Untreated MS Research Articles

Fibrinolysis associated proteins and lipopolysaccharide bioactivity in plasma and cerebrospinal fluid in multiple sclerosis

The coagulation cascade and fibrinolysis have links with neuroinflammation and increased activation of the coagulation system has been reported in MS patients. We quantified levels of D-dimer, tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1) and the bioactivity of bacterial lipopolysaccharide (LPS) in cerebrospinal fluid (CSF) and plasma from newly diagnosed untreated MS patients and controls. These molecules showed multiple correlations with each other as well as with age, HLA-DRB1*15:01, body-mass-index and CSF IgG. Our results confirm previous findings of increased plasma PAI-1 and LPS in MS patients compared to controls indicating changes in platelet function and gut permeability in MS.

Interferon-beta exposure in-utero and the risk of infections in early childhood.

Knowledge within the field of multiple sclerosis treatment during pregnancy is vital to ensure the most optimal clinical practice. Immunomodulatory treatment in pregnancy could in theory affect the normal development and maturation of the immune system of the fetus with a potential increased risk of infections, consequently. We therefore set out to investigate whether exposure to interferon-beta in utero affected the risk of acquiring infections in early childhood. This retrospective matched cohort study utilized data from the Danish Multiple Sclerosis Registry linked with national Danish registries to identify all children born of mothers with MS in Denmark from 1998 to 2018. The study included 510 children exposed to interferon-beta in utero. The children were matched 1:1 on various of demographic characteristics with children born to mothers with untreated MS and 1:3 with children born to mothers without MS. Each child was followed for up to five years. Using individual-level data, we investigated all-cause mortality, rate of hospital admissions due to infections, and redeemed prescriptions of antibiotics. The primary statistical model applied was a negative binomial regression analysis. We found no differences in childhood mortality, for hospital admissions the rate ratio compared to healthy controls was 0.79 (0.62-1.00). Regarding antibiotic prescriptions, the results were similar (RR 1.00 (0.90-1.11). Furthermore, we found no certain dose-response relationship between interferon-beta exposure duration and hospital admission rate (P=0.47) or redeemed antibiotic prescription (P=0.71). Exposure to interferon-beta during gestation has little to no impact on the risk of acquiring significant infections during the first five years of childhood.

Central and peripheral myeloid-derived suppressor cell-like cells are closely related to the clinical severity of multiple sclerosis

Multiple sclerosis (MS) is a highly heterogeneous demyelinating disease of the central nervous system (CNS) that needs for reliable biomarkers to foresee disease severity. Recently, myeloid-derived suppressor cells (MDSCs) have emerged as an immune cell population with an important role in MS. The monocytic-MDSCs (M-MDSCs) share the phenotype with Ly-6Chi-cells in the MS animal model, experimental autoimmune encephalomyelitis (EAE), and have been retrospectively related to the severity of the clinical course in the EAE. However, no data are available about the presence of M-MDSCs in the CNS of MS patients or its relation with the future disease aggressiveness. In this work, we show for the first time cells exhibiting all the bona-fide phenotypical markers of M-MDSCs associated with MS lesions, whose abundance in these areas appears to be directly correlated with longer disease duration in primary progressive MS patients. Moreover, we show that blood immunosuppressive Ly-6Chi-cells are strongly related to the future severity of EAE disease course. We found that a higher abundance of Ly-6Chi-cells at the onset of the EAE clinical course is associated with a milder disease course and less tissue damage. In parallel, we determined that the abundance of M-MDSCs in blood samples from untreated MS patients at their first relapse is inversely correlated with the Expanded Disability Status Scale (EDSS) at baseline and after a 1-year follow-up. In summary, our data point to M-MDSC load as a factor to be considered for future studies focused on the prediction of disease severity in EAE and MS.

