[show abstract][hide abstract] ABSTRACT: In this study we evaluated the percentages of CD3(-)CD56(bright), CD3(-)CD56(dim), CD3(-)CD56(bright)perforin(+) and CD3(-)CD56(dim)perforin(+) Natural Killer (NK) cells in peripheral blood from untreated secondary progressive (SP) and primary progressive (PP) multiple sclerosis (MS) patients and age and sex matched healthy subjects. Both PPMS patients and SPMS patients showed increased percentages of circulating CD3(-)CD56(dim)perforin(+) NK cells than healthy subjects. The increased percentage of CD3(-)CD56(dim) NK cells expressing perforin in patients affected by the progressive forms of MS suggests a possible role of this NK cell subpopulation in the pathogenesis of the disease.
Journal of neuroimmunology 10/2013; · 2.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: Objective: Multiple Sclerosis (MS) is an inflammatory, chronic, degenerative and demyelinating disease of the Central Nervous System (CNS). It is known that HLA-DR2 haplotype is a predisponent genetic factor to MS. The activation of myeline reactive T cells in the periphery lead to demyelination in CNS, through the induction of expression of adhesion molecules that allow their entry in the CNS through the Blood-Brain Barrier (BBB). We investigated the TR-β repertoire of T cells specific for epitope MBP85-99, immunodominat in HLA-DR2+ patients.
Methods: We focused on DR2+ subjects: 7 untreated patients with diagnosis of active MS; 6 pazients with Clinically Isolated Syndrome (CIS); 6 MS patients in remission phase (REM); 7 healthy control subjects (HS). PBMC were cultured in vitro with or without MBP85-99 and with MBP111-129. We used the “Immunoscope” technique (a TRBV-TRBJ spectratyping) to investigate the TR-β repertoire of T cells. Magnetic separation of IL17+ and IFN+ populations was performed and their TR-β repertoire was analyzed. We performed the sequencing analysis of the more interesting TRBV-TRBJ rearrangements.
Results: In a first analysis we identified 20 MBP85-99 specific TRBV-TRBJ rearrangements shared among MS DR2+ patients. The MBP111-129 specific rearrangements showed expansions unshared with the MBP85-99 epitope. Shared TCRs were studied in the IL17+ cells. As control, the same rearrangements were checked in the IFN+ cells and in polarized T cells from HS. The TCR repertoire usage in Th17 and Th1 cells is mutually exclusive. In one patient we founded 1 rearrangement specific to Th17 cells, while 25% of the investigated TCRs were specific for IFN+ cells. Longitudinal analyses in one patient with active MS and after 6 (REM) and 12 (relapse) months of therapy showed a correlation between TCR usage and the treatment efficacy. We performed the sequencing of the shared TCRs to confirm the similar response to MBP85-99 in the MS patients, not detectable in the CIS and healthy subjects.
Conclusions: These data hint that the repertoire of TCR specific for MBP85-99 may be broader in CIS than in MS patients. In REM and during efficient treatment we observe a contraction of TCRs, in particular the ones secreting IFNgamma and IL17. We could be able to monitor the identified specific TCRs in the peripheral blood of the MS patients during the various relapsing-remitting phases of the disease and during therapy.
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.; 08/2013
[show abstract][hide abstract] ABSTRACT: The aim of the study was to evaluate the type-1 immune response by analyzing T-bet expression in circulating T and B cells in Primary Progressive (PP) and Secondary Progressive (SP) Multiple Sclerosis (MS) patients. We found higher percentages of circulating CD4+T-bet+ and CD8+T-bet+ T cells in SPMS and PPMS than in remitting-relapsing MS patients and controls. Moreover, in SPMS, we observed a positive correlation between the percentages of circulating CD4+T-bet+ or CD8+T-bet+ T cells and disease severity. The increased percentages of Th1 and Tc1 cells suggest that MS progressive forms, unlike RRMS, are characterized by a permanent peripheral type-1 immune activation.
Journal of neuroimmunology 05/2012; 249(1-2):112-6. · 2.84 Impact Factor
[show abstract][hide abstract] ABSTRACT: Swine-origin H1N1 influenza virus (S-OIV) appeared in 2009 with a higher incidence rate among children. Although fever was the most common symptom, some complicated cases occurred. We evaluated the percentages of effector T cells, B cells, and regulatory T cells in peripheral blood from 5 children infected by S-OIV (1 with acute necrotizing encephalitis, 2 with pneumonia, and 2 without complications), 5 children with seasonal influenza, and 5 healthy children. We found higher percentages of T-bet(+) CD4(+)CD8(+) T cells, monocytes, and B cells, granzyme B(+) and perforin(+) CD4(+), and CD8(+) T cells in affected children with both seasonal and H1N1 influenza than in controls, whereas both groups demonstrated similar percentages of CD4(+)CD25(+)Foxp3(+) regulatory T cells. In infected children with complications we observed high percentages of perforin(+) and interferon-γ(+) CD4(+) and CD8(+) T cells associated with low percentages of T regulatory cells. Our data suggest a dysregulation of antipathogen type I immune responses in complicated S-OIV infections.
