells. These studies indicate that the population of Ag-reactive cells to be large and relatively refractory to antibody therapy. The implication of these results to therapy of human autoimmune disease is discussed.

Topics: Adjuvants, Immunologic; Animals; Antibodies, Monoclonal; Antigens, Differentiation, T-Lymphocyte; Chronic Disease; Drug Administration Schedule; Encephalomyelitis, Autoimmune, Experimental; Female; Freund's Adjuvant; Guinea Pigs; Immunization, Passive; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred Strains; Myelin Basic Protein; Recurrence; T-Lymphocytes

Acute and chronic relapsing forms of experimental allergic encephalomyelitis (EAE) can be induced in SJL/J mice following transfer of myelin basic protein (MBP)-sensitized T cells which have been challenged in vitro with MBP. In this study, addition of specific anti I-A antibody during the culture blocked the antigen-specific proliferation of T cells and inhibited the transfer of both acute and relapsing EAE. Treatment of T cell recipients with anti I-As antibody daily for 10 days suppressed the induction of acute EAE. Further treatment of mice with anti I-As antibody reduced the number of relapses and improved their conditions. We conclude that MBP-sensitized T cells interact with Ia positive cells, both in vitro and in vivo, to induce acute and chronic relapsing EAE, respectively. The mechanism of this interaction and its role in the disease process are discussed.

Topics: Acute Disease; Animals; Antibodies, Monoclonal; Autoantibodies; Cells, Cultured; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Galactosylceramides; Histocompatibility Antigens Class II; Immunization, Passive; Immunosuppressive Agents; Mice; Mice, Inbred Strains; Myelin Basic Protein; Recurrence; T-Lymphocytes

Optic nerve and spinal cord tissue from untreated guinea pigs with chronic relapsing experimental autoimmune encephalomyelitis, guinea pigs with experimental autoimmune encephalomyelitis in which the disease was treated with injections of myelin basic protein (MBP) combined with galactocerebroside (GC), and normal guinea pigs, has been studied morphologically, immunocytochemically and morphometrically. MBP/GC treatment induced widespread proliferation of oligodendrocytes and extensive central nervous system (CNS) remyelination in tissue from both sites. Whereas some oligodendrocytes within lesions from treated animals appeared to be derived from surviving cells which underwent mitosis, the frequent occurrence of nests of oligodendrocytes at the periphery of nerve fiber fascicles in optic nerve among perivascular astrocytic elements, raises the possibility that remyelinating oligodendrocytes might possess progenitors located in these regions. Observations from multiple sclerosis lesions showed that oligodendrocyte proliferation and CNS remyelination occur in human subcortical white matter, but to a lesser degree than that seen in the CNS of MBP/GC/treated guinea pigs. Immunocytochemical examination of CNS tissue from experimental autoimmune encephalomyelitis animals confirmed the morphologic identification of oligodendroglia. Preliminary morphometric analysis confirmed the impression of an increase in oligodendroglial cells in MBP/GC-treated animals. This increase was somewhat obscured statistically by a concomitant rise in the number of fibrous astrocytes. In view of the ability of oligodendrocytes to proliferate and produce new myelin in multiple sclerosis, the possibility is raised that an experimental immunologic approach similar to that employed here might have a beneficial effect in the human disease.

Topics: Animals; Autoimmune Diseases; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Galactosylceramides; Guinea Pigs; Immunohistochemistry; Male; Multiple Sclerosis; Myelin Basic Protein; Myelin Sheath; Neuroglia; Oligodendroglia; Optic Nerve; Recurrence; Spinal Cord

Topics: Animals; Cell Division; Central Nervous System; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Galactosylceramides; Guinea Pigs; Myelin Basic Protein; Myelin Sheath; Neuroglia; Oligodendroglia

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.

