A COMPARATIVE STUDY OF ORGAN DOSES ASSESSMENT FOR PATIENTS UNDERGOING CONVENTIONAL X-RAY EXAMINATIONS: PHANTOM EXPERIMENTS VS. CALCULATIONS.

A series of phantom experiments were performed with the aim of estimating organ doses for patients undergoing conventional X-ray chest and pelvis examinations. The experiments were performed using physical phantoms corresponding to an adult and a 5-year-old child. Mean organ doses and entrance surface dose were measured using TL-dosemeters. The measured organ doses were compared with the data obtained by calculations using available software tools (EDEREX and PCXMC 2.0) based on the computational MIRD-5 stylized models. The differences between calculated and measured doses for organs located fully or partly in the primary radiation beam did not exceed ±33% with the probability of 95% for the tube voltage 60-140 kV both for an adult and a 5-year-old child phantom. This study suggests that EDEREX and PCXMC 2.0 can be used to estimate organ and effective dose for adult as well as pediatric patients undergoing conventional X-ray examinations.

The selective sphingosine 1-phosphate receptor modulator BAF312 redirects lymphocyte distribution and has species-specific effects on heart rate.

BACKGROUND AND PURPOSE: BAF312 is a next-generation sphingosine 1-phosphate (S1P) receptor modulator, selective for S1P(1) and S1P(5 ) receptors. S1P(1) receptors are essential for lymphocyte egress from lymph nodes and a drug target in immune-mediated diseases. Here, we have characterized the immunomodulatory potential of BAF312 and the S1P receptor-mediated effects on heart rate using preclinical and human data. EXPERIMENTAL APPROACH: BAF312 was tested in a rat experimental autoimmune encephalomyelitis (EAE) model. Electrophysiological recordings of G-protein-coupled inwardly rectifying potassium (GIRK) channels were carried out in human atrial myocytes. A Phase I multiple-dose trial studied the pharmacokinetics, pharmacodynamics and safety of BAF312 in 48 healthy subjects. KEY RESULTS: BAF312 effectively suppressed EAE in rats by internalizing S1P(1) receptors, rendering them insensitive to the egress signal from lymph nodes. In healthy volunteers, BAF312 caused preferential decreases in CD4(+) T cells, T(naïve) , T(central memory) and B cells within 4-6 h. Cell counts returned to normal ranges within a week after stopping treatment, in line with the elimination half-life of BAF312. Despite sparing S1P(3) receptors (associated with bradycardia in mice), BAF312 induced rapid, transient (day 1 only) bradycardia in humans. BAF312-mediated activation of GIRK channels in human atrial myocytes can fully explain the bradycardia. CONCLUSION AND IMPLICATIONS: This study illustrates species-specific differences in S1P receptor specificity for first-dose cardiac effects. Based on its profound but rapidly reversible inhibition of lymphocyte trafficking, BAF312 may have potential as a treatment for immune-mediated diseases.

Cerebrospinal fluid levels of a proliferation-inducing ligand (APRIL) are increased in patients with neuropsychiatric systemic lupus erythematosus.

OBJECTIVES: A proliferation-inducing ligand (APRIL) and B-cell activation factor (BAFF) are B-cell stimulation and survival molecules. We have investigated whether APRIL and/or BAFF activity is enhanced in the systemic and/or intrathechal compartment of patients with neuropsychiatric systemic lupus erythematosus (NPSLE). In particular, the association between fatigue and APRIL/BAFF activity was investigated. METHODS: Twenty-eight NPSLE patients were evaluated clinically, with sampling of cerebrospinal fluid (CSF) and blood, and magnetic resonance imaging (MRI). CSF and blood samples from 13 multiple sclerosis (MS) patients and 17 patients with other neurological diseases (OND) were used as controls. Protein levels of BAFF and APRIL were quantified in CSF and plasma, mRNA expression levels of BAFF and APRIL were determined in peripheral blood (PBMC) and CSF mononuclear cells (CSF-MC). The Fatigue Severity Scale (FSS) was used to quantify the degree of fatigue. RESULTS: NPSLE patients had higher levels of APRIL in CSF as compared to OND (p < 0.01). No corresponding increase in APRIL mRNA levels was detected in CSF-MC. BAFF levels in plasma were higher in NPSLE than in OND (p < 0.001). BAFF mRNA expression in PBMC was also higher in NPSLE patients than in controls (p < 0.05). FSS scores in patients with NPSLE correlated significantly with APRIL levels in CSF. CONCLUSIONS: Protein levels of APRIL in CSF were increased in NPSLE as compared to OND. Moreover, there was a positive correlation between CSF APRIL levels and fatigue. Our results suggest that APRIL and possibly also BAFF may be involved in the pathogenesis of NPSLE and in SLE-related fatigue.

