CSF antibodies to myelin basic protein and oligodendrocytes in multiple sclerosis and other neurological diseases

Cerebrospinal fluid (CSF) from 18 multiple sclerosis (MS) patients, 13 subacute sclerosing panencephalitis (SSPE) patients, 22 other neurological disease (OND) patients, and 7 neurotic patients as controls were tested in an 125I-labeled anti-human F(ab')2 binding assay for the presence of antibodies to normal human brain cells from tissue culture, human fibroblasts, plasma membranes of MS and normal human brain, myelin basic protein (MBP) and bovine oligodendrocytes. Antibodies to MBP and to oligodendrocytes were found in the CSF of MS, SSPE and OND patients. Absorption of CSF with bovine CNS myelin significantly diminished binding activity to oligodendrocytes. Antibodies in the CSF against MBP and oligodendrocytes, on which some myelin determinants are expressed, seem to be a common feature of diseases in which demyelination is a component.     

亚低温治疗对实验性脑出血大鼠死亡率及脑组织钙含量的影响

目的本研究观察３２℃亚低温对实验性脑出血大鼠２４ｈ内死亡率和脑组织钙含量的影响。方法１３４只大鼠分成两部分：（１）６８只大鼠用于死亡率观察;（２）６６只大鼠用于脑组织钙含量测定。每一部分分成假手术对照组、常温脑出血组及亚低温脑出血组。结果常温组２４ｈ内死亡率为３６．６７％,亚低温组为４．５５％;脑组织钙含量常温组较对照组和亚低温组为高。结论亚低温治疗能显著减少实验性脑出血大鼠２４ｈ内死亡率,减少脑出血后脑组织钙含量。     

Frequency of T cells specific for myelin basic protein and myelin proteolipid protein in blood and cerebrospinal fluid in multiple sclerosis

T cell sensitization to two myelin components, myelin basic protein (MBP) and myelin proteolipid protein (PLP), may be important to the pathogenesis of multiple sclerosis (MS). Using the limiting dilution assay, we demonstrated that the blood of MS patients had an increased frequency of MBP-reactive T cells compared with normal subjects and patients with other neurological diseases (OND) and rheumatoid arthritis. There was no difference in T cell frequency to a synthetic peptide, PLP139-151, or Herpes simplex virus. Within cerebrospinal fluid (CSF), 37% of IL-2/IL-4-reactive T cell isolates from MS patients responded either to MBP or PLP139-151 while only 5% of similar isolates from OND patients responded to these myelin antigens. The mean relative frequency of MBP-reactive T cells within CSF from MS patients was significantly higher than that of OND patients (22 x 10(-5) cells versus 1 x 10(-5) cells) and was similar to that of MBP reactive T cells within the central nervous system of rats with experimental autoimmune encephalomyelitis. These results lend new support to the hypothesis that myelin-reactive T cells mediate disease in MS.     

IgG reactivity against citrullinated myelin basic protein in multiple sclerosis.

An increased level of citrullinated myelin basic protein (MBP-C8) has been reported in the brains of multiple sclerosis (MS) patients. However, the involvement of the immune response to post-translational modified MBP in the pathophysiology of MS remains speculative. The aim of this study was to compare the levels of immunoglobulin G antibodies to several MBP epitopes, before and after citrullination, in the cerebrospinal fluid (CSF) and sera of MS patients using enzyme-linked immunosorbent assay (ELISA). We analyzed antibody reactivity against various MBP-peptides in the CSF and sera of 60 MS patients, and 30 patients with other neurological diseases (OND) as controls. The peptides tested were: MBP(75-98) (peptide 1), native (peptide 2) and citrullinated (peptide 3) MBP(108-126) (ARG(122)-->Cit(122)), and native (peptide 4) and citrullinated (peptide 5) MBP(151-170) (ARG(159, 170)-->Cit(159, 170)). All selected peptides could support an immune reactivity in CSF and sera of MS and OND patients. A higher reactivity against peptide 4 was found in the CSF of MS patients compared with OND patients (P<0.0001), but not against citrullinated peptides (peptides 3 and 5). However, we observed that the citrullination state of peptide 2 modified the patterns of immune reactivity more markedly in MS patients (P<0.0001) than in OND patients (P<0.02). Although some MBP epitopes could be a potential target in MS, our data did not demonstrate any difference of antibody response to MBP peptides in their citrullinated forms.     

