Cerebrospinal fluid IgG profiles and oligoclonal bands in Chinese patients with multiple sclerosis

OBJECTIVES: To investigate the significance of oligoclonal bands (OCBs) and intrathecal IgG fractions (IgGIF) for the diagnosis of multiple sclerosis (MS) in northern China. MATERIALS AND METHODS: OCBs in cerebrospinal fluid from 30 patients with MS, 34 with other inflammatory neurological diseases (IND) and 22 with non-inflammatory neurological diseases (NIND) were detected using isoelectric focusing. IgGIF was calculated based on corresponding formula. RESULTS: There was no significant difference in the frequencies of positive OCBs and elevated IgGIF between the MS group and the IND group. Compared with NIND, the MS and IND groups had a significantly higher incidence of OCBs and elevated IgGIF. The sensitivity, specificity and positive result likelihood ratio of OCBs for the diagnosis of MS were 63.3%, 74.2% and 2.5 respectively; those of IgGIF were 36.7%, 84.5% and 2.4. CONCLUSIONS: The two parameters, OCBs and IgGIF are of less diagnostic value for MS in China.     

Identification of IgG subclasses'' oligoclonal bands in multiple sclerosis CSF

IgG subclasses' oligoclonal bands in unconcentrated CSF from MS patients were detected by isoelectric focusing in agarose gel with subsequent immunoblotting using mouse monoclonal antibodies to human IgG subclasses and double-antibody avidin-biotin-alkaline phosphatase system. All MS CSF showed presence of oligoclonal bands specific to the IgG1 subclass; in addition, several of these samples also had oligoclonal bands specific to IgG3, IgG2, or IgG4, in order of decreasing frequency. Since the CSF of a greater number of MS patients showed oligoclonal bands specific to the IgG1 and IgG3 subclasses, the findings are consistent with those reported in patients with chronic viral infections and autoimmune diseases.     

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.     

Oligoclonal IgG bands in Japanese patients with multiple sclerosis. A comparative study between isoelectric focusing with IgG immunofixation and high-resolution agarose gel electrophoresis

A low prevalence of oligoclonal IgG bands (OB) has been reported as a unique feature of Japanese multiple sclerosis (MS) as compared with Western MS. We compared the frequency of OB between isoelectric focusing (IEF) and agarose gel electrophoresis (AGE) in 59 Japanese patients with clinically definite MS [39 with conventional form of MS (CMS) and 20 with optic-spinal form of MS (OSMS)]. The frequency of IEF-OB in total MS was 54%, whereas that of AGE-OB was only 17%. When OSMS was excluded, the frequency increased to 77% by IEF, whereas it remained 23% by AGE. Our study strongly suggests that IEF is highly effective for detecting OB in Japanese MS patients.     

寡克隆区带和IgG指数在多发性硬化中的诊断意义

目的研究寡克隆区带（OCBs）和IgG指数（IgGI）对多发性硬化（MS）诊断的敏感性及其影响因素。方法用等电聚焦结合银染色法检测30例MS、40例神经系统炎性疾病（NID）和22例神经系统非炎性疾病（NNID）患者CSF中OCBs，并计算IgG I。结果MS组和NID组比较OCBs阳性率、IgG I异常率均无显著性差异（P〉0．05）；MS组、NID组与NNID组比较。差异均有显著性（P〈0．05）；传统型MS和脊髓型MS比较，差异均无显著性（P〉0．05）。OCBs对MS诊断的敏感性、特异性和阳性结果似然比分别为63．3％、77．7％和2．8；IgG I分别为40．0％、76．7％和1．7。结论本地区MSOCBs阳性率和IgG I异常率较低，可能与遗传背景、疾病类型和药物应用有关，OCBs和IgG I对MS诊断具有相对特异性。     

Identification of light chain type bands in CSF and serum oligoclonal IgG from patients with multiple sclerosis

The light (L) chain types (kappa and lambda) of oligoclonal IgG bands of matching CSF and serum from 10 MS patients were identified in immunofixation after isoelectric focusing in polyacrylamide gel. Each specimen showed 10-15 oligoclonal bands in pH region of 7.5-9.3. In 7 MS CSF and 5 sera a greater number of oligoclonal IgG bands were of kappa (kappa)-type whereas in 3 CSF and 2 sera the majority was of lambda (lambda)-type. In 3 sera a clearcut correlation of bands with either type of L chain was not observed due to diffuse staining background. Only a small number of oligoclonal IgG bands in 7 of 10 CSF and serum pairs had identical isoelectric points and the same type of L chain. The results show that the individual MS patient had oligoclonal IgG bands in serum, differ with respect to number, isoelectric point and L chain type from the oligoclonal IgG profile seen in the patient's CSF.     

Prevalence of cerebrospinal fluid oligoclonal IgG bands in Greek patients with clinically isolated syndrome and multiple sclerosis

Lower prevalence of cerebrospinal fluid oligoclonal IgG bands (IgG-OCBs) has been reported in multiple sclerosis (MS) patients from Southern Europe compared to other western countries.     