RGMA and IL21R show association with experimental inflammation and multiple sclerosis.

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)=1.33). Polymorphisms of RGMA also correlated with changes in the expression of interferon-gamma (IFN-gamma) and TNF in cerebrospinal fluid of MS patients. In IL21R, there was one positively associated (OR=1.14) and two protective (OR=0.87 and 0.68) haplotypes. One of the protective haplotypes correlated to lower IFN-gamma 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.

Analysis of cerebrospinal fluid and cerebrospinal fluid cells from patients with multiple sclerosis for detection of JC virus DNA.

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 follow-up. 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.

Induction of systemic TNFalpha in natalizumab-treated multiple sclerosis.

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 alpha (TNFalpha) mRNA and number of cells secreting TNFalpha and interferon gamma 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.

Evaluation of conversion coefficients from measurable to risk quantities for external exposure over contaminated soil by use of physical human phantoms.

Conversion coefficients from measurable quantities such as air kerma free-in-air or personal dose equivalent to effective dose were determined by phantom experiments. Heterogenic anthropomorphic phantoms representing children of one and five years age, and a Rando phantom representing an adult were exposed in the open field contaminated by different levels of radiocesium in the upper soil layer, in a forest site and inside a wooden house. LiF thermoluminescent (TL) detectors were used inside the phantoms for the estimation of organ doses and effective dose. Personal dosimeters similar to those used in radiation protection for individual dose measurements were placed onto the phantom surface (chest area). The ratios of dose values in separate organs to air kerma free-in-air varied from 0.69 to 1.15 for the children phantoms, and from 0.55 to 0.94 for the adult phantom, respectively, when irradiated in the open field. Body size (weight) was found to be the most important factor influencing the values of the conversion coefficients. The differences observed can reach approximately 40% when comparing conversion factors from air kerma free-in-air to effective dose for adults and newborns. For conversion coefficients from personal dose to effective dose, these differences can reach approximately 15%. The dependences of the various conversion coefficients on body mass were quantified by regression analysis. The results were compared with those calculated for a plane mono-energetic photon source having an energy of 700 keV and being located in the ground at a depth of 0.5 g cm(-2). Calculated and measured conversion coefficients from air kerma free-in-air to effective dose agreed within 12%.

No signs of immunoactivation in the cerebrospinal fluid during treatment with infliximab.

Neuroinflammatory (demyelinating) disease is a rare but feared complication of treatment with anti-tumour necrosis factor (TNF)alpha in patients with polyarthritis. In this study, blood and cerebrospinal fluid markers of inflammation were analysed in 10 people with polyarthritis before and during treatment with infliximab. An increased systemic expression of interferon (IFN)gamma was detected. Systemic administration of IFNgamma is known to exacerbate multiple sclerosis. However, the present study failed to detect signs of inflammation in the cerebrospinal fluid samples-that is, pleocytosis, oligoclonal immunoglobulin G bands, increased expression of IFNgamma, TNFalpha or interleukin 10, or increased levels of nitric oxide oxidation products. Our initial hypothesis, that the few cases of clinical neuroinflammatory disorders observed during treatment of polyarthritis with anti-TNFalpha represent the extreme end of a commonly occurring minor intrathecal immune activation, which in most cases does not give any overt neurological dysfunction, was not supported. Induction of systemic IFNgamma production may still be relevant in neuroinflammation associated with treatment with anti-TNFalpha.

External and internal irradiation of a rural Bryansk (Russia) population from 1990 to 2000, following high deposition of radioactive caesium from the Chernobyl accident.

In 1990, a joint Nordic-Russian project was initiated in order to make independent estimations of the effective dose to selected groups of inhabitants in a highly contaminated area around the city of Novozybkov in the western Bryansk region of Russia. The inhabitants were living in six villages with initial contamination levels of (137)Cs between 0.9 and 2.7 MBq m(-2). Some villages had been decontaminated, others not. Both school children and adults participated in the study. The external irradiation of 100-130 inhabitants was determined during 1 month in September-October each year from 1990 to 2000 (except 1999), using individual thermoluminescent dosemeters. The body burden of (137,134)Cs was determined by in vivo measurements in about 500 inhabitants annually from 1991 to 2000, and for a subgroup also with analysis of the (137)Cs concentration in urine. The mean effective dose (E) from external and internal irradiation due to (137,134)Cs deposition varied between 2.5 and 1.2 mSv per year between 1990 and 2000. The total mean E decreased, on average, by 9% per year, while the mean external dose decreased by 16% per year. The dose rate from internal radiation decreased more slowly than the dose rate from external radiation, and also showed an irregular time variation. The contribution from the internal dose to the total E was 30-50%, depending on the village. Predictions for the long-term changes in the effective dose to people living in the areas are presented. The cumulated E for the 70 years following the accident was estimated to be about 90 mSv with the assumption that both internal and external dose decrease by 2% per year after year 2000. The highest E during a life-time received by single individuals living in the area may amount to around 500 mSv considering the individual variations in E.

