Changes of the MS functional composite and EDSS during and after treatment of relapses with methylprednisolone in patients with multiple sclerosis.

OBJECTIVES: The Multiple Sclerosis Functional Composite (MSFC) comprises quantitative functional measures of leg, hand/arm and cognitive function. We examined the responsiveness of the MSFC compared with the Expanded Disability Status Scale (EDSS) during treatment of relapses in patients with multiple sclerosis (MS). PATIENTS AND METHODS: 27 patients received 1000 mg intravenous methylprednisolone (i.v.-MP) for 5 days, followed by oral methylprednisolone for 14 days. The MSFC and the EDSS-score were assessed on day 0, before the first corticosteroid treatment, on day 5, after the last course of i.v. MP, and on day 20 after the treatment was finished. Before the first administration of the MSFC, patients were trained for the paced auditory addition test (PASAT) performing three test trials. In order to analyse practice effects, 10 MS patients without an acute exacerbation were tested three times under the same conditions as the treated group. RESULTS: The median EDSS-score was 2.5 in both groups. On day 5 it remained unchanged in all treated patients, on day 20 a decrease of 0.5 EDSS point occurred in five patients, and in two patients an improvement with a decrease of more than 0.5 point was observed. There was no statistically significant difference between the EDSS-scores on day 0, 5 and 20. The mean MSFC-score in the treated group was -0.14 +/- 0.63 on day 0, 0.17 +/- 0.66 on day 5, and 0.42 +/- 0.59 on day 20. On the last study day, 26 patients improved compared with day 0. The differences between the MSFC-scores at the three points of time were statistically significant for the treated group (P < 0.001), but not for the control group. CONCLUSION: During and after treatment of relapses in patients with MS, the MSFC appears to be more sensitive in detecting changes in function than the EDSS.

Differential release of beta-chemokines in serum and CSF of patients with relapsing-remitting multiple sclerosis.

OBJECTIVE: beta-chemokines were recently demonstrated in active MS-lesions. We tested whether MCP-1 and RANTES can also be detected in CSF and serum of patients with MS and whether release is associated with inflammatory disease activity. MATERIALS AND METHODS: CSF and serum from 34 patients with newly diagnosed relapsing-remitting MS (RR-MS), 17 patients with viral meningitis (VM) and 19 patients with non-inflammatory neurological diseases (NIND) were investigated by ELISA. RR-MS patients underwent lumbar puncture and Gd-enhanced MRI examinations within 2 days. RESULTS: MCP-1 was strong intrathecally released in all patients. Compared to NIND CSF-levels were increased in VM (P<0.001) and were decreased in RR-MS (P<0.05). RANTES was only detected in serum in all patients. Levels were higher in VM and RR-MS compared to NIND (P<0.05). A total of 14/34 RR-MS patients exhibited active Gd-enhancing lesions on MRI. They had lower MCP-1 levels in CSF (P<0.001) and serum (P<0.05) and higher serum levels of RANTES (P<0.05) as compared to patients without active lesions. CONCLUSIONS: MCP-1 and RANTES are differentially released during acute attacks of RR-MS, which might reflect different immunregulatory roles of these beta-chemokines in RR-MS.

Immunomodulatory effects of interferon-beta-1b in patients with multiple sclerosis.

The mechanisms by which IFN beta-1b acts in the treatment of patients with multiple sclerosis (MS) are not completely known. Immunomodulatory effects of IFN beta-1b were investigated in patients with relapsing-remitting (RR) MS in vivo and in vitro. Compared to baseline and controls, defined as patients with RR-MS without immunomodulatory therapy, the expression of TGF beta-1-mRNA by peripheral blood mononuclear cells (PBMC) was persistently increased at week 6, month 3 and month 6 (p < or = 0.05), that of the TGF beta-1 receptor type II from day 5 up to month 6 (p < 0.01). The expression of TNF alpha-mRNA decreased from day 1 to month 3 compared to day 0 and the controls (p < 0.01). The in vitro investigations performed on isolated peripheral blood lymphocytes demonstrated that these effects were dose-dependent. The mRNA and protein expression of TNF alpha-R-I (55 kD-receptor) was only temporarily elevated at the beginning of the therapy in vivo. The expression of TNF alpha-R-I-mRNA increased dose-dependently after stimulation with IFN beta-1b for 24 h in vitro. Serum levels of soluble vascular cell adhesion molecule (sVCAM) were increased during the whole time of in vivo treatment (p < 0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p < 0.01) in the MS patients treated with IFN beta-1b in vivo. No persistent, significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 nor in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFN beta-1b induces the mRNA expression of TGF beta-1 and TGF beta-R-II by PBMC, decreases that of TNF alpha and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in blood. These might be other mechanisms by which IFN beta-1b mediates its positive effects in the treatment of MS patients.

