Suppression of chronic allergic encephalomyelitis: relevance to multiple sclerosis.

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

Multiple sclerosis: distribution of T cell subsets within active chronic lesions.

The distribution of T cells and T cell subsets was examined within the human central nervous system in active lesions from seven patients with chronic multiple sclerosis. The monoclonal antibodies anti-T11, anti-T4, and anti-T8 were used to detect total (whole) T cells, helper T cells, and suppressor-cytotoxic T cells, respectively, and a monoclonal antibody against human Ia was used for macrophages and B cells. Lesion progression was associated with large numbers of T4+ cells at the lesion margin and these extended great distances into the adjacent normal-appearing white matter. T8+ cells were most commonly concentrated around the lesion margin and displayed a preferential perivascular distribution. Within the lesion center, only a few T cells were found. Ia+ macrophages were most numerous within the centers of active lesions and were always present in the adjacent normal white matter. The monoclonal antibodies to T cells did not cross-react with glial cells including oligodendrocytes. These results indicate that T4+ cells are actively involved in lesion extension and Ia+ cells, in demyelination.

Autoimmune encephalomyelitis: simultaneous identification of T and B cells in the target organ.

Monoclonal antibodies to guinea pig T cells and antibodies to guinea pig immunoglobulin G were used in immunofluorescence studies to identify T and B cells in central nervous system tissue from guinea pigs with acute autoimmune encephalomyelitis. T cells appeared before B cells and were distributed within the white matter parenchyma, while B cells remained in perivascular spaces.

Field testing of a minimal record of disability in multiple sclerosis: the United States and Canada.

The IFMSS Minimal Record of Disability (MRD) in Multiple Sclerosis was field tested at eight medical centers in the U.S. and Canada. The goals were to conduct a qualitative and quantitative evaluation of the MRD. Assessment were completed on 249 patients with definite MS by neurologists and allied health professionals. Effective administration required some study and practice. Refinement of some unclear wording and awkward format will improve ease of administration. The MRD fit well into clinic routines and was accepted by staff and patients. Scoring presented few problems and these were related to overlap among the MRD scales, poor wording, and content not appropriate to MS. Quantitative evaluation of the MRD indicated that Incapacity Status primarily reflects disability in mobility and self-care when used as a composite score. Heterogeneity of content in Incapacity Status suggests that summed scores be used cautiously. Both Incapacity and Environmental Status had high levels of reliability and high correlations with established measures of impairment in MS. Inter-rater agreement of the ISS and ESS were also high. Once some necessary revisions are made, the MRD should be well on its way to achieving the IFMSS goal of developing a brief, reliable, valid, and appropriate instrument acceptable to a wide variety of workers in MS.

A prospective study of depression and immune dysregulation in multiple sclerosis.

This study examined psychologic distress and immune function in patients with chronic-progressive multiple sclerosis participating in a placebo-control trial of cyclosporine. Immune measures included percentages and absolute numbers of CD2+, CD4+, CD8+, Leu-11-b+, HLA-DR (IA+), and transferrin-receptor-positive cells, which were evaluated by immunofluorescence using monoclonal antibodies. Distress was measured with self-report scales. The Expanded Disability Status Scale assessed neurologic disability. Subjects were followed up for 2 years, and their high-depressed and low-depressed times were compared. Times of greater depression were associated with lower CD8+ cell numbers and CD8+%, and a higher CD4/CD8 ratio. CD4+ cell numbers and percent were also higher when subjects were depressed, but only in the placebo group. There were no differences in Expanded Disability Status Scale when subjects were more depressed. Evaluation of a single subject revealed that Ia+ and transferrin-receptor-positive lymphocytes increased 3 months before distress increased. It was concluded that distress is associated with immune dysregulation in multiple sclerosis, although the mechanisms of this association have yet to be delineated.

Antioligodendrocyte antibodies in cerebrospinal fluid of multiple sclerosis and other neurologic diseases.

Thirty-one concentrated cerebrospinal fluid (CSF) pools--10 multiple sclerosis (MS), and 21 controls with other neurologic diseases--were tested for antibodies against oligodendrocytes by indirect immunofluorescence on frozen brain sections and by complement fixation (CF) against isolated oligodendrocytes. Oligodendrocytes staining was found in 4 of 10 MS and 21 control CSF specimens. CF was detected in 4 of 10 MS and 3 of 21 controls. Positive results correlated with high albumin:IgG ratios. Using F(ab')2 fragments, specific antibodies against oligodendrocytes were detected in both MS and control specimens. In sera from MS and controls, oligodendrocyte staining was found in 16 of 23 MS samples and 11 of 30 controls. CF was present in one MS serum only. Therefore, antioligodendrocyte antibodies in CSF are not specific indicators for multiple sclerosis.

