Detection and isolation of immune complexes in multiple sclerosis cerebrospinal fluid

Immune complexes were studied in the cerebrospinal fluid (CSF) of 20 multiple sclerosis (MS) and 20 other neurological disease (OND) patients using polyethylene glycol precipitation; ten samples from each group were also examined using gel chromatography followed by ELISA. Polyethylene glycol detected predominantly IgG and IgM complexes in 13 of 20 MS samples and four of 20 OND samples. Intact MS complexes ranged in size from 230 to 340 kDa and contained 64 and 53 kDa antigens. Gel chromatography detected IgA complexes in eight of ten MS samples and one of ten OND samples; these complexes appeared to consist of polymeric IgA rather than true antigen. Chromatography detected IgG complexes in nine of ten MS and four of ten OND samples. Intact MS complexes ranged from 240 to 320 kDa and contained 200 and 150 kDa antigens. This study suggests that immune complexes are a very frequent finding in the CSF of MS patients and are in sufficient quantity to visualize on gel electrophoresis.     

Oligoclonal IgG in tears

We examined tears from patients with MS and systemic or eye diseases, and normal controls. Tears were isoelectrofocused on agarose gel, silver-stained, and immunofixated for IgG. In the cathodal portion of the gel (pH greater than or equal to 8.3), oligoclonal bands were detected in 14 of 21 (67%) MS tear samples. In most cases, these bands were not present in serum. Only 1 of 26 non-MS subjects showed two faint IgG tear bands, which were strongly present in serum. It appears that oligoclonal IgG bands can be detected in tears as well as in CSF of MS patients.     

CSF isoelectrofocusing in a large cohort of MS and other neurological diseases

The present study was performed in order to confirm the diagnostic value of isoelectrofocusing (IEF) in a large multiple sclerosis (MS) cohort and to evaluate the various neurological diseases probably to present a similar IEF profile. The cerebrospinal fluid (CSF) of 1292 patients with neurological diseases was studied by IEF. After a follow-up of 2-36 months, we only included patients with a definite MS or confirmed diagnosis of other neurological diseases (OND). MS was diagnosed in 407 patients and OND in 593 patients. For patients in whom three or more oligoclonal bands (OCB) were detected, IEF results showed a sensitivity of 85% and a specificity of 92% for the diagnosis of MS. The positive and negative predictive values were 86.5 and 90%, respectively. Inflammatory and infectious disorders of the central nervous system represented the main affections associated with OCB, including human immunodeficiency virus encephalitis, Lyme disease and less frequently Sjogren syndrome. Furthermore, when OCB were observed, 10 or more bands were more frequently found in MS than in OND (P < 0.0001). IEF of the CSF is a reliable method for the diagnosis of MS. The absolute number of bands may help to discriminate between MS and OND.     

Oligoclonal bands in cerebrospinal fluid: a comparative study of isoelectric focusing, agarose gel electrophoresis and IgG index

OBJECTIVE: To compare the sensitivity and specificity of isoelectric focusing (IEF) with immunofixation, agarose gel electrophoresis (AGE) and the IgG index in detecting intrathecally synthesized IgG in multiple sclerosis (MS) and in other nervous system disorders. MATERIALS AND METHODS: Cerebrospinal fluid (CSF) and serum from 147 patients with various nervous system diseases, 20 of whom had MS, were compared with IEF, AGE and the IgG index. RESULTS: CSF-restricted oligoclonal bands (OCB) were found in 20 of 20 patients with MS using IEF and in 9 of 20 using AGE. OCB were found in 12 patients with other nervous system disorders (OND) using IEF and 4 using AGE. The mean IgG index was 0.50 in OND and 0.96 in MS (P< 0.0001). Of 20 MS patients, 9 had an IgG index above the defined cut-off value of 0.72. CONCLUSIONS: IEF is about twice as sensitive as AGE in detecting OCB in MS. IEF is also far superior to the IgG index in determining intrathecal IgG synthesis.     

