Serum Of Multiple Sclerosis Patients Research Articles

Interferon inhibitor factor predicting success of plasmapheresis in patients with multiple sclerosis

Interferons (IFN) are biological molecules with antiviral, antiproliferative, and immunomodulatory actions. Plasmapheresis (PP) combined with IFN therapy in 24 multiple sclerosis (MS) patients was associated with a rapid increase in detectable IFN levels. We describe the presence of a detectable factor in the serum of MS patients which decreases the efficacy of IFN therapy in these patients. We call this factor "interferon inhibitor factor" (IIF). Standard IFN assay indicates inhibition of WISH cell protection by IFN owing to the presence of IIF in patient's serum. Similar results were also obtained with human fibroblast and human leukocyte IFNs. The best results were obtained with an IFN mixture; results with 1:20 diluted patient's sera showed elevation of 120% greater than 1:10 dilution. With 1:40 dilution, an elevation of 1,041% was noticed. The IIF from patient sera collected during PP was purified and characterized. Native gel electrophoresis of IIF indicates a single protein band; further analysis on SDS gels indicates two bands at the 200 and 21-kD range. ELISA failed to reveal the presence of any anti-IFN antibodies. This study demonstrates the presence of IIF in MS patients' sera which are removed from the circulation by PP. Removal of IIF from circulation was associated with increased IFN levels and clinical improvement as measured by Kurtzke's disability status scale (KDSS).

Decreased vitamin B12 and folate levels in cerebrospinal fluid and serum of multiple sclerosis patients after high-dose intravenous methylprednisolone.

Twenty-one patients (15 women, 6 men) with definite multiple sclerosis (MS) were treated with 1000 mg intravenous methylprednisolone-succinate (MP) daily for 10 days. Before MP treatment there was a negative correlation (r = 0.59, P = 0.0084) between serum vitamin B12 and progression rate, defined as the ratio of the score on Kurtzke's Expanded Disability Status Scale and disease duration. A significant decrease was demonstrated in the cerebrospinal fluid (CSF) and serum levels of folate and in the CSF level of vitamin B12 after MP treatment. The decrease in serum B12 was not statistically significant. After MP treatment all median levels of vitamin B12 and folate were below the reference medians. We hypothesize that low or reduced vitamin B12/folate levels found in MS patients may be related to previous corticosteroid treatments. Otherwise a more causal relationship between low vitamin B12/folate and MS cannot be excluded. Further studies may be required to clarify the vitamin B12 and folate metabolism in patients with MS.

Increased interleukin-1 production by peripheral blood mononuclear cells in patients with multiple sclerosis

The production of interleukin-1 (IL-1) by peripheral blood mononuclear cells (MNC) was assessed in patients with relapsing multiple sclerosis (MS) in both the active and inactive phase, in chronic progressive MS patients, in other neurological diseases, and in healthy subjects. Production was determined by measuring the IL-1 concentration in cultures with MNC supernatants using enzyme-linked immunosorbent assay (ELISA). IL-1 in sera of MS patients and healthy subjects also was investigated. MNC IL-1α production was significantly higher in MS patients (180.2 ± 177.5 pg/ml) than in healthy subjects (66.2 ± 66.0 pg/ml) ( P < 0.05). Relapsing MS patients in the active phase had significantly higher MNC IL-1α concentrations (360.1 ± 130.0 pg/ml) than normal subjects ( P < 0.001), but MNC IL-1α production in patients with relapsing MS in the inactive phase (65.3 ± 52.8 pg/ml) or chronic progressive MS (80.9 ± 71.9 pg/ml) was not increased significantly. MNC IL-1β production in MS patients was not elevated significantly. IL-1α and -1β were not detected in sera of MS patients. The correlation between increased IL-1α production and the clinical course of MS suggests that activated MNC may play a role in the pathogenesis of MS.

Monoclonal anti-conjugated azelaic acid antibody production

We have previously reported the existence of anti-conjugated azelaic acid (Aze A) antibodies in the serum of patients with multiple sclerosis (MS). In order to demonstrate the specificity of these antibodies, we have produced a monoclonal antibody directed against Aze A conjugated by an acylation reaction to a protein. In competition experiments, with ELISA method, we demonstrated that a part of the antibodies, raised in rabbit after immunization by human immunoglobulins (Ig) of MS patients, recognized the antigen-combining site of our monoclonal anti-conjugated Aze A antibody. These results clearly demonstrate that a part of human Ig obtained from sera of MS patients shared common idiotopes with mouse monoclonal antibody raised against conjugated Aze A.

Linoleic acid levels in white blood cells, platelets, and serum of multiple sclerosis patients.

