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Publikationer
2011
Smoking and two human leukocyte antigen genes interact to increase the risk for multiple sclerosis
© 2011 Brain, a journal of neurology.
Osteopontin concentrations are increased in cerebrospinal fluid during attacks of multiple sclerosis
© 2011 Multiple Sclerosis Journal
Axonal Damage in Relapsing Multiple Sclerosis is Markedly Reduced by Natalizumab
© 2011 American Neurological Association
A Swedish national post-marketing surveillance study of natalizumab treatment in multiple sclerosis
© 2011 Multiple Sclerosis Journal
2010
Influence of Perineurial Cells and Toll-Like Receptors 2 and 9 on Herpes simplex Type 1 Entry to the central Nervous System in Rat Encephalitis
© 2010 PLoS ONE | www.plosone.org
TNF Production in Macrophages Is Genetically Determined and Regulates Inflammatory Disease in Rats
© 2010 The Journal of Immunology
IL12A, MPHOSPH9/CDK2AP1 and RGS1 are novel multiple sclerosis susceptibility loci
© 2010 Macmillan Publishers Limited
Cerebrospinal fluid CXCL13 in multiple sclerosis: a suggestive prognostic marker for the disease course
© 2010 Sage Publications
RGMA and IL21R show association with experimental inflammation and multiple sclerosis
© 2010 Macmillan Publishers Limited
Increased cerebrospinal fluid concentrations of the chemokine CXCL13 in active MS
© 2010 Neurology
Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis
© 2010 Macmillan Publishers Limited
2009
Cancer risk among patients with multiple sclerosis and their parents
© 2009 AAN Entrerprises
TLR Activation Induces TNF-α Production from Adult Neural Stem/Progenitor Cells
© 2009 The Journal of Immunology
Tobacco smoking, but not Swedish snuff use, increases the risk of multiple sclerosis
© 2009 Neurology
A resource for the simultaneous high-resolution mapping of multiple quantitative trait loci in rats: The NIH heterogeneous stock
© 2009 Cold Spring Harbor Laboratory Press
The effects of natalizumab on inflammatory mediators in multiple sclerosis: prospects for treatment-sensitive biomarkers
© 2009 European Journal of Neurology
Progressive Multifocal Leukoencephalopathy after Natalizumab Monotherapy
© 2009 The New England Journal of Medicine
Multiple loci comprising immune-related genes regulate experimental neuroinflammation
© 2009 Macmillan Publishers Limited
Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis
© 2009 Macmillan Publishers Limited
2008
Induction of systemic TNFa in Natalizumab-treated multiple sclerosis
© 2008 The Author(s)
Journal compilation © 2008 EFNS European Journal of Neurology
Progress and prospects in rat genetics: a community view
© 2008 Nature Publishing Group
Congenic Rats with Allelic Differences in the Class II Transactivator Gene Display Altered Susceptibility to Experimental Autoimmune Encephalomyelitis
© 2008 The Journal of Immunology
Analysis of cerebrospinal fluid and cerebrospinal fluid cells from patients with multiple sclerosis for detection of JC virus DNA
© 2008 Multiple Sclerosis
Increased IL-10 mRNA and IL-23 mRNA expression in multiple sclerosis: interferon-² treatment increases IL-10 mRNA expression while reducing IL-23 mRNA expression
© 2008 Multiple Sclerosis
Maternal smoking during pregnancy and multiple sclerosis amongst offspring
© 2008 European Journal of Neurology
The genetics of multiple sclerosis and its experimental models
© 2008 Department of Clinical Neuroscience, Karolinska Institute, Stockholm, Sweden
Host strain-dependent difference in susceptibility in a rat model of herpes simplex type 1 encephalitis and gene–environment studies.
© 2008 Journal of NeuroVirology
2007
Variation in interleukin 7 receptor a chain (IL7R)
influences risk of multiple sclerosis
© 2007 Nature Publishing Group http://www.nature.com/naturegenetics
Genetic analysis of neuropathic pain-like behaviorfollowing peripheral nerve injury suggests a role of the major histocompatibility complex in development of allodynia
© 2007 Published by Elsevier B.V. on behalf of International Association for the Study of Pain.
2006
Advanced Intercross Line Mapping of Eae5 Reveals Ncf-1 and
CLDN4 as Candidate Genes for Experimental Autoimmune
Encephalomyelitis
© 2006 by The American Association of Immunologists, Inc
Combined-cross analysis of genome-wide linkage scans for experimental
autoimmune encephalomyelitis in rat
© 2006 Elsevier Inc. All rights reserved.
Autoimmunity.
Basic and clinical science approach complex diseases — new
molecules and mechanisms
Current Opinion in Immunology 2006, 18:1–4 0952-7915
© 2006 Elsevier Ltd. All rights reserved.
Genetics of autoimmune neuroinflammation
Current Opinion in Immunology 2006, 18:1–7
© 2006 Elsevier Ltd. All rights reserved.
Cortical Demyelination Can Be Modeled in Specific Rat
Models of Autoimmune Encephalomyelitis and Is Major
Histocompatability Complex (MHC) Haplotype-Related
© 2006 by the American Association of Neuropathologists, Inc.
Genetically determined susceptibility to neurodegeneration is
associated with expression of inflammatory genes
© 2006 Elsevier Inc. All rights reserved.
Definition of a 1.06-Mb Region Linked to Neuroinflammation in Humans,
Rats and Mice
© 2006 by the Genetics Society of America DOI: 10.1534/genetics.106.057406
2005
MHC2TA is associated with differential MHC molecule
expression and susceptibility to rheumatoid arthritis,
multiple sclerosis and myocardial infarction
© 2005 Nature Publishing Group http://www.nature.com/naturegenetics
© 2005 Nature Publishing Group All rights reserved
Eae19, a New Locus on Rat Chromosome 15 Regulating Experimental
Autoimmune Encephalomyelitis
© 2005 by the Genetics Society of America
2004
Current Gene-Mapping Strategies in Experimental
Models of Multiple Sclerosis
© 2004 Blackwell Publishing Ltd. Scandinavian Journal of Immunology
T Cell Ig- and Mucin-Domain-Containing Molecule-3 (TIM-3)
and TIM-1 Molecules Are Differentially Expressed on Human
Th1 and Th2 Cells and in Cerebrospinal Fluid-Derived
Mononuclear Cells in Multiple Sclerosis
© 2004 The Journal of Immunology
2003
Discrete Gene Loci Regulate Neurodegeneration,
Lymphocyte Infiltration, and Major Histocompatibility
Complex Class II Expression in the CNS
© 2004 The Journal of Neuroscience
Abstracts/artiklar
2011
Smoking and two human leukocyte antigen genes interact to increase the risk for multiple sclerosis
Anna Karin Hedström, Emilie Sundqvist, Maria Bäärnhielm, Nina Nordin, Jan Hillert, Ingrid Kockum, Tomas Olsson and Lars Alfredsson.
Both genetic and environmental factors display low or modest associations with multiple sclerosis. Hypothetically, gene–environment interactions may exert much stronger effects. In this study, we investigated potential interactions between genetic risk factors and smoking in relation to risk of developing multiple sclerosis. A population-based case–control study involving incident cases of multiple sclerosis (843 cases, 1209 controls) was performed in Sweden. Cases and controls were classified according to their smoking status and human leukocyte antigen DRB1 as well as human leukocyte antigen A genotypes. Subjects with different genotypes and smoking habits were compared with regard to incidence of multiple sclerosis, by calculating odds ratios with 95% confidence intervals employing logistic regression. The potential interaction between different genotypes, as well as between genotype and smoking, was evaluated by calculating attributable proportion due to interaction. A significant interaction between two genetic risk factors, carriage of human leukocyte antigen DRB1*15 and absence of human leukocyte antigen A*02, was observed among smokers whereas such an interaction was absent among non-smokers. There were considerable differences in odds ratios between the various groups. Compared with non-smokers with neither of the genetic risk factors, the odds ratio was 13.5 (8.1–22.6) for smokers with both genetic risk factors. The odds ratio for smokers without genetic risk was 1.4 (0.9–2.1) and the odds ratio for non-smokers with both genetic risk factors was 4.9 (3.6–6.6). Among those with both genetic risk factors, smoking increased the risk by a factor of 2.8 in comparison with a factor of 1.4 among those without the genetic risk factors. The risk of developing multiple sclerosis associated with human leukocyte antigen genotypes may be strongly influenced by smoking status. The findings are consistent with our hypothesis that priming of the immune response in the lungs may subsequently lead to multiple sclerosis in genetically susceptible people.
© 2011 Brain, a journal of neurology.
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Osteopontin concentrations are increased in cerebrospinal fluid during attacks of multiple sclerosis
Lars Börnsen, Mohsen Khademi, Tomas Olsson, Per Soelberg Sørensen and Finn Sellebjerg
Background: The cytokine osteopontin (OPN) is a potential key player in the immunopathogenesis of multiple sclerosis
(MS) and a candidate biomarker for disease activity.
Objective: The objective of this study was to examine concentrations of OPN in the cerebrospinal fluid (CSF) across
the clinical spectrum of MS.
