Binding properties of cerebrospinal fluid IgG in multiple sclerosis and other neurological diseases

A solid phase radioimmunoassay (RIA) was used to detect antibodies to myelin or myelin basic protein (MBP) in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) or other neurological diseases (OND). When measured at the same IgG concentration, MS samples had higher binding values than OND against myelin, but not against MBP. Using F(ab')2 fragments purified from pools of MS and OND CSF there was no difference in binding to myelin between MS and OND samples. These results indicate that anti-MBP antibodies are nt a feature of MS and binding of CSF IgG to myelin is not due to specific antibody, but is probably the result of non-specific binding to Fc receptors.     

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.     

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 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.     

Detection of antibodies to myelin basic protein by solid-phase radioimmunoassay with [I]protein A

A solid phase radioimmunoassay (RIA) for the detection of antibodies to myelin basic protein (MBP) in sera has been developed employing MBP-coated flexible polyvinylchloride microtiter trays and [125I]protein-A as the radiolabel. [125I]Protein-A directly binds to the Fc region of serum IgG from several animal species and serves as an excellent reagent for detecting antibody. It can also be used to bind to a second antibody ligand, thereby making it useful even when it does not bind directly to the primary antibody Fc region. The use of one preparation of [125I]protein-A label allows sera from several species to be tested for antibodies to MBP simultaneously, thereby making this RIA technically simple, rapid and economical. This assay has been particularly useful in examining serum samples from animals with experimental autoimmune encephalomyelitis and hypomyelinogenisis congenita. In patients with multiple sclerosis low levels of antibody to MBP can be detected in cerebrospinal fluid (CSF) and acid-eluted extracts from brain plaque material; detection of low levels of antibody in human serum has not been possible due to non-specific binding of human serum Ig in this RIA.     

Detection of antibodies to central nervous system antigens by solid phase radioimmunoassay

A solid phase radioimmunoassay is described which employs 125I-protein-A to detect the presence of antibodies against a panel of cellular and soluble central nervous system (CNS) specific antigens coated onto polyvinylchloride Microtiter plates. Serum antibodies from rabbits immunized against myelin, myelin basic protein (MBP), glial acidic fibrillary protein (GFAP), astroglioma cells, and cerebellar cells were easily detected, and high specificity for each antiserum and antigen was also demonstrable. The assay is applicable to cerebrospinal fluid (CSF) from patients with neurological diseases to detect antibodies against CNS-specific antigens. The assay should be useful for examining cell lines derived from CNS tissue for the presence of brain proteins.     

Cerebrospinal fluid levels of myelin basic protein and creatin kinase BB as index of active demyelination

Radioimmunoassay-determined myelin basic protein (MBP) shed to CSF during active demyelination, has been found to be a useful but non-specific test for MS. CSF creatin kinase BB (CK-BB), as measured by radioimmunoassay, is increased in a variety of neurological diseases, and has been considered a useful indication of brain damage but not of demyelinating diseases. Taking into account that the mean concentration of CSF CK-BB should not be increased in patients during the acute phase of MS, we suggest that the CSF MBP/CK-BB ratio could be a more specific index to demyelination than CSF-MBP alone. We also defined a laboratory demyelination pattern (CSF MBP greater than mean control MBP + 2 S.D. and CK-BB less than MBP). CSF levels of MBP and CSF levels of CK-BB were determined by radioimmunoassay in 232 patients with several neurological disorders and 33 control subjects. Patients diagnosed as having MS with clinical exacerbation had significantly higher values of CSF-MBP/CSF-BB ratio than control subjects. Our study showed a significant presence of demyelination pattern in CSF of patients with MS. We conclude that the CSF MBP/CK-BB ratio and the CSF demyelination pattern may be new and reliable tests for the diagnosis of MS.     

