Presence of anti-CSL antibodies in the cerebrospinal fluid of patients: A sensitive and specific test in the diagnosis of multiple sclerosis

The carbohydrate-binding protein (lectin) CSL is an antigen involved in the stabilization of the myelin structure by interacting with the carbohydrate moiety of myelin glycoproteins. Since anti-CSL Fab fragments were able to produce destruction of CNS myelin in vitro, CSL was considered as a potential immunological target in multiple sclerosis. The presence of anti-CSL antibodies has been examined in the cerebrospinal fluid of 1388 different patients with various neurological diseases. It is concluded that the presence of anti-CSL antibodies in the cerebrospinal fluid of patients less than 50 years old constitutes a very sensitive and specific test for multiple sclerosis.     

Follow-up of patients with suspected multiple sclerosis: a clinical and electrophysiological study.

We re-examined 21 patients with suspected multiple sclerosis, seven classified as probable and 14 as possible cases. At the first investigation all patients but two had abnormal visual evoked potentials and somatosensory evoked potentials, or both. All but three had an increased intracerebral production of immunoglobulin G expressed by the cerebrospinal fluid IgG index. At follow-up two to four years later, 13 of 16 patients (81%) in whom both evoked potentials and IgG index were abnormal initially had entered into a higher multiple sclerosis diagnostic class. In the five patients in whom either evoked potentials or IgG index were normal the original diagnosis was unchanged.     

Suspected and clinically definite multiple sclerosis: the relationship between CSF immunoglobulins and clinical course.

CSF immunoglobulins were examined in 103 patients with clinically definite multiple sclerosis, 106 patients with either suspected or progressive possible multiple sclerosis and 72 patients with other neurological diseases. Raised CSF IgG index and oligoclonal banding were found in 71% and 75% of clinically definite multiple sclerosis patients respectively and both tests were abnormal in 11% of patients with other neurological diseases. The CSF IgG index and the presence of oligoclonal IgG did not relate to the severity or duration of established disease in these patients. In patients with suspected and progressive possible multiple sclerosis, both a raised IgG index and the presence of oligoclonal banding were found significantly more frequently than in the OND group. Abnormalities of these parameters were significantly correlated with the presence of an abnormal evoked response in these patients (chi 2 = 10.16 p less than 0.01). When 47 patients with suspected multiple sclerosis were studied prospectively the presence of oligoclonal banding at presentation was associated with development of further disease activity.     

The concentration of IgM in the cerebrospinal fluid of neurological patients

The level of IgM was determined by Particle Counting Immunoassay in the cerebrospinal fluid. In non-neurological patients (N = 20) the mean was 97.5 μg/l with the upper reference limit at 380 μg/1. The mean IgM index was 0.021 with the upper reference limit at 0.071. Of 21 patients with stroke, 5 had an IgM index exceeding the reference limit. High levels and indices of IgM were observed in most patients (N = 27) with infectious meningo-encephalitis. In this group, the IgM index was abnormal in about 30% of cases with a normal total protein content, and was more often increased than the IgG index. In multiple sclerosis patients (N = 80), the IgM index was increased in 32%. In this disease very high values of IgM index (> 0.13) were never associated with very high values of IgG index (> 1.8). A significantly higher proportion of males was found in the group of patients with very high values of IgM index (N = 11). No significant influence of the age of onset, the interval between onset and sampling and clinical state was observed. However, of 10 patients with a multiple sclerosis history exceeding 15 years none had an IgM index exceeding the upper reference limit. Four patients with multiple sclerosis had a high IgM index without either an increase of the IgG index or the presence of oligoclonal bands.     