Current gene-mapping strategies in experimental models of multiple sclerosis.

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 hypothesis-driven 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.

Identification of rat quantitative trait loci that regulate LPS-induced pro-inflammatory cytokine responses.

Bacterial lipopolysaccharides (LPSs) trigger innate immune effector functions, such as the production of pro-inflammatory cytokines. Here we utilized major histocompatibility complex (MHC)-congenic rats to dissect the genetic basis of strain-dependent variations of LPS-induced tumour necrosis factor-alpha (TNF-alpha) levels in a whole blood in vitro assay. PVG.1AV1 background was associated with a high response, ACI background with a medium response, and LEW.1AV1 and DA backgrounds were associated with low responses. To determine the location of regulating non-MHC genes, a genome-wide linkage analysis with 236 microsatellite markers was performed on 186 F2 progeny of high TNF-alpha responder PVG.1AV1 and MHC identical but low TNF-alpha responder LEW.1AV1 rats. A region on rat chromosome 1 displayed linkage to LPS-induced TNF-alpha responses (P = 3.3 x 10-5). In addition, a locus on chromosome 2 was linked to responses of both interleukin-6 (IL-6) (P = 2.3 x 10-5) and TNF-alpha (possible linkage, P = 8 x 10-3). Both chromosome regions have been linked to inflammatory diseases in rats, and so have the homologous regions in mice and humans. We therefore suggest that continued genetic dissection of the described in vitro phenotypes will give clues to both normal physiological regulation of LPS-induced TNF-alpha production and disease pathways.

Depletion of Vbeta5.2/5.3 T cells with a humanized antibody in patients with multiple sclerosis.

A potentially pathogenic expansion of T cells expressing T cell receptor (TCR) Vbeta5.2/5.3 has been demonstrated in patients with multiple sclerosis (MS). A humanized antibody (ATM-027) directed against these T cells has been developed to further investigate the role of this subpopulation of T cells in MS. The pharmacokinetics/dynamics and safety of ATM-027 (0.3-300 mg intravenously over 30 min) were investigated in 14 patients with MS. The effect of treatment on cytokine expression and autoreactivity to peptides of myelin basic protein (MBP) was also studied. ATM-027 was well tolerated and raised no safety concerns. Clearance of the antibody was low and elimination half-life was approximately 3 weeks. The majority of the target Vbeta5.2/5.3 expressing T cells were depleted for at least 18 months. The small remaining fraction of target cells showed a marked decrease in their TCR expression, which was recovered within 8 months. The numbers of peripheral blood mononuclear cells (PBMCs) with spontaneous expression of IFN-gamma was decreased at 72 h and 8 weeks after treatment, whilst no clear effects on TNF-alpha, IL-4, IL-10, TGF-beta expression were observed. There was also a significant decrease in the number of PBMCs producing IFN-gamma in response to MBP peptide 80-102. We conclude that long-term depletion of T cells expressing defined Vbeta subgroups in MS patients is feasible using selective immunotherapy. The selective depletion of Vbeta5.2/5.3 expressing T cells in this study resulted in a decrease in potentially disease promoting anti-MBP reactivity and pro-inflammatory cytokine production.

Polygenic control of autoimmune peripheral nerve inflammation in rat.

Experimental autoimmune neuritis (EAN) is the principal animal model for Guillain-Barré syndrome (GBS), an inflammatory disease of the peripheral nervous system. Little is known on the genetic regulation of these diseases. We provide the first genetic linkage analysis of EAN. Susceptibility to EAN in a rat F2 population segregated with high levels of anti-PNM IgG, as well as IgG2b and IgG2c isotype levels, which support that disease genes regulate preferential Th1/Th2 differentiation. Linkage analysis demonstrated co-localization of EAN loci with reported susceptibility loci for experimental arthritis and/or encephalomyelitis and a new region on chromosome 17. Further dissection of these loci may disclose disease pathways in GBS.