Gut microbiome of multiple sclerosis patients and paired household healthy controls reveal associations with disease risk and course

Changes in gut microbiota have been associated with several diseases. Here, the International Multiple Sclerosis Microbiome Study (iMSMS) studied the gut microbiome of 576 MS patients (36% untreated) and genetically unrelated household healthy controls (1,152 total subjects). We observed a significantly increased proportion of Akkermansia muciniphila, Ruthenibacterium lactatiformans, Hungatella hathewayi, and Eisenbergiella tayi and decreased Faecalibacterium prausnitzii and Blautia species. The phytate degradation pathway was over-represented in untreated MS, while pyruvate-producing carbohydrate metabolism pathways were significantly reduced. Microbiome composition, function, and derived metabolites also differed in response to disease-modifying treatments. The therapeutic activity of interferon-β may in part be associated with upregulation of short-chain fatty acid transporters. Distinct microbial networks were observed in untreated MS and healthy controls. These results strongly support specific gut microbiome associations with MS risk, course and progression, and functional changes in response to treatment.

Exploring Retinal Blood Vessel Diameters as Biomarkers in Multiple Sclerosis.

We aimed to determine whether retinal vessel diameters and retinal oxygen saturation in newly diagnosed patients with multiple sclerosis (pwMS) are different from those of a healthy population. Retinal blood vessel diameters were measured using imaging with a spectrophotometric non-invasive retinal oximeter. Twenty-three newly diagnosed untreated relapsing-remitting MS (RRMS) patients (mean age: 32.2 ± 7.5 years, age range = 18–50 years, 56.5% female) were measured and compared to 23 age- and sex-matched healthy controls (HCs) (mean age: 34.8 ± 8.1 years). Patients with Optic Neuritis were excluded. Retinal venular diameter (143.8 µm versus 157.8 µm: mean; p = 0.0013) and retinal arteriolar diameter (112.6 µm versus 120.6 µm: mean; p = 0.0089) were smaller in pwMS when compared with HCs, respectively. There was no significant difference in the oxygen saturation in retinal venules and arterioles in pwMS (mean: 60.0% and 93.7%; p = 0.5980) compared to HCs (mean: 59.3% and 91.5%; p = 0.8934), respectively. There was a significant difference in the median low contrast visual acuity (2.5% contrast) between the pwMS and the HC groups (p = 0.0143) Retinal arteriolar and venular diameter may have potential as objective biomarkers for MS.

The effect of miRNAs targeting TGFβ-Signaling in Multiple Sclerosis

Abstract Multiple sclerosis (MS) is a demyelinating disease of the central nervous system (CNS) that is mediated by a dysregulated immune system. It can lead to severe neurological issues and is one of the major causes for disability in young adults. The cause of the disease is unknown, the immune and neurodegenerative mechanisms underlying the pathophysiology of the disease are poorly understood. To this date there is no cure and current therapies are not able to completely slow down disease progression. In a large miRNA profiling study on naïve and effector/memory CD4 T cell in untreated MS patients, our lab found a number of miRNAs differently expressed. One of the main pathways affected by this dysregulation of miRNAs is the TGFβ signaling pathway. This impairment limits the differentiation of regulatory T cells. To study the effect of TGFβ-targeting miRNAs in a murine model of MS, neonatal mice were injected with miRNAs that were found to be upregulated in MS patients at day 0 and 21 of life and experimental autoimmune encephalomyelitis (EAE) was induced using conditions under which the controls would only get a mild form of the disease. Mice, injected with these miRNAs had an earlier onset of EAE and disease severity was significantly increased compared to control mice. Not only did an injection of TGFβ-targeting miRNAs caused mice to have reduced numbers of Tregs in the spleen and thymus compared to control mice, but these Tregs also showed a dramatic loss of TCR diversity in most Vβ genes as TCR Vβ deep sequencing of sorted Tregs revealed. Our findings contribute to the understanding of Treg dysfunction in MS patients and determining candidate markers for disease susceptibility and treatment efficacy. Supported by grants from NIH (R01 AI152435-01)

Effect of cladribine on COVID-19 serology responses following two doses of the BNT162b2 mRNA vaccine in patients with multiple sclerosis