Human immunology 05/2011; 72(8):632-5. · 2.55 Impact Factor
[show abstract][hide abstract] ABSTRACT: Circulating T cells and monocytes expressing T-bet, pSTAT1 and pSTAT3 increase in relapsing-remitting multiple sclerosis (RRMS) during relapse. Natalizumab (NZB) is an effective drug in RRMS, but exacerbation of the disease after its discontinuation has been described in some patients. The aim of this research was to study the effect of NZB treatment on circulating lymphomonocyte subpopulations expressing T-bet, pSTAT1, pSTAT3 and CD4+CD25+Foxp3+ regulatory T cells. Flow cytometry was used to evaluate the percentages of circulating CD4+ and CD8+ T cells, CD14+ monocytes and B cells expressing T-bet, pSTAT1, and pSTAT3, and CD4+CD25+Foxp3+ regulatory T cells from RRMS patients before and after 6-12 NZB infusions. In NZB-treated RRMS patients, the percentages of CD4+pSTAT1+ and CD8+pSTAT1+ T cells, CD14+pSTAT1+ monocytes, CD4+T-bet+, CD8+T-bet+ and CD4+pSTAT3+ T cells and CD14+pSTAT3+ monocytes increased after 12 drug infusions and were similar to those observed in untreated relapsing RRMS patients. Otherwise in vitro NZB exposure of peripheral blood mononuclear cells from untreated RRMS patients and controls had no effect. It was concluded that NZB treatment determines an accumulation of CD4+pSTAT1+, CD8+pSTAT1+, CD4+T-bet+, CD8+T-bet+ and CD4+STAT3+ T cells in peripheral blood that may account for the exacerbation of the disease observed in some patients after the discontinuation of the drug.
[show abstract][hide abstract] ABSTRACT: Facioscapulohumeral muscular dystrophy (FSHD) is an inherited disease, and although strongly suggested, a contribution of inflammation to its pathogenesis has never been demonstrated. In FSHD patients, we found by immunohistochemistry inflammatory infiltrates mainly composed by CD8(+) T cells in muscles showing hyperintensity features on T2-weighted short tau inversion recovery magnetic resonance imaging (T2-STIR-MRI) sequences. Therefore, we evaluated the presence of circulating activated immune cells and the production of cytokines in patients with or without muscles showing hyperintensity features on T2-STIR-MRI sequences and from controls. FSHD patients displaying hyperintensity features in one or more muscles showed higher CD8(+)pSTAT1(+), CD8(+)T-bet(+) T cells and CD14(+)pSTAT1(+), CD14(+)T-bet(+) cells percentages and IL12p40, IFNγ and TNFα levels than patients without muscles displaying hyperintense features and controls. Moreover, the percentages of CD8(+)pSTAT1(+), CD8(+)T-bet(+) and CD14(+)pSTAT1(+) cells correlated with the proportion of muscles displaying hyperintensity features at T2-STIR sequences. These data indicate that circulating activated immune cells, mainly CD8(+) T cells, may favour FSHD progression by promoting active phases of muscle inflammation.
Journal of Clinical Immunology 11/2010; 31(2):155-66. · 3.38 Impact Factor
[show abstract][hide abstract] ABSTRACT: A defect of CD4(+) regulatory T cells (Treg) seems to be involved in the pathogenesis of multiple sclerosis (MS). Besides Treg, CD8(+) T cells also can suppress the immune response. Forkhead box p3 (Foxp3) is known to program the acquisition of suppressive capacities in CD4(+) T cells and recent studies showed that in vitro antigen activation leads to Foxp3 expression in CD8(+) T cells, gaining of suppressive activity. By flow cytometry we found a lower percentage of circulating CD8(+)Foxp3(+) T cells in relapsing than in remitting patients with MS and in controls. No significant differences were observed in CD8(+)Foxp3(+) T cell percentage between healthy subjects and patients in remission. Our data suggest that peripheral CD8(+)Foxp3(+) T cells may play a role in the maintenance of tolerance in MS.