We found a small but statistically significant reduction in the linoleic acid concentration of white blood cells and platelets in MS patients. The percent linoleic acid concentration (mean +/- S.D.) in the white blood cells if 24 MS patients was 8.8 +/- 1.8% as compared with 11.4 +/- 4.9 in 24 age and sex-matched controls (p less than 0.05). Platelet levels were 8.5 +/- 2.4% and 10.6 +/- 3.8% respectively (P less than 0.05). Serum linoleic acid levels were not significantly different in the two groups. The possible role of linoleic acid in the pathogenesis of MS has yet to be defined.

Anti-glycolipid antibodies and their immune complexes in multiple sclerosis

Antibody titers against myelin constituents in sera and CSF of patients with multiple sclerosis (MS) were examined by a solid-phase radioimmunoassay. Anti-G M4 and anti-galactocerebroside antibody titers were significantly elevated in the CSF of MS patients, but not anti-G M1 and anti-myelin basic protein antibodies. In sera of MS patients, the titers of antibodies against these myelin constituents were not elevated. Total IgG level was also significantly elevated in the CSF, but not in the sera of MS patients. Immune complexes from the CSF of MS patients were dissociated by acid-ultrafiltration and assayed for antibodies to G M4, G M1, and galactocerebroside. Anti-G M4 and antigalactocerebroside antibody titers were significantly enhanced after acid dissociation and ultrafiltration. These data suggest that antibodies of the IgG class against G M4 and galactocerebroside are present in CSF of MS patients, and a significant number of them exist as immune complexes with their corresponding glycolipid antigens.

Absence of oligoclonal IgA in CSF and serum of multiple sclerosis patients

CSF and serum from patients with MS and other neurologic diseases were analyzed by isoelectric focusing (IEF) in polyacrylamide gel and immunofixation with specific anti human IgA sera. The IgA band patterns seen in pH 4.5–6.0 region were rather diffuse and lacked oligoclonal feature.

The effect of serum from multiple sclerosis patients in remission on the incubated rat brain slices

Addition of human serum to incubated rat cerebral slices induced increased generation of myelin-related, membranous fragments floating on 0.32 M sucrose. Sera from 20 healthy subjects and 19 patients with various neurological disorders were equally active in this respect. On the other hand, the myelin-degrading activity of sera from 20 multiple sclerosis patients in remission was found to be significantly elevated by about 50%. The present findings support the contention that the serum of multiple sclerosis patients possesses increased potency to induce myelin sheath alterations.

Measles antibodies and histocompatibility types in multiple sclerosis

Measles antibody titres and HLA antigens were determined in 71 White and 11 Coloured multiple sclerosis (MS) patients and 71 White and 11 Coloured age and sex-matched controls. Measles antibody titres were determined by the hemagglutination inhibition test and HLA antigens were determined serologically by a micro-lymphocytotoxicity test. Measles antibody titres were significantly higher in MS than in control cases and this was true for both female and male patients. No association was observed between the HLA antigens, especially HLA-A3 or HLA-B7 and measles antibody titres in the sera of MS patients or controls.

Multiple sclerosis: Immunochemical studies on the demyelinating serum factor

Sera from patients with multiple sclerosis (MS) frequently produce demyelination of central nervous system tissue cultures. The nature of the factors responsible for demyelination is not as yet clearly established. However, several authors previously reported, in in vivo and in vitro models, demyelinating activity in IgG fractions isolated from sera and cerebrospinal fluid of MS patients14,42,48. We found the demyelinating activity of MS sera to be extremely labile to conventional biochemical treatments. Therefore, we isolated IgG from MS sera by absorption with staphylococcal protein A. Protein A binds specifically IgG1. IgG2 and IgG4. With this method we were able to remove most of the IgG, leaving only a small percentage, most probably IgG3, in the sera. Isolated IgG fractions from several sera of MS patients in the presence of human complement actively demyelinated central nervous system tissue cultures. Although a small, but significant, decrease in demyelinating activity could be observed in most of the sera absorbed in most of the sera absorbed with protein A, the majority of the demyelinating activity could not be removed by this treatment. From these studies, it appears that at least the majority of demyelinating activity in MS sera is not associated with IgG1, IgG2 or IgG4. If IgG is responsible for demyelination, IgG3 will most likely be the active factor.

Biosynthesis of myelin and neurotoxic factors in the serum of multiple sclerosis patients.

The in vitro synthesis of myelin proteins has been studied by measuring the incorporation of [3H] lysine in developing rat brain slices. This incorporation system has been used to assay potentially gliotoxic and myelinolytic agents. A reduced incorporation of the labelled amino acid into myelin proteins occurs in the presence of anti-myelin anti-serum and anti-basic protein anti-serum. Diphtheria toxin has been found to inhibit the synthesis of myelin basic and proteolipid protein in the white matter slices of developing rats. Recent experiments with serum samples from multiple sclerosis patients in exacerbation suggest the presence of a factor which interferes with the synthesis of myelin in white matter slices.

