Immune complexes in serum and cerebrospinal fluid of multiple sclerosis patients and patients with other neurological diseases

Paired serum and cerebrospinal fluid (CSF) specimens from 30 multiple sclerosis (MS) patients and 30 patients with other neurological diseases (ONDs) were analyzed for the presence of immune complexes (ICs). With each of the 4 tests used, ICs were found more frequently in sera from both MS and OND patients than in sera from healthy blood donors. IC-positivity for MS and OND patient CSF varied from 10-33 % and from 10-17 % in different tests. The number of IC-positive sera or CSF in MS patients did not differ significantly from those in OND patients. For both MS and OND patients, the positivity pattern for serum and CSF specimens in each IC test was essentially unique. Furthermore, because several CSF IC-positive and serum IC-negative paired specimens were found, intrathecal IC formation may be independent of IC formation in peripheral blood. The presence of ICs in serum or CSF did not correlate with the clinical status of or laboratory data on the MS patients, nor was a correlation found with the diagnosis of the OND patients. In total, these results suggest that the presence or absence of ICs in MS or OND patients may simply reflect changes in the immunological regulation of individual patients.     

Nerve growth factor (NGF) in cerebrospinal fluid (CSF) from patients with various neurological disorders

It has been recently shown that NGF is not only involved in the survival and development of sympathetic and neural crest-derived sensory neurons, but also in some mechanisms of the immune system. For this reason, we studied the content of NGF in CSF samples from patients with diseases in which neuroimmonological mechanisms seem to be involved (multiple sclerosis, amyotrophic lateral sclerosis, Alzheimer disease, chronic relapsing polyradiculoneuritis, Guillain-Barré syndrome, and tumors of the nervous system), as well as from a number of normal control subjects. We setup an ELISA aimed at the beta subunit of NGF, obtaining good validation tests and a detection limit of 28 pg βNGF per ml. None of the samples was found to contain detectable levels of NGF and, when a concentration method for sample enrichment was used, only one patient was NGF-positive. This suggests that NGF is probably not involved in the neuroimmunological mechanisms underlying some inflammatory and degenerative diseases of the nervous system. Paper presented at the “International Quincke Symposium”, G?ttingen, September 18–21, 1991.     

Soluble immune complexes in cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases

The occurrence of soluble immune complexes (IC) in the cerebrospinal fluid (CSF) of 14 multiple sclerosis (MS) patients, four acute polyradiculoneuritis patients, 30 patients with other neurological diseases (OND) and 30 patients with disc prolapse (DP) was examined by a solid phase C1q-protein A binding assay (C1q-PABA) and a complement consumption test. IC-positive reactions were observed only in the C1q-PABA. The binding indices determined by the C1q-PABA differed significantly ( P < 0.01) when the MS or the OND patient groups were compared to the DP group. No significant ( P < 0.1) difference was observed between the indices in the MS and OND groups. Binding indices in C1q-PABA showed no correlation either to IgG concentration, total protein concentration or cell counts in CSF of MS patients. Three of the four polyradiculoneuritis patients were strongly IC-positive while the fourth patient was negative. Filtration and PEG-precipitation data indicated that a major part of the IgG-containing IC in CSF detected by C1q-PABA was of macromolecular nature.     

Immune complexes and the complement factors C4 and C3 in cerebrospinal fluid and serum from patients with chronic progressive multiple sclerosis

Immune complexes (IC) have been found in both serum and cerebrospinal fluid (CSF) in multiple sclerosis (MS). The complement system is known to play a major role as a mediator of inflammation in immune complex disease. Therefore, we have investigated paired samples of serum and CSF from 32 patients with progressive MS for IC, the levels of the complement factors C4 and C3, and presence of their activation products (AP). IC was found in serum from 17 of the 32 MS patients (53%) and in CSF from 9 of 31 MS patients (29%). No correlation was found between the occurrence of IC in serum and in CSF. The levels of C3 in serum and CSF from the MS patients did not differ from the levels in a control group, whereas the levels of C4 in MS-serum were elevated and the C4 levels in MS-CSF reduced. A low level of CSF-C4 correlated significantly to the occurrence of CSF-IC. AP of C4 and C3 in serum were seen in 11 of the 32 patients (34%), appearing significantly more frequently among patients with circulating IC. No C4- or C3AP could be identified in CSF.     