Human immunology 02/2010; 71(5):437-41. · 2.55 Impact Factor
[show abstract][hide abstract] ABSTRACT: Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is considered an auto-immune disorder. We evaluated expression of pSTAT1, T-bet, and pSTAT3 in circulating T-cells, B-cells, and monocytes and spontaneous production of interleukin-17 (IL17), interferon-gamma (IFN gamma), and interleukin-10 (IL10) by peripheral blood mononuclear cells (PBMCs) from 14 active CIDP patients compared with 6 patients with long-lasting remission and 20 controls. Active disease patients showed higher pSTAT1, T-bet, and pSTAT3 in CD4(+) T-cells than controls (p < 0.001, p = 0.0002, p = 0.0097, respectively) and remission patients (p < 0.001, p = 0.0036, p = 0.0008, respectively). pSTAT1, T-bet, and pSTAT3 were also higher in monocytes from active CIDP patients than controls (p = 0.0011, p = 0.0041, p = 0.0413, respectively) and remission patients (p = 0.0073, p = 0.0274, p = 0.0251, respectively). Moreover in CD8(+) T-cells, pSTAT3 expression was higher in active CIDP patients than in remission patients (p = 0.0345) and in controls (p = 0.0023). IL17 and IFN gamma production were significantly higher in active CIDP patients than in controls (p < 0.0395, p = 0.0010, respectively); IFN gamma levels were higher also in remission CIDP patients (p = 0.0073). IL10 levels were higher in active phase patients than in controls (p = 0.0334). Our data suggest that pSTAT1, T-bet, and pSTAT3 can be considered putative markers of disease activity and potential targets for specific therapies.
Journal of the Peripheral Nervous System 06/2009; 14(2):107-17. · 2.57 Impact Factor
[show abstract][hide abstract] ABSTRACT: Regulatory CD4+ CD25+Foxp3+ T cells are involved in the regulation of immune response and inhibit protective antitumor immunity. Celiac disease (CD), a food gluten-sensitive enteropathy, is considered a T-cell-mediated autoimmune disease and is generally associated with an overall increased risk of cancer in CD patients. We observed a higher percentage of circulating CD4+CD25+Foxp3+ T cells and an increased Foxp3 expression in CD4+CD25+ T cells from untreated than from treated CD patients. In co-culture, CD4+CD25+ T cells from both treated and untreated CD patients significantly suppressed the proliferation of autologous CD4+CD25(-) T cells similarly to values in healthy subjects. Our study suggests that Treg proportion and Foxp3 expression in circulating CD4+CD25+ T cells could justify the increased global risk of malignancy in CD population and support the efficacy of lifelong gluten-free diet in the reduction of the cancer risk.
Human immunology 05/2009; 70(6):430-5. · 2.55 Impact Factor
[show abstract][hide abstract] ABSTRACT: Not all patients with clinically isolated syndrome (CIS) suggestive of multiple sclerosis (MS) develop clinically defined MS (CDMS). At first clinical event we observed increased production of IL17, IFNgamma and IL10 by peripheral blood mononuclear cells from patients with CIS that remained high in remission. In CD4+ T cells pSTAT3 expression was higher in patients who subsequently converted to CDMS than in patients who did not and controls. The persistency of high levels of pSTAT3 in circulating CD4+ T cells from CIS patients after the first clinical event may favor the early conversion to CDMS.
Journal of Neuroimmunology 11/2008; 205(1-2):126-34. · 3.03 Impact Factor
[show abstract][hide abstract] ABSTRACT: Multiple sclerosis (MS) and celiac disease (CD) are considered to be T-cell-mediated autoimmune diseases. We describe a woman affected by a relapsing-remitting demyelinating disease of the central nervous system and occult CD (MS-CD), who showed during neurologic exacerbations a strong increased expression of T-bet, the key transcription factor for the development of Th1 cells, in circulating T, B cells and monocytes. Conversely, no difference of T-bet expression was observed in B cells from relapsing-remitting MS patients, either in relapse or in remission, and in controls. In the MS-CD patient, the interaction between MS- and CD-related inflammatory processes may result in an amplification of Th1 immune response.
Human Immunology 11/2008; 69(12):837-9. · 2.30 Impact Factor
[show abstract][hide abstract] ABSTRACT: We evaluated the spontaneous IL17, IFNgamma and IL10 production by peripheral blood mononuclear cells from patients affected by clinically isolated syndromes (CIS) suggestive of multiple sclerosis (MS) both in acute phase and in remission, relapsing remitting MS (RRMS) both in relapse and in remission, not-relapsing secondary progressive MS (SPMS) and controls. We observed higher IL17 levels in CIS patients both in acute phase and in remission than in SPMS patients and controls. On the contrary no difference in IL17 production was observed among RRMS patients and CIS, SPMS patients and controls. IFNgamma levels were significantly higher in CIS patients in acute phase than in CIS and RRMS patients in remission, SPMS patients and controls. Moreover, we observed higher IFNgamma spontaneous production in relapsing RRMS patients than in remitting RRMS and SPMS patients and controls. IL10 levels were significantly higher in remitting CIS and in relapsing RRMS patients than in SPMS patients and controls. There was no difference in IFNgamma, IL10 and IL17 levels between SPMS patients and controls. Our data suggest that IL17 might play a crucial role mainly in the early phase of MS, while IFNgamma seems to be involved both in the early phase and in the following relapses of the disease.