Expression of TNFalpha and TGFbeta1 in Guillain-Barré syndrome: correlation of a low TNFalpha-/TGFbeta1-mRNA ratio with good recovery and signs for immunoregulation within the cerebrospinal fluid compartment.

In the pathogenesis of Guillain-Barré syndrome (GBS) a dysregulation of cytokines is supposed. The protein concentration and mRNA expression of TNFalpha and TGFbeta1 were investigated in cerebrospinal fluid (CSF) and blood in 10 patients with GBS. TNFalpha-mRNA was increased at the beginning of the disease and demonstrated a decline during the time course (P = 0.001). The level of TNFalpha protein was elevated in only a few patients. TGFbeta1-mRNA (P = 0.001) and the active TGFbeta1 protein (P < 0.05) increased during the course of GBS, and the level of total TGFbeta1 protein was temporarily elevated (P = 0.005). A low ratio of TNFalpha-/TGFbeta-mRNA expression correlated to a good clinical course (P < 0.05). The results indicate an association of TNFalpha with disease activity. TGFbeta1 seems to terminate and limit the inflammatory reactions and to promote the healing course of GBS. In addition the investigations show that in GBS immunoregulatory mechanisms also take place in the CSF compartment itself and that CSF cells are involved in the production of pro-inflammatory as well as immunosuppressive cytokines.

Immunomodulatory effects of IFNbeta-1b on the mRNA-expression of TGFbeta-1 and TNFalpha in vitro.

The mechanisms by which IFNbeta-1b acts in the treatment of multiple sclerosis (MS) patients are not completely known. We investigated the influence of IFNbeta-1b on the mRNA-expression of the immunosuppressive cytokine TGFbeta-1 and the proinflammatory mediator TNFalpha in an in vitro model by the method of non-radioactive in situ hybridization. Peripheral blood lymphocytes (PBL) were isolated from eight patients with relapsing remitting form of MS during remission and from six healthy controls. They were stimulated with IFNbeta-1b in different concentrations for 24 h. In both groups a statistically significant dose-dependent increase of TGFbeta-1-mRNA and decrease of TNFalpha-mRNA was demonstrable in the cultured stimulated blood lymphocytes compared to unstimulated cells. Stimulations with lipopolysaccharide (LPS) led to an increase of both cytokine-mRNAs in the lymphocytes. These data suggest specific and dose-dependent effects of IFNbeta-1b and hint at immunomodulatory properties of this drug to regulate the cytokine dysbalance in MS. This might be one mechanism by which IFNbeta-1b mediates its beneficial effects on the course of the disease.

Immunomodulatory effects of interferon-beta-1b in vivo: induction of the expression of transforming growth factor-beta1 and its receptor type II.

The mechanisms by which interferon-beta-1b (IFNbeta-1b) acts in the treatment of patients with multiple sclerosis (MS) are not completely known. A total of 10 MS patients were treated with 8 million units of IFNbeta-1b every other day. Compared to baseline and control group the expression of TGFbeta-1-mRNA by PBMC was persistently increased at week 6, month 3 and month 6 (p < or = 0.04), that of the TGFbeta-1 receptor type II from day 5 up to month 6 (p < 0.01). The mRNA and protein expression of tumor necrosis factor-alpha (TNFalpha)-receptor (55 kDa) was only temporarily elevated at the beginning of the therapy. Serum levels of sVCAM were increased during the whole time of treatment (p < 0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p < 0.01). No persistently significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 or in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFNbeta-1b induces the expression of TGFbeta-1- and TGFbeta-R-II-mRNA by PBMC and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in serum. These might be other mechanisms by which IFNbeta-1b mediates its positive effects in the treatment of MS patients.