Diagnostic problems in “clinically definite” multiple sclerosis patients with normal CSF and multiple MRI abnormalities

Among patients who underwent cerebrospinal fluid (CSF) and magnetic resonance imaging (MRI) examination during a 5-year period, we found 18 patients at the multiple sclerosis center of the University of Rome and clinically definite multiple sclerosis, MRI white matter abnormalities, normal CSF examination, a disease duration of at least 1 year and an upper age of < 45 years at onset These patients were examined again with a variety of tests screening for different diseases mimicking multiple sclerosis. Alternative diagnoses reached after laboratory tests were: Lyme disease, two cases of vasculitis, mitochondrial encephalomyopathy, multiple ischemic lesions caused by atrial septum aneurysm and olivopontocerebellar atrophy. Hence, six of these 18 patients had a final diagnosis other than multiple sclerosis, while 12 remained with a final diagnosis of “MS with normal CSF”. Our study suggests that in patients with a clinical picture of multiple sclerosis and disseminated white matter MRI lesions but no CSF abnormalities, the classical clinical criteria may not be sufficiently specific, unless confirmed by a very prolonged clinical history with repeated MRI.     

Microzone electrophoresis of the unconcentrated and concentrated CSF

Summary Concerning the small methodical range of error and the little expenditure of work microzone electrophoresis of the non-concentrated CSF after staining with nigrosine and after evaluation on non-transparent acetate film should have priority to microzone electrophoresis of concentrated CSF. This paper reports a comparison of microzone electrophoresis of non-concentrated and concentrated CSF.

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Zusammenfassung Mikrozonenelektrophoresen des Nativliquors nach Färbung mit Nigrosin und Auswertung auf nichttransparenter Azetatfolie sind den Liquorelektrophoresen nach Einengung wegen der größeren methodischen Genauigkeit und dem geringen Arbeitsaufwand vorzuziehen. Berichtet wird über einen Vergleich von Mikrozonenelektrophoresen des Nativliquors und des Liquors nach Einengung.

     

Phagocytic activity of CSF monocytes in neurological diseases

Summary A standard latex phagocytosis test was used to determine the phagocytic activity of CSF monocytes under various pathological conditions. The mean values for groups of neurological diseases did not differ significantly, with the exception of tumour patients. Differences in the phagocytosis of CSF monocytes were seen if subdivisions within the groups of patients with multiple sclerosis (MS) and vertebral irritation syndrome (VIS) were evaluated separately. The active stage of MS and the intervertebral disc prolapse were accompanied by higher values. It is concluded that although the findings are of no use for diagnostic purposes they may be helpful in assessing prognosis and therapy.     

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.     

Antibodies against oligodendrocytes in serum and CSF in multiple sclerosis and other neurological diseases: 125I-protein A studies

Antibodies against oligodendrocytes were determined in pairs of unconcentrated CSF serum from 12 patients with multiple sclerosis (MS) and 25 control patients including 10 with aseptic meningoencephalitis (AM), using a 125I-protein A microassay. Antibody levels in serum and in CSF did not differ between MS and controls. Calculating the antibody index equal to (CSF/serum antibodies against oligodendrocytes):(CSF/serum albumin) in analogy to the CSF IgG index, thereby compensating for influence of serum antibody concentration as well as altered blood-brain barrier, no evidence was obtained for intrathecal antibody production in the patients with MS. Those with AM had higher antibody index values, probably reflecting intrathecal synthesis. Antibodies against oligodendrocytes seem to be a regular component of CSF and serum in neurological diseases; intrathecal antibody production is less frequent in MS than in AM.     