Persistent anti-myelin basic protein IgG antibody response in multiple sclerosis cerebrospinal fluid

Antibodies to myelin components, such as myelin basic protein (MBP), may play a role in pathogenesis of multiple sclerosis (MS) but results from determinations of anti-MBP antibodies are inconsistent. Enumeration of cells secreting antibodies represents a new approach to evaluate a specific antibody response regarding extent and localization, and reduces effects of e.g. antibody binding to target. Anti-MBP IgG antibody secreting cells were present in MS patients' cerebrospinal fluid (CSF) at a mean value of 1 per 833 cells, and they amounted to a mean value of about 2454 in the whole CSF compartment. Similar numbers were encountered in patients with other inflammatory neurological diseases (OIND). During follow-up, anti-MBP IgG antibody secreting cells persisted regarding frequency and numbers in MS, but decreased in OIND. Such cells were rarely detected in patients with tension headache. No correlations to clinical exacerbation of MS, disability or duration were discernable. In blood from MS and OIND patients, anti-MBP IgG antibody secreting cells were detected infrequently and at low numbers. The anti-MBP antibody response is strongly restricted to the IgG isotype. The anti-MBP IgG antibody response which is persistent and compartmentalized to the diseased organ, may be important for the development of MS.     

Myelin basic protein antibodies in the serum and CSF of multiple sclerosis and subacute sclerosing panencephalitis patients

A solid-phase radioimmunoassay was developed for the detection of myelin basic protein antibodies of immunoglobulin G (IgG) class. Purified basic protein of myelin (MBP) was adsorbed onto polystyrene beads, followed by incubation in dilutions of serum or cerebrospinal fluid (CSF). ¹²⁵I-labelled anti-human IgG was used to quantify antibodies bound to the solid-phase. The assay was optimized in tests with rabbit antibodies to MBP and with ¹²⁵I-labelled anti-rabbit IgG.
Serum and CSF specimens from 41 multiple sclerosis (MS), 16 subacute sclerosing panencephalitis (SSPE) and 58 control patients were tested for MBP antibodies. No statistically significant differences were found between MS and control patient groups, but the subgroup of acute MS patients had slightly elevated ( P 0.02) antibody levels in their CSF specimens. The SSPE patients had markedly elevated levels ( P 0.001) of antibodies to MBP in their CSF specimens.     

CSF antibodies to myelin basic protein and to myelin-associated glycoprotein in multiple sclerosis. Evidence of the intrathecal production of antibodies

Cerebrospinal fluid (CSF) from 40 multiple sclerosis (MS) patients was tested by solid-phase radioimmunoassay (RIA) for ability to bind 2 common structural components of myelin and oligodendroglia, i.e., to bind myelin basic protein (MBP) and myelin-associated glycoprotein (MAG). To prevent the effect of differences in CSF IgG concentration on binding activity, the CSF samples were tested at equal IgG concentration 1 mg/ml. The mean binding activity to MBP and MAG was significantly higher than in control neurotics, respectively P less than or equal to 0.05 and P less than or equal to 0.001. In 33% of MS cases, CSF antibody against both antigens was found. Indirect data were obtained that autoantibodies whose antigens are associated with myelin-oligodendrocyte unit are produced locally within the central nervous system (CNS). Anti-MAG and anti-MBP CSF antibody activity was significantly higher, P less than or equal to 0.01 for both antibody specificity, in MS cases characterized by high IgG Index, greater than or equal to 0.70 = means + SD in the neurotic group, versus MS cases characterized by normal IgG Index (less than or equal to 0.70). Correlation coefficient between antibody activity and IgG Index values was 0.785 for anti-MBP antibody, and 0.400 for anti-MAG antibody. The importance of intrathecally produced antibody to MBP and MAG lies in the fact that it indicates an active humoral autoimmune process against a myelin-oligodendrocyte unit in which more than one autoantigen is involved.     