Corticosteroids induce expression of transforming-growth-factor-beta1 mRNA in peripheral blood mononuclear cells of patients with multiple sclerosis.

The mechanisms by which corticosteroids act in the treatment of an acute relapse in multiple sclerosis (MS) are not completely known. We investigated the mRNA and protein expression of transforming-growth-factor-beta1 (TGFbeta1), a cytokine with anti-inflammatory and immunosuppressive potentials, in peripheral blood mononuclear cells (PBMC) and serum of 10 patients with an acute relapse of MS before, during and after the treatment with 500 mg prednisolone daily over 5 days. The expression of TGFbeta1-mRNA increased at day 3-5 and declined at day 8-10. Serum levels of TGFbeta1 demonstrated a comparable course. The present data suggest that corticosteroids induce the expression of TGFbeta1 in vivo. This is might be an other mechanism by which corticosteroids mediate immunosuppression.

Expression of tumor necrosis factor-alpha and transforming growth factor-beta 1 in cerebrospinal fluid cells in meningitis.

Meningitis is an acute inflammatory disease of the pia and arachnoid and the fluid in the subarachnoid space, in which a participation of cytokines can be expected. While tumor necrosis factor-alpha (TNF alpha) promotes inflammatory reactions, transforming growth factor-beta 1 (TGF beta 1) has antagonistic effects and suppresses the inflammation in the subarachnoid space. We investigated the protein concentration and mRNA expression of TNF alpha and TGF beta 1 in cerebrospinal fluid (CSF) by ELISA and intracellularly by non-radioactive in situ hybridization in 23 patients with bacterial or viral meningitis. A higher amount of both cytokines on protein and mRNA level, especially of TNF alpha, could be detected in bacterial infection. While an imbalance of both cytokines with a preponderance of TNF alpha- compared to TGF beta 1-mRNA was visible in CSF cells of patients with bacterial meningitis, a balance of TNF alpha- and TGF beta 1-mRNA or a higher expression of TGF beta 1-mRNA could be detected in viral meningitis. In the acute phase of the disease neutrophil granulocytes expressed more TNF alpha- and TGF beta 1-mRNA than lymphocytes and monocytes/macrophages, while these cell types were dominating the cytokine synthesis during the healing phase. These data indicate that immunomodulatory mechanisms take place in the CSF compartment itself, regulated by CSF cells in different but specific ways. In addition, TGF beta 1 seems to be involved in the down-regulation of the inflammatory activity and to be one factor in the cytokine network, which could contribute to a lower rate of complications and positive outcomes. Moreover this study favors the possibility to monitor the immunomodulatory mechanisms by non-radioactive in situ hybridization.

[Neuropsychiatric symptoms in vitamin B12 deficiency and microcarcinoidosis. The complications of chronic atrophic gastritis]. / Neuro-psychiatrische Symptome bei Vitamin-B12-Mangel und Mikrokarzinoidose. Komplikationen einer chronischen atrophischen Gastritis.

HISTORY AND FINDINGS: A 69-year-old woman reported marked restriction of voluntary movements of the hands in the preceding 6 months. She had also experienced loss of motivation, memory and concentration. Her skin was pale yellow, and scratches on her skin indicated marked pruritus. INVESTIGATIONS: Neurological examination revealed decreased vibratory sense in both legs. Haemoglobin concentration was 8.3 g/dl, mean corpuscular volume 114 fl, vitamin B12 level < 100 ng/l, folic acid level normal. Antibody titre against parietal cells was increased, vitamin B12 resorption diminished. Gastroscopy revealed small raised lesions, made up of hyperplastic cells which stained with chromogranin, indicating a diagnosis of microcarcinoid of the gastric mucosa. TREATMENT AND COURSE: On administration of cobalamine (1,000 micrograms i.m. daily for 2 weeks, twice weekly for 6 weeks, then once per week for the last 7 months) the blood picture returned to normal, but the microcarcinoids, the psychological symptoms and the apraxia of the hands were unchanged.