Autoantibodies to myelin basic protein are not present in the serum and CSF of MS patients

Myelin basic protein (MBP) is one of the main constituents of the CNS myelin sheaths, and an autoimmune response directed against MBP may be crucial in the demyelination process in patients with multiple sclerosis (MS). In this study sera and cerebrospinal fluid (CSF) from 25 MS patients, 25 patients with other neurological diseases and 16 healthy controls were examined for antibodies against MBP by using radio immunoblot, western blot, radio immunoassay and enzyme-linked immunosorbant assay. No evidence for the presence of antibodies to MBP was found in sera or CSFs in either the MS patients, or in the control groups tested.     

Serum and cerebrospinal fluid antibodies against myelin basic protein and their IgG subclass distribution in multiple sclerosis.

IgG class antibodies reactive with myelin basic protein (MBP) were determined by enzyme-linked immunosorbent assay (ELISA) in serum and cerebrospinal fluid (CSF) of 37 patients with multiple sclerosis and a control group of 32 patients with tension headache or psychoneurosis. Using standardised amounts of IgG from CSF and serum in ELISA, significantly higher mean antibody levels were found in CSF as well as in serum from the patients with multiple sclerosis. Ten (27%) of the multiple sclerosis CSF samples and 15 (41%) of the multiple sclerosis sera revealed anti MBP antibody levels exceeding 2 SD of the control group. Seven patients (19%) showed exclusive or higher levels of anti MBP antibodies in CSF, suggesting synthesis within the central nervous system. Analysis by ELISA for IgG subclasses of anti MBP antibodies revealed that they were restricted to IgG 1 in four patients and IgG 3 in one.     

海洛因海绵状白质脑病的脑脊液和血清髓鞘碱性蛋白及其抗体检测及意义

目的　通过对海洛因海绵状白质脑病 (HeroinSpongiformLeukoencephalopathy ,HSLE)患者血清和脑脊液髓鞘碱性蛋白 (myelinbasicprotein ,MBP)及其抗体 (Anti MBP)检测 ,探讨MBP与HSLE的关系。方法　采用酶联免疫吸附法 (ELISA)检测 1 8例HSLE患者、2 3例Heroin成瘾者 (吸毒但无HSLE临床症状者 ) ,1 7例多发性硬化患者(MS组 )、对照组 2 0例 (NC组 )的血清以及脑脊液 (CSF)中MBP及Anti MBP水平。结果　HSLE组、MS组CSF和血清MBP均值明显高于NC组和Heroin成瘾组 (P <0 .0 5) ,HSLE组与MS组CSF、血清MBP均值无统计学差异 (P >0 .0 5) ,Heroin成瘾组血清和CSFMBP均值与NC组间无统计学差异 (P >0 .0 5) ,4组血清和CSF的Anti MBP含量差异不显著 (P >0 .0 5)。结论　HSLE患者CSF及血清中MBP含量增高 ,MBP上升水平与髓鞘损伤程度有关 ,该病的髓鞘存在病理性损害 ,以及血脑屏障 (blood brainbarrier,BBB)的通透性改变。MBP检测可作为HSLE诊断及与海洛因成瘾者鉴别诊断的重要参数 ,它在HSLE病理过程中的作用机制有待进一步研究     

Myelin basic protein in the CSF of patients with multiple sclerosis, optic neuritis and various neurological diseases

We tested 104 patients for myelin basic protein (MBP) content in the CSF. Of these subjects 14 were selected as control group, 36 were affected by multiple sclerosis (MS), 14 by optic neuritis (ON) and 42 presented other not primarily demyelinating neurological diseases (ND ND). CSF MBP level was significantly higher in the MS group than in the other groups of patients, while no statistical difference was found between the MS patients with acute exacerbation and those in remission.     

