髓鞘碱蛋白对脑梗死和多发性硬化的诊断价值

目的探讨髓鞘碱蛋白（ＭＢＰ）对脑梗死（ＣＩ）和多发性硬化（ＭＳ）的诊断价值。方法对１１４例ＣＩ患者、２８例ＭＳ患者、２４名正常者血清以及部分患者脑脊液（ＣＳＦ）的ＭＢＰ进行了检测。结果ＣＩ、ＭＳ患者血清和ＣＳＦ的ＭＢＰ均显著高于正常对照组（Ｐ〈０．０１）,ＭＳ组又高于ＣＩ组（Ｐ〈０．０１）;ＭＳ组中病情活动期患者ＭＢＰ又高于缓解期。结论血清和ＣＳＦ的ＭＢＰ检测对ＭＳ和ＣＩ的鉴别诊断,以及判断ＭＳ病     

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.     

CSF T-cell subsets in multiple sclerosis: Relationship to cerebrospinal fluid myelin basic protein and clinical activity

Summary Cerebrospinal fluid myelin basic protein and cerebrospinal fluid and peripheral blood T-cell subsets have been studied in patients with multiple sclerosis and other inflammatory and non-inflammatory nervous system diseases. These biological parameters have been correlated with clinical disease activity. No changes in peripheral blood T-cell subsets were seen in multiple sclerosis patients. Low cerebrospinal fluid T8+ cells occurred only in multiple sclerosis, while high cerebrospinal fluid T4+ cells were detected both in clinically active multiple sclerosis and in inflammatory nervous system diseases. A close relationship was found between cerebrospinal fluid T4/T8 ratio and myelin basic protein in relapsing multiple sclerosis patients.Presented in part at the International Symposium on Neuroimmunology, 20–21 September 1988, Cagliari, Italy     

The relationship of interleukin-2 and soluble interleukin-2 receptors to intrathecal immunoglobulin synthesis in patients with multiple sclerosis

The in vivo relationship of interleukin-2 (IL-2) to the local humoral immune response within the central nervous system (CNS) in patients with multiple sclerosis (MS) is hitherto largely unknown. Intrathecal levels of IL-2 and soluble IL-2 receptors (sIL-2R) were correlated to the local CNS synthesis of immunoglobulin G, A, D, and M isotypes in 70 patients with clinically definite MS. Levels were also determined in 19 normal control subjects to establish normal reference limits. High cerebrospinal fluid levels of IL-2 and sIL-2R were detected mainly in patients with acute relapsing-remitting MS and were significantly higher than corresponding serum levels. Intrathecal levels of IL-2 significantly correlated with local CNS synthesis of IgD and IgM, while no correlation was found with either IgG or IgA. Similarly, intrathecal sIL-2R levels significantly correlated with local CNS production of IgD and IgM, but not IgG or IgA. These findings further extend previous reports and also suggest that IL-2 and sIL-2R are involved in the early intrathecal humoral immune response in MS.     

Serum soluble interleukin-2 receptor levels in chronic progressive, stable and steroid-treated multiple sclerosis

Serum levels of the soluble interleukin-2 receptor (sIL-2R), an indicator of T cell activation, were significantly elevated in chronic progressive MS (CPMS) patients, clinically stable MS patients and in patients with other neurological diseases (OND) as compared to healthy controls. Levels of sIL-2R in steroid treated CPMS patients were markedly lower than in untreated CPMS patients and were comparable to healthy controls. Thus, systemic T cell activation occurs in MS during clinically stable and progressive disease stages and in other neurological disorders. The ability of oral corticosteroids to depress serum sIL-2R levels in vivo may be one mechanism by which they exert their therapeutic effect.     