Sensitization of cerebrospinal fluid and peripheral blood lymphocytes to myelin basic protein in multiple sclerosis

Cerebrospinal fluid (CSF) and peripheral blood (PB) lymphocyte sensitization to rabbit myelin basic protein (MBP) in 44 multiple sclerosis (MS) patients, 21 patients with other neurological diseases (OND) and 14 persons with neurosis was studied with the antigen-active rosette forming cells (Ag-ARFC) assay. The frequency of sensitization of CSF lymphocytes to MBP in groups of MS patients in the relapse stage and the chronic progressive stage was higher than in the group of MS patients in the stable stage and the OND patients. None of the healthy subjects showed a positive reaction with MBP. In BP there were no differences in the incidence of sensitization to MBP between patients in various stages of the disease, but it was higher than in the group of patients with OND and neuroses. In the patients who had suffered from MS for less than 4 years, sensitization to MBP was more common in CSF lymphocytes than in BP lymphocytes. The results suggest that primary sensitization to MBP occurs in CSF, and is probably secondary to myelin damage. However at present it is difficult to determine the extent to which sensitization of CSF and PB lymphocytes to MBP play a role in further demyelination processes.     

Cell-mediated immunity to myelin-associated glycoprotein, proteolipid protein, and myelin basic protein in multiple sclerosis

Peripheral blood lymphocytes (PBL) from active and stable multiple sclerosis (MS) patients, patients with other neurologic diseases (OND), and control subjects were tested for sensitization to two myelin antigens not previously examined in multiple sclerosis, using a [3H]thymidine incorporation assay. The antigens investigated were myelin-associated glycoprotein (MAG) and proteolipid protein (PLP). In addition, sensitization to myelin basic protein (MBP) was also tested. Lymphocyte stimulation indices in active MS patients that were greater than 2 standard deviations above controls were as follows: 9/30 for MAG, 0/17 for PLP, and 8/81 for MBP. No control subjects responded to MAG or PLP, and only 1/29 control subjects responded to MBP. Three of the patients that responded to MAG also responded to MBP. Although the mean proliferative response to MAG and to MBP was greater in the population of active MS patients than in stable MS, ONDs, or controls, the difference was not statistically significant. The OND group was the only population which proliferated to PLP (6/16). The only statistically significant differences among the groups for all myelin antigens tested were the proportion of individuals with active MS vs. controls that responded to MAG (P less than 0.05), and OND vs. controls and active MS that responded to PLP (P less than 0.025). The greatest individual responses to the three antigens tested were to MBP in active MS patients. Elimination of the T8 (cytotoxic/suppressor) subset amplified the responses to myelin antigens in some patients and ONDs studied. These studies have demonstrated reactivity to MAG but not PLP in some patients with active MS, and reactivity to PLP in some patients with other neurologic diseases.     

Immunoblot detection of oligoclonal anti-myelin basic protein IgG antibodies in cerebrospinal fluid in multiple sclerosis

Migration properties and occurrence of antibodies against myelin basic protein (MBP) in paired CSF and serum specimens from patients with multiple sclerosis (MS) were demonstrated after agarose isoelectric focusing, immunoblot transfer, and immunoperoxidase staining. Oligoclonal IgG antibody bands directed against MBP were found in the CSF of 9 of 28 patients with MS (32%), but not in the CSF of any of 34 patients with other neurologic diseases. No serum showed anti-MBP antibody bands. The CSF anti-MBP antibodies migrated to the anodal region of the IgG area in a different fashion from oligoclonal IgG and anti-measles IgG antibodies, which were detected in parallel. The anti-MBP bands were transient in three of seven patients whom we studied consecutively. Enzyme-linked immunosorbent assay (ELISA) of serum and CSF for detection of IgG reactivity against MBP showed absorbance values above 2 standard deviations of controls in 44% of the MS patients and in 21% of those with other neurologic diseases. Results of this assay correlated partly with those of the immunoblot assay. ELISA positive and immunoblot negative results might be due to a broad polyclonal anti-MBP antibody response.     

吉兰-巴雷综合征患者脑脊液中S-100b、MBP的检测及意义

目的 研究吉兰-巴雷综合征(green-barre syndrome,GBS)患者脑脊液中S-100 b和髓鞘碱性蛋白(MBP)的含量变化,探讨S-100 b及MBP在GBS发病过程中的作用以及与病情严重程度的关系.方法 采用酶联免疫吸附法(ELISA)测定32例不同临床分级的GBS患者(GBS组),30例其他神经系统...     