Methods: Our research consisted of a cross-sectional study of patients from two randomized, placebo-controlled trials.
Concentrations of OPN and other blood and CSF markers were determined using an enzyme-linked immunosorbent
assay (ELISA). Samples were obtained from untreated patients with exacerbation of clinically isolated syndrome (CIS)
(n¼25) and relapsing–remitting MS (RRMS) (n¼41) of whom 48 participated in clinical trials, randomly allocated to
treatment with placebo or methylprednisolone (MP) and undergoing repeated sampling after 3 weeks. Furthermore, we
obtained CSF and blood samples from patients with primary progressive MS (PPMS, n¼9), secondary progressive MS
(SPMS, n¼28) and other neurological disorders (OND, n¼44), and blood samples from 24 healthy subjects.
Results: OPN concentrations were significantly increased in the CSF of patients with CIS (p¼0.02) and RRMS
(p<0.001) in exacerbation compared to patients with OND, and increased levels of OPN were associated with high
values of other biomarkers of inflammation. At 3-week follow-up CSF OPN concentrations had decreased significantly
from baseline regardless treatment with placebo or MP. Patients with PPMS had increased OPN levels in the CSF
(p¼0.004) and high CSF levels of OPN were associated with high degrees of disability.
Conclusions: OPN concentration in the CSF is a dynamic indicator of disease activity in RRMS, presumably reflecting
ongoing inflammation. Increased CSF OPN concentrations in PPMS may indicate ongoing inflammation even in these
patients.
© 2011 Multiple sclerosis journal
Axonal Damage in Relapsing Multiple Sclerosis is Markedly Reduced by Natalizumab
Martin Gunnarsson, Clas Malmeström,Markus Axelsson, Peter Sundström, Charlotte Dahle, Magnus Vrethem, Tomas Olsson, Fredrik Piehl, Niklas Norgren, Lars Rosengren, Anders Svenningsson, and Jan Lycke
Objective: The impact of present disease-modifying treatments (DMTs) in multiple sclerosis (MS) on nerve injury and
reactive astrogliosis is still unclear. Therefore, we studied the effect of natalizumab treatment on the release of 2
brain-specific tissue damage markers into cerebrospinal fluid (CSF) in MS patients.
Methods: CSF samples from 92 patients with relapsing forms of MS were collected in a prospective manner prior to
natalizumab treatment and after 6 or 12 months. In 86 cases, natalizumab was used as second-line DMT due to
breakthrough of disease activity. The levels of neurofilament light (NFL) and glial fibrillary acidic protein (GFAP) were
determined using highly sensitive in-house developed enzyme-linked immunosorbent assays.
Results: Natalizumab treatment led to a 3-fold reduction of NFL levels, from a mean value of 1,300 (standard
deviation [SD], 2,200) to 400 (SD, 270) ng/l (p < 0.001). The later value was not significantly different from that
found in healthy control subjects (350ng/l; SD, 170; n ¼ 28). Subgroup analysis revealed a consistent effect on NFL
release, regardless of previous DMT or whether patients had relapses or were in remission within 3 months prior to
natalizumab treatment. No differences between pre- and post-treatment levels of GFAP were detected.
Interpretation: Our data demonstrate that natalizumab treatment reduces the accumulation of nerve injury in
relapsing forms of MS. It is anticipated that highly effective anti-inflammatory treatment can reduce axonal loss,
thereby preventing development of permanent neurological disability.
© 2011 American Neurological Association
A Swedish national post-marketing surveillance study of natalizumab treatment in multiple sclerosis
Carolina Holmén, Fredrik Piehl, Jan Hillert, Anna Fogdell-Hahn, Malin Lundkvist, Elin Karlberg, Petra Nilsson, Charlotte Dahle, Nils Feltelius, Anders Svenningsson, Jan Lycke and Tomas Olsson
Background: A post marketing surveillance study was conducted to evaluate safety and efficacy of natalizumab in
Swedish multiple sclerosis (MS) patients since its introduction in August 2006 until March 2010.
Methods: Patients were registered in the web-based Swedish MS-registry at 40 locations and evaluated every 6 months.
Adverse events and clinical outcomes were recorded.
Results: One thousand one hundred and fifty-two patients were included (71.4% female) and 149 patients stopped
treatment; the main reason was planned pregnancy. Anti-natalizumab antibodies were found in 4.5% (52 patients) of
which 1.6% displayed persistent antibodies. Serious adverse events were rare, but included three cases with progressive
multifocal leukoencephalopathy (PML). There were seven fatal cases, probably unrelated to the natalizumab treatment.
For relapsing–remitting MS patients (n¼901), mean Expanded Disability Status Scale (EDSS, 10.7%), Multiple Sclerosis
Severity Scale (MSSS, 20.4%), Multiple Sclerosis Impact Scale (MSIS-29, physical 9.9%, psychological 13.3%) and
Symbol Digit Modalities Test (SDMT, þ10.7%) all showed significant improvements during 24 months of treatment with
natalizumab. The Swedish web-based MS quality registry proved to function as a platform for post-marketing MS drug
surveillance, providing long-term data regarding drug effects and adverse events beyond clinical trials.
Conclusions: Our results indicate that natalizumab is generally well tolerated and has sustained efficacy for patients
with active MS, though the risk of PML is still an important concern.
© 2011 Multiple sclerosis journal
2010
Influence of Perineurial Cells and Toll-Like Receptors 2 and 9 on Herpes simplex Type 1 Entry to the central Nervous System in Rat Encephalitis
Biborka Bereczky-Veress, Nada Abdelmagid, Fredrik Piehl, Tomas Bergström, Tomas Olsson, Birgit Sköldenberg, Margarita Diez
Herpes simplex encephalitis (HSE) is a rare disease with high mortality and significant morbidity among survivors. We have previously shown that susceptibility to HSE was host-strain dependent, as severe, lethal HSE developed after injection of human Herpes simplex type 1 virus (HSV-1) into the whiskers area of DA rats, whereas PVG rats remained completely asymptomatic. In the present study we investigated the early immunokinetics in these strains to address the underlying molecular mechanisms for the observed difference. The virus distribution and the immunological responses were compared in the whiskers area, trigeminal ganglia and brain stem after 12 hours and the first four days following infection using immunohistochemistry and qRT-PCR. A conspicuous immunopathological finding was a strain-dependent difference in the spread of the HSV-1 virus to the trigeminal ganglia, only seen in DA rats already from 12 hpi. In the whiskers area infected perineurial cells were abundant in the susceptible DA strain after 2 dpi, whereas in the resistant PVG rats HSV-1 spread was confined only to the epineurium. In both strains activation of Iba1+/ED1+ phagocytic cells followed the distribution pattern of HSV-1 staining, which was visible already at 12 hours after infection. Notably, in PVG rats higher mRNA expression of Tolllike receptors (Tlr) -2 and -9, together with increased staining for Iba1/ED1 was detected in the whiskers area. In contrast, all other Tlr-pathway markers were expressed at higher levels in the susceptible DA rats. Our data demonstrate the novel observation that genetically encoded properties of the host nerve and perineurial cells, recruitment of phagocyting cells together with the low expression of Tlr2 and -9 in the periphery define the susceptibility to HSV-1 entry into the nervous system.
© 2010 PLoS ONE | www.plosone.org
TNF Production in Macrophages Is Genetically Determined and Regulates Inflammatory Disease in Rats
Alan Gillett, Monica Marta, Tao Jin, Jonatan Tuncel, Patrick Leclerc, Rita Nohra, Stefan Lange, Rikard Holmdahl, Tomas Olsson, Robert A. Harris and Maja Jagodic
Dysregulation of TNF is an important pathophysiological phenotype for many diseases. Recently, certain genetically regulated loci have been identified to regulate several inflammatory diseases. We hypothesized that a region on rat chromosome 4 known to regulate experimental autoimmune encephalomyelitis, experimental arthritis and experimental autoimmune neuritis harbors a gene regulating central inflammatory molecules, such as TNF. We therefore mapped TNF production using linkage analysis in the 12th generation of an advanced intercross line between DA and PVG.AV1 rats, which differ in susceptibility to several inflammatory conditions. A single TNF-regulating quantitative trait locus with a logarithm of odds score of 6.2 was identified and its biological effect was confirmed in a congenic rat strain. The profound TNF regulation mapped in congenic strains to the macrophage population. Several TLR signaling cascades led to the same reduced proinflammatory phenotype in congenic macrophages, indicating control of a convergence point for innate inflammatory activity. The decreased TNF potential and reduced proinflammatory macrophage phenotype in congenic rats was also associated with reduced clinical severity in experimental autoimmune encephalomyelitis, pristane-induced arthritis and sepsis experimental models. Determination of genes and mechanisms involved in this genetically determined TNF regulation will be valuable in understanding disease pathogenesis and aid treatment development.
