Analysis of the IL28RA locus as genetic risk factor for multiple sclerosis.

Recently, we reported an association between a SNP in IL28RA and MS. Here, we performed a fine-mapping of the IL28RA locus by genotyping 10 haplotype-tagging SNPs in a Basque-Spanish population. In addition, based on shared genetic risk loci between autoimmune diseases, a psoriasis-associated SNP located at this locus, rs4649203, was genotyped in four independent populations, comprising a total of 2582 cases and 2614 controls. We did not find any consistent association between IL28RA and MS in these populations, suggesting that, although it may play a role in other autoimmune diseases, this gene is unlikely of general relevance to MS pathogenesis.

Network analysis of transcriptional regulation in response to intramuscular interferon-ß-1a multiple sclerosis treatment.

Interferon-ß (IFN-ß) is one of the major drugs for multiple sclerosis (MS) treatment. The purpose of this study was to characterize the transcriptional effects induced by intramuscular IFN-ß-1a therapy in patients with relapsing-remitting form of MS. By using Affymetrix DNA microarrays, we obtained genome-wide expression profiles of peripheral blood mononuclear cells of 24 MS patients within the first 4 weeks of IFN-ß administration. We identified 121 genes that were significantly up- or downregulated compared with baseline, with stronger changed expression at 1 week after start of therapy. Eleven transcription factor-binding sites (TFBS) are overrepresented in the regulatory regions of these genes, including those of IFN regulatory factors and NF-κB. We then applied TFBS-integrating least angle regression, a novel integrative algorithm for deriving gene regulatory networks from gene expression data and TFBS information, to reconstruct the underlying network of molecular interactions. An NF-κB-centered sub-network of genes was highly expressed in patients with IFN-ß-related side effects. Expression alterations were confirmed by real-time PCR and literature mining was applied to evaluate network inference accuracy.

IL28B polymorphisms are not associated with the response to interferon-ß in multiple sclerosis.

Recent studies have revealed an association between interleukin 28B (IL28B) and response to IFN-alpha treatment in hepatitis C patients. Here we investigated the influence of IL28B polymorphisms in the response to interferon-beta (IFNß) in multiple sclerosis (MS) patients. We genotyped two SNPs of the IL28B gene (rs8099917 and rs12979860) in 588 MS patients classified into responders (n=281) and non-responders (n=307) to IFNß. Combined analysis of the study cohorts showed no significant associations between SNPs rs8099917 and rs12979860 and the response to treatment. These findings do not support a role of IL28B polymorphisms in the response to IFNß in MS patients.

Time course transcriptomics of IFNB1b drug therapy in multiple sclerosis.

Despite its generalized use as drug therapy for multiple sclerosis (MS), the molecular mechanisms of action of interferon beta (IFNB) are still poorly understood. IFNB therapy is long-termed and clinical effects are not immediate, therefore reliable early biomarkers for IFNB activity should maintain a differential expression over time, but longitudinal studies at a transcriptional level have been rare. Microarrays were used to monitor 18 IFNB1b treated MS patients at four time points spanning a period of 1 year. Genes showing in the majority of patients the greatest and most consistent changes in their expression levels were studied. Interferon regulated genes were significantly overrepresented. Fifteen markers were differentially expressed during all three time points and followed a consistent time course pattern: EIF2AK2, IFI6, IFI44, IFI44L, IFIH1, IFIT1, IFIT2, IFIT3, ISG15, MX1, OASL, RSAD2, SN, XAF1 and the marker 238704_at. Except for the last one, these biomarkers were all formerly identified as being indicative for IFNB activity. Expression changes were both early detectable and long lasting and could thus be optimal biomarkers for IFNB activity in long-term studies. Other known biomarkers of IFNB activity were found to be differentially expressed just for certain periods after therapy onset: Interleukin-8 was a short lasting marker and changes in STAT1 were detected with delay.

[Therapy monitoring in patients with multiple sclerosis based on transcriptomic analysis]. / Monitoring der Therapie von Multiple-Sklerose-Patienten basierend auf Transkriptomuntersuchungen.

Multiple sclerosis (MS) is a multifactorial immune mediated disease; its course and response to therapy is influenced by various genes. A goal of the biomedical MS research is the discovery of sets of predisposing genes of which differential expression predict the disease course and outcomes of drug therapy. Such knowledge would have direct consequences for selection, refinement or development of treatments. The genome-wide RNA profile of an individual represents one component of the comprehensive determination of disease- or drug response-related elements; it serves as a large-scale approach. This work reviews the state of the art in MS research applying genomewide screening methods at the transcriptome level in peripheral immune cells of humans.

Monoclonal antibodies in the therapy of multiple sclerosis: an overview.

