Comparison of Antibodies Against Different Viruses in Cerebrospinal Fluid and Serum Samples from Patients with Multiple Sclerosis

The occurrence of a local production in the central nervous system (CNS) of antibodies against different selected viruses was analyzed by comparison of titers in serum and cerebrospinal fluid samples from groups of 50 patients with multiple sclerosis from Finland, Norway, and Sweden. Measles antibodies were determined in hemagglutination inhibition, hemolysis inhibition, and nucleocapsid complement fixation tests; mumps, parainfluenza virus type 1, and rubella virus antibodies were determined in hemagglutination inhibition tests; and herpes simplex virus type 1 antibodies were determined in passive hemagglutination tests. For reference purposes tests were also made for adenovirus antibodies in penton hemagglutination enhancement tests and poliovirus antibodies in neutralization enhancement tests. Among the 150 multiple sclerosis patients, a local production of antibodies against measles virus was found in the CNS in 57%, against rubella virus in 19%, mumps virus in 15%, herpes simplex virus type 1 in 11%, and parainfluenza virus type 1 (Sendai) in 3%. A local production in the CNS of antibodies against any of the viruses studied was found in 71% of multiple sclerosis patients. These included 48, 16, and 7% that produced antibodies to one, two, and three or more viruses, respectively.     

Cells of Cerebrospinal Fluid of Multiple Sclerosis Patients Secrete Antibodies to Myelin Basic Protein In Vitro

To characterize the role of B lymphocytes in the pathogenesis of Multiple Sclerosis (MS), we have isolated mononuclear cells from cerebrospinal fluid (CSF) and stimulated them with a polyclonal B-cell mitogen (pokeweed mitogen). This study has been done with MS patients selected for the occurrence of an acute attack in the course of the disease and with patients hospitalized for other neurological diseases. Five of the 11 MS patients had B lymphocytes producing in vitro antibodies (Abs) directed against purified human myelin basic protein (hMBP), as revealed by Western blot analysis. None of the 20 patients with other neurological diseases showed such a reactivity. The produced Abs recognized only 1 or 2 hMBP peptides without dominance for a certain peptide. This result emphasizes the presence of B cells producing Abs against MBP in CSF of MS patients and shows the interest of studying mononuclear cells of CSF as a good marker of the pathogenesis.     

Immunoglobulin Synthesis In Vitro by Cerebrospinal Fluid Cells in Patients with Multiple Sclerosis

Incubated cerebrospinal fluid (CSF) cells from patients with multiple sclerosis synthesized IgG and IgA in vitro. The synthesized IgG had an oligoclonal distribution and showed the same elcctrophoretic pattern as the IgG of the CSF. The amount synthesized was greater during exacerbations than during remissions. Blood lymphocytes from the same patients synthesized an IgG in vitro that showed a completely different electrophoretic pattern The amount IgG synthesized by the blood lymphocytes was less than the amount synthesized by the CSF cells. The results demonstrate that at least part of the oligoclonal IgG of the CSF of patients with multiple sclerosis is synthesized intrathecally and suggest that the CSF cells are antigenically stimulated in vivo.     

Molecular Biomarkers in the Cerebrospinal Fluid in Multiple Sclerosis

Neuroscience and Behavioral Physiology - Multiple sclerosis (MS) is a chronic autoimmune disease affecting the spinal cord and brain. Detection of the disease at its initial stages is a difficult...     

Axon Reactive B Cells Clonally Expanded in the Cerebrospinal Fluid of Patients with Multiple Sclerosis

Demyelination and axonal loss have been described as the histological hallmarks of inflammatory lesions of multiple sclerosis (MS) and are the pathological correlates of persistent disability. However, the immune mechanisms underlying axonal damage in MS remain unknown. Here, we report the use of single chain-variable domain fragments (scFv) from clonally expanded cerebrospinal fluid (CSF) B cells to show the role of an anti-axon immune response in the central nervous system (CNS) in MS. The cellular and subcellular distribution of the antigen(s) recognized by these CSF-derived clonal scFv antibodies (CSFC-scFv Abs) was studied by immunochemical staining of brain tissues obtained at autopsy from patients with MS. Immunochemistry showed specific binding of CSFC-scFv Abs to axons in acute MS lesions. The stained axons showed three major types of axonal pathological changes: 1) linear axons, axonal ovoid formation, and axonal transection were seen in the myelinated white matter adjacent to the lesion; 2) accumulation of axonal ovoid formations and Wallerian degeneration were seen at the border between demyelinated lesions and the adjacent white matter; and 3) Wallerian degeneration occurred at the center and edge of acute demyelinated lesions. These findings suggest a B cell axonal specific immune response in the CNS in MS.     