© 2010 The Journal of Immunology
IL12A, MPHOSPH9/CDK2AP1 and RGS1 are novel multiple sclerosis susceptibility loci
The International Multiple Sclerosis Genetics Consortium (IMSGC)
A recent meta-analysis identified seven single-nucleotide polymorphisms (SNPs) with suggestive evidence of association with multiple sclerosis (MS). We report an analysis of these polymorphisms in a replication study that includes 8,085 cases and 7,777 controls. A meta-analysis across the replication collections and a joint analysis with the discovery data set were performed. The possible functional consequences of the validated susceptibility loci were explored using RNA expression data. For all of the tested SNPs, the effect observed in the replication phase involved the same allele and the same direction of effect observed in the discovery phase. Three loci exceeded genome-wide significance in the joint analysis: RGS1 (P value¼3.55109), IL12A (P¼3.08108) and MPHOSPH9/CDK2AP1 (P¼3.96108). The RGS1 risk allele is shared with celiac disease (CD), and the IL12A risk allele seems to be protective for celiac disease. Within the MPHOSPH9/CDK2AP1 locus, the risk allele correlates with diminished RNA expression of the cell cycle regulator CDK2AP1; this effect is seen in both lymphoblastic cell lines (P¼1.18105) and in peripheral blood mononuclear cells from subjects with MS (P¼0.01). Thus, we report three new MS susceptibility loci, including a novel inflammatory disease locus that could affect autoreactive cell proliferation. Genes and Immunity advance online publication, 17 June 2010; doi:10.1038/gene.2010.28
© 2010 Macmillan Publishers Limited
Cerebrospinal fluid CXCL13 in multiple sclerosis: a suggestive prognostic marker for the disease course
Mohsen Khademi, Ingrid Kockum, Magnus L Andersson, Ellen Iacobaeus, Lou Brundin, Finn Sellebjerg, Jan Hillert, Fredrik Piehl and Tomas Olsson
Background: Levels of CXCL13, a potent B-cell chemoattractant, are elevated in the cerebrospinal fluid (CSF) during
multiple sclerosis (MS) and are associated with markers of MS activity. Levels decrease upon effective treatments.
Objective: Here we validate the potential role of CSF CXCL13 as a biomarker for aspects of MS in a large amount of
clinical material, the majority collected at early diagnostic work-up.
Methods: CXCL13 was measured by ELISA in 837 subjects: relapsing–remitting MS (RRMS; n¼323), secondary progressive
MS (SPMS; n¼40), primary progressive MS (PPMS; n¼24), clinically isolated syndrome (CIS; n¼79), other
neurological diseases (ONDs; n¼181), ONDs with signs of inflammation or viral/bacterial infections (iONDs; n¼176)
and healthy controls (n¼14).
Results: Subjects with viral/bacterial infections had extremely high CXCL13 levels compared to all included groups
(p<0.0001). CXCL13 was otherwise significantly higher in MS compared to the remaining controls (p<0.0001), and CIS
(p<0.01). A significant and positive correlation between CXCL13 and relapse rate, the results obtained for the
Expanded Disability Status Scale (EDSS) and the number of lesions detected by MRI was demonstrated. CXCL13 was
increased in CIS conversion to clinically definite MS (p<0.001). Oligoclonal immunoglobulin band (OCB)-positive CIS or
MS had significantly increased CXCL13 levels compared to OCB-negative CIS or MS (p<0.001 and p<0.0001,
respectively).
Conclusion: CXCL13 was associated with disease exacerbations and unfavourable prognosis in RRMS. Increased
CXCL13 was not specific for MS since subjects with viral/bacterial infections exhibited even higher levels. High levels
predicted CIS conversion to MS. We suggest that measurement of CSF CXCL13 can be part of the armamentarium in
the diagnostic and prognostic work-up in MS and be of help in future treatment decisions.
© 2010 Sage Publications
RGMA and IL21R show association with experimental inflammation and multiple sclerosis
R Nohra, AD Beyeen, JP Guo, M Khademi, E Sundqvist, MT Hedreul, F Sellebjerg, C Smestad,
AB Oturai, HF Harbo, E Wallström, J Hillert, L Alfredsson, I Kockum, M Jagodic, J Lorentzen and T Olsson
Rat chromosome 1 harbors overlapping quantitative trait loci (QTL) for cytokine production and experimental models of inflammatory diseases. We fine-dissected this region that regulated cytokine production, myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), anti-MOG antibodies and pristane-induced arthritis (PIA) in advanced intercross lines (AILs). Analysis in the tenth and twelfth generation of AILs resolved the region in two narrow QTL, Eae30 and Eae31. Eae30 showed linkage to MOG-EAE, anti-MOG antibodies and levels of interleukin-6 (IL-6). Eae31 showed linkage to EAE, PIA, anti-MOG antibodies and levels of tumor necrosis factor (TNF) and IL-6. Confidence intervals defined a limited set of potential candidate genes, with the most interesting being RGMA, IL21R and IL4R. We tested the association with multiple sclerosis (MS) in a Nordic case–control material. A single nucleotide polymorphism in RGMA associated with MS in males (odds ratio (OR)¼1.33). Polymorphisms of RGMA also correlated with changes in the expression of interferon-g (IFN-g) and TNF in cerebrospinal fluid of MS patients. In IL21R, there was one positively associated (OR¼1.14) and two protective (OR¼0.87 and 0.68) haplotypes. One of the protective haplotypes correlated to lower IFN-g expression in peripheral blood mononuclear cells of MS patients. We conclude that RGMA and IL21R and their pathways are crucial in MS pathogenesis and warrant further studies as potential biomarkers and therapeutic targets. Genes and Immunity advance online publication, 14 January 2010; doi:10.1038/gene.2009.111
© 2010 Macmillan Publishers Limited
Increased cerebrospinal fluid concentrations of the chemokine CXCL13 in active MS
F. Sellebjerg, L. Börnsen, M. Khademi, M. Krakauer, T. Olsson, J.L. Frederiksen, P.S. Sørensen
Background: Accumulating evidence supports a major role of B cells in multiple sclerosis (MS)
pathogenesis. How B cells are recruited to the CNS is incompletely understood. Our objective was
to study B-cell chemokine concentrations in MS, their relationship with disease activity, and how
treatment with methylprednisolone and natalizumab affected the concentration in CSF.
Methods: Using a cross-sectional design, CSF and blood samples were obtained from cohorts of
patients with clinically isolated syndromes (CIS), relapsing-remitting MS (RRMS), primary progressive
MS (PPMS), or secondary progressive MS (SPMS), and noninflammatory neurologic disease
control subjects. Some patients with RRMS were studied before and after treatment with
methylprednisolone or natalizumab.
Results: In CSF, concentrations of CXCL13, but not CXCL12, were higher in patients with CIS,
RRMS, SPMS, and PPMS than in controls. CSF concentrations of CXCL13 correlated with the
CSF B-cell count, with markers of immune activation, and with disease activity in patients with
CIS and RRMS. CSF concentrations of CXCL13 decreased after treatment with high-dose methylprednisolone
and natalizumab. High CSF concentrations of CXCL13 correlated with low expression
of messenger RNA encoding the immunoregulatory cytokines interleukin 10 and
transforming growth factor 1, but not with the expression of T-helper type 1 (Th1) and Th17
factors.
Conclusion: The chemokine CXCL13 may play a major role in recruitment of B cells and T-cell
subsets expressing the chemokine receptor CXCR5 to the CNS in multiple sclerosis (MS), and
may be a useful biomarker for treatment effects in MS. Furthermore, CXCL13 or its receptor
CXCR5 should be considered as therapeutic targets in MS.
© 2010 Neurology
Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis
J Öckinger, P Stridh, AD Beyeen, F Lundmark, M Seddighzadeh, A Oturai, PS Sørensen, ÅR Lorentzen, EG Celius, V Leppä, K Koivisto, PJ Tienari, L Alfredsson, L Padyukov, J Hillert, I Kockum, M Jagodic and T Olsson
Multiple sclerosis (MS) is a complex disorder of the central nervous system, causing inflammation, demyelination and axonal damage. A limited number of genetic risk factors for MS have been identified, but the etiology of the disease remains largely unknown. For the identification of genes regulating neuroinflammation we used a rat model of MS, myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), and carried out a linkage analysis in an advanced intercross line (AIL). We thereby redefine the Eae18b locus to a 0.88Mb region, including a cluster of chemokine genes. Further, we show differential expression of Ccl2, Ccl11 and Ccl11 during EAE in rat strains with opposite susceptibility to EAE, regulated by genotype in Eae18b. The human homologous genes were tested for association to MS in 3841 cases and 4046 controls from four Nordic countries. A haplotype in CCL2 and rs3136682 in CCL1 show a protective association to MS, whereas a haplotype in CCL13 is disease predisposing. In the HLA-DRB1*15 positive subgroup, we also identified an association to a risk haplotype in CCL2, suggesting an influence from the human leukocyte antigen (HLA) locus. We further identified association to rheumatoid arthritis in CCL2, CCL8 and CCL13, indicating common regulatory mechanisms for
complex diseases.
© 2010 Macmillan Publishers Limited
2009
Cancer risk among patients with multiple sclerosis and their parents
S. Bahmanyar, S.M. Montgomery,J. Hillert, A. Ekbom, T. Olsson
Background: We investigated cancer risk among patients with multiple sclerosis (MS) and whethervariation by age at MS diagnosis helps to elucidate mechanisms underlying the previously reportedreduced cancer risk. We also studied cancer risk among parents to ascertain if MS susceptibility genes may confer protection against cancer in relatives.