Topics: Animals; Chronic Disease; Cross-Sectional Studies; Encephalomyelitis, Autoimmune, Experimental; Enzyme-Linked Immunosorbent Assay; Guinea Pigs; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Myelin Basic Protein; Recurrence

We showed previously by using imprint electroimmunofixation that the oligoclonal IgG in sera and CSF from chronic relapsing EAE in guinea pigs were specific to spinal cord and Mycobacterium tuberculosis. We now show that most oligoclonal IgG bands are directed predominantly against isolated myelin basic protein (MBP). Activity to the latter could be removed from sera or CSF by absorption with MBP but not with histone or lysozyme. The oligoclonal IgG reacted weakly with isolated proteolipid apoprotein, and lacked reactivity to myelin-associated glycoprotein. When the oligoclonal IgG activity to myelin proteins was removed from the sera by absorption with a preparation of delipidated myelin before imprint electroimmunofixation, a few bands in some sera still reacted with whole spinal cord homogenate. These results indicate that, in some sera, a part of the oligoclonal IgG was directed against non-myelin proteins or lipids. In contrast to chronic relapsing EAE, CSF oligoclonal IgG from patients with multiple sclerosis showed no reactivity against human brain homogenate, whole myelin, delipidated myelin, and MBP in imprint electroimmunofixation.

Topics: Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunoglobulin G; Multiple Sclerosis; Myelin Basic Protein

Using two methods of immunization (A and B), 5/10 (A) and 27/45 (B), BALB/c by J mice immunized with human myelin proteolipid protein developed a demyelinating disease with a spectrum of chronic progressive to relapsing-remitting courses. Demyelinative lesions were seen histopathologically in all clinically affected animals that were examined. Many of the clinically unaffected animals also had histopathologic evidence of demyelination. Some of the animals had evidence of multiple ages of foci of activity. Evidence is presented that contamination with myelin basic protein could not account for the disease. This is a new model for multiple sclerosis in mice.

Topics: Animals; Brain; Chronic Disease; Demyelinating Diseases; Disease Models, Animal; Female; Glial Fibrillary Acidic Protein; Humans; Hypersensitivity, Delayed; Immunization; Mice; Mice, Inbred BALB C; Myelin Basic Protein; Myelin Proteins; Myelin Proteolipid Protein; Neurons; Spinal Cord

Antibody responses to the myelin/oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) were determined in the sera of Hartley guinea pigs with chronic relapsing experimental allergic encephalomyelitis (CREAE) using an enzyme-linked immunoassay. The sera were also tested for in vivo demyelinating activity by infusion into the subarachnoid space of normal rats. In contrast to the MBP titres, the anti-MOG antibody titres showed good correlation with the in vivo demyelinating activity of the sera (r = 0.91, P less than 0.001). This result suggests that antibodies directed against MOG may be involved in the pathogenesis of demyelination in CREAE.

Topics: Animals; Autoantibodies; Chronic Disease; Demyelinating Diseases; Encephalomyelitis, Autoimmune, Experimental; Glycoproteins; Guinea Pigs; Humans; Myelin Basic Protein; Myelin Proteins; Neuroglia; Oligodendroglia

Adherence of red blood cells from SJL mice suffering of chronic relapsing experimental allergic encephalomyelitis was studied to myelin basic protein coated microtiter plates. Control animals received either bovine serum albumine or "protein-antigen free" adjuvant using the same immunization protocol. Characteristic changes in adherence were found in bovine or human myelin basic protein injected animals compared to the bovine serum albumine immunized group. After a nonspecific increase in adherence between Days 2 to 6 observed in all 3 groups, in the encephalitogen challenged animals on Days 13-14 a marked decrease in red blood cell adherence was detected which maintained at this decreased level during the clinically active stage of the disease and reappeared with the relapse of EAE. No such decreased adherence of red blood cells was observed in BSA immunized animals or in adherence of cells from myelin basic protein injected animals to other basic type protein such as histone. Thus, decreased adherence of red blood cells in animals with EAE appears to be an interestingly unique measure of the disease activity.