T Cells Recognizing Multiple Peptides of Myelin Basic Protein are Found in Blood and Enriched in Cerebrospinal Fluid in Optic Neuritis and Multiple Sclerosis

The cause of multiple sclerosis (MS) is unknown. Recently reported abnormal T-cell responses to several myelin proteins and myelin basic protein (MBP) peptides in peripheral blood constitute one line of evidence that autoimmune mechanisms could be involved in the pathogenesis of the disease. Monosymptomatic unilateral optic neuritis (ON) is a common first manifestation of MS and important to examine for a possible restriction of the T-cell repertoire early in the disease. T-cell activities to MBP and the MBP amino acid sequences 63–88, 110–128 and 148–165 were examined by short-term cultures of mononuclear cells from cerebrospinal fluid (CSF) and blood in the presence of these antigens, and subsequent detection and counting of antigen-specific T cells that responded by interferon-gamma (IFN-γ) secretion. Most patients with MS and ON had MBP and MBP peptide-reactive T cells in CSF, amounting to mean values of between about 1 per 2000 and 1 per 7000 CSF cells and without immunodominance for any of the peptides. Numbers were 10-fold to 100-fold lower in the patients' blood. Values were similar in ON and MS, and no evidence was obtained for a more restricted T-cell repertoire in ON. The MBP peptide-recognizing T-cell repertoire was different in CSF than in blood in individual patients with ON and MS, thereby giving further evidence for an autonomy of the autoimmune T-cell response in the CSF compartment. No relations were observed between numbers of autoreactive T cells and presence of oligoclonal IgG bands in CSF or abnormalities on magnetic resonance imaging of the brain in ON or clinical variables of MS. The high numbers of MBP and MBP peptide-reactive T cells could play a role in the pathogenesis of ON via secretion of effector molecules, one of them being IFN-γ, as well as in the transfer of ON to MS.     

Tetanus Toxoid Reactive T Lymphocytes in the Cerebrospinal Fluid of Multiple Sclerosis Patients

Cerebrospinal fluid (CSF) lymphocytes of 6 multiple sclerosis (MS) patients were cultured with tetanus toxoid (TT) and irradiated autologous antigen presenting cells (APC) followed by propagation of the responding T-cells in Interleukin-2 containing medium. TT-reactive cell lines were recovered from 4 of the 6 CSF samples, even though the patients had not been TT booster immunized in recent years. These findings suggest an active circulation of antigen reactive lymphocytes from the systemic immune compartment(s) into the CSF even without recent activation by booster immunization. Since immune reactions to TT are very unlikely to be pathogenic in MS, these findings also indicate that the presence of CSF lymphocytes reactive to a particular antigen does not necessarily imply a causal role.     

Cerebrospinal Fluid Cytokine Expression Profile in Multiple Sclerosis and Chronic Inflammatory Demyelinating Polyneuropathy

Background: Cerebrospinal fluid (CSF) analysis in patients with particular neurologic disorders is a powerful tool to evaluate specific central nervous system inflammatory markers for diagnostic needs, because CSF represents the specific immune micro-environment to the central nervous system. Methods: CSF samples from 49 patients with multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP), and non-inflammatory neurologic disorders (NIND) as controls were submitted to protein expression profiles of 47 inflammatory biomarkers by multiplex Luminex bead assay to investigate possible differences in the inflammatory process for MS and CIDP. Results: Our results showed differences in CSF cytokine levels in MS and CIDP; in particular, IL12 (p40) was significantly highly expressed in MS in comparison with CIDP and NIND, while SDF-1α and SCGF-β were significantly highly expressed in CIDP cohort when compared to MS and NIND. IL-9, IL-13, and IL-17 had higher expression levels in NIND if compared with the other groups. Conclusions: Our study showed that, despite some common pathogenic mechanisms, central and peripheral nervous system demyelinating diseases, such as MS and CIDP, differ in some specific inflammatory soluble proteins in CSF, underlining differences in the immune response involved in those autoimmune diseases.     

Identification of Different Measles Virus-Specific Antibodies in the Serum and Cerebrospinal Fluid from Patients with Subacute Sclerosing Panencephalitis and Multiple Sclerosis