Long-term persistence of intrathecal viral antibody responses in postinfectious diseases of the central nervous system and in Rett syndrome

Twenty patients: seven with herpes simplex virus encephalitis (HSVE), six with other severe central nervous system (CNS) infections and 7 with Rett syndrome were studied to determine whether they showed any intrathecal synthesis of virus-specific or total IgG in CNS. The study of the postinfectious patients was made a mean of 20 years after the primary infection in childhood. Four of seven patients with HSVE had an elevated IgG index and four showed intrathecal viral antibody production which was both specific (against HSV) and non-specific. One patient with congenital syphilis and one with tuberculotic meningitis showed non-specific intrathecal viral antibody synthesis. In three of seven patients with Rett syndrome intrathecal antibody production was observed. The clarification of the mechanism of polyclonal immunoactivation in postinfectious diseases would be of interest since similar persistent immunoactivation is a common feature in multiple sclerosis. In Rett syndrome the immunoactivation may also have pathogenetic significance.     

Myelin encephalitogenic protein fragments in cerebrospinal fluid of persons with multiple sclerosis.

With a double-antibody radioimmunoassay performed on unconcentrated cerebrospinal fluid, eight of 14 patients in an acute phase of multiple sclerosis had levels of 3.4 to 15.4 ng per milliliter of the P1 fragment (residues 43-88) of myelin encephalitogenic protein. Encephalitogenic protein-P1 was found only in the acute phase and was present in six of seven persons in the first week of an exacerbation and absent in 29 multiple sclerosis patients who were stable or had a gradually progressive course. Six of 117 controls had detectable cerebrospinal fluid encephalitogenic protein-P1. Only in two of these, one with a recent cerebral infarction and one with diabetic nephropathy who was in coma, were the levels in the range encountered in patients in the acute phase of multiple sclerosis. Although not entirely specific for multiple sclerosis, the presence of material in the cerebrospinal fluid of multiple sclerosis patients cross-reacting with encephalitogenic protein-P1 appears to be a characteristic of acute exacerbations.     

An endogenous lectin found in rat astrocyte cultures has a role in cell adhesion but not in cell proliferation

The presence of an endogenous cerebellar soluble lectin (CSL) has been demonstrated in cultured rat astrocytes by using immunocytochemical techniques. In these cells, the location of lectin CSL was found intracellularly as well as on the external surface of the plasma membrane of the cell bodies and processes, especially in the zones of contact between cells. This suggested that CSL could have a role in adhesion of astrocytes to sister cells. Kinetics of adhesion of astrocytes to culture dishes precoated with CSL showed a rapid binding of these cells. In confluent astrocyte cultures, anti-CSL Fab fragments affected the shape and organization of astrocytes (retraction of the cytoplasm), but they did not detach cells from the substratum. These results indicated that CSL has adhesive properties for astroglial cells and is probably involved 1) in adhesion of astrocytes to sister cells; 2) in binding of protoplasmic regions of astrocyte membrane to the substratum. Further support for these roles came from demonstration of the presence in cultures of glycoprotein ligands recognized by this lectin. The problem of the mitogenic properties of the lectin was also questioned. The addition of CSL to confluent astroglial cultures was able to stimulate only by 40% the proliferation of these cells at an optimal concentration of 5 micrograms CSL lectin/ml of culture medium. This indicated that CSL is not a powerful growth factor for astrocytes.     

Cerebrospinal fluid oligoclonal IgG bands in patients with spinal arteriovenous malformation and structural central nervous system lesions

OBJECTIVE: To investigate the incidence and characteristics of patients with structural central nervous system (CNS) lesions and cerebrospinal fluid oligoclonal IgG bands. DESIGN: A retrospective study. METHOD: The medical records of patients with cerebrospinal fluid oligoclonal IgG bands were evaluated for the presence of structural CNS lesions, their location and cause, and for clinical characteristics. SETTING: Cerebrospinal fluid oligoclonal IgG bands were examined in the Neuroimmunology Laboratory, Hadassah University Hospital, Jerusalem, Israel. PATIENTS: Two hundred seventy of 570 patients with positive cerebrospinal fluid oligoclonal IgG bands were available for analysis. Twenty patients had structural CNS lesions. RESULTS: Twenty (7.5%) of the 270 patients had structural CNS lesions: 3 patients had spinal arteriovenous malformation; 5 patients had tumors; 9 patients had compressive cervical myelopathy. Traumatic leukomalacia, Arnold-Chiari malformation type 1, and CNS hemosiderosis were present in 1 patient each. In 2 patients (1 patient with recurrent meningioma and 1 patient with posttraumatic encephalomalacia) the presence of a structural CNS lesion was followed by the development of multiple sclerosis. In all 3 patients with spinal arteriovenous malformation, oligoclonal IgG identification prolonged the time to diagnosis and therapy, which varied from a few weeks to 3 years. CONCLUSIONS: Structural CNS lesions, responsible for the neurological disorder, were present in 20 patients (7.5%) with cerebrospinal fluid oligoclonal IgG bands. The mechanism underlying oligoclonal IgG presence in spinal arteriovenous malformation and the coexistence of multiple sclerosis and structural CNS lesions is unknown, but may be related to recurrent tissue damage with repeated presentation of CNS antigens to the immune system.     