[show abstract][hide abstract] ABSTRACT: In this study we observed higher serum leptin levels in relapsing-remitting multiple sclerosis (RRMS) patients during remission than in controls. The expression of leptin receptor (ObR) was higher in CD8+ T cells and monocytes from RRMS patients in relapse than in patients in remission and in controls. Relapsing patients showed high levels of pSTAT3 and low expression of SOCS3 and leptin administration induced an up-regulation of pSTAT3 only in monocytes from patients in relapse. Our data suggest that ObR may be involved in the development of clinical relapses in RRMS patients and suggest a rationale for potential targeting of the leptin axis during MS.
Journal of Neuroimmunology 01/2008; 192(1-2):174-83. · 3.03 Impact Factor
[show abstract][hide abstract] ABSTRACT: High dose glucocorticoid (GC) treatment has been demonstrated to have a short-term beneficial effect on functional recovery in relapsing multiple sclerosis (MS) patients but the exact mechanism of action of GCs in MS is unclear. We found that high dose intravenous GCs strongly reduced T-bet and pSTAT1 expression in CD4+, CD8+, CD14+ circulating cells in RRMS patients in relapse. pSTAT1and T-bet reduction was associated with the decline of IFNgamma production by PBMCs. A significant increase of AV-positive CD4+ and CD8+ T cells was detectable after GC treatment without any variation in the percentage of annexin V-positive monocytes. By in vitro analysis, patients during relapse, either before or after GC treatment, exhibited a lower proportion of apoptotic lymphocytes than remitting patients and controls. Our study suggests that GCs can modulate T-bet and STAT1 expression and that IFNgamma signalling inhibition contributes to anti-inflammatory action of GCs in the treatment of relapses of MS patients.
[show abstract][hide abstract] ABSTRACT: Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system, and it is considered to be a T helper 1 (Th1) cell-mediated autoimmune disease. T-bet has been identified as a key transcription factor for the development of Th1 cells and the induction of interferon (IFN)-gamma production. T-bet is induced during T-cell activation by the IFN-gamma signal transducer and activator of transcription (STAT)-1 signalling pathway. In this study we found an up-regulation of T-bet and pSTAT1 in peripheral blood CD4+ and CD8+ T cells and monocytes from relapsing-remitting MS patients in relapse compared with patients in remission and with healthy subjects. The increased expression of pSTAT1 strongly correlated with T-bet expression in CD4+ and CD8+ cells and monocytes from patients in relapse and was associated with an increased production of IFN-gamma by peripheral blood mononuclear cells (PBMCs). pSTAT3 was also up-regulated in CD4+ and CD8+ cells and monocytes from patients in relapse and was associated with an increased production of interleukin (IL)-10 but not of IL-6. pSTAT1, pSTAT3, and T-bet expression strongly correlated with Gd-DTPA-enhanced lesions on brain and spinal cord magnetic resonance imaging. Our data show for the first time that there is an up-regulation of type 1 immunity-correlated transcription factors such as STAT1 and T-bet in peripheral blood subpopulations of MS patients in the active phase of disease. The evaluation of T-bet and pSTAT1 expression in peripheral blood CD4+, CD8+ T cells and monocytes could be used as a marker of disease activity in relapsing-remitting MS.
Journal of Neuroscience Research 11/2006; 84(5):1027-36. · 2.97 Impact Factor
[show abstract][hide abstract] ABSTRACT: Although interferon (IFN) beta is a widely used disease-modifying therapy in multiple sclerosis (MS), the mechanisms responsible for its effects are not fully understood. Some studies demonstrated that IFNbeta induces nerve growth factor (NGF) secretion by astrocytes and by brain endothelial cells. In this study, we determined the production of various neurotrophins (brain-derived neurotrophic factor, BDNF; NGF; glial cell line-derived neurotrophic factor; neurotrophin 3; neurotrophin 4) by peripheral blood mononuclear cells (PBMCs) in relapsing-remitting (RR) and secondary progressive (SP) MS patients during IFNbeta treatment. There were no main variations in neurotrophin production either among all MS patients globally considered or in the group of SPMS subjects. Instead, in the group of RRMS patients who did not present clinical exacerbation of disease up to the end of the study, we found a significant increase in BDNF production as from 6 months after starting therapy.