Backgroundmultiple sclerosis (MS) patients are treated with immunomodulatory treatments that can influence their ability to develop a protective antibody response to the SARS-CoV-2 vaccine.Vaccine efficacy is important for treatment decision and for patients’ reassurance.The main objective is to assess antibody response to SARS-CoV-2 vaccine in MS patients treated with cladribine. MethodsSerology response was tested in 97 participants, 67 MS patients and 30 healthy controls (HCs), using two independent methods, 2–3 weeks following the second dose of the BNT162b2 vaccine. ResultsHCs (n = 30) and MS patients treated with cladribine (n = 32) had 100% positive serology response against the SARS-CoV-2 spike protein following the second vaccine dose (mean S1/S2-IgG and RBD-IgG:284.5 ± 104.9, 13,041±9411 AU/mL and 226.3 ± 121.4, 10,554±11,405 AU/mL respectively). Comparable findings were observed for untreated MS patients, and interferon beta-1a-treated MS patients (mean S1/S2-IgG: 282.1 ± 100.1, 276.9 ± 94.31 AU/mL respectively). No correlation was found between lymphocyte counts, treatment duration, or time between cladribine dose and vaccination, and serology response or antibody titers. Conclusion and relevanceCladribine treated MS patients are able to produce antibodies to the SARS-CoV-2 mRNA vaccine. In the era of the COVID-19 pandemic, it is reassuring and important for both patients and physicians and will allow to develop consensus guidelines.

Anti-CD20 therapy corrects a CD8 regulatory T cell deficit in multiple sclerosis

Objective: To determine the effect of long-term anti-CD20 B-cell-depleting treatment on regulatory T cell immune subsets that are subnormal in untreated MS patients. Methods: 30 clinically stable MS patients, before and over 38 months of ocrelizumab treatment, were compared to 13 healthy controls, 29 therapy-naïve MS, 9 interferon-β-treated MS, 3 rituximab-treated MS, and 3 rituximab-treated patients with other autoimmune inflammatory diseases. CD8, CD28, CD4, and FOXP3 expression in peripheral blood mononuclear cells was quantitated with flow cytometry. Results: CD8+ CD28− regulatory cells rose from one-third of healthy control levels before ocrelizumab treatment (2.68% vs 7.98%), normalized by 12 months (13.5%), and rose to 2.4-fold above healthy controls after 18 months of ocrelizumab therapy (19.0%). CD4+ FOXP3+ regulatory cells were lower in MS than in healthy controls (7.98%) and showed slight long-term decreases with ocrelizumab. CD8+ CD28− and CD4+ FOXP3+ regulatory T cell percentages in IFN-β-treated MS patients were between those of untreated MS and healthy controls. Interpretation: Long-term treatment with ocrelizumab markedly enriches CD8+ CD28− regulatory T cells and corrects the low levels seen in MS before treatment, while slightly decreasing CD4+ FOXP3+ regulatory T cells. Homeostatic enrichment of regulatory CD8 T cells provides a mechanism, in addition to B cell depletion, for the benefits of anti-CD20 treatment in MS.

Urokinase, CX3CL1, CCL2, TRAIL and IL-18 induced by interferon-β treatment.

To identify serum proteins associated with MS and affected by interferon beta treatment. Plasma samples from 29 untreated relapsing-remitting MS patients and 15 healthy controls were investigated with a multiplexed panel containing 92 proteins related to inflammation. Follow-up samples were available from 13 patients at 1 and 3months after initiation of treatment with interferon beta-1a. Ten proteins were differentially expressed in MS patients. Five of these were altered by treatment with IFN-β 1a: uPA, CX3CL1, CCL2, TRAIL and IL18. CCL2 and TRAIL were confirmed to be modulated with interferon beta treatment in MS. As novel findings, we now report that uPA and CX3CL1 were differentially expressed in MS and increased after IFN-beta-1a treatment. Conflicting results have been reported on how interferon beta affects IL-18.