Multiple sclerosis: relevance of class I and class II MHC-expressing cells to lesion development.

Expression of Class I (HLA-ABC) and Class II (HLA-Dr; Ia) major histocompatibility (MHC) antigens on endothelial cells and astrocytes was investigated in multiple sclerosis (MS) lesions of variable disease activity and in normal central nervous system (CNS) using immunocytochemical techniques. Findings were correlated to lesion pathology and to the presence and distribution of T cells, T cell subsets, and interleukin-2 (IL-2) receptor-bearing cells. HLA-ABC was present on virtually all endothelial cells in normal and pathologic tissue samples. Ia was absent from controls and was detectable on about 10% of CNS endothelial cells in MS. In normal CNS, astrocytes were Ia-negative and rarely expressed HLA-ABC. In MS, Class I and II MHC-positive astrocytes were found, and both displayed a high frequency in active lesions. Class I-reactive glia were primarily associated with T cell infiltrates and were less common in older lesions in which macrophages predominated. In contrast, Class II-positive astrocytes were found in all active MS lesions independent of the composition of inflammatory cells. Expression of HLA-ABC and Ia molecules on astrocytes in MS lesions could indicate their involvement in local presentation of antigen to cytotoxic (T8+) and helper/inducer (T4+) T cells, respectively. The observed distinct distribution patterns of HLA-ABC and Ia-positive astrocytes might suggest that cytotoxic T8+ cells are operative early during lesion development in MS. This could be followed by a more extensive Class II MHC-restricted helper T cell-mediated immune response which leads to selective destruction of myelin via activated macrophages.

The pathogenesis and therapy of multiple sclerosis is based upon the requirement of a combination of myelin antigens for autoimmune demyelination.

It is postulated that the pathogenesis of demyelination in multiple sclerosis (MS) might lie in the cooperative effect of a T cell response against one myelin antigen (e.g. myelin basic protein--MBP) and a B cell response against a second myelin component which may act as a hapten or a carrier for the primary antigen. The hypothesis is based upon recent experiments in guinea pigs in which the encephalitogenicity of MBP was enhanced by the myelin glycolipid, galactocerebroside. This pathogenetic mechanism might be analogous to antibody-dependent, cell-mediated demyelination. Based upon this assumption, therapeutic trials in MS should take into consideration the possibility that instead of MBP alone, MBP might be more effective in combination with a lipid hapten.

Increase in anti-astrocyte antibodies in the serum of guinea pigs during active stages of experimental autoimmune encephalomyelitis.

From previous studies on the induction and treatment of experimental autoimmune encephalomyelitis (EAE) in guinea pigs and mice, antibodies have been implicated during both demyelination and remyelination. In the present study, sera from guinea pigs with acute, chronic and myelin basic protein/galactocerebroside (MBP/GC)-treated chronic EAE were evaluated for the presence of anti-glial cell antibodies by immunocytochemical techniques. Antigen specificity was confirmed by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The majority of sera from acute and chronic active EAE animals displayed intense labelling of astrocytes and only weak staining of oligodendrocytes when tested on sections of normal guinea pig brain tissue. In contrast, sera from animals with chronic EAE treated with MBP/GC gave strong labelling of oligodendrocytes and only minor staining of astrocytes. By immunoblotting, astrocyte staining was shown to be due to the presence of antiglial fibrillary acidic protein (GFAP) antibodies. The intense oligodendrocyte staining observed in sections reacted with sera from MBP/GC-treated guinea pigs corresponded well with high titers of serum anti-GC and anti-MBP antibodies measured by an ELISA. It was concluded that the presence of antibodies against astrocytes was possibly related to astrocytic antigens within the disease-inducing emulsion, at least during the initial phases of EAE, and not to their release from the central nervous system of affected animals.

On the presence of Ia-positive endothelial cells and astrocytes in multiple sclerosis lesions and its relevance to antigen presentation.

Using a monoclonal antibody in combination with the 4-step modified peroxidase-antiperoxidase (PAP) technique, Ia expression was demonstrated on endothelial cells and astrocytes in MS lesions of different ages. On endothelial cells, Ia antigen was found most frequently in grey and white matter parenchyma of acute MS brain and showed lower, comparable numbers in active and silent chronic MS brain tissue. Ia+ astrocytes were most numerous in acute MS lesions. In active chronic MS, Ia+ astrocytes predominated at the lesion edge, where they frequently displayed an atypical, rounded configuration. Positive astrocytes showed somewhat lower numbers within the lesion center and in normal white matter close to the lesion edge. Their frequency was significantly lower in normal white matter remote from the lesion. In silent chronic MS, Ia+ astrocyte processes were detectable only within the lesion center. Grey matter astrocytes displayed no staining with anti-Ia antibody. In normal brain tissue, no Ia antigen could be detected. The presence of Ia molecules on some endothelial cells and astrocytes in MS brain tissue suggests a role in antigen presentation perhaps relevant to the initiation and perpetuation, respectively, of the inflammatory process.

