Oligoclonal immunoglobulin D in the cerebrospinal fluid of neurologic patients.

Oligoclonal immunoglobulin D (IgD) bands were observed in 20 of 110 CSF samples investigated by our method for isoelectric focusing (IEF) of IgD in unconcentrated CSF. These were mostly from demyelinating disorders, viral CNS infections, and CNS tumors. The patterns of expression of IgD were similar to that of IgG, except in cases with tumors where oligoclonal IgD may occur independently of IgG. The study shows for the first time the presence of oligoclonal IgD in CSF from neurologic patients.

The occurrence of oligoclonal IgG in tears from patients with MS and systemic immune disorders.

Using isoelectric focusing (IEF) and immunoperoxidase staining of proteins transferred to nitrocellulose membranes, we have examined the IgG band pattern in tears and matched serum and CSF specimens of 28 patients with MS, 4 patients with optic neuritis (ON), 30 individuals with systemic, inflammatory, or other neurologic diseases, and 5 patients with tension headache. We found no evidence of positive oligoclonal IgG in tears in any MS or ON patients, while 10 out of 16 cases with systemic immune disorders or infections of the CNS had positive tear oligoclonal bands. We are thus not able to support the hypothesis that tears from MS patients reveal abnormalities in their humoral immune response.

A new qualitative method for detecting IgD in unconcentrated cerebrospinal fluid.

We report a method which is capable of demonstrating the isoelectric focusing (IEF) pattern of immunoglobulin D in unconcentrated cerebrospinal fluid (CSF) samples containing as little as 0.1-0.5 ng of total IgD. The method used was an immuno-sandwich technique, with alkaline phosphatase enzyme amplification. Oligoclonal and polyclonal IgD patterns were seen in CSF samples. No cross-reactivity with other immunoglobulins (IgG, IgA and IgM) was detected.

Oligoclonal immunoglobulin G in cerebrospinal fluid of myasthenia gravis patients.

Immunoglobulin G (IgG) band patterns were investigated in cerebrospinal fluid (CSF) from 19 patients with myasthenia gravis (MG) in a search for abnormalities indicating central nervous system (CNS) involvement in this disorder. Using the isoelectric focusing (IEF) technique and antiserum immunoblotting against IgG, we found no evidence of the presence of oligoclonal IgG in CSF from most of MG patients. In 2 cases, the positive findings of oligoclonal IgG in CSF may have reflected a manifestation of an associated disease, which has already been associated with immune abnormalities within the CNS. Further investigations with more sophisticated techniques are required to give additional insight into humoral immune events within the CNS in MG patients.

IgG paraproteins in neurological diseases: lack of association with neurotropic viral/bacterial antigens.

In this study we tested the hypothesis that IgG paraproteins in neurological diseases might be regarded as an anti-idiotypic response raised by a nervous system antigen associated with certain neurotropic agents. After initial ELISA screen, positive samples from 34 neurological patients who had paraproteins in their CSF and serum on routine IEF investigation, were tested by immunoblot technique for the presence of specific monoclonal immunoglobulin G (IgG) against eight different neurotropic antigens. Only one patient had specific monoclonal IgG against herpes simplex virus. The results of this study did not confirm our hypothesis. Further study of IgM/IgA paraproteins is indicated.

Spleen versus CNS immunoglobulin G in multiple sclerosis: an isoelectric focusing study.

Immunoglobulin G (IgG) band patterns were investigated in a peripheral immune compartment, the spleen, and the central nervous system (CNS) compartment in 6 multiple sclerosis (MS) subjects and 3 controls in a search for systemic humoral immune response that may be related to the localized immune response seen in MS. Using the isoelectric focusing (IEF) technique with immunoperoxidase staining of proteins transferred to nitrocellulose membranes, we were not able to demonstrate any qualitative abnormalities of spleen or serum IgG in the MS cases, whilst all matched CNS tissues and CSF specimens were clearly positive for oligoclonal IgG bands. Our results suggest that there is no systemic oligoclonal IgG secretion in MS. This further supports the presence of a compartmentalized IgG intrathecal immune response in MS and emphasizes the organ specificity of MS.