Methods: Cox proportional hazards regression, adjusted for age, sex, area, and socioeconomic index, estimated cancer risk among 20,276 patients with MS and 203,951 individuals without MS, using Swedish general population register data. Similar analyses were conducted among 11,284 fathers and 12,006 mothers of patients with MS, compared with 123,158 fathers and 129,409 mothers of controls.
Results: With an average of 35 years of follow-up, there was a decreased overall cancer risk among patients with MS (hazard ratio 0.91, 0.87–0.95). Increased risks were observed for brain tumors (1.44, 1.21–1.72) and urinary organ cancer (1.27, 1.05–1.53). Parents of patients with MS did not have a notably increased or decreased overall cancer risk.
Conclusions: The reduction in cancer risk in patients with multiple sclerosis (MS) may result from behavioral change, treatment, or we speculate that some immunologic characteristics of MS disease activity improve antitumor surveillance. The lack of association among parents indicates that a simple inherited characteristic is unlikely to explain the reduced cancer risk among patients with MS. MS is associated with increased risk for some cancers, such as of urinary organs and brain tumors (although surveillance bias may be responsible).
© 2009 AAN Entrerprises
TLR Activation Induces TNF-α Production from Adult Neural Stem/Progenitor Cells
Ruxandra Covacu, Lisa Arvidsson, Åsa Andersson, Mohsen Khademi, Helena Erlandsson-Harris, Robert A. Harris, Mikael A. Svensson, Tomas Olsson, and Lou Brundin
Adult neural stem cells (NSCs) are believed to facilitate CNS repair and tissue regeneration. However, it is not yet clear how these cells are influenced when the cellular environment is modified during neurotrauma or neuroinflammatory conditions. In this study, we determine how different proinflammatory cytokines modulate the expression of TLR2 and TLR4 in NSCs and how these cells respond to TLR2 and TLR4 agonists. Primary cultures of neural stem/progenitor cells isolated from the subventricular zone of brains from adult Dark Agouti rats were exposed to 1) supernatants from activated macrophages; 2) proinflammatory cytokines IFN-γ, TNF-α, or both; and 3) agonists for TLR2 and TLR4. Both TLR2 and TLR4 were expressed during basal conditions and their mRNA levels were further increased following cytokine exposure. TLR4 was up-regulated by IFN-γ and this effect was reversed by TNF-α. TLR2 expression was increased by supernatants from activated macrophages and by TNF-α, which synergized with IFN-γ TLR agonists induced the expression of TNF-α mRNA. Importantly, TNF-α could be translated into protein and released into the supernatants where it was quantified by cytokine ELISA. In conclusion, we demonstrate that NSCs constitutively express TLR2 and TLR4 and that their expression is increased as a consequence of exposure to proinflammatory mediators. Additionally, activation of these receptors can induce production of proinflammatory cytokines. These findings suggest that NSCs may be primed to participate in cytokine production during neuroinflammatory or traumatic conditions.
© 2009 The Journal of Immunology
Tobacco smoking, but not Swedish snuff use, increases the risk of multiple sclerosis
Anna K. Hedström, Maria Bäärnhielm, Tomas Olsson and Lars Alfredsson
Objective: The aim of this study was to estimate the influence of tobacco smoking and Swedish snuff use on the risk of developing multiple clerosis (MS).
Methods: A population-based case-control study was performed in Sweden, using incident cases of MS (902 cases and 1,855 controls). A case was defined as a subject from the study base who had received a diagnosis of MS, and controls were randomly selected from the study base. The
incidence of MS among smokers was compared with that of never-smokers. We also investigated whether the use of Swedish snuff had an impact on the risk of developing MS.
Results: Smokers of both sexes had an increased risk of developing MS (odds ratio [OR] 1.4, 95% confidence interval [CI] 1.2–1.7 for women, and OR 1.8, 95% CI 1.3–2.5 for men). The increased risk was apparent even among subjects who had previously smoked moderately (5 pack-years)
prior to index, and the risk increased with increasing cumulative dose (p0.0001). The increased risk for MS associated with smoking remained up to 5 years after stopping smoking. In contrast, taking Swedish snuff for more than 15 years decreased the risk of developing MS (OR 0.3, 95%
CI 0.1–0.8).
Conclusions: Smokers of both sexes run an increased risk of developing multiple sclerosis (MS), and the risk increases with cumulative dose of smoking. However, the use of Swedish snuff is not
associated with elevated risk for MS, which may indicate that nicotine is not the substance responsible for the increased risk of developing MS among smokers.
© 2009 Neurology
A resource for the simultaneous high-resolution mapping of multiple quantitative trait loci in rats: The NIH heterogeneous stock
Martina Johannesson, Regina Lopez-Aumatell, Pernilla Stridh, Margarita Diez,Jonatan Tuncel, Gloria Blázquez, Esther Martinez-Membrives, Toni Cañete,Elia Vicens-Costa, Delyth Graham, Richard R. Copley, Polinka Hernandez-Pliego, Amennai D. Beyeen, Johan Öckinger, Cristina Fernández-Santamaría, Percio S. Gulko, Max Brenner, Adolf Tobeña, Marc Guitart-Masip, Lydia Giménez-Llort, Anna Dominiczak, Rikard Holmdahl, Dominique Gauguier, Tomas Olsson, Richard Mott, William Valdar, Eva E. Redei, Alberto Fernández-Teruel, and Jonathan Flint
The laboratory rat (Rattus norvegicus) is a key tool for the study of medicine and pharmacology for human health. A
large database of phenotypes for integrated fields such as cardiovascular, neuroscience, and exercise physiology exists
in the literature. However, the molecular characterization of the genetic loci that give rise to variation in these traits
has proven to be difficult. Here we show how one obstacle to progress, the fine-mapping of quantitative trait loci
(QTL), can be overcome by using an outbred population of rats. By use of a genetically heterogeneous stock of rats,
we map a locus contributing to variation in a fear-related measure (two-way active avoidance in the shuttle box) to a
region on chromosome 5 containing nine genes. By establishing a protocol measuring multiple phenotypes including
immunology, neuroinflammation, and hematology, as well as cardiovascular, metabolic, and behavioral traits, we
establish the rat HS as a new resource for the fine-mapping of QTLs contributing to variation in complex traits of
biomedical relevance.
© 2009 Cold Spring Harbor Laboratory Press
The effects of natalizumab on inflammatory mediators in multiple sclerosis: prospects for treatment-sensitive biomarkers
M. Khademia, L. Bornsenb, F. Rafatniaa, M. Anderssonc, L. Brundinc, F. Piehla,c, F. Sellebjergb and T. Olsson
Background: Natalizumab affects systemic cytokine expressions and clinical course in
relapsing–remitting multiple sclerosis (RRMS). We analyzed levels of inflammatory
cytokines in cerebrospinal fluid (CSF) cells and peripheral blood mononuclear cells
(PBMCs), levels of matrix metalloproteinase (MMP)-9 and osteopontin (OPN) in
CSF, and clinical outcome measures in 22 natalizumab-treated RRMS patients.
Methods: mRNA levels of cytokines in cells were detected with real-time RT-PCR.
Protein levels of OPN and MMP-9 were measured by ELISA.
Results: Natalizumab reduced CSF cell counts (P < 0.0001). Tumor necrosis factor
(TNF) and interferon-c (IFN-c) mRNAs were significantly increased in PBMCs. In
contrast, expressions of IFN-c and interleukin (IL)-23 were decreased but IL-10 increased
in the CSF cells. OPN and MMP-9 were reduced in the CSF. Patients being in
remission at baseline showed the same deviations of mediators as those in relapse after
natalizumab treatment. The open label clinical outcome measures were either stable or
improved during therapy.
Conclusions: Natalizumab attenuates pro-inflammatory mediators intrathecally and
the reduced pro-inflammatory milieu may allow increased production of the antiinflammatory
mediator IL-10. The increased systemic cytokines may impede the
improvement of certain clinical measures like fatigue. The affected mediators seem to
be sensitive to an immune-modifying treatment which could be used as biomarkers for
this therapy.
© 2009 European Journal of Neurology
Progressive Multifocal Leukoencephalopathy after Natalizumab Monotherapy
Hans Lindå, Anders von Heijne, Eugene O. Major, Caroline Ryschkewitsch, Johan Berg, Tomas Olsson, and Claes Martin
We describe progressive multifocal leukoencephalopathy (PML) caused by infectionwith human polyomavirus JC virus in a patient with multiple sclerosis who wastreated with natalizumab. The first PML symptoms appeared after 14 monthly infusions of the drug. Magnetic resonance imaging (MRI) showed a presumed multiplesclerosis lesion, and JC virus DNA was not detected on polymerase-chain-reaction(PCR) assay of cerebrospinal fluid. The patient’s symptoms worsened, and the diagnosisof PML was established with a more sensitive quantitative PCR assay after 16 infusions of natalizumab. Plasma exchange was used to accelerate clearance ofnatalizumab. Approximately 3 weeks after plasma exchange, an immune-reconstitution inflammatory syndrome appeared. JC virus DNA was no longer detectable on quantitative PCR assay, and the patient’s symptoms improved.