Natalizumab induced blood eosinophilia: A retrospective pharmacovigilance cohort study and review of the literature

ObjectiveTo describe frequency of natalizumab related eosinophilia and clinical symptoms of eosinophilic disease in our monocentric cohort. MethodsComparison of clinical characteristics of 115 natalizumab treated and 116 untreated RRMS patients and review of literature. Results38% of natalizumab treated patients had eosinophilia, which occurred significantly more frequently compared to untreated MS patients (3%, p-value<0.001). In symptomatic patients, mean eosinophil counts were significantly higher than in asymptomatic patients and symptoms developed within one year. DiscussionEosinophilia is a side effect of natalizumab and mostly asymptomatic. However, few patients develop within one year after start of natalizumab an eosinophilic disease as severe side effect.

MRI activity in MS and completed pregnancy: Data from a tertiary academic center.

ObjectiveTo evaluate postpartum MRI activity in patients with MS and a completed pregnancy and to compare these results to an age-matched untreated nonpregnant MS cohort.MethodsPatient with MS from a tertiary care MS center between 2006 and 2015, with prepartum and postpartum neurologic follow-ups and MRI scans were analyzed. Clinical activity and inflammatory brain MRI activity (new T2-hyperintense or gadolinium-enhancing [Gd+] lesions) were assessed peripartum. The results were compared with untreated reproductive-age patients with MS from the placebo arm of the clinical trials.ResultsA total of 123 pregnancies in 123 women (median Expanded Disability Status Scale 1.0) were analyzed. Approximately 7.2% relapsed during pregnancy and 48.7% relapsed postpartum. Of pregnancies with prepartum and postpartum gadolinium (Gd)-enhanced MRI (n = 112), 8% had Gd+ lesions prepartum and 33% had new Gd+ lesions postpartum. Overall, 54.4% had either new T2 or Gd+ lesions postpartum. Seventy-nine percent of subjects with postpartum relapse had new MRI activity compared with 37.1% without relapse (p < 0.001). Twenty-five percent had both clinical and radiographic activity and only 24.9% maintained no evidence of disease activity status postpartum. There was no association between postpartum MRI activity and disease-modifying treatments (DMTs) (p > 0.5). MRI and clinical outcomes were also assessed for 126 nonpregnant untreated female patients with MS. Comparing pregnancy and no pregnancy groups, there was no difference in MRI activity at follow-up.ConclusionsThere was a high level of inflammatory radiographic disease activity which was related to relapses in postpartum patients with MS. Further studies are needed to determine whether hormonal fluctuations vs extended time off DMTs may be the underlying cause of our observations.

IL-11 induces surface marker expression and migration of T-lymphocytes

Abstract Objective To study the phenotypic and migratory changes of PBMCs following IL-11 stimulation Background Multiple sclerosis (MS) is presumably an autoimmune disease in which immune cells mediate neuroinflammation and neurodegeneration in central nervous system. Cytokines play a significant role in orchestrating the inflammatory response during the disease. Our lab has previously shown that in untreated MS patients, IL-11 is upregulated in the CSF and serum in comparison to matched healthy controls (HCs). Previous study has shown that CD4+IL11+ cells are accumulated in the CSF and the brain MS lesions, suggesting that they play a role in the lesion formation. Methods 14 untreated RRMS patients and age-, sex- and race-matched HCs were enrolled in the study upon signing an Institutional Review Board consent forms. Results By using flow cytometry we characterized IL-11-secreting CD4+ lymphocytes and show that they have increased expression of CCR6, CCR4, ICAM-1, IL-11RA, GM-CSF, and IFNγ in comparison to CD4+IL-11− cells (p values&amp;lt;0.03). In vitro studies further showed that stimulation of PBMCs with recombinant human IL-11, upregulated the expression of CCR4 and CCR6 (p values&amp;lt;0.03, 1.2 and 1.4 fold changes) in CD4+ gated cells. In a trans-well system we observed that IL-11 can act as a chemoattractant for anti-CD3/CD28 mAb stimulated PBMCs and increase their migration (p value=0.04, migration index=1.2) in both patients and HCs. Conclusion We observed that IL-11 induces migratory phenotype in CD4+ lymphocytes in patients with RRMS, which can be explored as a therapeutic target in early RRMS.