[Findings of cerebrospinal fluid in the diagnosis of multiple sclerosis]. / Likvorski nalazu dijagnostici multiple skleroze.

In order to define the cerebrospinal fluid (CSF) profile indicative of multiple sclerosis (MS), it is essential that each laboratory specifies the percentage of clinically definite MS patients with as well as the ranges of obtained values for each CSF parameter in these patients. CSF of 92 patients with clinically defined MS were analysed for cell count, concentrations of total protein, albumin and IgG, blood-CSF barrier function as assessed by CSF/serum albumin quotient, quantitative measurements of intrathecal IgG production (IgG index and IgG daily synthesis rate) and the presence of CSF oligoclonal IgG. For the detection of CSF oligoclonal IgG the isoelectric focusing (IEF) of unconcentrated CSF on agarose with transfer of proteins to cellulose nitrate and immunoperoxidase staining, was performed. CSF oligoclonal IgG bands not matched in parallel sera were detected in 96% of MS patients. Quantitative measurements of intrathecal IgG synthesis were significantly less sensitive than IEF. The concentration of total proteins in CSF was normal in 48% of patients, slightly elevated in 7% of patients and moderately elevated in 4% of patients with clinically definite MS. CSF cell count was normal in 71% of patients and slightly increased in the remaining 29%. The differential cell count was normal in all patients. CSF albumin and CSF/serum albumin quotient were elevated in 58 identical MS patients of 92 tested, indicating slight or, rarely moderate dysfunction of CSF-brain barrier in these patients. In 99% of MS patients, the total protein was <980 mg/L, the albumin <740 mg/L and the total cell count <20 cells per mm3. According to our data, the CSF findings that support the diagnosis of clinically definite MS are: 1) the presence of intrathecally produced CSF oligoclonal IgG by IEF; 2) normal or slightly to moderately elevated level of CSF proteins and albumin; 3) normal CSF-brain barrier function or slight to, rarely, moderate CSF-brain barrier dysfunction as assesed by CSF/serum albumin quotient, and 4) normal cell count or slight pleocytosis. If this profile is not found in a patient suspected of MS, the diagnosis should be questioned or a complication should be expected.

Oligoclonal IgG and free chains in multiple sclerosis demonstrated by thin-layer polyacrylamide gel isoelectric focusing and immunofixation.

A modified technique of isoelectric focusing on thin-layer polyacrylamide gel followed by immunofixation with monospecific antisera was used to identify individual cerebrospinal fluid (CSF) and serum proteins and to define the oligoclonal reaction observed in multiple sclerosis (MS). "Normal" IgG gave about 20 to 30 bands at pH 3.5 to 9.5, IgA about 10 bands at pH 3.5 to 6.4, beta-trace protein a smear at pH 3.5 to 8.5, and gamma-trace protein 1 or 2 bands at pH 8.0, 9.5 or both. Up to 11 oligoclonal IgG bands migrating between pH 6.5 and 9.5 were found in CSF from 26 of 27 consecutive patients with MS and also in 20 of the corresponding sera, although at lower numbers and concentrations. In 26 patients, 1 or more of the bands corresponding to normal polyclonal IgG were stronger in CSF than in serum. These data support the hypothesis that two colonies of lymphocytes are activated intrathecally, one of them synthesizing oligoclonal and the other polyclonal IgG. Up to 11 mostly faint bands of free light chains, predominantly of lambda type and migrating between pH 3.5 and 9.5, were found in 8 of 9 CSF specimens from patients with MS.

Specificity of immunoglobulins synthesized within the central nervous system in neurosyphilis.