Topics: Animals; Cell Adhesion; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Erythrocytes; Immunization; Mice; Mice, Inbred Strains; Myelin Basic Protein; Time Factors

We present a young child with an illness characterized by remissions and exacerbations and signs of disseminated lesions in the central nervous system. Visual evoked response and brainstem auditory evoked potential gave abnormal findings showing lesions also of the optic nerves and brainstem. The CSF IgG index was slightly elevated and myelin basic protein was high. There was also intrathecal antibody production against different viruses, the highest against measles. The HLA type was A3, B7, Dr2. The T4/T8 ratio reflected an immunological active disease. All these signs could have been diagnosed as multiple sclerosis. However, there were also lower motoneuron lesions and metabolic studies showed disturbances in pyruvate metabolism as in Leigh's disease. Cerebral tomography and magnetic resonance imaging showed scattered lesions in the nuclei lentiformis and capsula interna. We would like to stress the importance of careful etiological studies in cases of MS in childhood.

Topics: Brain Diseases, Metabolic; Child; Chronic Disease; Diagnosis, Differential; Humans; Immunoglobulin G; Leigh Disease; Male; Myelin Basic Protein; Pyruvate Dehydrogenase Complex; Recurrence; T-Lymphocytes, Helper-Inducer

Chronic relapsing experimental allergic encephalomyelitis was induced in SJL mice by adoptive transfer of long-term cultured T cell lines. The T cells which were activated with myelin basic protein (MBP) derived from various species, all induced chronic relapsing experimental allergic encephalomyelitis with a similar high incidence. During the relapsing stage, lymphocytes obtained from the spleen responded well to MBP and were capable of transferring experimental allergic encephalomyelitis, whereas thymus lymphocytes did not respond to MBP. There was no difference in the proliferative response of splenocytes to MBP when splenocytes were isolated either from mice with clinical relapse or from mice that did not relapse. Pathological examination revealed a transient appearance of inflammatory cells during the acute stage. Similar cell infiltrates were also observed at the relapsing stage. The I-region associated (Ia) antigens appeared on vessels and astrocytes in the acute inflammatory lesions which coincided with the appearance of inflammatory cell infiltrates. Ia antigen expression diminished with the disappearance of inflammatory cells. During the relapsing stage, the Ia antigens were also expressed on the vessels and astrocytes in the fresh lesions. Our data indicate that MBP-reactive T cells persist at least in the spleen, for a long time. They may be reactivated by certain mechanisms probably in the central nervous system associated with the Ia-antigen expression, which facilitates the effector phase again. The initial event that triggers the Ia-expression is not known as yet.

Topics: Acute Disease; Animals; Cell Division; Cell Line; Central Nervous System; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Histocompatibility Antigens Class II; Histocytochemistry; Immunoenzyme Techniques; Mice; Myelin Basic Protein; Recurrence; Spleen; T-Lymphocytes; Thymus Gland

Of 66 patients (31 female and 35 male) with demyelinating inflammatory polyradiculoneuropathy (DIP), 12% (8/66) had a chronic relapsing and/or progressive course (CR-DIP) and 88% (58/66) had an acute monophasic illness (acute Guillain-Barré syndrome or GBS). Ten (15%) of the 66 had one or more associated putative autoimmune diseases; of these ten, five had CR-DIP and five had GBS. Cell-mediated immune responsiveness (CMI) of 30 cases with DIP was tested in vitro by lymphocyte transformation. Peripheral nervous system neuritogenic protein (NP) and central nervous system encephalitogenic myelin basic protein were the challenge antigens. Eighteen (60%) of the 30 patients had CMI to NP of human peripheral nervous system origin when a stimulation index (SI) of 2 or more was evaluated as positive; eight 27% (1) had CMI to NP when a positive SI was 3 or more. Of the 44 control patients with other neuropathies, only two (4.6%) demonstrated CMI to NP (SI, greater than or equal to 2). The in vitro response of patients with DIP to myelin basic protein (7/30) was not significantly different from that of the control population (16/44). The high incidence of DIP associated with autoimmune diseases and the CMI to NP in this group suggest that DIP may be an autoimmune disease with NP as one possible major antigen.