Discontinuos distribution of IgG oligoclonal bands in cerebrospinal fluid from multiple sclerosis patients

To test the effect of sampling on the detection of immunoglobulin (Ig) cerebrospinal fluid (CSF) abnormalities, we analyzed the first and last 1 ml fraction of 10 ml obtained during a single CSF removal from 27 multiple sclerosis (MS) patients and six patients with other neurological diseases. IgG index, hyperbolic function, and IgG synthesis rate decreased between the first and the last CSF aliquot. Discordant results were found in (15%) MS patients. In (7.5%) clinically definite MS patients, the number of CSF oligoclonal bands (OCB) decreased between the first and the last fraction. In one of the two patients, the three OCB visualized in the first fraction were not found in the last. We conclude that fractionated sampling may partially account for the absence of OCB in the CSF of some definite MS patients.     

Discontinuous distribution of IgG oligoclonal bands in cerebrospinal fluid from multiple sclerosis patients

To test the effect of sampling on the detection of immunoglobulin (Ig) cerebrospinal fluid (CSF) abnormalities, we analyzed the first and last 1 ml fraction of 10 ml obtained during a single CSF removal from 27 multiple sclerosis (MS) patients and six patients with other neurological diseases. IgG index, hyperbolic function, and IgG synthesis rate decreased between the first and the last CSF aliquot. Discordant results were found in 4/27 (15%) MS patients. In 2/27 (7.5%) clinically definite MS patients, the number of CSF oligoclonal bands (OCB) decreased between the first and the last fraction. In one of the two patients, the three OCB visualized in the first fraction were not found in the last. We conclude that fractionated sampling may partially account for the absence of OCB in the CSF of some definite MS patients.     

Kappa light chain predominance in serum and cerebrospinal fluid from human immunodeficiency virus type 1 (HIV-1)-infected patients

We measured kappa/lambda light chain ratios of Ig and IgG in 41 serum and 34 cerebrospinal fluid (CSF) samples from 47 patients at different clinical stages of human immunodeficiency virus type 1 (HIV-1) infection and in serum and CSF samples from control subjects. Both ratios were more elevated in HIV-1 seropositive subjects than controls. The elevation was more evident in samples from asymptomatic seropositive patients (ASP) than those from patients with acquired immunodeficiency syndrome (AIDS). In addition, there was a statistically significant elevation of Ig kappa/lambda ratios in ASP CSF compared to serum. We also delineated the light chain composition of oligoclonal IgG bands (OCB) by isoelectric focusing followed by immunofixation in CSF and serum samples from selected ASP and patients with AIDS who had neurological involvement. Five of six AIDS and all seven ASP samples had IgG OCB exclusively or predominantly of the kappa type. Four IgG OCB of the lambda type and one free lambda chain band were seen in CSF from a pediatric AIDS patient. The presence of an abnormally elevated kappa/lambda ratio correlated with the presence of IgG kappa OCB (p less than 0.02). We conclude that HIV-1 infection is associated with a kappa light chain predominance and with OCB mainly composed of kappa light chains.     

Immunoglobulin class and light chain type of oligoclonal bands in CSF in multiple sclerosis determined by agarose gel electrophoresis and immunofixation.

Agarose gel electrophoresis and immunofixation of CSF and serum from 39 patients with multiple sclerosis (MS) revealed oligoclonal IgG in the CSF in all cases and oligoclonal IgA and IgM in 1 patient each. IgG kappa bands only were found in 10 patients, while no patient had IgG lambda bands alone. IgG kappa bands predominated in 20 patients and IgG lambda bands in 5, while 4 patients had the same number of IgG kappa and IgG lambda bands. Twenty-seven patients also displayed IgG bands with kappa and lambda present simultaneously. Bands of free lambda chains were found in 7 patients, while free kappa chain bands were not seen. One or 2 faint IgG bands in 4 patients constituted the only serum abnormality. In 4 additional MS patients selected on the basis of normal findings on agarose gel electrophoresis of the CSF, immunofixation did not reveal oligoclonal Ig, while isoelectric focusing showed bands in 1. Immunofixation is recommended for proving the presence of oligoclonal Ig in CSF and for characterizing oligoclonal Ig into classes and types of light chains.     