The myelin basic protein gene is not a major susceptibility locus for multiple sclerosis in Italian patients

To verify whether multiallelic polymorphism adjacent to the gene encoding for myelin basic protein is associated with or linked to multiple sclerosis in Italians, we studied 54 sporadic patients, 55 control subjects and 18 families with two or more affected individuals. Allelic typing was carried out by analysis of fragment length polymorphisms after DNA amplification by the polymerase chain reaction. The presence of linkage with the disease was tested according to either autosomal dominant or autosomal recessive modes of inheritance, and with or without the introduction of liability classes accounting for the age of the individuals. Furthermore sib-pair analysis was performed in 11 siblings. No evidence for association or linkage between the myelin basic protein gene polymorphism and multiple sclerosis was found. Our data indicate that in the Italian population the myelin basic protein gene does not play a major role in conferring genetic susceptibility to multiple sclerosis, and suggest that the latter is a heterogeneous phenomenon, possibly influenced by the different ethnic origin of the populations which have been investigated.     

Myelin basic protein in cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases

Summary Cerebrospinal fluid (CSF) from 105 patients was analyzed by radio-immunoassay for the presence of material cross-reactive with peptide 89–169 of bovine myelin basic protein (BP).In a group of 72 multiple sclerosis patients, 52 showed higher BP content than the control group, i.e. more than 2 ng/ml CSF. Increased BP or BP fragments could be detected in CSF from almost all patients who recently (within 2 weeks) had had an acute episode, or after deterioration in the progressive form of the disease. Fifteen to 30 days after the onset of exacerbation or in a stable period, BP content decreases and in the slowly progressive form was in the range of the control group with one exception.BP content was also elevated in the CSF of patients with other neurological diseases. The presence of BP in the CSF from patients with isolated retrobulbar neuritis is of particular interest. Thus the presence of material cross-reactive with BP fragment 89–169 is not specific for multiple sclerosis, but is a useful parameter in diagnosis and evaluation of MS.

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Zusammenfassung Mit Hilfe eines Radioimmunoassays wurde der Liquor von 105 Patienten auf Substanzen untersucht, die mit dem Fragment 89–169 des bovinen basischen Myelinproteins (BP) kreuzreagieren. Von 72 Patienten mit Multipler Sklerose hatten 52 einen höheren BP-Gehalt im Liquor als die Kontrollgruppe, d.h. mehr als 2 ng/ml. Dies war bei fast all jenen Patienten der Fall, bei denen der Liquor innerhalb von 2 Wochen nach einem akuten Schub bzw. zum Zeitpunkt einer deutlichen Verschlechterung des Zustandes bei der chronisch-progredienten Form gewonnen wurde. Der BP-Gehalt des Liquors nimmt zwischen dem 15. und 30. Tag nach Schubbeginn deutlich ab. In der schleichenden progredienten Form wurde mit einer Ausnahme kein erhöhter BP-Gehalt gefunden.Das basische Myelinprotein ist aber auch im Liquor von Patienten mit anderen neurologischen Erkrankungen zu finden. Hervorzuheben ist das Auftreten von BP im Liquor von Patienten mit Retrobulbär-neuritis.Der Nachweis von mit Bruchstück 89–169 kreuzreagierenden Substanzen im Liquor ist infolgedessen zwar nicht spezifisch für die Multiple Sklerose, ist aber ein wertvoller Parameter in der Diagnose und Einschätzung der MS.

     

In vivo relationship of interleukin-2 and soluble IL-2 receptor to blood-brain barrier impairment in patients with active multiple sclerosis

Interleukin (IL)-2 has well-recognized effects on cerebral endothelial cells and, therefore, may mediate disruption of the blood-brain barrier in patients with multiple sclerosis (MS). To evaluate the in vivo relationship of the IL-2 system to blood-brain barrier impairment in MS, levels of IL-2 and soluble IL-2 receptors (sIL-2R) in cerebrospinal fluid (CSF) and serum samples from 50 patients with active MS and 49 controls were correlated with values of the CSF to serum albumin ratio. Intrathecal levels of IL-2 and sIL-2R were significantly higher in MS compared with the control groups and correlated with albumin ratios in MS patients. Intrathecal levels of IL-2 and sIL-2R also correlated with the degree of barrier damage in these patients. It is suggested that intrathecal levels of IL-2 and sIL-2R are related to barrier impairment in MS and may be important in understanding some of the pathological changes of this condition.     

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.     

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.     