Cerebrospinal fluid from four patients with serologically and clinically verified neurosyphilis contained oligoclonal bands when analysed by isoelectric focusing. By antiserum immunofixation these bands consisted by IgG, and by antigen immunofixation and autoradiography the bands were shown to contain treponemal antibodies. Treponemal antibodies synthesized within the CNS were also detected in polyclonal CSF IgG fractions in two of the patients. In one patient with neurosyphilis followed over 2.5 years, local synthesis of IgM and IgA disappeared within 1 year after treatment, while IgG synthesis persisted. In this patient, besides treponemal antibodies, antibodies against herpes simplex virus were also detected in oligoclonal and polyclonal IgG fractions of CSF. The presence of treponemal antibodies synthesized within the CNS was identified in 2 of 16 controls with negative serological tests for syphilis. This might reflect a previous contact with apathogenic treponemes, and the intrathecal antibody synthesis might be a result of an unspecific activation of memory lymphocytes in CNS.

Measles IgM antibodies in cerebrospinal fluid and serum in subacute sclerosing panencephalitis.

Using a direct enzyme-linked immunosorbent assay (ELISA), we examined serum and cerebrospinal fluid (CSF) from six patients with subacute sclerosing panencephalitis (SSPE) and control subjects for presence of measles-virus-specific IgM antibodies. All samples from the SSPE patients contained demonstrable titers of measles antibodies. The levels of measles IgM antibodies were higher in CSF diluted 1:5 than in serum diluted 1:50, reflecting a local production of IgM antibodies in the central nervous system. Antibody titers remained constant over the course of SSPE in three of the patients followed for three to six months. The IgM ELISA had high sensitivity as well as specificity and was not complicated by false-positive reactions owing to the presence of rheumatoid factor.

Short-term high dose steroid therapy does not affect the hypothalamic-pituitary-adrenal axis in relapsing multiple sclerosis patients. Clinical assessment by the insulin tolerance test.

Ten patients in acute exacerbation of multiple sclerosis were treated with 1000 mg of methylprednisolone for 7 days, followed by abrupt cessation of therapy. The function of hypothalamic-pituitary-adrenal (HPA) axis was assessed by the response of ACTH and cortisol to insulin tolerance test (ITT). ITT was performed 1 day before and 1, 3, 8, 13 and 23 days after the termination of the therapy (days 0, 8, 10, 15, 20 and 30 of the study, respectively). The response of these hormones to insulin-induced hypoglycemia prior to therapy was normal. There was no suppression of the ACTH response to hypoglycemia after the methylprednisolone therapy based on the 100% rise of ACTH after ITT. Cortisol response during ITT was suppressed at day 8 (1 day after ending of therapy) but recovered on day 10 (3 days after ending of therapy). In conclusion, 7 day-therapy with 1000 mg methylprednisolone does not result in the permanent suppression of the HPA axis, suggesting that no regular supplemental corticosteroid coverage is required. The observed transitory suppression of the HPA axis recovered spontaneously after the therapy.

CNS tumours: oligoclonal immunoglobulin D in cerebrospinal fluid and serum.

Oligoclonal immunoglobulin D bands were detected by isoelectric focusing in 7 out of 25 unconcentrated cerebrospinal fluid (CSF) samples obtained from patients with tumours of the central nervous system (CNS). The tumours were confirmed by clinical and histological findings. Two patients with CNS malignancy had intrathecal synthesis of oligoclonal bands both IgD and IgG. Four patients with a variety of CNS tumours had a systemic IgD immune response but no oligoclonal IgG bands. One patient with the most malignant tumour histology had a systemic IgD response as well as local synthesis of IgG. The study reveals several new aspects regarding CNS tumours: they are immunologically active and are capable of invoking oligoclonal immunoglobulin production both within the CNS and systemically. Multiple immunoglobulin activation can be found in malignant CNS tumours, and systemic IgD production may occur independently from IgG synthesis and may represent an immune response to a neoantigen produced in the CNS compartment.