Chronic experimental allergic encephalomyelitis in strain 2 guinea pigs. Absence of resistance to nervous system changes and sex dependence of clinical disease.

Experimental autoimmune (allergic) encephalomyelitis (EAE) was induced in Strain 2 guinea pigs, a strain usually regarded as resistant to EAE. The development of disease in groups of juvenile male or female Strain 2/N guinea pigs was irregular with clinical EAE manifesting only in some groups of females. All animals examined morphologically (12 females and 9 males), between 3 and 18 months postinoculation, showed extensive changes in the central nervous system, although 12 of these had displayed no neurologic signs. This silent central nervous system disease, reminiscent of similar phenomena in man, indicates that Strain 2/N is fully competent immunologically at the level of the target organ.

Monoclonal anti-T cell antibodies are applicable to the study of inflammatory infiltrates in the central nervous system.

Monoclonal anti-human T cell antibodies were tested by a modified PAP technique on frozen sections of human central nervous system (CNS) tissue from inflammatory and non-inflammatory conditions. It was found that whole T cells and T cell subsets--T4+ (helper-inducer) and T8+ (suppressor/cytotoxic) T cells--could be differentiated specifically from other mononuclear cells and that these markers did not cross-react with human CNS tissue elements, particularly oligodendrocytes, under the same conditions. The lack of cross-reactivity between monoclonal antibodies to T cells and oligodendrocytes was further confirmed by double-labelling with an anti-galactocerebroside serum. It is concluded that in inflammatory conditions like multiple sclerosis, monoclonal antibodies promise considerable elucidation of immunopathogenetic events.

Multiple sclerosis. Distribution of T cells, T cell subsets and Ia-positive macrophages in lesions of different ages.

Using monoclonal antibodies in combination with the PAP technique, total (T11+) T cells, helper-inducer (T4+) T cells, suppressor-cytotoxic (T8+) T cells and Ia+ cells (macrophages and B cells) were localized in frozen sections of multiple sclerosis (MS) lesions with varied disease activity. In acute MS, T11+, T4+, T8+ cells and Ia+ macrophages were found in large numbers throughout the lesion but were virtually absent from normal white matter. In active chronic MS lesions, the numbers of T11+, T4+ and T8+ cells increased from the center towards the edge of the lesion. T11+ and T4+ cells penetrated deeply into the normal-appearing white matter adjacent to the lesion, while T8+ cells were more confined to the lesion edge. Ia+ macrophages displayed a reverse distribution pattern to that of T cells. They showed the highest density in the lesion center and their numbers decreased slightly towards the lesion edge. Small numbers of T11+, T4+, T8+ and Ia+ cells were always present in normal white matter. In silent chronic MS lesions, the numbers of both T cells and Ia+ cells were significantly lower than in active chronic MS. While T11+ and T4+ cells were found throughout the central nervous system (CNS), T8+ cells were virtually absent from the lesion center. Ia+ macrophages were also present in small numbers throughout the CNS and, sometimes, showed some accumulation at the lesion edge. Thus, T cells and T cell subsets have been demonstrated to be involved in lesion pathogenesis in MS in that lesion progression was associated with T4+ cells while ongoing demyelination depended upon the presence of Ia+ macrophages.

Lymphocyte responsiveness to oligodendrocytes in multiple sclerosis.

Lymphocyte responsiveness to oligodendrocytes has been examined with a modification of the antigen-reactive early T cell test in a total of 206 blood samples [104 multiple sclerosis (MS); 62 other neurological diseases (OND); 14 non-neurologic diseases; and 26 normals]. Oligodendrocyte reactivity was detectable more frequently in MS (49%) than in OND (23%) and normals (8%) but was absent from patients with non-neurologic diseases. Percentages of lymphocytes binding oligodendrocyte antigens were determined by direct and indirect immunofluorescence and immunocyto-adherence. Comparable results were obtained from all 3 systems. Percentages of oligodendrocyte antigen-binding lymphocytes were higher in MS than in OND and normals. With the present battery of techniques, lymphocyte responsiveness to oligodendrocytes has been shown to be more common in MS but since it is also found among OND, it is not MS-specific and most likely represents an epiphenomenon associated with white matter destruction.

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

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

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

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

T and B lymphocytes in the cerebrospinal fluid of various neurological diseases.