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     

Intrathecal synthesis of autoantibodies to myelin basic protein in multiple sclerosis

A solid phase radioimmunoassay was used to detect anti-myelin basic protein (MBP) antibodies in the CSF and serum of multiple sclerosis (MS) patients and controls. CSF and serum samples were assayed prior to acid hydrolysis in order to detect free anti-MBP as well as after acid hydrolysis to measure the total (free and bound) amount of antibody. An anti-MBP index controlling for serum levels as well as the degree of breakdown of the blood brain barrier was used to estimate intrathecal synthesis of anti-MBP. MS patients with acute exacerbations or chronically progressive disease have significantly elevated levels of both free and total CSF anti-MBP. The anti-MBP index is also significantly increased in MS patients with both forms of active disease. Anti-MBP antibodies are intrathecally produced in MS patients with active disease.     

Neutralization of anti-myelin basic protein by cerebrospinal fluid of multiple sclerosis patients in clinical remission

Autoantibodies to myelin basic protein (anti-MBP) can be detected in the cerebrospinal fluid (CSF) of MS patients using a solid phase radioimmunoassay. Acute relapses of MS are characterized by an elevated, above unity, free (F) to bound (B) anti-MBP ratio; while in contrast, patients with chronic progressing disease have a low, below unity, free to bound ratio of anti-MBP. As patients with acute relapses enter into remission, the F/B anti-MBP ratio gradually decreases and eventually the level of these autoantibodies becomes undetectable. Neutralization of free anti-MBP was observed in intra- and interpatient experiments in which CSF from MS patients in remission was reacted with CSF of patients with acute relapses. Inhibition of anti-MBP neutralization was produced by CSF from MS patients with chronic progressing disease.     

Immunoreactive myelin basic protein in cerebrospinal fluid of patients with peripheral neuropathies

The presence of myelin basic protein (MBP)-like material in cerebrospinal fluid (CSF) usually reflects breakdown of central nervous system myelin. Immunoreactive MBP levels were measured in 70 CSF specimens from 66 patients with a variety of peripheral neuropathies. Immunoreactive MBP was present in CSF in 70% (16 of 23) of patients with biopsy-proved chronic demyelinating polyneuropathies, whereas in neuropathies producing primarily axonal damage it was present in only 22% (5 of 23). In Guillain-Barré syndrome, 45% (9 of 20) of patients had immunoreactive MBP in the CSF. We conclude that MBP-like material is present in the CSF of most patients with longstanding demyelinating polyradiculoneuropathies and probably reflects detection of peripheral nervous system P1 protein.     

Anti-MOG and anti-MBP antibody subclasses in multiple sclerosis.

In a subset of multiple sclerosis (MS) patients antibodies against myelin antigens seem to be important in the demyelinating process. In this study we investigated IgM, IgA and IgG serum antibodies against the myelin oligodendrocyte glycoprotein (MOG) and the myelin basic protein (MBP) in 261 MS patients. Seventy-two per cent had anti-MOG antibodies, 59% were anti-MBP seropositive. The dominating antibody was anti-MOG IgM. A significant relationship between IgA and a progressive disease course was found. The predominance of IgGI together with the significantly associated occurrence of IgG3 against MOG corresponds to the prevailing IgGI and IgG3 isotypes in other autoimmune diseases. Patients who actually suffered from a relapse were significant more often anti-MOG and anti-MBP IgG3 seropositive than those in remission. However, patients treated either with intravenous immunoglobulins or interferon-beta showed a significant reduction of anti-MOG IgG3 antibodies.     