多发性硬化患者脑脊液sIL-2R水平变化的研究

目的：探讨可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）在多发性硬化（ＭＳ）发病机理中的作用及其临床意义。方法：采用双抗体夹心ＥＬＩＳＡ法对４４例ＭＳ患者脑脊液ｓＩＬ－２Ｒ水平进行了检测。结果：ＭＳ组脑脊液ｓＩＬ－２Ｒ水平显著高于正常对照组,脑脊液ｓＩＬ－２Ｒ水平变化与ＭＳ患者疾病活动状态及病残程度密切相关。结论：ＭＳ患者体内Ｔ淋巴细胞处于活化状态,ｓＩＬ－２Ｒ在ＭＳ发病机理中发挥着重要作用,检测脑脊液ｓ     

Myelin basic protein gene polymorphism is not associated with chronic progressive multiple sclerosis

In the present study a tetranucleotide (TGGA)n repeat polymorphism 5' to the myelin basic protein (MBP) gene was evaluated in a group of HLA-class II-typed, chronic progressive multiple sclerosis (MS) patients. This polymorphism has been reported by others to be associated with MS. Contrary to these reports we observed similar allele frequencies in patients and controls. Our results indicate that there is no association between MS and a polymorphism 5' to the MBP gene.     

HLA-DM polymorphisms do not associate with multiple sclerosis: an association study with analysis of myelin basic protein T cell specificity

The MHC region on 6p harbors at least one susceptibility gene for multiple sclerosis (MS). Within this region, HLA-DM loci are of interest being involved in class II antigen processing. We investigated the association of HLA-DM polymorphisms with MS. Sixty-three patients with MS and 46 healthy controls from continental Italy were typed for HLA-DM polymorphisms and HLA-DRB1 alleles. Besides, among the donors characterized for the DM polymorphisms, we considered 6 MS patients previously studied for the fine specificity of their MBP-specific T lymphocyte lines (TLL). The frequencies of allelic variants at the DMA and DMB loci were similar between MS patients and controls, even when HLA-DRB1^*1501 positive and negative donors were analyzed separately. Patients with predominant responses to different MBP epitopes did not differ for their HLA-DM haplotype while patients with predominant responses to the same MBP epitope could present different HLA-DM haplotypes. HLA-DM polymorphisms do not associate with MS and may not affect specific patterns of T cell responses to MBP.     

Linkage analysis of candidate myelin genes in familial multiple sclerosis

Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system. A complex genetic etiology is thought to underlie susceptibility to this disease. The present study was designed to analyze whether differences in genes that encode myelin proteins influence susceptibility to MS. We performed linkage analysis of MS to markers in chromosomal regions that include the genes encoding myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG), oligodendrocyte myelin glycoprotein (OMGP), and myelin oligodendrocyte glycoprotein (MOG) in a well-characterized population of 65 multiplex MS families consisting of 399 total individuals, 169 affected with MS and 102 affected sibpairs. Physical mapping data permitted placement of MAG and PLP genes on the Genethon genetic map; all other genes were mapped on the Genethon genetic map by linkage analysis. For each gene, at least one marker within the gene and/or two tightly linked flanking markers were analyzed. Marker data analysis employed a combination of genetic trait model-dependent (parametric) and model-independent linkage methods. Results indicate that MAG, MBP, OMGP, and PLP genes do not have a significant genetic effect on susceptibility to MS in this population. As MOG resides within the MHC, a potential role of the MOG gene could not be excluded. Received: January 12, 1999 / Published online: August 17, 1999     

Specificity of autoantibodies to epitopes of myelin proteins in multiple sclerosis

An autoimmune response to one or more myelin-protein components is thought to be part of the pathogenesis of multiple sclerosis (MS). The immunodominant-autoantibody epitope may be localized on a linear peptide segment, on a conformation-sensitive epitope, or on an epitope resulting from post-translational modifications. Primary, secondary, and tertiary structures of myelin proteins may determine the specific site for binding of autoantibodies. A myelin protein-specific autoantibody can bind to either a linear or conformational epitope, whereas all of the T cell epitopes are linear. At present, the conformational epitopes of myelin proteins have not been identified; most of the methods used to identify the myelin-protein epitopes corresponding to the pathogenesis of multiple sclerosis are involved in the linear epitope mapping. Polymorphism or mutations may cause inappropriate expression of the myelin proteins with alterations to their linear and/or conformational epitopes, and make them susceptible to autoantibody binding, especially if these changes occur at the surface of the protein. This review focuses on the specificity of autoantibodies to the epitopes of myelin proteins and correlates this to the structures of proteins. Factors that influence the expression of myelin-protein epitopes such as the alpha-helical or beta-sheet structure of the protein, the tri-proline site, and the post-translational modifications as well as physicochemical properties of amino acid changed are included.     

