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.     

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.     

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。     

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.     

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.     

寡克隆区带和IgG指数对多发性硬化的诊断价值

目的　探讨寡克隆区带 (OCB)和IgG指数对多发性硬化 (MS)诊断的敏感性及特异性。方法　收集 4 8例MS、6 8例神经系统炎性疾病 (NID)及 110例非炎性疾病 (NNID) 3组患者的脑脊液 (CSF)和血清标本 ,分别进行OCB的检测 (等电聚焦 )和IgG指数的计算。并对其阳性结果似然比 (PRLR)进行分析。结果　MS组与NID组比较 ,CSF中OCB阳性率和IgG指数异常率的差异均没有显著性 (均P >0 0 5 ) ;但MS组、NID组与NNID组比较 ,差异均有极显著性 (均P <0 0 0 0 1)。MS组CSF中OCB和IgG指数的敏感性分别为 39 6 %、6 0 4 % ;特异性分别为 80 3%、72 1% ;PRLR分别为 2 0、2 2。当用于判断有无IgG鞘内合成时 ,特异性分别为 97 2 %、92 7% ;PRLR分别为 13 5、7 3。结论　CSF中OCB阳性和IgG指数升高强烈提示有中枢神经系统局部IgG合成 ,对MS有一定的辅助诊断价值     

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.     

Relationships between abnormal IgG index,oligoclonal bands,acute phase reactants and some clinical data in multiple sclerosis

Summary The IgG-index and acute phase reactants were measured and oligoclonal bands were looked for in 30 patients with clinically definite multiple sclerosis (MS) and were compared with the clinical data. IgG-index was found elevated in 77% against 22% in a comparable material of patients with other neurological diseases. Oligoclonal bands were found in 74 and 17%, respectively. No correlation was found between these parameters and age, duration of illness, disability, coefficient of progression or age of onset. A statistical evaluation defines the specificity, sensitivity and validity of the methods used. CRP could not be demonstrated in 28, the rest had normal values. C3A was normal in all cases, but appeared to be elevated a little with increasing age, and to be positively correlated to the albumin-index, indicative of a defective blood-brain barrier. No correlation was found between CRP, C3A and the clinical data.

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Zusammenfassung Bei 30 Patienten mit klinisch sicherer Multipler Sklerose (MS) wurden IgG-Index und akute Fase Reaktanten gemessen und die Anwesenheit von oligoklonalen Bändern untersucht. Diese Daten wurden mit klinischen Daten verglichen. Der IgG-Index ist um 77% erhöht gegenüber 22% in einem Vergleichsmaterial aus Patienten mit anderen neurologischen Krankheiten. Oligoklonale Bänder wurden bei 74% bzw. 17% gefunden. Kein Zusammenhang wurde zwischen diesen Parametern und Alter, Krankheitsdauer, Invalidität, Progressionskoeffizient oder Alter bei Erkrankungsbeginn gefunden. Bei statistischer Untersuchung wurden Spezifizität, Sensitivität und Validität der verwendeten Methoden bestätigt. CRP war nicht nachweisbar bei 28, und die übrigen 2 zeigten Werte innerhalb des Normbereichs. C3A war in allen Fällen normal, war jedoch leicht erhöht in höherem Alter und auch bei höherem Albumin-Index als Parameter einer geschädigten Bluthirnschranken-Funktion. Kein Zusammenhang zwischen CRP, C3A und den klinischen Daten.

     

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。     

Antinuclear antibodies positivity is not rare during multiple sclerosis and is associated with relapsing status and IgG oligoclonal bands positivity