OCB、抗MBP抗体及抗MOG抗体在中枢神经系统炎性脱髓鞘疾病诊断中的意义

目的：通过观察中枢神经系统炎性脱髓鞘疾病患者寡克隆区带（OCB）、抗髓鞘碱性蛋白（MBP）抗体及抗髓鞘少突胶质细胞糖蛋白（M OG ）抗体的阳性率,为临床诊断提供一定的参考。方法入组91例中枢神经系统炎性脱髓鞘疾病患者为观察组（其中MS 30例,NMO 61例）及50例神经系统非炎性病变患者为对照组。首先分析观察组与对照组间OCB、脑脊液及血清中抗MBP抗体、抗MOG抗体阳性率的差异,再分析观察组中MS亚组及NMO亚组间上述指标阳性率的差异。结果2组比较,OCB、血清抗MBP抗体及血清抗MOG抗体阳性率比较差异有统计学意义（P＜0.05）,脑脊液抗MBP抗体及脑脊液抗MOG抗体阳性率比较差异无统计学意义（P＞0.05）。MS亚组与NMO亚组,OCB、血清抗MBP抗体阳性率比较差异有统计学意义（P＜0.05）,脑脊液抗MBP抗体、血清抗MOG抗体及脑脊液抗MOG抗体阳性率比较差异无统计学意义（P＞0.05）。结论 OCB阳性对诊断MS具有重要意义。血清抗MBP抗体及抗MOG抗体可为中枢神经系统炎性脱髓鞘疾病的诊断提供重要实验室依据,血清抗MBP抗体在MS与NMO的鉴别诊断中具有一定价值。     

Autoantibodies against amyloid and glial-derived antigens are increased in serum and cerebrospinal fluid of Lewy body-associated dementias

There is increasing evidence that in Lewy body-associated dementias (encompassing Parkinson's disease with dementia (PDD) and dementia with Lewy bodies (DLB)), the adaptive immune system is altered and the degenerative process includes glial cells in addition to neuronal structures. We therefore aimed to determine levels of autoantibodies against amyloid and glial-derived structures in these dementia types. Using a newly developed Enzyme-linked immunosorbent assay (ELISA), we measured levels of IgG autoantibodies against neuronal and glial structures in serum and cerebrospinal fluid of a total of 91 subjects (13 PDD, 14 DLB, 11 Alzheimer's disease (AD), 11 frontotemporal dementia (FTD), 11 vascular dementia patients (VaD), and 31 healthy controls). Autoantibody levels against α-synuclein, amyloid-β?? (Aβ??), myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), and S100B were determined. In all groups, autoantibody levels were about three magnitudes higher in serum than in CSF. Serum autoantibody levels against α-synuclein, Aβ??, MOG, MBP, and S100B were higher in PDD/DLB compared to tau-associated dementias (AD, FTD), VaD, and controls, respectively, with most of them reaching highly significant p-values. In cerebrospinal fluid (CSF), levels of antibodies against oligodendrocyte-derived antigens (MOG, MBP) were significantly increased in PDD/DLB. Increased levels of autoantibodies against both neuronal- and glial-derived antigens in serum and CSF of Lewy body-associated dementias indicate an altered activity of the adaptive immune system in these dementia types. The potential of neural-derived IgG autoantibodies as part of a biomarker panel for the diagnosis of Lewy body-associated dementias should be further evaluated.     

Effect of methylprednisolone on CSF IgG parameters, myelin basic protein and anti-myelin basic protein in multiple sclerosis exacerbations

Clinical exacerbations of multiple sclerosis (MS) are characterized by elevated levels of cerebrospinal fluid (CSF) myelin basic protein (MBP). The purposes of this study were to determine whether anti-MBP antibodies are present in increased titer in CSF of MS patients with exacerbations, and whether they can be suppressed by the administration of immunosuppressive dosages of methylprednisolone (MP). A solid phase radio-immunoassay (RIA) was used to detect free and total anti-MBP antibodies before and after acid hydrolysis of CSF. In MS exacerbations, the majority of elevated anti-MBP is in the free form. With the exception of subacute sclerosing panencephalitis (SSPE) and some cases of post infectious encephalomyelitis, anti-MBP antibodies are not present in either MS patients in remission or in non-MS controls. Anti-MBP levels remained elevated over a 10 day period when patients are managed by bed rest only or when treated with intravenous (IV) ACTH. IV administration of MP in "high" (160 mg/day) or "mega" (2 g/day) dosages produces a highly significant reduction of both MBP (p less than 0.01) and anti-MBP (p less than 0.001) levels. Total intrathecal IgG synthesis is also significantly suppressed by IV-MP but not by ACTH.     

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.