A seasonal pattern of clinical activity remains in untreated MS patients in Montpellier and CLIMB cohort. (781)

A seasonal pattern of clinical activity remains in untreated MS patients in Montpellier and CLIMB cohort. (781)

Gut microbiome variation is associated to Multiple Sclerosis phenotypic subtypes.

ObjectiveMultiple sclerosis (MS) is a heterogenous, inflammatory disease of the central nervous system. Microbiota alterations in MS versus healthy controls (HC) are observed, but results are inconsistent. We studied diversity, enterotypes, and specific gut microbial taxa variation between MS and HC, and between MS subgroups.MethodsAmplicon sequencing of the 16S ribosomal RNA V4 region (Illumina MiSeq) was used to evaluate alpha and beta diversity, enterotypes, and relative taxa abundances on stool samples. MS subgroups were based on phenotype, disease course modifiers, and treatment status. Results were controlled for recently identified confounders of microbiota composition.ResultsNinety‐eight MS patients and 120 HC were included. Microbial richness was lower in interferon‐treated (RRMS_I, N = 24) and untreated relapsing–remitting MS during relapse (RRMS_R, N = 4) when compared to benign (BMS, N = 20; Z = −3.07, Pcorr = 0.032 and Z = −2.68, Pcorr = 0.055) and primary progressive MS (PPMS, N = 26; Z = −2.39, Pcorr = 0.062 and Z = −2.26, Pcorr = 0.071). HC (N = 120) and active untreated MS (RRMS_U, N = 24) showed intermediate microbial richness. Enterotypes were associated with clinical subgroups (N = 218, χ 2 = 36.10, P = 0.002), with Bacteroides 2 enterotype being more prevalent in RRMS_I. Butyricicoccus abundance was lower in PPMS than in RRMS_U (Z = −3.00, Pcorr = 0.014) and BMS (Z = −2.56, Pcorr = 0.031), lower in RRMS_I than in BMS (Z = −2.50, Pcorr = 0.034) and RRMS_U (Z = −2.91, Pcorr = 0.013), and inversely correlated with self‐reported physical symptoms (rho = −0.400, Pcorr = 0.001) and disease severity (rho = −0.223, P = 0.027).InterpretationThese results emphasize the importance of phenotypic subcategorization in MS‐microbiome research, possibly explaining previous result heterogeneity, while showing the potential for specific microbiome‐based biomarkers for disease activity and severity.

Treatment choices and neuropsychological symptoms of a large cohort of early MS.

ObjectiveTo assess clinical characteristics, distribution of disease-modifying treatments (DMTs), and neuropsychological symptoms in a large cohort of patients with early-stage MS.MethodsThe German National MS Cohort is a multicenter prospective longitudinal cohort study that has recruited DMT-naive patients with clinically isolated syndrome (CIS) and relapsing-remitting MS (RRMS) since 2010. We evaluated their baseline characteristics and the prevalence of neuropsychological symptoms.ResultsOf 1,124 patients, with a 2.2:1 female-to-male ratio and median age at onset of 31.71 years (interquartile range [IQR]: 26.06–40.33), 44.6% and 55.3% had CIS and RRMS, respectively. The median Expanded Disability Status Scale (EDSS) score at baseline was 1.5 (IQR: 1.0–2.0). A proportion of 67.8% of patients started DMT after a median time of 167.0 days (IQR 90.0–377.5) since the first manifestation. A total of 64.7% and 70.4% of the 762 patients receiving early DMT were classified as CIS and RRMS, respectively. Fatigue, depressive symptoms, and cognitive dysfunction were detected in 36.5%, 33.5%, and 14.7% of patients, respectively.ConclusionBaseline characteristics of this large cohort of patients with early, untreated MS corroborated with other cohorts. Most patients received early DMT within the first year after disease onset, irrespective of a CIS or RRMS diagnosis. Despite the low EDSS score, neuropsychological symptoms affected a relevant proportion of patients.