Medical News

<h3>`Associated agent,' lymphotoxin among clues to multiple sclerosis</h3> More and more tantalizing clues are accumulating as increasingly sophisticated research probes the mystery of multiple sclerosis. In the East, investigators in Philadelphia have extended earlier work by others, indicating that serum and brain tissue from multiple sclerosis patients can harbor a "multiple sclerosis-associated agent," apparently a virus. In the West, researchers at the University of California, Irvine, College of Medicine are purifying and characterizing a "lymphotoxic factor" from the serum of multiple sclerosis patients. Both the multiple sclerosis-associated agent and the lymphotoxic factor seem to be easier to detect during periods when the disease is exacerbated. The Eastern investigators are Werner Henle, MD; Paul Koldovsky, MD, PhD; Ursula Koldovsky, PhD; Gertrude Henle, MD; Rudolf Ackermann, PhD; and Gunter Haase, MD, working at the Joseph Stokes Jr Research Institute, Children's Hospital of Philadelphia, and the University of Pennsylvania School of Medicine.

Serum measles antibodies in multiple sclerosis.

Serum haemagglutination inhibition/HI/antibody titres to measles virus were examined in 80 multiple sclerosis patients, their 20 sibs, in 990 healthy controls, and 25 control patients. The measles HI titres were significantly raised in the serum of multiple sclerosis patients compared with healthy controls. There was no statistical difference between the levels of HI antibody titres in multiple sclerosis patients and their sibs. The measles HI titres decreased significantly in older age groups of healthy controls, whereas an analogous drop was not found in the multiple sclerosis group. The levels of serum HI antibody titres did not correlate with the sex of patients with multiple sclerosis or the activity of the disease. In the CSF of six multiple sclerosis patients low titres of HI antibodies were detected, whereas none of eight control patients had measurable traces of measles antibodies in the CSF. The significance of these findings in the aetiology of multiple sclerosis is briefly discussed.

Serum immunoglobulins in multiple sclerosis patients.

Of the several hypotheses of the etiology of multiple sclerosis (MS), that of an autoimmune process has gained prominence (1, 2). However, attempts to demonstrate circulating antibodies in sera of MS patients against components of brain tissue have yielded negative results (3-5). A myelinolytic agent first thought to be specific for MS sera (6), has also been found in sera of patients with other neurologic diseases and even in those of normal controls (7). Seitelberger (8) recently attempted to reconcile experimental immunologic data and pathologic-anatomic findings in MS to arrive at a plausible hypothesis of its pathogenesis. He postulated a central nervous system (CNS) virus infection or other primary insult followed by entrance of CNS immunogenic proteins into the circulation with ensuing immunologic sequelae. Earlier, several authors had suggested a virus etiology (9, 10), and measles antibodies had been extensively studied in patients with MS (11, 12). Other carefully controlled virus studies (13) c...

Failure to demonstrate circulating antibody to alcoholic brain extracts in multiple sclerosis.

Since Sachs and Steiner (1934) claimed to have demonstrated complement-fixing antibodies to brain in the serum of multiple-sclerosis patients, attempts to confirm and extend their findings have resulted in sharply divided conclusions. Some have agreed, culminating in the report by Raskin (1955), who has reported almost double their percentage of positives ; others, however, have not been able to find such antibodies at all. To decide between these conflicting observations is obviously important in the study of the pathogenesis of multiple sclerosis, especially since a humoral myelotoxic factor has recently been described in serum from animals with experimental allergic encephalitis (Bornstein and Appel, 1959). The present work was originally planned to follow the method of Raskin (1955), who has reported the greatest success, but this was quickly found to lead to technical difficulties in that results, both positive and negative, were not reproducible. Use of completely fresh (unfrozen) sera did not remove these inconsis tencies, and a detailed examination of the methods used by various authors was therefore made. As a result we believe that, to a considerable degree, discrepancies can be traced to variations in technical method ; of these, differences in the amount of complement used is the most important. Materials

Antibrain Antibodies in Multiple Sclerosis

This study is based on the premise that the serum of a multiple sclerosis patient may contain antibrain antibodies and that the disease is of an allergic nature. The etiology of multiple sclerosis being unknown, it was thought desirable to explore the possibility of a specific reaction of the blood sera of multiple sclerosis patients with the antigens prepared from the multiple sclerosis brains. Specific serologic studies in multiple sclerosis have been reported by Sachs and Steiner,1Steiner,2Frick,* and others. Acute disseminated encephalomyelitis accompanied by production of antibrain antibodies has been induced in animals by injection of emulsions of normal homologous and heterologous brain or spinal cord tissues, with or without adjuvants. † The production of these antibodies in the monkey was reported by Kopeloff and Kopeloff.9Schwentker and River10produced antibody response in the rabbits by injections of emulsions of homologous brain. Hill11