© 2009 The New England Journal of Medicine
Multiple loci comprising immune-related genes regulate experimental neuroinflammation
M Marta1, P Stridh1, K Becanovic, A Gillett1, J Öckinger, JC Lorentzen, M Jagodic and T Olsson
A 58Mb region on rat chromosome 4 known to regulate experimental autoimmune encephalomyelitis (EAE) was geneticallydissected. High-resolution linkage analysis in an advanced intercross line (AIL) revealed four quantitative trait loci (QTLs), Eae24–Eae27. Both Eae24 and Eae25 regulated susceptibility and severity phenotypes, whereas Eae26 regulated severity and Eae27 regulated susceptibility. Analyses of the humoral immune response revealed that the levels of serum anti-myelin oligodendrocyte glycoprotein (MOG) immunoglobin G1 (IgG1) antibodies are linked to Eae24 and anti-MOG IgG2b antibodies are linked to both Eae24 and Eae26. We tested the parental DA strain and six recombinant congenic strains that include overlapping fragments of this region in MOG-EAE. Eae24 and Eae25 showed significant protection during the acute phase of EAE, whereas Eae25 and Eae26 significantly modified severity but not susceptibility. The smallest congenic fragment, which carries Eae25 alone, influenced both susceptibility and severity, and protected from the chronic phase of disease. These results support the multiple QTLs identified in the AIL. By demonstrating several QTLs comprising immune-related genes, which potentially interact, we provide a significant step toward elucidation of the polygenically regulated pathogenesis of MOGEAE and possibly multiple sclerosis (MS), and opportunities for comparative genetics and testing in MS case–control cohorts.
© 2009 Macmillan Publishers Limited
Genetic variants of CC chemokine genes in experimental autoimmune encephalomyelitis, multiple sclerosis and rheumatoid arthritis
J Öckinger, P Stridh, AD Beyeen, F Lundmark, M Seddighzadeh, A Oturai, PS Sørensen, ÅR Lorentzen, EG Celius, V Leppä, K Koivisto, PJ Tienari, L Alfredsson, L Padyukov, J Hillert, I Kockum, M Jagodic and T Olsson
Multiple sclerosis (MS) is a complex disorder of the central nervous system, causing inflammation, demyelination and axonal damage. A limited number of genetic risk factors for MS have been identified, but the etiology of the disease remains largely unknown. For the identification of genes regulating neuroinflammation we used a rat model of MS, myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), and carried out a linkage analysis in an advanced intercross line (AIL). We thereby redefine the Eae18b locus to a 0.88Mb region, including a cluster of chemokine genes. Further, we show differential expression of Ccl2, Ccl11 and Ccl11 during EAE in rat strains with opposite susceptibility to EAE, regulated by genotype in Eae18b. The human homologous genes were tested for association to MS in 3841 cases and 4046 controls from four Nordic countries. A haplotype in CCL2 and rs3136682 in CCL1 show a protective association to MS, whereas a haplotype in CCL13 is disease predisposing. In the HLA-DRB1*15 positive subgroup, we also identified an association to a risk haplotype in CCL2, suggesting an influence from the human leukocyte antigen (HLA) locus. We further identified association to rheumatoid arthritis in CCL2, CCL8 and CCL13, indicating common regulatory mechanisms for complex diseases.
© 2009 Macmillan Publishers Limited
2008
Induction of systemic TNFa in Natalizumab-treated multiple sclerosis
M. Khademi, D. Stol, T. Olsson and E. Wallström
The mRNA expression of eight different cytokines in peripheral blood mononuclear cells in 19 individuals with multiple sclerosis was determined at baseline and after 6 months of open-label treatment with natalizumab. Cellular expression of tumor necrosis factor a (TNFa) mRNA and number of cells secreting TNFa and interferon c protein significantly increased over the 6 months. Kurtzke EDSS scores improved because of the resolution of relapses, but not fatigue severity scores. The observed increases in systemic proinflammatory cytokines by natalizumab treatment are discussed in relation to fatigue and systemic immunity.
2008 EFNS European Journal of Neurology
Progress and prospects in rat genetics: a community view
Timothy J Aitman, John K Critser, Edwin Cuppen, Anna Dominiczak, Xose M Fernandez-Suarez, Jonathan Flint, Dominique Gauguier, Aron M Geurts, Michael Gould, Peter C Harris, Rikard Holmdahl1, Norbert Hubner, Zsuzsanna Izsvák, Howard J Jacob, Takashi Kuramoto, Anne E Kwitek, Anna Marrone, Tomoji Mashimo, Carol Moreno, John Mullins, Linda Mullins, Tomas Olsson, Michal Pravenec, Lela Riley, Kathrin Saar, Tadao Serikawa, James D Shull, Claude Szpirer, Simon N Twigger, Birger Voigt& Kim Worley
The rat is an important system for modeling human disease.Four years ago, the rich 150-year history of rat research was transformed by the sequencing of the rat genome, ushering in an era of exceptional opportunity for identifying genes and pathways underlying disease phenotypes. Genome-wide association studies in human populations have recently provided a direct approach for finding robust genetic associations in common diseases, but identifying the precise genes and their mechanisms of action remains problematic. In the context of significant progress in rat genomic resources over the past decade, we outline achievements in rat gene discovery to date, show how these findings have been translated to human disease, and document an increasing pace of discovery of new disease genes, pathways and mechanisms.Finally, we present a set of principles that justify continuing and strengthening genetic studies in the rat model, and further development of genomic infrastructure for rat research.
© 2008 Nature Publishing Group
Congenic Rats with Allelic Differences in the Class II Transactivator Gene Display Altered Susceptibility to Experimental Autoimmune Encephalomyelitis
Karin Harnesk, Maria Swanberg, Johan Öckinger, Margarita Diez, Olle Lidman, Erik Wallström, Anna Lobell, Tomas Olsson and Fredrik Piehl
Presentation of Ag bound to MHC class II (MHC II) molecules to CD4 T cells is a key event in adaptive immune responses. Genetic differences in MHC II expression in the rat CNS were recently positioned to allelic variability in the CIITA gene (Mhc2ta), located within the Vra4 locus on rat chromosome 10. In this study, we have examined reciprocal Vra4-congenic strains on the DA and PVGav1 backgrounds, respectively. After experimental nerve injury the strain-specific MHC II expression on microglia was reversed in the congenic strains. Similar findings were obtained after intraparenchymal injection of IFN- in the brain. Expression of MHC class II was also lower on B cells and dendritic cells from the DA.PVGav1-Vra4- congenic strain compared with DA rats after in vitro stimulation with IFN-.We next explored whether Vra4 may affect the outcome of experimental autoimmune disease. In experimental autoimmune encephalomyelitis induced by immunization with myelin oligodendrocyte glycoprotein, DA.PVGav1- Vra4 rats displayed a lower disease incidence and milder disease course compared with DA, whereas both PVGav1 and VGav1.DAVra4 rats were completely protected. These results demonstrate that naturally occurring allelic differences in Mhc2ta have profound effects on the quantity of MHC II expression in the CNS and on immune cells and that this genetic variability also modulates susceptibility to autoimmune neuroinflammation.
© 2008 The Journal of Immunology
Analysis of cerebrospinal fluid and cerebrospinal fluid cells from patients with multiple sclerosis for detection of JC virus DNA
E Iacobaeus, C Ryschkewitsch, M Gravell, M Khademi, E Wallstrom, T Olsson, L Brundin and EO Major
Objective 1) To determine whether JC virus (JCV) DNA was present in the cerebrospinal fluid (CSF)
and blood from patients with multiple sclerosis (MS) in comparison with controls and 2) to find out
if our clinical material, based on presence of JCV DNA, included any patient at risk for progressive
multifocal leukoencephalopathy (PML).
Methods The prevalence of JCV DNA was analyzed in CSF and plasma from 217 patients with MS,
86 patients with clinically isolated syndrome (CIS), and 212 patients with other neurological diseases
(OND). In addition, we analyzed CSF cells, the first report of JCV DNA in CSF cells in a single sample,
and peripheral blood cells in a subgroup of MS (n = 49), CIS (n = 14) and OND (n = 53).
Results A low copy number of JCV DNA was detected in one MS cell free CSF sample and in one
MS CSF cell samples. None of these had any signs of PML or developed this disease during followup.
In addition, two OND plasma samples were JCV DNA positive, whereas all the other samples
had no detectable virus.
Conclusion A low copy number of JCV DNA may occasionally be observed both in MS and other
diseases and may occur as part of the normal biology of JC virus in humans. This study does not
support the hypothesis that patients with MS would be at increased risk to develop PML, and consequently
screening of CSF as a measurable risk for PML is not useful.
