transforming-growth-factor-beta and Multiple-Sclerosis--Chronic-Progressive

Mitoxantrone is an antineoplastic agent also used for the treatment of multiple sclerosis (MS). However, despite its efficacy, few data are available on its mechanism of action. The current study was designed to evaluate the short-term (1 month) and long-term (12 months) in vivo effects of mitoxantrone on pro- and anti-inflammatory cytokine production by the peripheral blood mononuclear cells (PBMC) of secondary progressive MS patients.. Eighteen patients with secondary progressive MS underwent mitoxantrone therapy (at a dose of 12 mg/m(2) once every 3 months) over a 1-year period. Blood samples were obtained at baseline, after 1 month and after 12 months of treatment. The production of cytokines in the PBMC was measured by enzyme-linked immunosorbent assay.. There were no significant effects of mitoxantrone on proinflammatory cytokines [interleukin (IL) 6 and IL-12p40] and anti-inflammatory cytokines (IL-10 and transforming growth factor-beta) in our patients. Patients who showed no signs of therapeutic response were characterized by a higher basal PBMC production of IL-6 compared with that of the responding patients (p < 0.05) and mitoxantrone reduced this production after 12 months of treatment (p < 0.05). In the responding patients, IL-10 was significantly increased by mitoxantrone after 12 months of treatment (p < 0.05).. These findings provide additional information useful in the selection of the patient population suitable for mitoxantrone treatment and suggest that most probably the therapeutic action of mitoxantrone in MS is not entirely mediated by its immunosuppressant effects.

Topics: Adult; Antineoplastic Agents; Cells, Cultured; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Immunosuppressive Agents; Inflammation Mediators; Interleukin-10; Interleukin-12 Subunit p40; Interleukin-6; Leukocytes, Mononuclear; Male; Middle Aged; Mitoxantrone; Multiple Sclerosis, Chronic Progressive; Time; Transforming Growth Factor beta; Treatment Outcome; Up-Regulation

We compared the patterns of the pro-inflammatory cytokines, interferon-gamma (IFN-gamma), interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha), and the anti-inflammatory cytokines, interleukin-10 (IL-10) and tumor growth factor-beta (TGF-beta) from peripheral blood of male and female patients with relapsing-remitting (RR) and secondary progressive (SP) forms of multiple sclerosis (MS). The relationships between pro-inflammatory cytokines and disability (expanded disability status scale, EDSS) were also examined. Peripheral blood anti-coagulated with heparin was obtained from 47 MS patients (30 women and 17 men) and activated with phorbol-12-myristate 13 acetate (PMA) and ionomycin in the presence of brefeldin A and stained for flow cytometry with fluorescently labeled antibodies against intracellular IFN-gamma, TNF-alpha, IL-2, IL-4 and IL-10. The T cells were delineated with peridinin chlorophyll protein (Per-CP) labeled anti-CD3 antibody. The stained samples were analyzed on a flow cytometer to assess the intracellular pro-inflammatory cytokine patterns. The levels of interleukin-10 (IL-10) and tumor growth factor-beta (TGF-beta) were measured in plasma using enzyme-linked immunoassay. The percentage of TNF-alpha-producing CD3 positive cells was significantly higher (P=0.045) in men (mean+/-S.D., 39+/-13%) than in women (mean+/-S.D., 29+/-13%) RR-MS patients. The percentage of CD3 positive cells producing IFN-gamma was significantly correlated with EDSS in females but not in males (Spearman rank correlation r(S)=0.49, P=0.018). The secretion of the pro-inflammatory cytokines, IFN-gamma and TNF-alpha, is influenced by gender in MS patients and may contribute to the sexual dimorphism of MS.