High serum levels of BMP-2 correlate with BMP-4 and BMP-5 levels and induce reduced neuronal phenotype in patients with relapsing-remitting multiple sclerosis

Blockage of bone morphogenetic protein (BMP) signaling is required for differentiation of neurons and oligodendrocytes from neural stem cells (NSCs). Sera of untreated relapsing-remitting multiple sclerosis (RR-MS) patients expressed significantly higher levels of BMP-2 compared to sera of healthy controls. BMP-2 levels correlated with BMP-4 and -5 levels only in sera of untreated MS patients. Furthermore, sera of untreated patients inhibited the neuronal differentiation of RA-treated P19 cells, which was associated with induction of phospho-SMAD signaling pathway. These results suggest that BMP-2 sera levels may play a role in the failure of remyelination and neuro-regeneration in RR-MS.

Cytotoxic activity and calcium signalling in natural killer cells in multiple sclerosis

Objectives Natural killer (NK) cell populations are affected in MS, with reduced circulating numbers and cytotoxic activity, altered cytokine production, and profile between CD56Dim and CD56Bright NK cell populations. Calcium permeable cation channels, transient receptor potential melastatin3 (TRPM3), are expressed on NK cells. However, the role of receptor/calcium interaction in NK cells has not been investigated in MS patients. Furthermore, the role of calcium mobilisation mediating NK lysis through receptors such as TRPM3, has not been studied in MS. Methods We examined NK cell cytotoxic activity and intracellular calcium mobilisation in untreated and alemtuzumab treated MS patients. We examined TRPM3 surface expression as a mechanism for altered NK function in MS. 22 MS (10 untreated, 12 treated alemtuzumab) and 22 healthy controls (HC) were examined. NK cytotoxic activity and calcium signalling were examined under stimulation with ionomycin, pregenelone sulphate (PregS), 2-Aminoethoxydiphenyl borate (2-ABP) and thapsigargin (TG). Receptor expression (TRPM3, CD69, CD107a) was determined on CD56Dim and CD56Bright NK cells. Results There was no difference in NK cytotoxic activity. CD69 expression was significantly lower on CD56Dimcells of untreated MS group when compared to HC (p-value 0.031) and treated MS group (p=0.036) in unstimulated conditions. This effect was abrogated in treated MS group in unstimulated and stimulated conditions. Calcium signalling was increased in CD56Bright cells of treated MS group compared to untreated MS group under stimulation with PregS, 2-ABP and TG. No difference in calcium signalling was observed in CD56Dimcells. TRPM3 expression was increased in untreated MS group compared to HC with PregS alone. Conclusions Altered NK cell profiling may be implicated in MS. Furthermore, alemtuzumab may mediate therapeutic effect via altered expression of CD56Dim and CD56Bright cells. Alemtuzumab alters calcium signalling in CD56Brightcells, however intracellular calcium signalling alone appears insufficient to explain all changes in NK cell profiling in alemtuzumab treated MS patients.

Untreated relapsing remitting multiple sclerosis patients show antibody production against latent Epstein Barr Virus (EBV) antigens mainly in the periphery and innate immune IL-8 responses preferentially in the CNS