Circulating immune complexes in serum and in cerebrospinal fluid of patients with multiple sclerosis

We studied circulating immune complexes (IC) in the serum and cerebrospinal fluid (CSF) of patients with clinically defined multiple sclerosis (MS), in order to establish a correlation with the clinical course of the disease and to investigate the molecular composition of the IC isolated from patients in active phase of the disease. Serum IC levels were found to be significantly increased in patients from the progressive and active relapsing-remittent subgroups with both the CIC-conglutinin and C1q-binding methods. High levels of IC in CSF were detected only in the subgroup consisting of the relapsing-remittent patients in disease exacerbation when IC were determined by the C1q-binding test. No significant increase in serum or in CSF were found using the mRF-I test. The preliminary results of a qualitative investigation on serum IC in MS indicated that they are heterogeneous in nature, their size is mainly of the intermediate type, and they contain IgG, IgM, complement components and beta 2-microglobulins, the latter presenting an observation both new and interesting for studies on serum IC in MS patients.     

T and B lymphocytes in the cerebrospinal fluid of various neurological diseases

Summary Cerebrospinal fluids (CSF) from 66 patients with a variety of neurological disorders were studied for total protein content, absolute amount of albumin, IgA, IgG and IgM, as well as their quotients (fraction to total protein ratio), cell numbers and B cell and T cell levels. In addition, the percentage of B cells and T cells in the blood was determined in 34 patients and serum immunoglobulin levels were estimated in 51 patients. In noninflammatory diseases of the CNS, the percentage of B cells was slightly higher and T cell levels were lower in the CSF in comparison to corresponding blood values. The B cell to T cell ratio in viral meningitis was altered in the CSF. An apparent increase in the T cell level led to a decrease of B cell values. Similar changes were also found in optic neuritis. The percentage of T cells was higher in relapsing multiple sclerosis than in the chronic progressive form. There were less striking changes in the B cell to T cell ratios in the CSF of other inflammatory diseases of the CNS.

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Zusammenfassung Von 66 Patienten wurden im Liquor außer Zellzahl, den B-und T-Zellen noch der Gesamteiweißgehalt sowie die absoluten Werte von Albumin, IgA, IgG, IgM und deren relativer Anteil am Gesamteiweiß untersucht. Im Blut war die Bestimmung der B- und T-Zellen bei 34 Patienten, die der Immunglobuline bei 51 Patienten möglich. Bei nicht entzündlichen Erkrankungen des zentralen Nervensystems liegt der prozentuelle Anteil der T-Zellen im Liquor zumeist etwas niedriger, der der B-Zellen etwas höher als die entsprechenden Blutwerte. Bei viralen Miningitiden kommt es zu einer deutlichen Verschiebung dieses B:T-Zellen-Verhältnisses, und zwar zu einem Ansteigen der T-Zellen bei daraus resultierendem B-Zellen-Abfall. Ähnliche Relationsveränderungen sind bei retrobulbären Neuritiden, die als Erstsymptome einer MS auftreten, zu finden. MS-Fälle mit schubförmiger Verschlechterung weisen einen höheren Anteil an T-Zellen auf als MS-Fälle mit chronisch progredientem Verlauf. Eitrige Meningitiden, Myelitiden und Polyneuropathie-Syndrome zeigen die Tendenz des T-Zellen-Anstieges und B-Zellen-Abfalles im Liquor in geringerer Ausprägung.

     

CSF myelin basic protein in multiple sclerosis

Cerebrospinal fluid (CSF) from 221 patients with multiple sclerosis (MS) and 85 patients with other neurological disorders (OND) was examined using a competitive radioimmunoassay for myelin basic protein (MBP) immunoreactivity. MBP was found in 46 of 55 MS patients (84%) examined within six weeks of relapse but in only 11 of 85 patients (13%) with OND. There was a significant correlation between the concentration of MBP in the CSF and relapse severity in patients seen within four weeks of the onset of symptoms (p less than 0.01). Of 44 patients in remission, MBP was detected in 12, and these patients had a significantly higher tendency to subsequent relapse (p less than 0.05). In 72 patients with progressive disease the presence of MBP in the CSF reflected the confidence of clinical diagnosis. The results of this study suggest that measurement of MBP in the CSF gives an objective method of monitoring disease activity in patient with MS.     