Matched serum and cerebrospinal fluid (CSF) samples from eight cases of subacute sclerosing panencephalitis (SSPE) and 15 cases of multiple sclerosis (MS) were characterized in neutralization, hemolysis-inhibition (HLI), hemagglutination-inhibition (HI) with Tween 80—ether-treated antigen, complement-fixation (CF), and immunodiffusion tests. CF tests were carried out with crude virus material, purified nucleocapsids, and small particle hemagglutinin as antigens. A certain diversity in the relative content of antibodies against different virus products in various sera was found. There was a high degree of correlation between titers of neutralizing and HLI antibodies, but a less strict correlation between titers of HLI and HI antibodies. Serum samples from two cases of MS and one case of SSPE contained high titers of HLI and neutralizing antibodies in the presence of only low titers of HI antibodies demonstrable with Tween 80—ether-treated antigen. The major fraction of antibodies detected in CF and immunodiffusion tests reacted with nucleocapsids. There was a tendency of nucleocapsid CF antibody titers, as compared to neutralization and HLI antibody titers, to be higher in samples from patients with SSPE than from cases of MS. No significant differences were found between antibody titers recorded in neutralization, HLI, and HI tests carried out with two different measles virus strains, Edmonston and a strain (LEC) derived from a case of SSPE. Comparison of antibodies against measles virus products and, as a reference, against a group-specific vertex capsomer antigen of adenovirus in matched serum and CSF samples revealed a production of measles virus-specific antibodies within the central nervous system of all cases of SSPE and 8 out of 15 cases of MS.     

Lymphocyte Subpopulations in the Cerebrospinal Fluid and Peripheral Blood in Patients with Multiple Sclerosis

The cerebrospinal fluid (CSF) and blood of patients with multiple sclerosis (MS) were studied with respect to the frequency of lymphocytes with intra-cellular immunoglobulins of different Ig classes as well as the relative frequency of B and T lymphocytes. An increased number of Ig-positive cells were found in CSF (mean, 0.52%) when compared with blood (mean, 0.18%). In CSF there was a striking dominance of IgG-positive cells, very few IgA-positive cells, and almost no IgM-positive cells. The distribution in blood was approximately normal. The ratios between x ⁻ and λ-positive cells in CSF were all outside the range in blood. In CSF there were fewer B cells (mean, 4.7%) and more T cells (mean, 74.2%) when compared with blood (mean, 11.5% and 61.8%, respectively). The values for MS blood were approximately the same as for normal controls. The increased number of IgG-containing cells in the CSF are in agreement with earlier studies, which showed a local immunoglobulin synthesis. The increased proportion of T lymphocytes in CSF of MS patients may indicate that these cells play a role in the pathogenesis of MS.     

Lymphocyte Subpopulations in the Cerebrospinal Fluid and Peripheral Blood in Patients with Multiple Sclerosis

The cerebrospinal fluid (CSF) and blood of patients with multiple sclerosis (MS) were studied with respect to the frequency of lymphocytes with intracellular immunoglobulins of different Ig classes as well as the relative frequency of B and T lymphocytes. An increased number of Ig-positive cells were found in CSF (mean, 0.52%) when compared with blood (mean, 0.18%). In CSF there was a striking dominance of IgG-positive cells, very few IgA-positive cells, and almost no IgM-positive cells. The distribution in blood was approximately normal. The ratios between χ- and λ-positive cells in CSF were all outside the range in blood. In CSF there were fewer B cells (mean, 4.7%) and more T cells (mean, 74.2%) when compared with blood (mean, 11.5% and 61.8%, respectively). The values for MS blood were approximately the same as for normal controls. The increased number of IgG-containing cells in the CSF are in agreement with earlier studies, which showed a local immunoglobulin synthesis. The increased proportion of T lymphocytes in CSF of MS patients may indicate that these cells play a role in the pathogenesis of MS.     

多发性硬化患者淋巴细胞亚群的研究

目的 观察多发性硬化患者外周血及脑脊液中淋巴细胞亚群的水平。方法 用碱性磷酸酶抗磷酸酶法检测了56例多发性硬化（MS）活动期患者外周血和脑脊液（CSF）的淋巴细胞亚群。结果：活动期MS患者外周血CD4^ 、CD8^ 细胞较对照组减少，CD25^ 细胞、CD4^ /CD8^ 比值较对照组升高（P＜0．05）。CSF中CD4^ 、CD25^ 细胞、CD4^ ／CD8^ 比值较对照组升高，CD8^ 细胞降低（P＜0．05＜0．05），且CSF中淋巴细胞亚群均高于自身外周血中的相应细胞（P＜0．05）。经肾上腺皮质类固醇激素治疗后，随病情缓解，外周血、CSF中的淋巴细胞亚群均有不同程度的改善。结论 淋巴细胞亚群可能参与MS的发病，并与MS的缓解－复发有关。     

多发性硬化患者淋巴细胞亚群的研究

目的观察多发性硬化患者外周血及脑脊液中淋巴细胞亚群的水平. 方法用碱性磷酸酶抗磷酸酶法检测了56例多发性硬化(MS)活动期患者外周血和脑脊液(CSF)的淋巴细胞亚群.结果:活动期MS患者外周血CD4+、CD8+细胞较对照组减少,CD25+细胞、CD4+/CD8+比值较对照组升高(p<0.05).CSF中CD4+、CD25+细胞、CD4+/CD8+比值较对照组升高,CD8+细胞降低(p<0.05<0.05),且CSF中淋巴细胞亚群均高于自身外周血中的相应细胞(p<0.05).经肾上腺皮质类固醇激素治疗后,随病情缓解,外周血、CSF中的淋巴细胞亚群均有不同程度的改善. 结论淋巴细胞亚群可能参与MS的发病,并与MS的缓解-复发有关.     