寡克隆带和IgG鞘内合成率对多发性硬化的诊断价值

目的探讨寡克隆带(OCBs)和IgG鞘内合成率(IgGSyn)对多发性硬化(MS)诊断的敏感性、特异性,以及定性和定量指标的相关性。方法选取30例MS(MS组)、40例神经系统炎性疾病(NID组)和22例神经系统非炎性疾病(NNID组)患者,应用速率散射比浊法测定血清和脑脊液(CSF)中免疫球蛋白G(IgG)、白蛋白(Alb)水平,等电聚焦结合银染色法检测CSF中OCBs,计算IgGSyn,并对其敏感性、特异性和阳性结果似然比(PRLR)进行分析。结果OCBs阳性率和IgGSyn异常率MS组与NID组比较差异无显著性;MS组、NID组与NNID组比较差异有极显著性(均P<0.01)。MS组和NID组中,OCBs阳性者与阴性者IgGSyn值差异无显著性。对MS诊断的敏感性、特异性和PRLR,OCBs分别为63.3%、77.7%和2.8;IgGSyn为46.7%、75.2%和1.9。结论OCBs和IgGSyn检测结果的不完全一致性提示中枢神经系统内存在不同的体液免疫反应机制,综合分析OCBs和IgGSyn,对MS诊断具有参考价值。     

IgG1,3 and 4 oligoclonal bands in multiple sclerosis and other neurological diseases

Cerebrospinal fluid (CSF)samples from 10 patients with Multiple Sclerosis (MS) and 7 with other neurological diseases (OND) were studied in order to detect oligoclonal restriction of IgG subclasses 1,3 and 4. Agarose isoelectric focusing (AGA-IEF) followed by Western capillary blotting and immunoperoxidase staining with specific monoclonal antibodies were used. All MS samples showed oligoclonal IgG1 and 6 of them also had IgG3 or IgG4 bands. In the OND group only patients with subacute sclerosing panencephalitis (SSPE) and Guillain-Barré disease (GBD) showed CSF oligoclonal patterns for IgG subclasses. Our results demonstrate that in MS CSF other IgG subclasses beside IgG1 may display an oligoclonal pattern. The finding of more than one subclass in the same band indicates a microheterogeneous composition of these oligoclonal bands.

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Sommario Allo scopo di rilevare un'eventuale restrizione oligoclonale per le sottoclassi IgG1,3 e 4 sono stati studiati campioni di liquido cerebrospinale (LCS) ottenuti da 10 pazienti con Sclerosi Multipla (SM) definita e da 7 pazienti con altre malattie neurologiche (AMN). La metodica utilizzata è stata l'isoelectric focusing in agarosio seguita da Western capillary blotting su nitrocellulosa e colorazione con immunoperossidasi mediante anticorpi monoclonali. Tutti i campioni di SM mostrarono IgG1 oligoclonali e sei di essi presentarono contemporaneamente bande di IgG3 o di IgG4. Tra le AMN solo i pazienti con Panencefalite Sclerosante Subacuta e sindrome di Guillain-Barré mostrarono bande oligoclonali formate da sottoclassi di IgG. I nostri dati dimostrano che nel LCS di SM altre sottoclassi di IgG, oltre alle IgG1, possono essere oligoclonali. La presenza di più di una sottoclasse nella stessa banda depone per una microeterogeneità di queste bande oligoclonali.

     

Clinical and CSF findings in multiple sclerosis patients with or without IgG oligoclonal bands at isoelectric focusing examination of CSF and serum proteins

The IgG oligoclonal bands in CSF can be found in high percentage of MS patients but the existence of a limited number of cases with CSF normal IgG profile is known as well. In the present study 63 out of 70 patients with definite MS and 24 out of 35 with probable MS had oligoclonal bands in the CSF at the isoelectric focusing examination. The 18 patients with normal CSF IgG pattern did not show any statistically significant difference as concerns the age at onset and the duration of the disease, the functional disability, the course of the disease and the quantitative CSF parameters (IgG index, IgG synthesis and serum/CSF albumin quotient). The oligoclonal pattern does not seem of value to discriminate different groups of MS patients but it remains essentially of great diagnostic importance in this disease.     

多发性硬化脑脊液寡克隆区带及24小时鞘内IgG合成率检测的比较

采取垂直平板聚丙烯酰胺凝胶电泳法、火箭免疫电泳法对30例多发性硬化病人进行脑脊液寡克隆区带及24小时鞘内IgG合成率的检测,结果发现寡克隆区带、合成率与多发性硬化的发病年龄、病程、病程进展类型、病残程度无统计学上的相关性,寡克隆区带阳性率46.7％,合成率增高占60％,两者合计异常率可达73.3％,提示我国多发性硬化寡克隆区带阳性率、合成率增高均较国外低,因而在我国对多发性硬化的实验室诊断两者同时检测更有意义。     

Light chain composition of CSF oligoclonal IgG bands in multiple sclerosis and subacute sclerosing penecephalitis

The light chain composition of MS and SSPE CSF oligoclonal IgG bands was examined using isoelectric focusing and sensitive peroxidase-anteperoxidase (PAP) staining technique specific for gamma heavy chains (γ), kappa light chains (κ), or lambda light chains (λ) and a radioimmunoassay (RIA) for γ, κ, or λ. Many bands in the 7 MS and 4 SSPE CSF examined were monoclonal, staining for either IgG-κ or IgG-λ. By staining, all MS CSF were κ predominant; SSPE CSF were variously κ orλ predominant. RIA confirmed the κ predominance of MS CSF. Three MS and 2 SSPE CSF contained bands staining for λ alone, i.e. free light chains. Analysis of RIA data confirmed these findings in 2 MS cases. The difference in light chain predominance of MS and SSPE CSF may reflect differences in the antigenic target, or the age of patient at the time when band-synthesizing clones are triggered. Six of 7 MS and all 4 SSPE CSF contained oligoclonal bands staining for γ and for both κ and λ, probably representing artifacts of IEF. No predominant immunochemical differences between bands in MS and SSPE were detected.