CSF antibodies to myelin basic protein and oligodendrocytes in multiple sclerosis and other neurological diseases

Cerebrospinal fluid (CSF) from 18 multiple sclerosis (MS) patients, 13 subacute sclerosing panencephalitis (SSPE) patients, 22 other neurological disease (OND) patients, and 7 neurotic patients as controls were tested in an 125I-labeled anti-human F(ab')2 binding assay for the presence of antibodies to normal human brain cells from tissue culture, human fibroblasts, plasma membranes of MS and normal human brain, myelin basic protein (MBP) and bovine oligodendrocytes. Antibodies to MBP and to oligodendrocytes were found in the CSF of MS, SSPE and OND patients. Absorption of CSF with bovine CNS myelin significantly diminished binding activity to oligodendrocytes. Antibodies in the CSF against MBP and oligodendrocytes, on which some myelin determinants are expressed, seem to be a common feature of diseases in which demyelination is a component.     

CSF and serum ganglioside antibody patterns in MS

The authors determined CSF and serum IgG and IgM antibodies to seven gangliosides in 48 patients with multiple sclerosis. Differing ganglioside antibody patterns in CSF but not serum allowed to reclassify 93% of MS patients correctly when compared to patients with Guillain-Barré syndrome or neuroborreliosis. This suggest that the antibody patterns are neither random nor alike in inflammatory diseases of the nervous system. CSF ganglioside antibody titres were found to be different for patients with relapsing remitting (RRMS; n = 35) and chronic progressive (CPMS; n = 13) multiple sclerosis. Our study reveals characteristic ganglioside antibody patterns in MS and confirms previous evidence of disturbed immunoregulation in MS.     

Glutamate levels in cerebrospinal fluid in amyotrophic lateral sclerosis: a reappraisal using a new HPLC method with coulometric detection in a large cohort of patients.

Glutamate is involved in the degeneration of motor neurons in amyotrophic lateral sclerosis (ALS). However, the aetiology of ALS appears heterogeneous, leading to the possibility that patient subgroups with different pathophysiology may exist. The concentration of glutamate in cerebrospinal fluid (CSF) is measured using a new HPLC method with coulometric detection in a large cohort of ALS patients and controls: 377 ALS patients, 88 neurological patients and 18 normal controls. In ALS patients, and only in these subjects, the existence of two groups was observed, one with normal glutamate concentrations and one (40.8% of ALS patients) with high glutamate concentrations. High glutamate concentrations were correlated with a spinal onset of the disease, more impaired limb function and a higher rate of muscle deterioration. These results suggest that elevations of CSF glutamate concentrations could reflect the intensity of cell insult in the spinal cord. It remains to be determined if the group of patients with high CSF glutamate concentrations represents a specific subgroup of patients in terms of mechanism of disease, or only in terms of the spatial extent of motor neuron insult.     

Cerebrospinal fluid markers in neurological disorders

Cerebrospinal fluid (CSF) markers are a useful tool for determining disease progression or activity in some neurological disorders which need parameters both for evaluating treatments and investigating pathobiological evolution in researchoriented follow-up. A number of CSF proteins are reviewed with data on biological properties, analytical methods, clinical usefulness of: myelin basic protein, S-100 protein, glial fibrillary acidic protein, neural-cell adhesion molecule, neuron-specific enolase and others.

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Sommario I marcatori liquorali sono un utile strumento per determinare la progressione della malattia ele fasi di attività in alcuni disordini neurologici che necessitano di parametri utili a valutare l’efficacia di un trattamento, o a studiare l’evoluzione patobiologica in follow-up finalizzati alla ricerca. Gli autori passano in rassegna un certo numero di proteine liquorali soffermandosi sulle loro proprietà biologiche, sui metodi analitici, sull’utilizzo a scopi clinici. Vengono prese in considerazione: la proteina basica della mielina, la proteina S-100, la proteina acida gliale fibrillare, la proteina di adesione neuronale N-CAM, la enolasi neurono-specifica e alcune altre.