© 2008 Multiple Sclerosis
Increased IL-10 mRNA and IL-23 mRNA expression in multiple sclerosis: interferon-² treatment increases IL-10 mRNA expression while reducing IL-23 mRNA expression
M Krakauer, P Sorensen, M Khademi, T Olsson and F Sellebjerg
Background Interferon (IFN)-β therapy in multiple sclerosis (MS) has been suggested to promote a
deviation from T lymphocyte production of pathogenic Th1 cytokines to less detrimental Th2 cytokines,
but this is still controversial. We studied patterns of in vivo blood mononuclear cell (MNC)
and whole blood cytokine and transcription factor mRNA expression before and during IFN-β therapy
in MS.
Methods Twenty patients with relapsing–remitting MS were sampled before and after 3 months of
treatment with IFN-β along with 15 healthy volunteers. An additional 39 patients and 50 healthy
volunteers served to confirm initial findings. mRNA was analyzed by real-time reverse transcriptase
polymerase chain reaction (PCR).
Results We found elevated expression of interleukin (IL)-23 and IL-10 in untreated MS patients.
IFN-β therapy increased IL-10 and decreased IL-23 expression independently of any Th1 or Th2
cytokines. The largest changes in cytokine mRNA levels occurred early (~9–12 h) after an IFN-β
injection.
Conclusion We found no evidence of a Th1- or Th2-mRNA-promoting effect of IFN-β therapy. The
therapeutic effect of IFN-β is more likely attributable to the induction of the regulatory cytokine
IL-10. The elevated IL-23 mRNA levels in MS patients are noteworthy in view of the newly discovered
IL-23-driven Th17 T-cell subset, which is crucial in animal models of MS. Since IFN-β therapy
resulted in decreased IL-23 mRNA levels, the Th17 axis could be another target of IFN-β therapy.
© 2008 Multiple Sclerosis
Maternal smoking during pregnancy and multiple sclerosis amongst offspring
S. M. Montgomery, S. Bahmanyar, J. Hillert, A. Ekbom and T. Olsson
Introduction: An association between parental smoking and multiple sclerosis (MS)
in offspring has been reported. This study examined whether maternal smoking during
pregnancy is associated with MS in offspring.
Methods: Swedish general population
registers provided prospectively recorded information on maternal smoking during
pregnancy. The study identified 143 cases with MS diagnosed by 2006 and 1730
matched controls. Subjects were born since 1982 and individually matched by year of
birth, age, sex and region of residence. Conditional logistic regression assessed the
association of maternal smoking with MS in offspring with adjustment for socioeconomic
index.
Results: Maternal smoking during pregnancy was not associated with
MS in offspring, with an odds ratio (and 95% confidence interval) of 0.96 (0.65–1.44).
When stratified by paediatric or later MS onset there was no association with maternal
smoking in either stratum.
Conclusion: It is unlikely that smoking during pregnancy
represents a risk for early-onset MS amongst offspring.
© 2008 European Journal of Neurology
The genetics of multiple sclerosis and its experimental models
Tomas Olsson and Jan Hillert
Purpose of review
We review the current thinking on the genetic aetiology of multiple sclerosis, new
aspects of the influence of genes within the human leucocyte antigen (HLA) complex,
how discrete non-HLA genes regulating multiple sclerosis start to be unequivocally
demonstrated, and how knowledge from the genetics of experimental models of multiple
sclerosis can be used.
Recent findings
Different alleles of the class II molecules display a hierarchy of risks including protective
effects in multiple sclerosis. In addition, HLA class I genes are associated with the
disease. Experimental multiple sclerosis models are regulated by many genes with small
effects, more than about 100. Studies in humans suggest a similar situation. Indeed, the
two genes unequivocally established, IL2RA and IL7RA, display a low odds ratio in the
order of 1.3. We envisage that many more genes of this kind may be unravelled in the
years to come.
Summary
There are hierarchies of influences from class II genes. Class I genes regulate disease as
well. The unambiguous demonstration of non-HLA genes also suggests that many other
genes impacting multiple sclerosis can be deciphered in the years to come. A complete
knowledge of the most important of these may pave the way for more selective therapy
and gene–environment studies.
© 2008 Department of Clinical Neuroscience, Karolinska Institute, Stockholm, Sweden
Host strain-dependent difference in susceptibility in a rat model of herpes simplex type 1 encephalitis and gene–environment studies.
Biborka Bereczky-Veress, Olle Lidman, Farideh Sabri, Ivan Bednar, Fredrik Granath, Tomas Bergström, Christian Spenger, Alf Grandien, Tomas Olsson, Fredrik Piehl, Margarita Diez, Birgit Sköldenberg
Herpes simplex encephalitis (HSE) is characterized by severe focal brain in flammation leading to substantial loss of nervous tissue. The authors estab lished a model of Herpes simplex virus type 1 (HSV)-1–induced acute en cephalitis in the rat by injecting into the whiskers’ area a virus strain isolated from a fatal human HSE case. The model might resemble natural propagation of HSV-1 in humans; spreading from the mouth and lips via the trigeminal nerve to trigeminal ganglia and subsequently entering the central nervous system (CNS). HSV-1 infected Dark Agouti (DA) rats developed a well-synchronized disease and died 5 days after inoculation. HSV-1 detection by quantitative polymerase chain reaction (qPCR), virus isolation and immunohistochemistry, magnetic resonance imaging, and histopathological examination verified dra matic encephalitis mainly in the brainstem, but also in the olfactory bulb and other segments of the brain of diseased rats. In contrast, Piebald Virol Glaxo (PVG) rats were completely resistant to disease, displaying a more rapid clearance of peripheral infection and no evidence of virus entering into neither the trigeminal ganglia nor the CNS. These results suggest a regulation of susceptibility to HSV-1–induced encephalitis at the level of peripheral infection and subsequent neuronal uptake/transport of the virus. This provides a basis for future positioning of genetic polymorphisms regulating HSE and for dissection of important pathogenetic mechanisms of this severe human disease.
© 2008 Journal of NeuroVirology
2007
Variation in interleukin 7 receptor a chain (IL7R) influences risk of multiple sclerosis
Frida Lundmark, Kristina Duvefelt, Ellen Iacobaeus, Ingrid Kockum, Erik Wallström, Mohsen Khademi, Annette Oturai, Lars P Ryder, Janna Saarela, Hanne F Harbo, Elisabeth G Celius, Hugh Salter, Tomas Olsson3 & Jan Hillert
Multiple sclerosis is a chronic, often disabling, disease of the central nervous system affecting more than 1 in 1,000 people in most western countries. The inflammatory lesions typical of multiple sclerosis show autoimmune features and depend partly on genetic factors. Of these genetic factors, only the HLA gene complex has been repeatedly confirmed to be associated with multiple sclerosis, despite considerable efforts. Polymorphisms in a number of non-HLA genes have been reported to be associated with multiple sclerosis, but so far confirmation has been difficult. Here, we report compelling evidence that polymorphisms in IL7R, which encodes the interleukin 7 receptor a chain (IL7Ra), indeed contribute to the non-HLA genetic risk in multiple sclerosis, demonstrating a role for this pathway in the pathophysiology of this disease. In addition, we report altered expression of the genes encoding IL7Ra and its ligand, IL7, in the cerebrospinal fluid compartment of individuals with multiple sclerosis.
© 2007 Nature Publishing Group http://www.nature.com/naturegenetics
Genetic analysis of neuropathic pain-like behaviorfollowing peripheral nerve injury suggests a role of the major histocompatibility complex in development of allodynia
Cecilia A. Dominguez, Olle Lidman, Jing-Xia Hao, Margarita Diez, Jonatan Tuncel, Tomas Olsson, Zsuzsanna Wiesenfeld-Hallin, Fredrik Piehl, Xiao-Jun Xu
Neuropathic pain is a common consequence of damage to the nervous system. We here report a genetic analysis of development of neuropathic pain-like behaviors after unilateral photochemically-induced ischemic sciatic nerve injury in a panel of inbred rat strains known to display different susceptibility to autoimmune neuroinflammation. Pain behavior was initially characterized in Dark-Agouti (DA; RT1av1), Piebald Virol Glaxo (PVG; RT1c), and in the major histocompatibility complex (MHC)-congenic strain PVG-RT1av1. All strains developed mechanical hypersensitivity (allodynia) following nerve injury. However, the extent and duration of allodynia varied significantly among the strains, with PVG displaying more severe allodynia compared to DA rats. Interestingly,the response of PVG-RT1avRT1 was similar to that of DA, suggesting regulation by the MHC locus. This notion was subsequently confirmed in an F2 cohort derived from crossing of the PVG and PVG-RT1av1strains, where allodynia was reduced in homozygous or heterozygous carriers of the RT1av1 allele in comparison to rats homozygous for the RT1c allele. These results indicate that certain allelic variants of the MHC could influence susceptibility to develop and maintain neuropathic pain-like behavior following periph- eral nerve injury in rats.
© 2007 Published by Elsevier B.V. on behalf of International Association for the Study of Pain.