With the generation of monoclonal antibodies (mAbs), a new therapeutical concept has gained importance. MAbs aim against selective antigens and so have changed our treatment strategies from non-specific to specific. Four therapeuticals have gained importance in the therapy of multiple sclerosis (MS): One has already been approved for therapy (natalizumab), whereas the other three are either in clinical trials or are about to enter phase III studies. Currently, two phase III studies that evaluate the efficacy of alemtuzumab have begun with recruitment (MS CARE I and II). Another mAb (daclizumab) under study is directed to the interleukin-2alpha chain (CD25). Results of clinical trials are promising by reporting reduction of relapses and progression in relapsing remitting and secondary progressive MS accompanied by reduction of new lesions in magnetic resonance imaging. A multicenter randomized controlled trial of daclizumab in MS is going to be initiated. Trials with a humanised antibody directed against the cell surface molecule CD20 are under development. Although the future will emphasise this trend to mAbs, the risks should not be ignored as has been shown in recent news. Still, mAbs have the possibility to revolutionise therapeutical concepts in the treatment of immune-mediated diseases, and will therefore be a useful addition to current therapeutic concepts.

MS therapy research applying genome-wide RNA profiling of peripheral blood.

MS is a multifactorial disease; its course and response to therapy is influenced by various genes. A goal of biomedical MS research is the discovery of sets of predisposing genes where differential expression predicts the disease course and outcome of drug therapy. Such knowledge would have direct consequences for selection, refinement and development of treatments. The genome-wide RNA profile of an individual represents one component of the comprehensive determination of disease and drug response-related elements; it serves as a large-scale approach. This work reviews the state of the art in MS therapy research applying genomewide screening methods at the transcriptome level in peripheral blood cells of humans.

Detection of apoptotic cells in cerebrospinal fluid of patients suffering from neurological disease.

Apoptotic elimination of pathogenic immune cells is considered one of several regulatory mechanisms in inflammatory diseases. To explore the potential relationship between detection of apoptotic cells in the cerebrospinal fluid (CSF) and different types of neurological diseases, we examined cellular apoptosis at the stage of DNA fragmentation, defined by morphological criteria and a molecular biology technique (in situ tailing). During a first phase, 3446 CSF samples derived from admitted patients suffering of inflammatory (IND) and non-inflammatory neurological diseases (NIND) were analysed in the course of routine clinical diagnostics. First, all specimens were inspected for cells displaying atypical morphology following established morphological criteria of intact lymphocytes or apoptosis. In a second phase, 76 additional CSF samples collected from individuals according to investigated clinical groups were analysed in parallel by means of in situ tailing, which indicates the advanced degree of apoptotic demise through labelling of controlled DNA fragmentation. No apoptotic processes were detected by either analytical method in CSF of clinically distinct diseases, amongst others multiple sclerosis (MS). This indicates that the detection of apoptotic cells in CSF during clinical routine diagnostics does not have sufficient explanatory power for the investigated conditions. Furthermore, based on immunohistochemistry, the proportion of CSF lymphocytes expressing the pro-apoptotic receptor Fas (CD95) tended to be higher in NIND patients compared to patients with other IND and MS, but the difference was not statistically significant. In contrast, expression of the anti-apoptotic protein Bcl-2 did not differ between investigated patient groups.

Frequency, heterogeneity and encephalitogenicity of T cells specific for myelin oligodendrocyte glycoprotein in naive outbred primates.

Auto-reactive T cells present in healthy subjects remain in a state of unresponsiveness, but may trigger autoimmunity under various situations. Although myelin oligodendrocyte glycoprotein (MOG) is a potential target antigen in multiple sclerosis (MS), MOG-reactive T cell responses are present in the blood of both healthy subjects and MS-affected individuals. To investigate the disease-inducing potential and regulation of these autoreactive T cells in healthy outbred populations, we have characterized MOG-reactive T cell clones obtained by limiting dilution from peripheral blood of unimmunized C. jacchus marmosets. We report an extraordinarily high prevalence of circulating MOG-reactive T cells in these naive animals (2.6 +/- 1.4 / 10(5) PBMC), and a broadly diverse repertoire of epitope recognition encompassing at least three regions within the extracellular domain of MOG. Adoptive transfer of a MOG21-40-specific T cell clone resulted in mild clinical experimental allergic encephalomyelitis, characterized pathologically by rare foci of inflammation and minimal demyelination. We conclude that MOG-reactive T cells are present in healthy primates at a highly prevalent frequency, and are potentially capable of triggering central nervous system autoimmunity. Expansion of these autoreactive T cells must be tightly controlled to maintain immune homeostasis in healthy individuals.