Cerebrospinal Fluid BAFF and APRIL Levels in Neuromyelitis Optica and Multiple Sclerosis Patients During Relapse

Background

BAFF (B-cell activating factor of the tumor necrosis factor family) and APRIL (a proliferation-inducing ligand) are two of the major survival factors for B cells. Many studies have shown that BAFF levels were elevated in MS patients. However, whether the levels of CSF BAFF/APRIL increased in NMO patients was still unclear.

Objective

To measure the CSF BAFF and APRIL concentration of in NMO patients, and explore their relationship with disease activity in NMO.

Methods

CSF BAFF and APRIL was measured by an enzyme-linked immunosorbent assay (ELISA) in NMO (n?=?22), MS (n?=?18) patients and controls (n?=?14).

Results

Concentration of BAFF and APRIL in NMO patients were significantly higher than MS and controls. CSF BAFF and APRIL levels in controls were also lower than MS. Both NMO and MS revealed an increased disease disability with increased CSF BAFF. CSF APRIL was associated with EDSS scores in NMO, but not found in MS.

Conclusions

BAFF/APRIL system considered important for aggressive B cells and T-cell responses, and may stimulates B cells and T cell activation in acute relapse of NMO and MS. In NMO patients, CSF BAFF and APRIL may be key factors of B cell immune response and reflect disease severity.     

多发性硬化症发作期外周血和脑脊液淋巴细胞亚群的分析

为分析多发性硬化症 (MS )患者发作期淋巴细胞亚群及给予甲基强的松龙 (MP )治疗后的变化 ,流式细胞仪测定 2 6例处于复发期MS患者外周血 (PB )和脑脊液 (CSF )及 8例MS患者予MP治疗后PB淋巴细胞CD3+ 、CD4 + 、CD8+ 、CD4 5RA+ 、CD4 + /CD4 5RA+ 、CD4 + /CD2 9+ 、CD19+ 、CD5 + /CD19+ 的百分率。结果发现MS患者PB中CD8+ 、CD4 5RA+ 和CD4 + /CD4 5RA+ 百分率降低 ,CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值升高 ;CSF中CD3+ 、CD4 + 、CD4 + /CD2 9+ 百分率和CD4 + /CD8+ 比值高于PB ;淋巴细胞亚群与临床伤残程度和距此次发作的时间无关 ;MP治疗不影响PB淋巴细胞亚群变化。表明MS患者淋巴细胞通过血脑屏障有选择性 ,淋巴细胞亚群的变化在MS发病机制中起作用     

Lymphocyte Populations in the Cerebrospinal Fluid and Peripheral Blood of Patients with Multiple Sclerosis and Optic Neuritis

The cerebrospinal fluid (CSF) and peripheral blood (PB) of patients with multiple sclerosis (MS), optic neuritis (ON), and aseptic meningitis (AM) were studied with respect to the percentage of B cells (using membrane Ig fluorescence), T cells, and T-cell subpopulations (using a rosetting technique or monoclonal antibodies). In the PB of all three patient groups there were normal B-cell levels but a significant decrease in T cells compared with PB of normal individuals. In the CSF the B cells were reduced but the T cells elevated when compared with the PB of the patients, and these differences were statistically significant. The level of total T cells was not influenced by disease activity in MS or ON, but there was a significant reduction of suppressor cells in PB during disease activity in MS patients. In CSF there were also fewer suppressor cells during active disease, but the reduction was not statistically significant. Differences in B and T cells in CSF and PB indicate that the immune reactions within the central nervous system are at least partially isolated from the rest of the immune system. The changes in the T-cell subpopulations in MS support the evidence for an immunoregulatory defect in this disease.     

Autoreactive T Cells in Multiple Sclerosis

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) characterized by infiltration of T lymphocytes and macrophages into white matter leading to demyelination [1-2]. This pathology is frequently associated with disability of neurological function, in particular sensory deficits, visual problems and paralysis. The acute MS plaques are markered by the presence of activated T cells expressing the IL-2 receptor as well as activated, class II MHC positive macrophages [3-4]. In addition, cytokines such as TNF and oligoclonal immunoglobulin have been found in the brain and cerebrospinal fluid (CSF) of patients with MS [5-7]. This active inflammatory process is confined to the CNS, not affecting either the peripheral nervous system or other organs. Although it is generally accepted that this CNS inflammatory process causes demyelination and the resulting neurologic disability in MS, the mechanism(s) by which the inflammation is initiated and maintained is unknown.