IntroductionAs an immune-mediated disease of the central nervous system, multifaceted aspects of a humoral immune response are widely described during multiple sclerosis (MS). However, the prevalence of different auto-antibodies, such as antinuclear antibodies (ANA), during MS is very variable and their clinical relevance remains controversial. Our aim was to evaluate the prevalence and clinical correlations of ANA positivity in South Tunisian MS patients.Material and methodsWe performed ANA screening using indirect immunofluorescence (IIF) on HEp-2 cells (Biosystems®) in 82 MS patients. For ANA positive samples (titer ≥1/160), anti-ds-DNA detection (IIF on Crithidia luciliae (Biosystems®)) and extractable nuclear antigen typing (immunodot (Euroimmun®)) were performed.ResultsANA were positive in 35/82 MS patients (42.7%). The titer was ≥ 1/320 in 16/35 patients. The antigenic specificity of ANA was identified in 7/35 patients. None of the patients had extra-neurological manifestations. No correlation was found between ANA and age, gender, MS course, disease duration, disability, annual relapse rate nor IgG index. ANA positivity was more frequent in patients with IgG oligoclonal bands (OCB) (47.1%) than in patients without IgG OCB (16,6%) (p = 0.049). Regarding disease activity, ANA positivity was significantly more frequent in patients with relapse (52.6%) than in patients in remission (25.9%) (p = 0.031).ConclusionOur results showed that ANA positivity in MS disease is not rare. This positivity was not associated with clinical expression of any connective tissue disease. ANA occurrence in MS was associated with IgG OCB+ profile and relapsing status, probably reflecting an ongoing immune dysregulation.     

Identification of the κ and λ light chains within the CSF immunoglobulin region in multiple sclerosis and subacute sclerosing panencephalitis by immunofixation after isoelectric focusing

Summary The distribution of the and light chains within the cerebrospinal fluid (CSF) IgG oligoclonal bands of 15 multiple sclerosis (MS) and 3 subacute sclerosing panencephalitis (SSPE) patients has been studied by isoelectric focusing (IF) followed by direct immunofixation. The oligoclonal bands demonstrated by IF were both homogeneous and heterogeneous with regard to the light chain type; in the MS patients there was a prevalence of heterogeneous bands and of homogeneous bands of type, while in SSPE a slight prevalence of homogeneous bands was found. The homogeneous bands were preferentially disposed in a more cathodal position in respect to IgG bands; in some cases bands probably constituted only of free light chains of type were found. Finally, in some cases, the oligoclonal IgG pattern was more distinct after immunofixation than after basal IF.

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Zusammenfassung Die Verteilung von leichten - und -Ketten in den oligoklonalen IgG-Fraktionen des CSF ist bei 15 MS- und 3 SSPE-Patienten durch isoelektrische Fokussierung (IF) und anschließende direkte Immunofixierung untersucht worden.Die durch IF hervorgehobenen oligoklonalen Fraktionen waren sowohl homogen als auch heterogen hinsichtlich des Typs der leichten Ketten; bei den MS-Patienten waren die heterogenen und homogenen IgG-Fraktionen des Typs vorherrschend, während bei den SSPE-Patienten die homogenen IgG-Fraktionen des Typs leicht vorherrschend waren.Die homogenen IgG-Fraktionen des Typs waren im Vergleich zu den IgG-Fraktionen des Typs vorzugsweise in einer kathodennaheren Position angetroffen worden; in einigen Fällen wurden IgG-Fraktionen gefunden, die wahrscheinlich nur aus freien leichten Ketten des Typs bestehen.Schließlich waren in einigen Fällen die oligoklonalen IgG-Fraktionen nach der Immunofixierung deutlicher als nach der IF.

     

Optic neuritis: oligoclonal bands increase the risk of multiple sclerosis

ABSTRACT- In 1974 we examined 30 patients 0.5–14 (mean 5) years after acute unilateral optic neuritis (ON), when no clinical signs of multiple sclerosis (MS) were discernable. 11 of the patients had oligoclonal bands in the cerebrospinal fluid (CSF). Re-examination after an additional 6 years revealed that 9 of the 11 ON patients with oligoclonal bands (but only 1 of the 19 without this CSF abnormality) had developed MS. The occurrence of oligoclonal bands in CSF in a patient with ON is - within the limits of the present observation time - accompanied by a significantly increased risk of the future development of MS. Recurrent ON also occurred significantly more often in those ON patients who later developed MS.     

CSF oligoclonal bands, MRI, and the diagnosis of multiple sclerosis

In this retrospective study, the results from investigations (MRI, evoked potentials, alkaline oligoclonal bands [OBs] in CSF) in 94 patients with clinical suspicion of demyelinative disease were evaluated to assess their impact on diagnosis. Forty-three patients were diagnosed as having definite MS, 10 probable MS, and 9 possible MS. MRI findings strongly suggestive of MS were evident in 52/62 (84%) patients, while 47/62 (76%) patients demonstrated OBs in their CSF. In 63% of patients both abnormalities were present. Patients with no OBs in their CSF were on the average older, were more often male, had experienced their first symptoms at a later age, and suffered more often from the chronic-progressive form of the disease than those with a positive CSF finding.     