Topics: Acute Disease; Adolescent; Adult; Aged; Autoimmune Diseases; Central Nervous System; Child; Child, Preschool; Chronic Disease; Demyelinating Diseases; Female; Humans; Immunity, Cellular; Infant; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Myelin Basic Protein; Peripheral Nerves; Polyradiculoneuropathy

Mononuclear cells extracted from regional lymph nodes, blood, spleen and central nervous system of guinea pigs with chronic relapsing experimental allergic encephalomyelitis (r-EAE), adjuvant immunized and untreated controls were cultured for 16 h in microtitre plates, and culture supernatants were then used to measure IgG and IgM, as well as IgG class anti-myelin antibody production by enzyme-linked immunosorbent assays. Increased synthesis of these immunoglobulins and antibodies was found during the course of r-EAE both in intra- and extrathecal compartments. Long-term cultures carried out for 7 days gave similar results but anti-myelin, anti-myelin basic protein and IgG synthesis was most pronounced intrathecally. Agarose isoelectric focusing of supernatants from these cultures showed oligoclonal IgG. These findings indicate in vivo synthesis of autoantibodies within the target for immune attack and a partial sequestration of the immune response to this compartment.

Topics: Animals; Autoantibodies; Brain; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunoglobulins; In Vitro Techniques; Lymphocytes; Myelin Basic Protein; Myelin Sheath; Recurrence

The current report describes a new technique for producing chronic experimental allergic encephalomyelitis (EAE)+ accompanied by demyelination in adult strain 13 guinea pigs. The disease is induced by a combination of passive transfer of lymph node cells sensitized to myelin basic protein (BP) and active challenge of the recipients with homologous spinal cord in Freund's complete adjuvants. The clinical-pathologic spectrum ranges from a progressively fatal form of chronic EAE leading to death in 4-7 wk, through a remitting-relapsing form, to a chronic-stable form lasting many months. In all of these forms large subpial plaques of demyelination occur in the spinal cord with active phagocytosis of myelin debris, especially at the edges. The axons are swollen, but remain intact throughout. The histologic appearances of the lesions suggest that lysis of myelin occurs before phagocytosis, one of the hypotheses proposed for the pathogenesis of lesions occurring in humans with multiple sclerosis.

Topics: Acute Disease; Animals; Axons; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunization, Passive; Multiple Sclerosis; Myelin Basic Protein; Myelin Sheath; Spinal Cord

Chronic relapsing experimental allergic encephalomyelitis has been induced in juvenile strain 13 guinea pigs with isologous spinal cord in Freund's complete adjuvant. Retention of antigen at the injection site and in the draining lymph nodes was studied by immunocytochemical staining with antiserum to myelin basic protein (BP). Antigen was detected in the skin more than 370 days after immunization, whereas it could be detected in the nodes only 200 days postinjection. Amputation of the hind feet to remove the antigenic depots prevented subsequent episodes of clinical EAE. Therefore, continuous antigenic stimulation by the antigen at the local skin site is more important in the induction of relapses than antigen in the draining lymph nodes.

Topics: Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Lymph Nodes; Male; Myelin Basic Protein; Recurrence; Skin

Topics: Animals; Chronic Disease; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Therapy, Combination; Encephalomyelitis, Autoimmune, Experimental; Female; Freund's Adjuvant; Galactosylceramides; Guinea Pigs; Male; Mice; Mice, Inbred Strains; Multiple Sclerosis; Myelin Basic Protein; Myelin Sheath; Rats; Rats, Inbred Lew; Rats, Inbred Strains; Recurrence; Spinal Cord

Topics: Animals; Chronic Disease; Drug Evaluation, Preclinical; Drug Therapy, Combination; Enterovirus Infections; Galactosylceramides; Hepatitis, Viral, Animal; Maus Elberfeld virus; Mice; Mice, Inbred BALB C; Murine hepatitis virus; Myelin Basic Protein; Myelin Sheath

The fine structure is described of a new model of chronic relapsing experimental allergic encephalomyelitis in the SJL/J mouse induced by the single adoptive transfer of myelin basic protein-sensitized lymph node cells. The neuropathology of the condition compared favorably with that seen in other species, and unlike a similar disease in the same strain of mouse induced by active sensitization with a central nervous system emulsion, there was little axonal pathology. Typical of mouse experimental allergic encephalomyelitis was the consistent involvement of polymorphonuclear leukocytes and extravasated material in central nervous system lesions. Remissions showed remyelination to be the major feature in the central nervous system, and clinical relapses were matched pathologically by fresh waves of inflammation and demyelination. Unusually large, apparently organized, sinusoidal collections of lymphoid cells (some of them displaying evidence of proliferation) were seen in the periventricular areas of the brain. The ability to induce chronic relapsing demyelination by passive means indicates that an antigen depot is not necessary for the perpetuation of the disease which is possibly transferred by memory cells in the inoculum. This model has virtue in its applicability to the pathogenesis and therapy of multiple sclerosis.