脑脊液、血清免疫球蛋白及脑脊液寡克隆区带对自身免疫性脑炎的诊断意义

目的：探讨脑脊液、血清免疫球蛋白及脑脊液寡克隆区带（OCB）对自身免疫性脑炎（AE）的诊断意义。方法前瞻性收集2014年3月～2016年3月 AE 患者12例，以同期病毒性脑炎（VE）28例，多发性硬化（MS）16例为对照。 AE 患者予以 AE 抗体筛查，测定3组 CSF 中 IgG 及血清 IgG 、IgA 、IgM 浓度，计算 IgG 指数，检测血清及脑脊液 OCB 。所有入组患者均予以 MR 及脑电图等检查。结果 AE 及 MS 组 IgG 指数及 CSF － IgG 均高于 VE 组，AE 组高于 MS 组（P ＜0．05）；AE 及 MS 组血清 IgG 均高于 VE 组，IgM 低于 VE 组（P ＜0．05），AE 组与 MS 组差异无统计学意义（P ＞0．05）；3组患者血清 IgA 水平差异无统计学意义（P ＞0．05）；脑脊液 OCB 阳性率，VE 组7．14％，MS 组62.50％，AE 组91．67％，AE 组高于 MS 组、VE 组（χ2＝13．75，P ＜0．05）。 IgG 指数＞0．7百分率，VE 7．14％，MS 组62．50％，AE 组91．66％，3组比较差异有统计学意义（χ2＝25．61，P ＜0．05）。3组 MR影像学表现，VE 多累及颞叶、额叶，AE 多累及颞叶、顶叶、枕叶、脑岛，多呈双侧对称性或多发性。 MS多分布在脑室周围白质、视神经、脊髓、脑干和小脑。结论 AE 患者鞘内蛋白合成增加，脑脊液 OCB及 IgG 指数对 AE 早期的诊断有一定意义。     

多发性硬化病人血清和脑脊液壳三糖苷酶活性的研究

目的探讨多发性硬化(MS)病人血清和脑脊液(CSF)壳三糖苷酶(CTTS)活性以及CSF免疫活化和炎症标志物。方法选择三所医院178例MS病人,其中复发缓解型MS(RRMS)120例,继发进展型MS(SPMS)32例,原发进展型MS(PPMS)26例,并选取40例其他神经疾患(OND)和30非神经疾患病人作为对照组,检测血清和CSF中CTTS活性及CSF单核细胞数(MNC)和鞘内IgG产物。结果 MS病人与OND组和对照组比较,CSF中CTTS活性明显升高,但血清不升高。RRMS和SPMS组CTTS指数高于对照组,但PPMS组正常。在伴有MNC升高或CSF寡克隆IgG区带的MS病人,CTTS指数高于无此表现者。结论 RRMS和SPMS病人CCTS指数升高,CCTS指数与CSF炎症或免疫活化标志物有关。     

Intrathecal production of measles-specific IgA in subacute sclerosing panencephalitis

Objective - We measured measles-specific IgA in matched pairs of cerebrospinal fluid (CSF) and sera of patients with subacute sclerosing panencephalitis (SSPE), multiple sclerosis (MS), other central nervous system (CNS) infectious diseases (INF) and other neurological diseases (OND) by using enzyme linked immunosorbent assay. Materials and methods - CSF and sera from 23 patients with SSPE, 15 with MS, 14 with INF, and 15 with OND were included in the study. Results - The ratios of measles-specific IgA in CSF to serum were increased in SSPE patients compared to patients with MS, INF or OND. Conclusion - The data indicate a local production of measles-specific IgA in the CNS of SSPE patients.     