IgG ratios and oligoclonal IgG in multiple sclerosis and other neurological disorders

The findings are reported of various CSF abnormalities, including IgG indices and oligoclonal IgG, in 160 patients with multiple sclerosis of differing diagnostic certainty and 146 patients with other neurological disorders.

An abnormal IgG index, defined as the ratio of IgG/albumin in CSF to that in serum, has been found in 77.7% of definite MS cases, falling to a figure of 32.1% in the single lesion group. A tendency, reported previously, for IgG levels to be higher in disabled patients, particularly those with a short history or early onset, has been confirmed.

Oligoclonal IgG, on the other hand, has been found in 56% of definite MS cases, less frequently than in most other reported series. Analysis of the literature suggests considerable variability in the finding of oligoclonal IgG in other than definite MS, and in other neurological disorders. The possibility that subjective factors are partly responsible for this variability, rather than discrepancies in patient selection requires consideration, and suggests that CSF electrophoresis and IgG estimations are complementary aids in the diagnosis of multiple sclerosis.

Differences have been expressed regarding the relationship of oligoclonal IgG to clinical parameters of the disease. Further sequential analysis of the development and variability of the oligoclonal pattern in MS is required.     

Isotachophoresis evaluation of synthesis of intrathecal IgG subfractions in multiple sclerosis

Summary Cerebrospinal fluid (CSF) and serum samples from patients with multiple sclerosis and other neurological diseases were examined by capillary isotachophoresis (ITP). The percentage and the rate of synthesis of CSF IgG which migrated slowly with ITP were calculated.CSF specimens of most patients with multiple sclerosis contained increased percentages of slowly migrating IgG (slow IgG), corresponding to IgG oligoclonal bands in the high-alkaline region on isoelectric focusing. The patients with multiple sclerosis were found to have increased intrathecal synthesis of slow IgG, which correlated closely with the rate of intrathecal CNS IgG synthesis calculated by Tourtellotte's formula.

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Zusammenfassung Liquor und Serum von Patienten mit Multipler Sklerose und anderen neurologischen Erkrankungen wurden mittels Kapillar-Isotachophoresis (ITP) untersucht. Es wurde berechnet, welchen Anteil das IgG im Liquor, welcher langsam mit der ITP wanderte, ausmachte und wie seine Synthesegeschwindigkeit war. Bei den meisten Patienten mit Multipler Sklerose enthielt der Liquor einen erhöhten Anteil von langsam wandernden IgG, welches oligoclonalen Banden im hochalkalischen Bereich beim isoelectric focusing entsprach. Es wurde gefolgert, daß die Patienten mit Multipler Sklerose eine erhöhte intrathekale Synthese des langsamen IgG aufwiesen, wobei dies eng korrelierte mit dem Anteil der intrathekalen Liquor-IgG-Synthese, wie sie nach der Tourtellotteschen Formel errechnet werden kann.

     

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。     

Comparison of two methods for the detection of oligoclonal bands in a large number of clinically isolated syndrome and multiple sclerosis patients

Objective

A novel oligoclonal band (OB) assay which consists of isoelectric focusing (IEF) and IgG immunodetection by alkaline phosphatase-labeled anti IgG antibody was reported to be very sensitive. It also accurately predicted conversion to MS in patients with CIS. The aim of our study was to compare sensitivity of a novel and the standard procedure with peroxidase immunodetection in a large number of CIS and MS patients.

Methods

OB were determined in serum and CSF samples in 161 patients (104 females), 47 with CIS and 114 with MS with median age 38 years (range 19–68) using both methods.

Results

Eighty-three percent of patients had CSF OB with the standard and 89% with the novel method. Median number of OB was 5 (range 0–17) with the peroxidase and 8 (range 0–18) with the alkaline phosphatase method; p = 0.001. Twenty-one percent of patients had ≥10 OB with the standard and 37% with the novel method of the detection; p = 0.021. Subjective impression of band clarity showed that 20% of patients had sharper and stronger bands when the peroxidase and 65% when the alkaline phosphatase method was used; p < 0.0001.

Conclusion

The alkaline phosphatase method is more sensitive than the peroxidase method and at the same time cheaper, easy to perform and less time consuming.     

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。