2006
Advanced Intercross Line Mapping of Eae5 Reveals Ncf-1 and CLDN4 as Candidate Genes for Experimental Autoimmune Encephalomyelitis
Kristina Becanovic, Maja Jagodic, Jian Rong Sheng, Ingrid Dahlman, Fahmy Aboul-Enein, Erik Wallstrom, Peter Olofsson, Rikard Holmdahl, Hans Lassmann and Tomas Olsson
Eae5 in rats was originally identified in two F2 intercrosses, (DA X BN) and (E3 X DA), displaying linkage to CNS inflammation and disease severity in experimental autoimmune encephalomyelitis (EAE), respectively. This region overlaps with an arthritis locus, Pia4, which was also identified in the (E3 X DA) cross. Two congenic strains, BN.DA-Eae5 and BN.DA-Eae5.R1, encompassing the previously described Eae5 and Pia4, were established. DA alleles within the chromosome 12 fragment conferred an increase in disease susceptibility as well as increased inflammation and demyelination in the CNS as compared with BN alleles. To enable a more precise fine mapping of EAE regulatory genes, we used a rat advanced intercross line between the EAEsusceptible DA strain and the EAE-resistant PVG.1AV1 strain. Linkage analysis performed in the advanced intercross line considerably narrowed down the myelin oligodendrocyte glycoprotein-EAE regulatory locus (Eae5) to a 1.3-megabase region with a defined number of candidate genes. In this study we demonstrate a regulatory effect of Eae5 on MOG-EAE by using both congenic strains as well as fine mapping these effects to a region containing Ncf-1, a gene associated with arthritis. In addition to structural polymorphisms in Ncf-1, both sequence polymorphisms and expression differences were identified in CLDN4. CLDN4 is a tight junction protein involved in blood-brain barrier integrity. In conclusion, our data strongly suggests Ncf-1 to be a gene shared between two organ-specific inflammatory diseases with a possible contribution by CLDN4 in encephalomyelitis. The Journal of Immunology, 2006, 176: 6055–6064.
© 2006 by The American Association of Immunologists, Inc.
Combined-cross analysis of genome-wide linkage scans for experimental autoimmune encephalomyelitis in rat
Maja Jagodic, Tomas Olsson
Center for Molecular Medicine, Department of Clinical Neuroscience, Neuroimmunology Unit, Karolinska Institutet, SE-17176 Stockholm, Sweden
Received 8 December 2005; accepted 23 August 2006
Unbiased genetic analysis of experimental autoimmune encephalomyelitis (EAE) can provide insights into the pathogenesis of multiple
sclerosis. To date five genome-wide scans using F2 crosses between different inbred rats have been performed with the aim of defining EAEregulating
quantitative trait loci (QTLs) as the starting point for identification of the underlying genes. We here report the first combined-cross
analysis of three F2 crosses previously performed in our group. The majority of QTLs was shared between the different strain combinations and
was therefore reproduced by the combined-cross analysis. Consequently, combined-cross analysis improved the power to detect QTLs with
modest effects and narrowed QTL confidence intervals. The findings also demonstrate a lack of power in previous F2 crosses and encourage
future use of larger populations. Moreover, the allelic states of shared QTLs could be established, thus providing critical information for narrowing
QTLs and identifying the key polymorphism by subsequent haplotype analysis.
© 2006 Elsevier Inc. All rights reserved.
Autoimmunity. Basic and clinical science approach complex diseases — new molecules and mechanisms
Editorial overview
Tomas Olsson and Roland Martin
Current Opinion in Immunology 2006, 18:1–4
0952-7915
© 2006 Elsevier Ltd. All rights reserved.
Genetics of autoimmune neuroinflammation
Tomas Olsson, Maja Jagodic, Fredrik Piehl and Erik Wallström
Detection of gene variants affecting the risk for multiple sclerosis provides insights into mechanisms central for autoaggressive neuroinflammation. Major histocompatibility complex (MHC) class II genes, and probably also MHC class I genes, regulate both human multiple sclerosis and rodent experimental autoimmune encephalomyelitis. However, the functional understanding of the MHC regulation requires further experimentation. Genome scans in human multiple sclerosis have failed to demonstrate significant non-MHC loci with genome-wide significance, but approximately 50 such loci have been described in different rodent experimental autoimmune encephalomyelitis models. Positional cloning of individual rodent genes is difficult, but genes or small genome regions now emerge. Association studies in large human cohorts are needed to confirm the human relevance of rodent genes and such cohorts will also be used for single nucleotide polymorphism-based whole-genome screening. It is realistic to assume that several non-MHC genes regulating autoimmune neuroinflammation, including target tissue responses, will be pinpointed in the next ten years. At the moment there are a few hot candidates, including MHC2TA, PRKCA and IL7R.
Current Opinion in Immunology 2006, 18:1–7
This review comes from a themed issue on Autoimmunity
Edited by Tomas Olsson and Roland Martin 0952-7915
© 2006 Elsevier Ltd. All rights reserved.
Cortical Demyelination Can Be Modeled in Specific Rat Models of Autoimmune Encephalomyelitis and Is Major Histocompatability Complex (MHC) Haplotype-Related
Maria K. Storch MD, Jan Bauer PhD, Christopher Linington PhD, Tomas Olsson MD, PhD, Robert Weissert MD, PhD, and Hans Lassmann MD
In recent years, a number of histopathologic studies revealed the
presence of cortical demyelination in multiple sclerosis (MS). The
underlying mechanisms responsible for cortical demyelination are
unresolved. Recently, the presence of cortical lesions in autoimmune
encephalomyelitis (EAE) induced in marmosets and Lewis rats has
been demonstrated. So far, it is not known whether cortical
demyelinated lesions are also present in other models of EAE. In
this study, we analyzed a large spectrum of different rat strains
actively immunized with myelin oligodendrocyte glycoprotein
(MOG), a model strongly mimicking MS for cortical demyelination.
By using sets of rat strains with the constant EAE-permissive LEW
nonmajor histocompatability complex (MHC) genome, but different
MHC haplotypes, we demonstrated that considerable cortical
demyelination was only found in LEW.1AR1 (RT1r2) and LEW.1W
(RT1u) strains. These rat strains have the isotypes and alleles
RT1.BuDu in the MHC II region and RT1.Cu in the nonclassic
MHC I region in common. Because cortical demyelination was most
prominent in LEW.1AR1 rats, an additional strong influence is
promoted by the RT1.Aa MHC class I allele. Demyelination was
accompanied by microglia infiltration and deposition of immunoglobulins
on myelin sheaths. Our study shows that extensive cortical
demyelination can be reproducibly induced in certain rat strains by
active immunization with MOG. Furthermore, our findings suggest
that cortical demyelination in EAE depends on particular combinations
of MHC I and class II isotypes and alleles. The mechanisms for
this influence and any similar effects in humans will be important
to define.
© 2006 by the American Association of Neuropathologists, Inc.
Genetically determined susceptibility to neurodegeneration is associated with expression of inflammatory genes
Maria Swanberg, Kristina Duvefelt, Margarita Diez, Jan Hillert, Tomas Olsson, Fredrik Piehl, and Olle Lidmana
Axonal damage, a core feature of neurological diseases, induces a retrograde reaction in neurons and surrounding glia. We determined transcriptional profiles of this reaction using Affymetrix oligonucleotide arrays. Gene expression was examined in spinal cord tissue prior to injury and following ventral root avulsion in two inbred rat strains, where the degree of neurodegeneration differs. Stringent statistical analysis revealed 278 regulated genes, whereof 245 were regulated by the injury and 68 differed between strains. Principal component analysis disclosed a common injury response pattern significantly modified by genetic background. Notably, inflammatory genes comprised the largest group of genes induced by injury and these transcripts prevailed in the strain most susceptible to neurodegeneration. In addition, levels of the strain regulated genes C1qb and Timp1 correlated with degree of neurodegeneration in a cohort of genetically heterogeneous animals. These results suggest a link between the inflammatory response elicited by nerve injury and subsequent neurodegeneration.
© 2006 Elsevier Inc.
Definition of a 1.06-Mb Region Linked to Neuroinflammation in Humans, Rats and Mice
Johan Öckinger, Pablo Serrano-Fernández, Steffen Möller, Saleh M. Ibrahim, Tomas Olsson and Maja Jagodic
Unbiased identification of susceptibility genes might provide new insights into pathogenic mechanisms
that govern complex inflammatory diseases such as multiple sclerosis. In this study we fine mapped Eae18a, a
region on rat chromosome 10 that regulates experimental autoimmune encephalomyelitis (EAE), an animal
model for multiple sclerosis. We utilized two independent approaches: (1) in silico mapping based on
sequence similarity between human multiple sclerosis susceptibility regions and rodent EAE quantitative
trait lociand(2) linkagemappingin anF10(DA3PVG.AV1) rat advanced intercrossed line.Thelinkagemapping
defines Eae18a to a 5-Mb region, which overlaps one intergenomic consensus region identified in silico.
The combined approach confirms experimentally, for the first time, the accuracy of the in silico method.
Moreover, the shared intersection between the results of both mapping techniques defines a 1.06-Mb region
containing 13 candidate genes for the regulation of neuroinflammation in humans, rats, and mice.