Target epitopes of myelin basic protein specific T cell lines in multiple sclerosis

A highly efficient new method for the generation of antigen specific T cell lines (TCL) is now available. By this method we established 134 myelin basic protein (MBP) TCL from the peripheral blood of 9 patients with definite multiple sclerosis (n=69) and 8 healthy donors (n=65). The yield of MBP reactive TCL in the two groups was comparable. So far 22 MBP specific TCL from 7 patients and 24 from 7 healthy individuals have been tested for their proliferative response to a panel of four synthetic peptides representing MBP residues 7–26, 80–99, 139–153 and 148–162. Although the peptide sequences did not encompass the whole MBP, the pattern of reactivity to these peptides in patients and controls seems to be similar. Further, when multiple TCL from the same donor were analysed, no dominant recognition emerged. Paper presented at the National Congress at Sorrento in 1991 and selected by the Editorial Board of the Journal     

A functional basis for the association of HLA class II genes adn susceptibility to multiple sclerosis: cellular immune responses to myelin basic protein in a multiplex family

This study has examined the cellular response to myelin basic protein (MBP) ina multiplex family with multiple sclerosis (MS). A total of 81 MBP-specific T cell liens (TCLs) were derived from three affected siblings and four healthy siblings. No difference was observed in estimated precursor frequenceis of MBP-specific TCLs or peptide specificity of TCLs when comparing affected and unaffected siblings. MPB-specific TCLs from affected siblings, however, were restricted to the DRw15/DQw6 allele more frequently than those from unaffected siblings (P < 0.02). These data suggest that restriction of autoantigen-specific T cells may be the functional basis for disease susceptibility related to HLA class II inheritance.     

牛脊髓髓鞘碱性蛋白的提取及初步鉴定

目的探索一条从牛脊髓中提取牛髓鞘碱性蛋白(M BP)的行之有效的方法,为实验性自身免疫性脑脊髓炎(EAE)动物模型的建立及研究工作奠定基础。方法组织匀浆器将新鲜的牛脊髓捣碎,用London法抽提,考马司亮蓝法鉴定粗提品的含量,SDS-PAGE观测其分子量,M BP免疫W istar大鼠,以鉴定其抗原性。结果粗提品蛋白含量达10m g/m l,粗提品诱导EAE症状典型,发病率高。结论采用London法可从牛脊髓中获取大量的M BP,与其它方法比较产量高,诱导EAE效果好,且本法简便,所以有一定的推广价值。     

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.     

A study of human T-cell lines generated from multiple sclerosis patients and controls by stimulation with peptides of myelin basic protein

We generated T-cell lines from the peripheral blood of controls and of patients with multiple sclerosis (MS) by stimulation with overlapping synthetic peptides representing the entire sequences of all four isoforms of human myelin basic protein (MBP). The T-cell lines reacted to a wide range of epitopes in the major isoforms of MBP and to epitopes that were present only in the minor isoforms. Many MS patients and controls had T-cells responding to one or more cryptic MBP epitopes, as indicated by the generation of a peptide-specific T-cell line(s)by stimulation with synthetic peptides but not by stimulation with whole MBP. About one-third of the peptide-generated lines were cytotoxic. Although we have shown that this technique of peptide stimulation is effective in generating human antiviral cytotoxic CD8⁺ T-cell lines, all the cytotoxic MBP-specific lines generated by this method were predominantly CD4⁺. Our study did not reveal any significant differences, between MS patients and controls, in reactivity to epitopes within any of the isoforms of MBP.