Anti-myelin basic protein and anti-proteolipid protein specific forms of multiple sclerosis

Human myelin basic protein (hMBP) and proteolipid protein (PLP) were used as antigens in a solid-phase radioimmunoassay to determine relative frequencies of anti-MBP and anti-PLP in cerebrospinal fluid (CSF) of optic neuritis and multiple sclerosis (MS) patients. Forty-nine of 55 patients with optic neuritis had increased CSF anti-MBP and the remaining 6 had increased anti-PLP. Of 385 MS patients, MS relapse: 173 of 180 patients had increased anti-MBP, 5 of the remaining 7 patients had elevated anti-PLP, and 2 had neither of these autoantibodies. Progressive MS: 111 of 116 patients had increased anti-MBP in either free and/or bound form, of the remaining 5 patients 4 had increased anti-PLP, and 1 had neither anti-MBP nor anti-PLP. MS remission: 15 of 87 patients had somewhat increased anti-MBP, none had anti-PLP. IgG was purified by affinity chromatography from necropsy central nervous system (CNS) tissue samples of 4 individual patients with clinically definite and neuropathologically confirmed MS. Three of these 4 patients who had increased levels of CSF anti-MBP also had increased anti-MBP titers in CNS tissue-extracted IgG. The fourth patient who had anti-PLP in CSF also had anti-PLP in brain tissue IgG. These autoantibodies were not detected simultaneously in any patient. These results suggest that there are at least two immunologically distinct forms of MS, i.e., a common form highly associated with anti-MBP and more frequent prominent inflammatory characteristics in CSF and CNS, and an infrequent form associated with anti-PLP in CSF and tissue, and less abundant inflammation. Anti-MBP purified from CNS tissue IgG by antigen-specific affinity chromatography was reacted with synthetic peptides of hMBP. The anti-MBP epitope on the hMBP molecule was restricted between residues 75 and 106. The PLP epitope for anti-PLP has not as yet been determined. These observations have theoretical implications for anticipated future specific immunotherapy of MS.     

Neuromyelitis optica immunoglobulins as a marker of disease activity and response to therapy in patients with neuromyelitis optica

ObjectiveTo determine whether neuromyelitis optica (NMO) immunoglobulin (IgG) antibody status in NMO/Devic's disease patients followed prospectively is persistent or can change relative to the clinical status and/or response to therapy.DesignA cross-sectional group of patients with NMO, relapsing extensive longitudinal transverse myelitis (RLETM) or optico-spinal multiple sclerosis (OSMS) were evaluated for the presence of NMO IgG antibodies. Repeated evaluation was made in all NMO/RLETM patients and in a subgroup of OSMS patients.SettingBaird Multiple Sclerosis Center, Buffalo, New York, an academic multiple sclerosis center.ResultsOut of a consecutive cohort of 38 patients evaluated for the presence of NMO IgG, 12 had NMO and 26 had OSMS. Five of the 12 NMO/RLETM patients were NMO IgG positive at the time of their initial evaluation. Four of these patients were repeatedly tested for NMO IgG: two of these became negative and two remained positive. One patient who was initially negative became positive during an acute event and again became negative during the stable disease phase following treatment. A positive test result was associated with active disease, whereas a negative NMO IgG result was consistently found in stable, long-term treated patients. None of the OSMS patients were positive for NMO IgG even during acute attacks.ConclusionsNMO IgG antibodies are associated with active NMO/RLETM. A well-controlled stable disease usually under effective immunosuppressive therapy can transform the NMO IgG to a negative status. Repeated NMO IgG testing should be considered as a useful biological marker for monitoring NMO/RLETM disease and or response to therapy.     