Potential role of transforming growth factor-beta 1 in terminating the immune response in patients with Guillain-Barré syndrome.

T-cell activation and proinflammatory cytokines seem to be important in promoting the disease activity in Guillain-Barré syndrome (GBS). Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional peptide with potent immunosuppressive activity, and can therefore be considered a putative disease-limiting cytokine. We determined levels of soluble TGF-beta 1 in the serum of 12 patients with GBS in serial investigations during the course of the disease, in 12 patients with other noninflammatory neurological diseases (OND), and in 12 healthy control subjects. Levels of biologically active and total TGF-beta 1 were significantly increased in patients with GBS compared with patients with OND and healthy controls. During the course of GBS, levels of TGF-beta 1 peaked in the plateau phase before onset of recovery. During the recovery phase levels of TGF-beta 1 decreased but still exceeded significantly the levels in patients with OND and healthy controls. The differences were more marked with biologically active than with total TGF-beta 1. The temporal relationship between increased serum levels of TGF-beta 1 and the end of the progressive phase indicates that TGF-beta 1 has a role in terminating the pathological immune response in GBS. These findings suggest that TGF-beta 1 may be important in recovery from GBS.

Detection of transforming growth factor beta 1 mRNA in cerebrospinal fluid cells of patients with meningitis by non-radioactive in situ hybridization.

Meningitis is a serious disease mostly caused by viral or bacterial infections. In complicated cases it may lead to brain damage and death. The infection and cell damage result in a cellular and immunological response. Following this, a high secretion of cytokines can be expected. Cytokines, especially tumour necrosis factor alpha (TNF-alpha) and interleukin-1 (IL-1), promote the inflammatory reactions in the subarachnoid space. Transforming growth factor beta 1 (TGF-beta 1) has antagonistic effects on TNF-alpha and IL-1-mediated processes. Therefore, it suppresses inflammatory reactions. To observe the expression of TGF-beta 1 in transcellular signalling in the inflammatory processes of meningitis, we investigated TGF-beta 1 mRNA in cells in the cerebrospinal fluid of three patients with meningitis by non-radioactive in situ hybridization. All patients fulfilled the usual clinical criteria of meningitis. In one case Neisseria menigitidis could be identified as the pathogenic agent. In the remainder, no agent could be isolated. In all cytological preparations of the cerebrospinal fluid of these patients a high level of TGF-beta 1 mRNA was detectable in the cell populations. It was possible to distinguish between the different cell types of the cerebrospinal fluid and to attach the mRNA expression to them. On the one hand, this makes it possible to investigate pathogenesis and defence mechanisms in bacterial and aseptic meningitis on a cellular level; on the other hand, it may open new perspectives in the control of disease development, prognosis, diagnosis and supporting therapy.

[Expression and secretion of alpha-2-macroglobulin by dust-stimulated alveolar macrophages]. / Expression und Sekretion von Alpha-2-Makroglobulin durch staubstimulierte Alveolarmakrophagen.

In dust-induced, fibrosing lung processes macrophages are increased activated by foreign body reactions. The release of monokines and proteolytic enzymes, which depends on phagocytosis, may lead to destruction of the extracellular matrix with the consequence of degradation and restitution. Also the transcellular signaling or cell-matrix-interaction may finally result in development of fibrosis. However the proteolytic effect of elastase and collagenase can be inhibited by alpha-2-macroglobulin. Alpha-2-macroglobulin is a protease-inhibitor, which is synthesized by macrophages and has a wide spectrum of inhibitory abilities. Our interest was focused on observation of the production of alpha-2-macroglobulin by alveolar macrophages after stimulation with inorganic dusts of different chemical and physical properties. Rat alveolar macrophages were isolated by bronchoalveolar lavage and exposed to crocidolite, quarz or welder steam dust in vitro. The expression and secretion of alpha-2-macroglobulin was examined by non radioactive in situ hybridization, indirect immunofluorescence and radial immunodiffusion according to Mancini. The stimulated rat alveolar macrophages showed an increased expression of alpha-2-macroglobulin-mRNA and also an enhanced synthesis of alpha-2-macroglobulin-protein. Besides only small differences between the substances used for stimulation were demonstrated.