© 2006 Genetics Society of America
2005
MHC2TA is associated with differential MHC molecule expression and susceptibility to rheumatoid arthritis, multiple sclerosis and myocardial infarction
Maria Swanberg, 0lle Lidman, Leonid Padyukov, Per Eriksson, Eva Åkesson, Maja Jagodic,Anna Lobell, Mohsen Khademil, Ola Börjesson, Cecilia M Lindgren, Pia Lundman, Anthony J Brookes, Juha Keres, Holger Luthman, Lars Alfredsson, Jan Hillert, Lars Klareskog, Anders Hamsten, Fredrik Piehl & Tomas Olsson
Antigen presentation to T cells by MHC molecules is essential for adaptive immune responses. To determine the exact position of a gene affecting expression of MHC molecules, we finely mapped a previously defined rat quantitative trait locus regulating MHC class II on microglia in an advanced intercross line. We identified a small interval including the gene MHC class II transactivator (Mhc2ta) and, using a map over six inbred strains combined with gene sequencing and expression analysis, two conserved Mhc2ta haplotypes segregating with MHC class 11 levels. In humans, a -168A-+G polymorphism in the type III promoter of the MHC class II transactivator (MHC2TA) was associated with increased susceptibility to rheumatoid arthritis, multiple sclerosis and myocardial infarction, as well as lower expression of MHC2TA after stimulation of leukocytes with interferon-y. We conclude that polymorphisms in Mhc2ta and MHC2TA result in differential MHC molecule expression and are associated with susceptibility to common complex diseases with inflammatory components.
© 2005 Nature Publishing Group
Two genes encoding immune-regulatory molecules (LAG3 and IL7R) confer susceptibility to multiple sclerosis
Z Zhang, K Duvefelt, F Svensson, T Masterman, G Jonasdotti3, H Salter, T Emahazion, D Hellgren, G Falk, T Olsson, J Hillert and M Anvret
Multiple sclerosis (MS) is a T-cell-mediated disease of the central nervous system, characterized by damage to myelin and
axons, resulting in progressive neurological disability. Genes may influence susceptibility to MS, but results of association
studies are inconsistent, aside from the identification of HLA class II haplotypes. Whole-genome linkage screens in MS have
both confirmed the importance of the HLA region and uncovered non-HLA loci that may harbor susceptibility genes. In this twostage
analysis, we determined genotypes, in up to 672 MS patients and 672 controls, for 123 single-nucleotide polymorphisms
(SNPs) in 66 genes. Genes were chosen based on their chromosomal positions or biological functions. In stage one, 22 genes
contained at least one SNP for which the carriage rate for one allele differed significantly (Po0.08) between patients and
controls. After additional genotyping in stage two, two genes—each containing at least three significantly (Po0.05) associated
SNPs—conferred susceptibility to MS: LAG3 on chromosome 12p13, and IL7R on 5p13. LAG3 inhibits activated T cells, while
IL7R is necessary for the maturation of T and B cells. These results imply that germline allelic variation in genes involved in
immune homeostasis—and, by extension, derangement of immune homeostasis—influence the risk of MS.
© 2005 Nature Publishing Group
Eae19, a New Locus on Rat Chromosome 15 Regulating Experimental Autoimmune Encephalomyelitis
Jian Rong Sheng, Maja Jagodic, Ingrid Dahlman, Kristina Becanovic, Rita Nohra, Monica Marta, Ellen Iacobaeus, Tomas Olsson and Erik Wallström
Center for Molecular Medicine, Department of Clinical Neuroscience, Neuroimmunology Unit, Karolinska Institutet, SE-17176 Stockholm, Sweden
Multiple sclerosis (MS) and its animal model, myelin oligodendrocyte glycoprotein-induced experimentalautoimmune encephalomyelitis (MOG-EAE), share a complex genetic predisposition with contributions from the major histocompatibility complex class II genes and many other genes. Linkage mapping in F2 crosses between the susceptible DA rat strain and the resistant ACI or BN rat strains in various models of autoimmune neuroinflammation have repeatedly displayed suggestive linkage to a region on rat chromosome 15. A direct study of this region was undertaken in congenic strains by transferring resistant ACI alleles to the susceptible DA background. Phenotypic analysis demonstrated lower maximal and cumulative EAE scores in the DA.ACI–D15Rat6-D15Rat71 (C15), DA.ACI–D15Rat6-D15Rat48, D15Rat126-D15Rat71 (C15R3b), and DA.ACI–D15Rat23-D15rat71 (C15R4) strains compared to the parental DA rat strain.Linkage analysis was then performed in a (DA X PVG.AV1)F7 advanced intercross line, resulting in a LOD score of 4.7 for the maximal EAE score phenotype at the peak marker D15Rat71 and a confidence interval of 13 Mb, overlapping with the congenic fragment defined by the C15R3b and the C15R4 strains. Thus,a new MOG-EAE locus with the designation Eae19 is identified on rat chromosome 15. There are 32 confirmed or predicted genes in the confidence interval, including immune-responsive gene 1 and neuronal ceroid lipofuscinose gene 5. Definition of loci such as Eae19 enables the characterization of genetically regulated, evolutionary conserved disease pathways in complex neuroinflammatory diseases.
© 2005 Genetics Society of America
2004
Current Gene-Mapping Strategies in Experimental Models of Multiple Sclerosis
K. Becanovic, M. Jagodic, E. Wallström & T. Olsson
Neuroimmunology Unit, Section forNeuroscience Research, Department of Clinical
Neuroscience, Karolinska Institutet, Karolinska
Hospital, Stockholm, Sweden
Received 3 March 2004; Accepted in revised form
6 April 2004
Both family-based linkage analyses and population-based association studies
have failed to identify disease-regulatory non-human leucocyte antigen genes
of importance in multiple sclerosis (MS). Instead, investigators have employed
experimental models, which offer major advantages in genetic studies. We
summarize the current main methodologies used and the status of both the
human and experimental approaches. Why is it important to find genes regulating
MS? There is an immense number of cellular and molecular interactions
defined in the immunological field and it is very difficult to unravel those that
are critical to an inflammatory disease, such as MS, by classical hypothesisdriven
research. Unbiased genetics defines evolutionary conserved gene polymorphisms
and pathways regulated by these genes, which are central in the
pathogenesis. These, in turn, are of interest as therapeutic targets and pharmacogenetic
markers.
© 2004 Blackwell Publishing Ltd. Scandinavian Journal of Immunology 60, 39–51
T Cell Ig- and Mucin-Domain-Containing Molecule-3 (TIM-3) and TIM-1 Molecules Are Differentially Expressed on Human Th1 and Th2 Cells and in Cerebrospinal Fluid-Derived Mononuclear Cells in Multiple Sclerosis
Mohsen Khademi, Zsolt Ille´s, Alexander W. Gielen, Monica Marta, Naruhiko Takazawa, Claire Baecher-Allan, Lou Brundin, Jan Hannerz, Claes Martin, Robert A. Harris, David A. Hafler, Vijay K. Kuchroo, Tomas Olsson, Fredrik Piehl, and Erik Wallström
T cell Ig- and mucin-domain-containing molecules (TIMs) comprise a recently described family of molecules expressed on T cells. TIM-3 has been shown to be expressed on murine Th1 cell clones and has been implicated in the pathogenesis of Th1-driven experimental autoimmune encephalomyelitis. In contrast, association of TIM-1 polymorphisms to Th2-related airway hyperreactivity has been suggested in mice. The TIM molecules have not been investigated in human Th1- or Th2-mediated diseases. Using real-time (TaqMan) RT-PCR, we show that human Th1 lines expressed higher TIM-3 mRNA levels, while Th2 lines demonstrated a higher expression of TIM-1. Analysis of cerebrospinal fluid mononuclear cells obtained from patients with multiple sclerosis revealed significantly higher mRNA expression of TIM-1 compared with controls. Moreover, higher TIM-1 expression was associated with clinical remissions and low expression of IFN- mRNA in cerebrospinal fluid mononuclear cells. In contrast, expression of TIM-3 correlated well with high expression of IFN- and TNF-. These data imply the differential expression of human TIM molecules by Th1 and Th2 cells and may suggest their differential involvement in different phases of a human autoimmune disease.
© 2004 The Journal of Immunolog.
2003
Discrete Gene Loci Regulate Neurodegeneration, Lymphocyte Infiltration, and Major Histocompatibility Complex Class II Expression in the CNS
Olle Lidman, Maria Swanberg, Linn Horvath, Karl W. Broman, Tomas Olsson and Fredrik Piehl
Departments of Clinical Neuroscience, Neuroimmunology Unit and 2Medicine, Rheumatology Research Unit, Karolinska Institute, Karolinska Hospital,
S-17176, Stockholm, Sweden, and 3Department of Biostatistics, Johns Hopkins University, Baltimore, Maryland 21205
Neurodegeneration and inflammation are fundamental aspects of many neurological diseases. A genome-wide scan of the response to ventral root avulsion (VRA) in a rat F2 cross discloses specific gene regions that regulate these processes. Two gene loci displayed linkage to neurodegeneration and T cell infiltration, respectively, and a single locus displayed extreme linkage to VRA-induced major histocompatibility complex class II expression on microglia. The demonstration that polymorphic genes in different loci control neurodegeneration and CNS inflammation has implications for various experimental rodent nervous system paradigms and potentially for genetically regulated susceptibility to a variety of human CNS diseases.
© 2003 The Journal of Neuroscience