Relation between benign course of multiple sclerosis and low-grade humoral immune response in cerebrospinal fluid.

The prognosis in multiple sclerosis (MS) is related to the presence of an abnorma humoral immune response within the central nervous system: 14/17 MS patients (82%) without oligoclonal CSF IgG displayed no or slight disability after a mean duration of MS of 17 years, while 53% of 88 patients with oligoclonal CSF IgG had a benign course after a mean duration of 13 years (p less than 0.05). A benign course also was more often accompanied by a normal CSF IgG index. MS patients without oligoclonal CSF IgG had elevated CSF/serum ratios of albumin in 6%, and of the complement factors C3 in 0% and C4 in 6%, as against 20%, 27% and 37%, respectively, in MS patients with oligoclonal CSF IgG.     

A prospective study on the predictive value of CSF oligoclonal bands and MRI in acute isolated neurological syndromes for subsequent progression to multiple sclerosis.

A prospective study in patients with a clinical acute isolated brainstem or spinal cord disorder was undertaken. The aim was to evaluate the predictive value of IgG intrathecal synthesis (through the detection of oligoclonal bands in CSF) and MRI lesions at presentation, for the subsequent progression to multiple sclerosis. Forty four patients took part in this study: 22 had a brainstem disorder and 22 a spinal cord disorder. After a mean period of 26 (SD 22) months, 30 patients (68.2%) developed clinically definite multiple sclerosis. The remaining 14 patients were followed up for more than seven years. Twenty six (59.1%) patients had oligoclonal bands in CSF, with a sensitivity of 80.0%, specificity of 85.7%, and a predictive value of 92.2%. Magnetic resonance imaging showed disseminated white matter lesions in 22 patients (50.0%), with a sensitivity of 60.0%, a specificity of 71.4%, and a predictive value of 81.7%. The difference between patients with multiple sclerosis and patients without the disease was statistically significant for the findings of an IgG intrathecal synthesis (P < 0.001). It was only borderline for the MRI findings (P = 0.052). Thus the detection of an intrathecal synthesis at presentation seemed to be a better prognostic indicator of the progression to multiple sclerosis in patients affected by acute isolated brainstem or spinal cord syndromes.     

Autoreactive IgG to intracellular proteins in sera of MS patients.

IgG binding to multiple protein constituents in lysates of Jurkat cells was detected by Western blot in sera of patients with multiple sclerosis (MS) and systemic lupus erythematosus (SLE). The distribution patterns of bands with sera tested against protein lysates from normal Jurkat cells or from Jurkat cells exposed to apoptosis or oxidative stress inducing conditions were similar in most patients, but with inter-individual differences. The number of bands with sera of both patient populations far exceeded those (0 or 2 bands) detected with sera of healthy controls. Proteinase K, RNase and DNase pre-treatment of cell lysates suggested a protein nature for all of the antigens and a ribonucleoprotein (RNP) nature for some of the antigens recognized by serum IgG of MS and SLE patients. Only two MS patients had positive anti-nuclear antibody (ANA) titers, while all of them had positive Western blots. In addition to similarities, dissimilarities were also recognized between the humoral immune responses in MS and SLE. No IgG molecules were detected against phosphorylated proteins in the sera of MS patients, while multiple phosphoproteins were recognized by IgG molecules of SLE patients in immunoprecipitation experiments. These data suggest that in addition to ANA, the sera of MS patients contain autoantibodies directed against multiple intracellular proteins. The protein recognition patterns of immunoglobulins in MS share similarities, but also have distinct features when compared to those in SLE. The biological significance of these autoantibodies in MS remains to be understood.     