多发性硬化症髓鞘素组分特异性抗体分泌细胞

本文用酶联免疫斑点法（Ｅｌｉｓｐｏｔ）检测了２３例临床确诊多发性硬化症（ＭＳ）和１２例无菌性脑膜炎（ＡＭ）患者外周血（ＰＢ）和脑脊液（ＣＳＦ）中髓鞘素碱性蛋白（ＭＢＰ）、髓鞘素结合糖蛋白（ＭＡＧ）和含脂质蛋白（ＰＬＰ）特异性ＩｇＧ抗体分泌细胞。两组患者ＣＳＦ中该３种抗体分泌细胞均呈明显增多趋势，ＭＳ组尤著，但两组ＰＢ中该类细胞数均很少。指示对髓鞘素组分的Ｂ细胞免疫应答主要局限于与中枢神经系统（ＣＮ     

Highly related immunoglobulin light chain sequences in different multiple sclerosis patients

Although immunoglobulin G and free light (L) chains of oligoclonal origin in cerebrospinal fluid (CSF) are the most common immunologic abnormalities in multiple sclerosis (MS), it is unknown whether homologous CSF L chain sequences are present in different individuals with MS. Using Southern blotting, a particular kappa (κ) L chain variable region (V) probe was recently found to hybridize to V_K cDNA from CSF B cells from almost one half of the MS patients tested but only 10% of normal or other neurologic disease controls [Zhou, S.-R., Maier, C.C., Mitchell, G.W., LaGanke, C.C., Blalock, J.E., Whitaker, J.N., 1998. A cross-reactive idiotope in cerebrospinal fluid cells in multiple sclerosis: further evidence for the role of myelin basic protein. Neurology 50, 411–417.] Here, we report that this likely results from remarkable sequence similarity in certain V_κ from CSF B cells from different individuals with MS. The high degree of sequence homology even extended to all three complementarity determining regions (CDR) which in part form an antibody combining site. In addition, marked sequence homology was observed between the light chains from the MS patients and those from certain mouse antibodies against myelin basic protein (MBP). The results establish, in principle, that the same or very similar κ light chain variable regions can be shared between CSF B lymphocytes from different individuals with MS as well as with certain antibodies against MBP.     

CSF myelin basic protein in multiple sclerosis

Cerebrospinal fluid (CSF) from 221 patients with multiple sclerosis (MS) and 85 patients with other neurological disorders (OND) was examined using a competitive radioimmunoassay for myelin basic protein (MBP) immunoreactivity. MBP was found in 46 of 55 MS patients (84%) examined within six weeks of relapse but in only 11 of 85 patients (13%) with OND. There was a significant correlation between the concentration of MBP in the CSF and relapse severity in patients seen within four weeks of the onset of symptoms (p less than 0.01). Of 44 patients in remission, MBP was detected in 12, and these patients had a significantly higher tendency to subsequent relapse (p less than 0.05). In 72 patients with progressive disease the presence of MBP in the CSF reflected the confidence of clinical diagnosis. The results of this study suggest that measurement of MBP in the CSF gives an objective method of monitoring disease activity in patient with MS.     

Cerebrospinal fluid levels of myelin basic protein-like material and soluble interleukin-2 receptor in multiple sclerosis

The presence, level and disease activity relationships of soluble interleukin-2 receptor (sIL-2R) in the cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients are unresolved. We measured CSF immunoreactive myelin basic protein (MBP), a marker of acute myelin damage, and sIL-2R levels in the CSF from 11 patients with active relapsing remitting (RR) MS, five with stable RR MS, eight with chronic progressive (CP) MS, five with other neurologic diseases, and three normal controls. No measurable (less than 100 units/ml) sIL-2R was present in any of the samples. Conversely, MBP levels were elevated in the active RR group compared to the other four groups. These results indicate that, at the sensitivity of assays currently available, levels of CSF sIL-2R do not correlate with the diagnosis or disease activity of MS.     