BackgroundMultiple sclerosis (MS) is an inflammatory and neurodegenerative disorder of the central nervous system (CNS). Reliable biomarkers are urgently needed for its diagnosis and management, and as clues to its pathogenesis, in which EBV is implicated. ObjectiveTo measure IgG antibodies against EBV nuclear antigen-1 (EBNA-1) and innate inflammation status in paired serum and cerebrospinal fluid (CSF) samples from untreated relapsing-remitting MS (RRMS) patients. Materials and methodsAnti-EBNA-1 IgG titers and IL-8, IL-1β, IL-6, IL-10, TNF-α and IL-12p70 cytokine levels were measured in 20 untreated RRMS-patients and 17 healthy controls. ResultsWe found higher serum anti-EBNA-1 IgG and IL-8 levels in RRMS-patients than in healthy controls. Interestingly, levels of IL-8 – relative to total protein – were much higher in the CSF, whereas the anti-EBNA-1 antibodies were significantly higher in the sera. More detailed analysis showed that anti-EBNA-1 antibodies relative to total IgG were also higher in the serum in the majority of RRMS patients compared to CSF. Levels of anti-EBNA-1 IgG and IL-8 showed a strong correlation between serum and CSF. ConclusionThese findings in newly diagnosed RRMS-patients imply anti-EBNA-1 antibody production mainly in the periphery and innate immune responses preferentially in the CNS. Both their potential as disease biomarkers and their implications for the pathogenesis of MS warrant further investigation.

The dynamics of mucosal-associated invariant T cells in multiple sclerosis.

BackgroundMultiple sclerosis (MS) is an autoimmune disease characterized by inflammatory demyelination, gliosis and axonal loss in the Central Nervous System. Although the etiology of the disease has remained enigmatic, recent studies have suggested a role of the innate-like T cells, called Mucosal Associated Invariant T cells (MAITs) in the pathophysiology. In the present study, we have analyzed the relative frequency of MAITs and the expression of the cell surface antigens in MAITs to seek a possible link to the disease.ResultsThere was little difference in the frequency of total MAITs between healthy donors (HDs) and untreated MS patients, whereas the latter harbored more CD8lo/neg (DN) MAITs concomitant with a decrease in CD8high MAITs and in CD4 MAITs compared with those in HDs. While the expression of CCR5, CCR6, CD95, CD127, and CD150 has increased in untreated subjects compared with that in HDs, CD45RO has declined in untreated subjects in both DN MAITs and CD8hi MAITs. FTY720 therapy has increased the relative frequency of total MAITs in a time-dependent fashion up to 2 years. Intriguingly, FTY720 therapy for 3 years reversed the above phenotype, engendering more CD8high MAITs accompanied with decreased DN MAITs. FTY720 therapy affected the cytokine production from CD4 T cells and also enhanced the relative frequency of cells producing both TNF-α and IFN-γ from MAITs, CD8 T cells, and CD4 T cells compared with that in untreated subjects.ConclusionsFTY 720 therapy enhanced the relative frequency of MAITs in MS patients in a time-dependent manner. Although the expression of CD8 in MAITs has been affected early by FTY720, longer treatment has reversed the phenotypic change. These data demonstrated that FTY720 induced dynamic change in the relative frequency and in the phenotype of MAITs in MS.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-016-2923-9) contains supplementary material, which is available to authorized users.

AKT isoforms modulate Th1-like Treg generation and function in human autoimmune disease.

Foxp3(+) regulatory T cells (Tregs) exhibit plasticity, which dictates their function. Secretion of the inflammatory cytokine IFNγ, together with the acquisition of a T helper 1 (Th1)-like effector phenotype as observed in cancer, infection, and autoimmune diseases, is associated with loss of Treg suppressor function through an unknown mechanism. Here, we describe the signaling events driving the generation of human Th1-Tregs. Using a genome-wide gene expression approach and pathway analysis, we identify the PI3K/AKT/Foxo1/3 signaling cascade as the major pathway involved in IFNγ secretion by human Tregs. Furthermore, we describe the opposing roles of AKT isoforms in Th1-Treg generation ex vivo Finally, we employ multiple sclerosis as an in vivo model with increased but functionally defective Th1-Tregs. We show that the PI3K/AKT/Foxo1/3 pathway is activated in ex vivo-isolated Tregs from untreated relapsing-remitting MS patients and that blockade of the pathway inhibits IFNγ secretion and restores the immune suppressive function of Tregs. These data define a fundamental pathway regulating the function of human Tregs and suggest a novel treatment paradigm for autoimmune diseases.