Gamma globulin subfractions and immunoglobulin G in the cerebrospinal fluid of patients with multiple sclerosis, subacute sclerosing panencephalitis and other diseases of the nervous system

CSF samples were obtained from 22 patients with multiple sclerosis, 14 patients with subacute sclerosis panencephalitis, and 10 with neuroses or headaches. Most samples were investigated simultaneously by a sensitive method of electrophoresis in polyacrylamide gel (PE) and radial immunodiffusion (RID). The absolute IgG concentration was in multiple sclerosis 6.8 +/- 3.3 mg/100 ml, and in SSPE 21.4 +/- 16.8 mg/100 ml and it was higher than in control subjects (p less than 0.01). In 84% of MS patients the Bauer index (IgG: total protein) was raised, 72% had relatively raised subfractions gamma 2-gamma 3 in PE, 64% had higher absolute concentration of GG (0.045 g/l or more) and 63% had increased proportion of one of gammaglobulin subfractions (over 4%). Of positive diagnostic importance was a rise of the IgG: total protein index, since 7 patients with SM (31%) with normal per cent gammaglobulin level (less than or equal to 13%) had an evident rise of this index. Raised value of the index IgG: total protein (greater than or equal to 10%) was observed also in 100% of SSPE cases, while in 92% the proportional value of one of gamma globulin subfractions was increased (over 4%), in 92% the subfractions gamma 3-gamma 4 were increased, in 85-86% of patients the relative (over 13%) and absolute (0.045 g/l or more) levels of gammaglobulins were increased. The index beta/gamma in MS and SSPE was 0.85 and 0.57 respectively, and was significantly lower than in patients with headaches. The authors discuss the diagnostic value of these findings.     

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.     

Intrathecal immune response in patients with neuroborreliosis: specificity of antibodies for neuronal proteins

Cerebrospinal fluid (CSF) and serum samples of 47 patients with serologically proven neuroborreliosis were examined by Western blotting for antibodies to a crude extract of human cortex (CNS) comprising a multitude (> 40) of protein bands. Intrathecal synthesis of total immunoglobulins was determined by the Reiber formula and of autoantibodies to CNS proteins by enzyme-linked immunoassay (ELISA) and by Western blotting. Employing ELISA, intrathecal synthesis of autoantibodies (IgG, IgM and/or IgA) was demonstrated in 40 of 47 patients with neuroborreliosis (85%), in 5 of 40 with multiple sclerosis (12%), and in 22 of 40 with viral meningoencephalitis (55%). Of 40, 35 and 15 patients with neuroborreliosis and an intrathecal synthesis of total IgG, IgM or IgA, 20 revealed an intrathecal production of IgG antibodies (50%), 24 of IgM antibodies (68%) and 6 of IgA autoantibodies (40%) in the CSF. The specificity of autoantibodies differed greatly between most patients. Of 24 different CNS proteins which elicited an immune response in various patients, identities could be determined only for the myelin basic protein (5 of 40) and for the three neurofilament proteins (NF-68, NF-150, NF-200) (13 of 40 patients). In this limited number of patients no significant correlation between individual clinical symptoms and certain autoantibodies could be detected. The higher frequency of intrathecally produced autoantibodies in patients with neuroborreliosis is assumed to result from mitogenic rather than specific activation of autoreactive B-cell clones byBorrelia burgdorferi. The pathogenic relevance of these autoantibodies remains to be determined.     

A characteristic ganglioside antibody pattern in the CSF of patients with amyotrophic lateral sclerosis.

Paired cerebrospinal fluid and serum samples of patients with amyotrophic lateral sclerosis (n = 35) revealed no consistent abnormalities of CSF cell count, CSF albumin, CSF IgG, CSF IgM, IgG or IgM index, or oligoclonal immunoglobulin band formation in the CSF. Determination of IgG and IgM CSF and serum antibodies to gangliosides GM1, GM2, GM3, AGM1, GD1a, GD1b, and GT1b showed a characteristic pattern which allowed the differentiation of amyotrophic lateral sclerosis from controls and from patients with other neurological disorders including multiple sclerosis. Specifically, patients with the disease had elevated CSF IgM antibodies to all gangliosides except AGM1. The lack of correlation between the CSF findings and corresponding serum antibodies suggests a chronic, compartmental, intrathecal immune response of low activity in amyotrophic lateral sclerosis. Whether this immune response is primary and of pathogenetic significance, or an epiphenomenon of neuronal degeneration, remains to be investigated.     