Detection of anti-Nogo receptor autoantibody in the serum of multiple sclerosis and controls

OBJECTIVES: A myelin-associated neurite outgrowth inhibitor Nogo-A plays a key role in inhibition of axonal regeneration. Axonal damage beginning at the early stage of multiple sclerosis (MS) is responsible for permanent neurological deficits, although its molecular mechanism remains unknown. The aim was to study the prevalence of autoantibodies against Nogo-A and Nogo receptor (NgR) in the serum of MS. METHODS: The antibodies were identified in the serum of 30 MS patients, 22 patients with non-MS other neurological diseases (OND), and 22 healthy control (HC) subjects by Western blot using recombinant human Nogo-A-specific segment (NAS), the shared segment of Nogo-A and -B (NAB), Nogo-66 (N66), the non-glycosylated form of NgR, the glycosylated NgR (NgR-Fc), and myelin oligodendrocyte glycoprotein (MOG). RESULTS: None showed immunoglobulin G (IgG) antibodies against NAS or NAB. In contrast, 30% of MS, 23% of OND and 32% of HC subjects exhibited anti-N66 IgG, while 27% of MS, 27% of OND and 18% of HC showed anti-MOG IgG. None of HC but 33% of MS and 14% of OND showed anti-non-glycosylated NgR IgG. Furthermore, 60% of MS, 18% of OND and 14% of HC showed anti-NgR-Fc IgG. CONCLUSIONS: Because IgG autoantibodies against N66, NgR and MOG are often detected in the serum of MS and controls, they do not serve as an MS-specific marker.     

Specificity of antibody-dependent lymphocyte cytotoxicity against cerebral tissue constituents in multiple sclerosis

In patients with multiple sclerosis (MS) there is not only an antibody-dependent lymphocyte cytotoxicity (ADLC) against basic protein of myelin (MBP), as was demonstrated earlier, but also against encephalitogenic peptide, cerebrosides and gangliosides. The reaction against cerebrosides and gangliosides is not specific for MS; it is also frequently positive in patients with other neurological diseases (OND), syphilis and rheumatoid arthritis. The ADLC against encephalitogenic peptide shows a very high specificity for MS. Out of 35 cases with OND, 23 had a positive result with MBP, but only one with encephalitogenic peptide. Patients with syphilis and rheumatoid arthritis reacted negatively without exception. According to this, the encephalitogenic peptide is a highly specific antigen for MS; MBP shows less specificity. The ADLC against the encephalitogenic peptide shows a markedly increased sensitivity in MS compared with MBP. Specificity and frequency of positive findings with encephalitogenic peptide support the pathogenic significance of the reaction in MS.     

Antibodies to myelin-oligodendrocyte glycoprotein in cerebrospinal fluid from patients with multiple sclerosis and controls

Myelin-oligodendrocyte glycoprotein (MOG) has been implicated as a target for antibody-mediated immune attack in experimental autoimmune encephalomyelitis (EAE) which has been used extensively as an experimental model of multiple sclerosis (MS). We have screened cerebrospinal fluid (CSF) and plasma from 30 patients with MS, 30 with other neurological diseases (OND) and 30 with tension headache for anti-MOG antibodies of IgG isotype by enzyme-linked immunosorbent assay (ELISA). Such antibodies were detected in CSF from seven of the patients with MS, compared to two with OND and one with tension headache. No anti-MOG IgG antibodies were demonstrable in plasma. Antibody specificity was confirmed by Western blot immunostaining. Antibody levels were higher in MS compared to OND and tension headache. No correlation was observed between anti-MOG IgG antibodies and total IgG levels in CSF. The significance of anti-MOG antibodies demonstrated in MS CSF remains 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-GM4 and anti-galactocerebroside antibody titers were significantly elevated in the CSF of MS patients, but not anti-GM1 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 GM4, GM1, and galactocerebroside. Anti-GM4 and antigalactocerebroside antibody titers were significantly enhanced after acid dissociation and ultrafiltration. These data suggest that antibodies of the IgG class against GM4 and galactocerebroside are present in CSF of MS patients, and a significant number of them exist as immune complexes with their corresponding glycolipid antigens.     

Multiple sclerosis: rat and human oligodendrocytes are not the target for CSF immunoglobulins

CSF Ig from 28 patients with MS and 25 patients with other neurologic diseases (OND) were examined for their capacity to bind to rat or human oligodendrocytes in culture. We have used a double-label approach combining CSF Ig with antibodies against galactocerebroside (GalC). As normal human IgG at a concentration higher than 250 micrograms/ml were found to bind nonspecifically to oligodendrocytes in culture, patients' CSFs were used unconcentrated, at a final IgG concentration never exceeding 115 micrograms/ml. In these experimental conditions, we have not been able to detect any fixation of CSF Ig from MS (or OND) patients to rat or human GalC+ oligodendrocytes.