Topics: Adolescent; Adult; Aged; Analysis of Variance; CD3 Complex; CD5 Antigens; Cytokines; Disability Evaluation; Female; Humans; Inflammation Mediators; Interferon-gamma; Interleukin-10; Interleukin-2; Interleukin-4; Male; Middle Aged; Multiple Sclerosis, Chronic Progressive; Multiple Sclerosis, Relapsing-Remitting; Sex Factors; T-Lymphocytes; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

One of the auto-antigens aberrantly targeted in Multiple sclerosis is myelin basic protein (MBP). In this study, chronic progressive multiple sclerosis (CPMS) patients receiving the experimental drug MBP8298, on a compassionate care trial, were examined before and after high dose peptide treatment for their circulating regulatory T-cell numbers and their responses to the common mitogens, phytohemagglutinin and poke-weed mitogen. Peripheral blood mononuclear cells (PBMCs) isolated from these patients before treatment displayed anergy upon stimulation with phytohemagglutinin; measured through reduced proliferation, IFN-γ and IL-17A secretion in an in vitro cell culture system. 6 Weeks and 6months after treatment their PBMCs displayed a reversal of anergy with phytohemagglutinin stimulation. There was also a marked increase in their CD4(+)CD25(+hi)FoxP3(+) T-cells regulatory T-cells. These results suggest that high dose MBP8298 treatment has a profound effect on the circulating T-cells of CPMS patients, capable of reversing peripheral anergy and establishing T regulation.

Topics: CD4 Lymphocyte Count; Clonal Anergy; Compassionate Use Trials; Dose-Response Relationship, Immunologic; Follow-Up Studies; Humans; Immunotherapy; Interferon-gamma; Interleukin-17; Interleukin-2 Receptor alpha Subunit; Multiple Sclerosis, Chronic Progressive; Myelin Basic Protein; Peptide Fragments; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Treatment Outcome

Reliable, and easy to measure, immunological markers able to denote disease characteristics in multiple sclerosis (MS) patients are still lacking. We applied a multivariate statistical analysis on results obtained by measuring-by real-time RT-PCR-mRNA levels of 25 immunological relevant molecules in PBMCs from 198 MS patients. The combined measurement of mRNA levels of IL-1beta, TNF-alpha, TGF-beta, CCL20 and CCR3 was able to distinguish MS patients from healthy individuals. CXCR5, CCL5, and CCR3 combined mRNA levels identify primary progressive MS patients while TNF-alpha, IL-10, CXCL10 and CCR3 differentiate relapsing MS patients. Our results indicate that multi-parametric analysis of mRNA levels of immunological relevant molecules in PBMCs may represent a successful strategy for the identification of putative peripheral markers of disease state and disease activity in MS patients.

Topics: Adult; Biomarkers; Chemokine CCL20; Chemokine CCL5; Chemokines, CC; Disease Progression; Female; Humans; Interleukin-1; Linear Models; Macrophage Inflammatory Proteins; Magnetic Resonance Imaging; Male; Middle Aged; Models, Immunological; Multiple Sclerosis; Multiple Sclerosis, Chronic Progressive; Multivariate Analysis; Receptors, CCR3; Receptors, Chemokine; Receptors, CXCR5; Receptors, Cytokine; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Glucocorticoids (GC) are commonly used to treat inflammatory disorders such as multiple sclerosis (MS) and may exert their immunosuppressive activity by inducing apoptosis in activated lymphocytes. However, unlike relapsing-remitting MS patients, those with progressive disease respond poorly to GC treatment. The data in this communication indicate that PLP peptide-specific T cell clones from progressive, but not relapsing-remitting MS patients are resistant to GC-induced apoptosis in vitro, in a fashion associated with expression of B-7 co-stimulatory molecules. Thus, failure to respond to GC treatment may reflect defect in apoptosis that develop during the progressive stages of chronic inflammatory disease.

Topics: Adult; Apoptosis; Autoantigens; Cell Cycle Proteins; Clone Cells; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Cyclins; Dexamethasone; Female; Gene Expression; Glucocorticoids; Humans; In Vitro Techniques; Interleukin-2; Male; Microtubule-Associated Proteins; Middle Aged; Multiple Sclerosis, Chronic Progressive; Myelin Proteolipid Protein; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; T-Lymphocytes; Transforming Growth Factor beta; Tumor Suppressor Protein p53; Tumor Suppressor Proteins