Passive transfer of demyelination by serum or IgG from chronic inflammatory demyelinating polyneuropathy patients

Chronic inflammatory demyelinating polyneuropathy (CIDP) is regarded as an autoimmune disorder, but no clearly defined autoimmune mechanism has been described. Although most patients respond to plasma exchange, no convincing role for autoantibodies has yet been demonstrated. In this study, we have successfully passively transferred disease using sera and purified IgG from 4 of 12 patients responsive to plasma exchange by bypassing the blood-nerve barrier by intraneural injection or opening it by activated T cells. The sera from CIDP patients or purified IgG produced marked conduction block and demyelination, but normal sera or IgG or that from patients with multiple sclerosis or other neuropathies did not. These observations strongly support an important role for anti-myelin/Schwann cell autoantibodies in the pathogenesis of CIDP at least in some patients.     

Autoantibodies to brain endothelial cells in the sera of patients with human T-lymphotropic virus type I associated myelopathy and other demyelinating disorders

We examined the sera of 21 cases of human T-lymphotropic virus type I (HTLV-1)-associated myelopathy (HAM), 30 cases of neuro-Beh?et (N-B) syndrome, and 36 cases of multiple sclerosis (MS) for the presence of autoantibodies to brain endothelial cells by enzyme-linked immunosorbent assay (ELISA) using cultured brain endothelial cells. The concentrations of immunoglobulin G (IgG) which bound to brain endothelial cells were significantly increased in the sera of patients with HAM before (P less than 0.001) and after (P less than 0.01) blocking Fc receptors compared to those of controls. The levels of IgG binding to brain endothelial cells were also significantly increased in the sera of patients with N-B syndrome (P less than 0.01), and MS (P less than 0.001) especially those in an exacerbation compared to those of controls regardless of blocking Fc receptors. These results suggest that IgG binding to brain endothelial cells may be mediated via an immunologically specific antigen-antibody interaction as a result of the blood-brain barrier (BBB) damage in cases of HAM, N-B syndrome and MS.     

OPTIC NEURITIS: STUDIES ON THE CEREBROSPINAL FLUID IN RELATION TO CLINICAL COURSE IN 61 PATIENTS

A prospective study on 61 previously healthy patients with acute mono-symptomatic optic neuritis is reported. Interest was focused on changes in the cerebrospinal fluid and the clinical course with possible development of multiple sclerosis. At the beginning of the disease, a mononuclear pleocytosis was noted in 51 per cent of the patients, an elevated IgG level in 18 per cent, and an oligoclonal IgG distribution in 41 per cent. These results are in sharp contrast to those in multiple sclerosis. Eleven patients have so far developed definite multiple sclerosis. At onset only five of these had cerebrospinal fluid findings indistinguishable from those in multiple sclerosis, with pleocytosis and bands on electrophoresis. In five more patients, who subsequently developed multiple sclerosis, the cerebrospinal fluid IgG was normal at onset, but an oligoclonal IgG appeared during the follow-up; there was no correlation in time between the appearance of new symptoms and cerebrospinal fluid changes. In six patients with normal cerebrospinal fluid and four patients with only mononuclear pleocytosis at the onset of disease, the IgG pattern became oligoclonal on electrophoresis during the follow-up period, although the patients had no further symptoms or signs of disease. It was concluded, therefore, that the cerebrospinal fluid was often normal in the first attack of what later proved to be multiple sclerosis, and that a normal fluid did not preclude a development into definite multiple sclerosis. Sometimes IgG bands appeared in previously normal cerebrospinal fluid, although the patients had not experienced new symptoms.