Topics: Animals; Brain; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Immunization, Passive; Lymph Nodes; Mice; Mice, Inbred Strains; Microscopy, Electron; Myelin Basic Protein; Nervous System; Optic Nerve; Recurrence; Sciatic Nerve; Spinal Cord; Spinal Nerve Roots

Juvenile strain-13 guinea pigs were challenged with isologous spinal cord in CFA. After recovery from the first EAE episode the animals were treated with guinea pig MBP inserted into liposomes, with cytochrome-c-liposomes, with MBP in saline or with MBP in IFA. Guinea pigs treated with MBP-liposomes showed a striking reduction in clinical signs and in the number and intensity of relapses. They displayed virtually no demyelinating lesions, and had comparatively little parenchymal inflammation in the spinal cord. Early T rosette levels showed an inverse correlation with the severity of histological lesions in the spinal cord but correlation with the clinical status at the time of rosette assay was less well defined.

Topics: Animals; Brain; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunity, Cellular; Injections; Liposomes; Myelin Basic Protein; Rosette Formation; Spinal Cord; Statistics as Topic

A chronic experimental allergic encephalomyelitis (EAE) has been produced in rabbits sensitized with bovine white matter proteolipid apoprotein. Eleven of 12 animals developed clinical disease one to six months after immunization with a single dose of the apoprotein. The clinical course was characterized by posterior ataxia, flaccid paralysis progressing to spastic paralysis, and incontinence. Spontaneous relapses and remissions were observed in 3 rabbits. Histologically, acute and chronic encephalomyelitis accompanied by primary demyelination were observed. Serum antibody production, assayed by both an enzyme-linked immunosorbent assay and an electroblot procedure, did not correlate with either the clinical course or the histopathological findings. Delayed hypersensitivity to proteolipid apoprotein was observed in all rabbits prior to the onset of clinical signs. The data suggest that lymphocytes specifically sensitized to the proteolipid may be involved in the pathogenesis of the demyelination in chronic EAE.

Topics: Animals; Antibodies; Antibody Formation; Apoproteins; Cattle; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Female; Myelin Basic Protein; Proteolipids; Rabbits

Topics: Animals; Autoantibodies; Autoantigens; Chronic Disease; Complement Fixation Tests; Encephalomyelitis, Autoimmune, Experimental; Enzyme-Linked Immunosorbent Assay; G(M1) Ganglioside; Galactosylceramides; Guinea Pigs; Myelin Basic Protein; Myelin Sheath; Precipitin Tests; Rats

Experimental allergic encephalomyelitis (EAE) in strain 13 guinea pigs is an acute monophasic disease induced with myelin basic protein (BP) in complete Freund's adjuvant (CFA). We report here that EAE can be effectively suppressed by transfer of a limited number of EAE-effector cells. Lymph node cells (LNC) from donors sensitized with BP/CFA are activated in vitro by incubation with BP and strain 2 peritoneal exudate cells (PEC). 5 X 10(7) of these cells are capable of inducing lethal EAE in recipients; 0.5 to 1 X 10(7) cells (a suboptimal transfer) effectively suppress EAE induction in animals subsequently sensitized with BP/CFA. Suppressed recipients develop an early mild disease from which they recover completely. In contrast, suboptimal transfer of BP-sensitized cells does not protect recipients against sensitization with whole central nervous system (CNS) tissue/CFA. About 50% of these recipients succumb to acute EAE; the survivors develop chronic EAE of varying intensity: a mild chronic form with recovery, a severe chronic form that is eventually fatal, and a chronic relapsing form that results in permanent neurologic deficit. Preliminary attempts at detecting suppressor cells in the suppressed animals were unsuccessful. Instead, PEC of all recipients of suboptimal transfers were capable of transferring EAE to naive animals.