     

Characterization of immune complexes in multiple sclerosis by an antigen-specific immune complex radioimmunoassay

Serum and cerebrospinal fluid samples from multiple sclerosis (MS) patients containing various levels of Clq-reactive immune complexes (IC) and samples from age- and sex-matched controls were tested by an antigen-specific IC radioimmunoassay which detects IC containing myelin membrane-related antigens. Positive reactivity in the assay was significantly associated with IC-containing MS sera (P less than 0.005) but such an association was not observed with MS cerebrospinal fluid. Analysis of longitudinal specimens revealed that the levels of the antigen-specific serum IC fluctuated with time. Significant correlation between serum levels of Clq-reactive IC and serum levels of IC containing myelin membrane-related antigens was observed (r = 0.62; P less than 0.001). Sucrose gradient centrifugation of sera from 1 patient showed that the IC had a peak density of 1.075 g/ml, indicating the presence of lipid material. The results suggest that serum IC of MS patients frequently contain myelin membrane-related antigens and that these antigens may be lipids or lipid-associated.     

A rapid and simple cannulation technique for repeated sampling of cerebrospinal fluid in freely moving rats

A cannulation technique for frequent sampling of cerebrospinal fluid (CSF) in unanaesthetized freely moving rats is described. A permanent stainless steel cannula, constructed in such a way that no loss of CSF occurs, is placed into the rat's cisterna magna and fixed to the skull by anchoring screws and dental cement. A special CSF outflow opening of the cannula is connected to polyethylene tubing for CSF sampling. Amounts of 50–150 μl CSF can be collected repeatedly without any sign of disturbing the animal. The technique lends itself not only to pilot studies in which within a short period of time a large amount of CSF is wanted, but also to experiments in which physiological conditions are required.     

Selective D1 agonism but not D2 antagonism is reflected in cAMP and cGMP levels in rat CSF

Cyclic adenosine 3′5′-monophosphate (cAMP) and cyclic guanosine 3′5′-monophosphate (cGMP) serve as second messengers in several cellular pathways within the central nervous system. In various neurological and psychiatric disorders with known deficits in neurotransmission function, CSF levels of cAMP and/or cGMP in patients were studied. Very little information is currently available on cAMP and cGMP levels in CSF of animals. Moreover, this is the first study on the effects of pharmacological treatment on cAMP and cGMP levels in rat CSF. Effect of systemic treatment with a D1 receptor agonist SKF82958 and a D2 receptor antagonist haloperidol on cAMP and cGMP levels, as well as baseline cAMP and cGMP levels in CSF of rats was determined. A significantly increased cAMP and cGMP level in cisternal CSF of rats systemically treated with the D1 receptor agonist SKF82958 was observed, while when treated with the D2 antagonist haloperidol, no effect on cAMP and only a slight decrease of cGMP was observed after treatment with the highest dose. Determining cAMP and/or cGMP in CSF of experimental animals can serve as a useful tool to study neural processes affected by disease and treatment.     

Intrathecal synthesis of different alpha-interferons in patients with various neurological diseases

CSF and sera from 238 newborns and children with various neurological diseases were assayed on bovine cells for the presence of alpha-interferon (IFN). An intrathecal synthesis of pH 2-resistant alpha-IFN was recovered in all newborns and in more than 90% of children with herpes encephalitis. It was also observed in one case of mumps encephalitis and in one case of encephalitis associated with Influenza A infection. An acid-labile alpha-IFN production was detected in CSF from more than one half of patients with viral meningitis or active congenital rubella and in those with neurological complications of systemic lupus erythematosus. This alpha-IFN subtype was also detected in CSF from only 2/37 children with measles encephalitis. In contrast, no alpha-IFN (less than 2 IU) in CSF was found among patients with subacute sclerosing panencephalitis, Guillain-Barré syndrome, Reye's syndrome, acute cerebellar ataxia, infantile spasms or facial paralysis of unknown origin.