Solid-phase immunofluorometric assay for quantification of CSF immunoglobulins. Determination of normal reference values

Because a highly sensitive method is required to quantify low concentrations of immunoglobulin (Ig) classes in cerebrospinal fluid (CSF), there have been a few papers reporting normal values of CSF IgG, IgA and IgM determined in the same samples. Enzyme immunoassay (EIA) is most frequently used, but has such drawbacks as susceptibility of enzyme to inhibition and denaturation and the requirement for additional incubation with a substrate. Therefore, solid-phase immunofluorometric assay was evaluated for quantification of CSF IgG, IgA and IgM in the nanogram range. We found this to be rapid and reproducible. The mean (SD) values of normal CSF samples obtained from 22 subjects with tension headache were 23.9 (7.6) micrograms/ml for IgG, 2.00 (0.90) microgram/ml for IgA and 197 (87) ng/ml for IgM. The normal mean (SD) values of indexes were 0.51 (0.10) for IgG, 0.25 (0.05) for IgA and 0.044 (0.017) for IgM. These values agreed quite well with those determined by EIA. The values of CSF albumin correlated significantly with those of CSF IgG or IgA, but did not with those of CSF IgM. Levels of each of the three Ig classes in CSF and serum were significantly correlated. When CSF/serum ratio was introduced, a significant correlation between the albumin ratio and each Ig ratio was found. These results suggest that the Ig content of normal CSF may depend upon that of serum and upon the characteristics of the Ig molecule.     

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.     

间接免疫荧光法研究EB病毒感染类型与多发性硬化发生的关系

目的 用间接免疫荧光法研究EB病毒(EBV)不同感染类型与多发性硬化(MS)发生的关系.方法 采用间接免疫荧光法检测20例MS患者和20例其他神经科疾病(OND)患者脑脊液(CSF)中抗EBV壳抗原(EBV-CA)IgG抗体、抗EBV-CA IgG抗体亲和力、抗EBV-CA IgM抗体、抗EBV早期抗原(EBV-EA)...     

Multiple sclerosis is associated with enhanced B cell responses to the ganglioside GD1a

The occurrence and role of autoantibodies to gangliosides and other lipid-containing components of the central nervous system in Multiple Sclerosis (MS) are unsettled. Using sensitive ELISAs, we measured IgG and IgM antibody titers and absorbances to the three major gangliosides GD1a, GD1b and GM1, and to sulfatides, cardiolipin and myelin proteins in paired serum and cerebrospinal fluid (CSF) from patients with untreated MS, optic neuritis (ON), acute aseptic meningo-encephalitis (AM) and other neurological diseases (OND). Twenty-three per cent of 30 MS (P<0.04) and 18% of 32 ON patients (P<0.05) presented elevated IgG antibody titers to GD1a in serum compared to 9% of patients with OND. Six (40%) of the patients with malignant MS had elevated serum IgG antibody titers to GD1a compared to one (6%) of the patients with benign MS (P<0.04). In CSF, elevated IgG antibody titers to GD1a were measured in 13% of MS and 20% of ON patients compared to 4% of patients with OND (P<0. 03 and P<0.02, respectively). The augmented IgG response to GD1a in serum also separated MS from Guillain-Barré syndrome. Compared to OND increased IgM absorbances to sulfatides and cardiolipin were observed in CSF of patients with MS, but also in AM. Elevated IgG antibody titers to myelin proteins were found more often in MS patients' serum and MS, ON and AM patients' CSF compared to OND. The data implicate that among the multitude of enhanced B-cell responses occurring in MS and ON, that directed to GD1a is common and more discriminative, and should be evaluated in future MS treatment studies.     