Topics: Animals; Ascitic Fluid; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunization, Passive; Immunosuppression Therapy; Lymph Nodes; Myelin Basic Protein; Spinal Cord; T-Lymphocytes, Regulatory

Possible sensitisation of peripheral blood mononuclear cells from multiple sclerosis (MS) patients was studied in the presence of both normal and MS brain subcellular fractions. Myelin, synaptosomal and microsomal fractions and myelin basic protein were prepared by sucrose density gradient centrifugation from normal and MS brain. These preparations were added to mononuclear cell microcultures from the peripheral blood of nine acute and six chronic MS patients, 10 patients with other neurological diseases and 10 normal subjects. There was no significant increment in the peripheral blood mononuclear cell transformation of any of the acute MS patients with any of the fractions. Therefore there is no evidence that immunity to antigens in any of the fractions of MS or normal brain is of primary pathogenetic importance. A small increment in peripheral blood mononuclear cell transformation in the presence of several fractions was observed in two of six chronic MS patients, which might indicate the development of a secondary immune response after prolonged disease.

Topics: Acute Disease; Adult; Antigens; Brain; Cells, Cultured; Chronic Disease; Female; Humans; Lymphocyte Activation; Lymphocytes; Male; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Subcellular Fractions

In order to obtain a clue to understand the induction mechanism of chronic form of EAE we studied the immunized site of EAE animals with immunocytochemical techniques. There were numerous vacuoles associated with inflammatory reactions in the subcutaneous tissue of immunized feet. Immunocytochemical staining with antiserum to myelin basic protein (MBP) disclosed remaining of MBP in and around the vacuoles. The MBP remained more in early stage and less in chronic stage; it remained even on 326th day of postimmunization. A possible role of the remained antigen in chronic EAE was discussed.

Topics: Animals; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Immunoassay; Myelin Basic Protein; Skin

Preliminary results are presented on the treatment of Strain 13 guinea pigs with chronic relapsing experimental autoimmune (allergic) encephalomyelitis (EAE) induced by a single sensitisation with whole spinal cord. Animals were treated at different stages of the disease with injection containing either myelin basic protein (MBP) alone in incomplete Freund's adjuvant (IFA), or MBP in combination with a lipid hapten of myelin, galactocerebroside (GC) in IFA. The rationale for this treatment stemmed from previous work which suggested that MBP was responsible for T cell sensitisation in EAE and that GC was important in producing demyelinating antibodies and that both myelin components were needed in the induction of disease. Although treatment with MBP alone caused some initial stabilisation of the disease process, subsequent relapses occurred in all animals. However, in animals given MBP and GC together, either early or late in the course of the disease, marked clinical improvement has been noted with little or no development of relapses over an observation period of more than one year post-treatment. In addition, evidence of extensive remyelination and oligodendroglial proliferation in CNS lesions has been found in MBP-GC-treated animals suggesting that this therapy might be beneficial for CNS repair and relevant to multiple sclerosis.

Topics: Animals; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Galactosylceramides; Guinea Pigs; Haptens; Immunotherapy; Male; Myelin Basic Protein; Spinal Cord

To study the distribution of myelin-associated glycoprotein (MAG) in human nervous tissue and in multiple sclerosis (MS) lesions, we used paraffin sections and our modification of the peroxidase-antiperoxidase technique. Sections of MS lesions also were treated with antiserum to basic protein (BP) and with histological stains for axons and myelin sheaths. In tissue from normal developing central nervous system, oligodendroglia, their processes, and wwly formed myelin sheaths were intensely stained by MAG antiserum. In adults, MAG was found in periaxonal regions of myelinated fibers of the central and peripheral nervous system. The most striking finding in MS lesions was the extension of decreased MAG immunostaining into white matter that appeared normal when treated with BP antiserum or luxol fast blue. In acute early MS lesions the decrease in MAG immunostaining extended far beyond the margin of acute demyelination, where the BP staining of degenerating sheaths often was increased. In chronic inactive plaques, this decrease in periaxonal MAG immunostaining was limited to relatively few fibers in a thin rim around each lesion. These observations suggest that in MS, immunoreactivity of periaxonal MAG is altered before myelin breakdown begins. Early in degeneration, myelin sheaths and their fragments often were more intensely stained by BP antiserum than normal sheaths; later the staining intensity decreased. In shadow plaques, BP antiserum stained some oligodendroglia. Their appearance and location among thinly myelinated axons suggested that these oligondendroglia were forming new sheaths around previously demyelinated axons.