Cerebrospinal fluids (CSF) from 66 patients with a variety of neurological disorders were studied for total protein content, absolute amount of albumin, IgA, IgG and IgM, as well as their quotients (fraction to total protein ratio), cell numbers and B cell and T cell levels. In addition, the percentage of B cells and T cells in the blood was determined in 34 patients and serum immunoglobulin levels were estimated in 51 patients. In noninflammatory diseases of the CNS, the percentage of B cells was slightly higher and T cell levels were lower in the CSF in comparison to corresponding blood values. The B cell to T cell ratio in viral meningitis was altered in the CSF. An apparent increase in the T cell level led to a decrease of B cell values. Similar changes were also found in optic neuritis. The percentage of T cells was higher in relapsing multiple sclerosis than in the chronic progressive form. There were less striking changes in the B cell to T cell ratios in the CSF of other inflammatory diseases of the CNS.

[Eosinophil leucocytes in csf after myelography (author's transl)]. / Liquoreosinophilie nach Myelographie

2 cases one of polyneuropathy and one of funicular myelosis, in which myelography was performed, are reported. After the insertion of the contrast medium an increase of eosinophil leucocytes was observed. In one case an additional increase of IgG and IgA, as well as a penetration of macromolecular proteins and an increase of the total protein, was detected. The second case showed an increase in haptoglobulin in addition to the cellular reaction described. After removal of the contrast medium, the CSF became normal in both cases. These changes were on one hand interpreted as a functional lesion of the barrier, and on the other hand as a special cellular and humoral immune reaction. This could be distinctly detected in the lumbar CSF while in the suboccipital CSF there was only an indication of it. This emphasizes the assumption of a pathological reaction in the section of the cerebrospinal region that was in contact with the contrast medium (allergen-hyperergic reaction of the vascular fibrous tissue of the leptomeninges). No marked changes were observed in the serum.

Acute experimental autoimmune encephalomyelitis. Differences between T cell subsets in the blood and meningeal infiltrates in susceptible and resistant strains of guinea pigs.

The percentages of early (active, high affinity-rosetting), late (total) T cells and TG cells (suppressor T cells) were determined longitudinally in the blood and meningeal infiltrating cells of Strain 13 (susceptibility) and Strain 2 (resistant) animals inoculated for acute EAE and in guinea pigs of both strains in which the disease was suppressed with myelin basic protein (MBP) in incomplete Freund's adjuvant (IFA). Reactivity of T cells to MBP and oligodendrocyte protein was tested using the antigen-reactive T cell test. After inoculation for acute EAE, a transient increase in circulating early T cells was found during the latent period in Strain 13 guinea pigs only, while both strains showed a decrease in early T cells later on. Low values of circulating TG cells were more apparent in Strain 13 than Strain 2. In infiltrating cells of the meninges, early T cell values were significantly higher in the meninges than in the blood (P less than 0.01) in Strain 13, but were only slightly elevated in Strain 2 animals. TG cell levels in meningeal infiltrates were slightly higher than corresponding blood levels in both strains. In animals which were given a suppressive regimen of MBP/IFA, circulating early T cells rose initially and showed normal values later on in both strains of guinea pigs. TG cell levels were slightly more increased in strain 13 than strain 2. In comparison to blood values, early T cells were higher in the CNS in Strain 13, and lower in the CNS in Strain 2. TG cell levels were increased over blood values in both strains. These quantitative discrepancies in T cell subset between Strain 13 and Strain 2 guinea pigs which had been inoculated for EAE might reflect a difference in the cell-mediated immune response to white matter antigens which might be related to the variation in susceptibility to EAE.

Interferon-gamma and Ia antigen are present on astrocytes in active chronic multiple sclerosis lesions.

Using immunocytochemical techniques, presence of interferon (IFN)-gamma, IFN-alpha and Class II major histocompatibility (MHC) antigen (HLA-DR, Ia) was evaluated in active central nervous system (CNS) lesions of 11 patients with chronic multiple sclerosis (MS) and the findings were correlated with a detailed analysis of inflammatory cells. Both IFN-gamma and Ia antigen were detectable on astrocytes at the edge of active chronic MS lesions and in the adjacent normal-appearing white matter, where CD4+ (helper/inducer) T cells and interleukin-2 receptor (IL-2R)-positive cells were also found. IFN-alpha was predominantly seen on Ia-positive macrophages and not on astrocytes. Astroglia in the inactive lesion center and in normal CNS parenchyma were unreactive for both types of IFN and Ia antigen. These findings suggested that similar to the situation in vitro, IFN-gamma can also induce Ia-expression on astrocytes in vivo. Via local antigen presentation, IFN-gamma might thus play an important role in the development of MS lesions, while IFN-alpha might be involved in local immunosuppression.