Serum IgG,IgA and IgM concentration in 1,038 patients with various neurological disorders

Summary Concentration of serum IgG, IgA and IgM in 1,038 unselected patients with various neurological diseases was determined. In 521 instances, the levels of CSF IgG, IgA and IgM were also established. In serum, the most frequent selective quantitative abnormality was found in the IgA concentration. In CSF, an increased level of IgG with normal IgA concentration and undetectable IgM was established about 8 times more frequently than isolated increase of CSF IgA. Selective quantitative abnormalities in serum IgG were observed in 35% of patients with subacute sclerosing panencephalitis as compared to 5% in instances in the definite MS group. Abnormal bands in the serum gamma-globulin field were most frequently seen in electropherograms from patients with subacute sclerosing panencephalitis, in cases of malignant lymphoproliferative disorders and in patients with definite MS. No correlation between the concentration of serum and CSF immunoglobulins could be established. Most frequent quantitative abnormalities in serum IgG, IgA and/or IgM were established in malignant lymphoproliferative disorders, in patients with myopathies including myositides and in subacute or chronic inflammatory CNS disorders. Highest incidence of increased CSF IgG, IgA and/or IgM concentration was detected in patients with inflammatory CNS disease, in instances of definite MS and in malignant lymphoproliferative disorders with CNS symptomatology. Increased CSF IgG and IgA concentration with detectable levels of IgM in patients with elevated CSF total proteins indicated alterations in the blood/CNS barrier structures.Supported by PHS grants NB05450 and 06793.     

Significance of CSF immunoglobulins in monitoring neurologic disease activity in Beh?et's disease

We examined the intrathecal production of immunoglobulins (Ig) G, A, and M in 16 patients with Beh?et's disease, 13 of whom have CNS involvement, and in 40 neurologic controls. Oligoclonal IgA and IgM bands were mainly detected in CSF samples from patients with active neuro-Beh?et's disease and were documented to disappear when neurologic manifestations remit. Oligoclonal IgG bands, however, were not related to disease activity and were also found in some neurologic controls. High immunoglobulin index values were detected in both active and quiescent diseases and were high in some patients with impaired blood-CSF barriers. The study presented here demonstrates that CSF oligoclonal IgA and IgM may be helpful in monitoring CNS disease activity in neuro-Beh?et's and could be useful in understanding the pathogenesis of this disease.     

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.     

Serum immunoglobulins in brain tumours and lumbar disc diseases

Changes of serum immunoglobulin (Ig) concentrations may occur in both brain tumours and lumbar disc diseases (LDD). The purpose of this study was to investigate the changes of pre- and post-operative serum Ig levels in brain tumours and LDDs. Serum IgG, IgA and IgM levels were measured in 127 patients with brain tumour, 100 patients with LDD and 20 healthy subjects without neurological disease. Increases in one or more of the pre-operative serum Ig levels were observed in the patients with both brain tumours and LDDs compared with controls. However pre-operative serum IgG level was highly increased in all brain tumour types and LDDs (p<0.001). Serum IgA levels and IgM levels in the post-operative stage were significantly decreased in patients with acoustic neurinoma (p<0.01, p<0.001, respectively). Post-operative serum IgG, IgA and IgM levels were significantly decreased (p<0.001) in patients with meningioma. Post-operative serum IgG and IgM levels were significantly decreased (p<0.001) in patients with glioma. Patients with LDD showed a significantly decline in post-operative serum IgA and IgM levels (p<0.001). We think that decline in post-operative serum Ig levels may be of prognostic value in the patients with brain tumours and LDDs.     

Features of intrathecal immunoglobulins in patients with multiple sclerosis

We have analyzed immunoglobulin (Ig) isotypes and IgG subclasses in cerebrospinal fluid (CSF) and serum of patients with multiple sclerosis (MS) and other neurological diseases to determine whether different Ig isotype patterns correlate with clinical or paraclinical findings and CSF B cell populations. Intrathecal IgG1 synthesis was elevated in MS patients. An increased intrathecal IgM production was found in patients with a higher cerebral MRI lesion burden, whereas other clinical and paraclinical parameters were not associated with a specific Ig isotype or subclass profile. Finally, intrathecal IgG production (IgG1 and IgG3) correlated with the presence of mature B cells and plasma blasts.