Topics: Acute Disease; Adolescent; Adult; Aged; Animals; Autolysis; Central Nervous System; Child; Child, Preschool; Chronic Disease; Glycoproteins; Histological Techniques; Humans; Immunoenzyme Techniques; Infant; Middle Aged; Multiple Sclerosis; Myelin Basic Protein; Myelin Sheath; Oligodendroglia; Rats; Staining and Labeling

The aim of this study was to investigate some of the conditions required for the consistent production of chronic relapsing experimental allergic encephalomyelitis (EAE). The concentration of mycobacterium/antigen was found to be a critical factor in producing relapses. Of the 3 antigens used to produce EAE--myelin, myelin basic protein (MBP) and whole spinal cord--only the latter produced a relapsing-remitting EAE. F1 crosses between female Hartley and male Strain 13 guinea pigs displayed an identical relapsing pattern to that of juvenile Strain 13 animals. The frequency of relapses was increased in Hartley guinea pigs when injected over the nuchal region.

Topics: Animals; Antigens; Antigens, Bacterial; Chronic Disease; Disease Models, Animal; Dose-Response Relationship, Immunologic; Encephalomyelitis, Autoimmune, Experimental; Female; Guinea Pigs; Hybridization, Genetic; Male; Mycobacterium tuberculosis; Myelin Basic Protein; Myelin Sheath; Recurrence; Spinal Cord

Experimental autoimmune encephalomyelitis (EAE) can be induced in genetically susceptible animals by injecting them with basic protein of myelin (BP) in a suitable adjuvant. EAE in adult Lewis rats is expressed clinically by acute paralysis and histologically by mononuclear cell infiltration of the central nervous system. Most rats spontaneously recover from EAE and show little or no damage to myelin. We report here that chronic progressive EAE with marked myelin lesions can be induced by a single injection of BP in complete Freund's adjuvant in intact 13-month old rats, or in 4-month old rats provided they have been splenectomized. Juvenile 2 1/2-month old rats recover spontaneously despite splenectomy. Thymectomy of young adult rats leads to relapsing EAE. These results illustrate that the clinical course of EAE is markedly influenced by age and integrity of immune organs. Furthermore, they provide an experimental model with features similar to those of chronic relapsing disease of the nervous system of man.

Topics: Aging; Animals; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Myelin Basic Protein; Rats; Recurrence; Splenectomy; Thymectomy

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.

Topics: Animals; Chronic Disease; Demyelinating Diseases; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Guinea Pigs; Leukocyte Count; Lymphocytes; Male; Myelin Basic Protein; Spinal Cord

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.

Topics: Animals; Chronic Disease; Disease Models, Animal; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Guinea Pigs; Immunotherapy; Leukocyte Count; Multiple Sclerosis; Myelin Basic Protein; T-Lymphocytes

Topics: Animals; Antibody Formation; Autoimmune Diseases; Brain; Chronic Disease; Encephalomyelitis, Autoimmune, Experimental; Freund's Adjuvant; Guinea Pigs; Hypersensitivity, Delayed; Immunologic Memory; Myelin Basic Protein; Recurrence; Spinal Cord; Tissue Extracts

Topics: Acute Disease; Antibodies; Brain Diseases; Chronic Disease; Encephalomyelitis; Female; Humans; Immunity, Cellular; Immunodiffusion; Iodine Radioisotopes; Lectins; Lymphocyte Activation; Male; Multiple Sclerosis; Myelin Basic Protein; Polyradiculopathy; Tritium

Topics: Animals; Brain; Chronic Disease; Demyelinating Diseases; Encephalomyelitis, Autoimmune, Experimental; Guinea Pigs; Lymphocytes; Macrophages; Microscopy, Electron; Myelin Basic Protein; Optic Nerve; Plasma Cells; Spinal Cord