Curative effects of recombinant human Interleukin-6 in DA rats with protracted relapsing experimental allergic encephalomyelitis.

We have studied the effects of treatment with recombinant human (rh)IL-6 on clinical, histological and immunological parameters of protracted relapsing (PR) experimental allergic encephalomyelitis (EAE) in DA rats. rhIL-6 (50 microg/rat subcutaneously/day) was given under three different regimens, as early prophylaxis, from 1 day prior to 14 days after immunization, in late prophylaxis, from day +7 until day 21 post-immunization (p.i.) and therapeutically to rats with clinical signs of EAE from day 14 to day 28 p.i. Although rhIL-6 failed to modulate the course of PR-EAE when administered as the early prophylactic regimen, it exerted clear-cut favourable effects on the course of the disease if was administered either as later prophylactic or as therapeutic treatment. Under these conditions, rhIL-6 accelerated recovery from EAE attacks and reduced/milded subsequent EAE episodes as compared to either PBS- or heat-inactivated rhIL-6-treated control rats. In agreement with this clinical effect, relative to PBS-treated rats, the animals injected with rhIL-6 exhibited lower numbers of MHC class II(+) and CD4(+) cells in their spinal cords. rhIL-6-treatment also profoundly modulated the endogenous cytokine network, the treated rats displaying increased numbers of spleen cells expressing mRNA transcripts of the anti-inflammatory cytokines IL-10 and TGF-beta along with simultaneously reduced numbers of mRNAs for TNF-alpha. In addition, upon ex vivo exposure to either myelin basic protein peptide 63-88 (MBP63-88) or to phytoaemagglutinin A, the numbers of IFN-gamma secreting splenocytes was also significantly reduced (ELISPOT analysis) in rhIL-6-treated rats as compared to PBS-treated controls.

Importance of the 3' untranslated region of ornithine decarboxylase mRNA in the translational regulation of the enzyme.

Translational regulation of ornithine decarboxylase (ODC), which catalyses the first step in the biosynthesis of polyamines, appears to be an important mechanism in the strong feedback control as well as in the hypotonic induction of the enzyme. However, the exact mechanisms are not yet understood. The ODC mRNA has long 5' and 3' untranslated regions (UTRs) which may be involved in the translational control of the enzyme. In the present study we have used a series of stable transfectants of Chinese Hamster ovary cells expressing ODC mRNAs with various truncations in the 5' and 3' UTRs to investigate the importance of these regions. It is demonstrated that neither the 5' UTR nor the 3' UTR appears to be involved in the polyamine-mediated feedback control of ODC synthesis. The hypotonic induction of ODC, on the other hand, was shown to be highly dependent on the presence of the 3' UTR, but not on the 5' UTR, of ODC mRNA. Cells expressing ODC mRNAs lacking the 3' UTR showed no, or only a very slight, induction of ODC whether the 5' UTR was present or not, whereas the cell lines expressing ODC mRNAs containing the 3' UTR (with or without the 5' UTR) markedly induced ODC after a hypotonic shock. The present finding of a role for the ODC mRNA 3' UTR in the hypotonic induction of ODC is the first demonstration of a specific effect of the 3' UTR in the regulation of ODC.

Long-term external radiation exposure of inhabitants in the western Bryansk region of Russia as a consequence of the Chernobyl accident.

The western Bryansk region in south-western Russia was highly contaminated with 137Cs and 134Cs due to the Chernobyl accident in 1986. In 1990, a joint Nordic-Russian project was initiated in order to make measurements and estimates of the absorbed doses to selected groups of inhabitants in this area. The participating individuals were living in small villages with contamination levels between 0.9 and 2.7 MBq m(-2). Only some villages had been decontaminated. Both school-children and adults participated in the study and the number of persons was between 100 and 130 each year, residing in 5 villages. Every year in September-October, from 1990 to 1998. we performed individual measurements of external absorbed doses, assessed with thermoluminescent (TL) dosemeters (LiF). The mean effective dose per year from external irradiation due to the Chernobyl accident of the inhabitants in the villages ranged between 0.8 and 2.9 mSv during the study period and decreased with an apparent half-time of 3.7-8.2 years, depending on village and group. The highest individual doses within one village were, on average higher by a factor of 3 than the mean value for that village. Under the conservative assumption of a decrease rate in the external effective dose of 2% per year after 1998, individuals in the most highly exposed village are assumed to receive a life-time effective dose of about 75 mSv (between 1986 and 2056) from external exposure to caesium radionuclides. The mean value for the villages under study was estimated to be around 65 mSv using the assumed rate of decrease.

Screening of several H-2 congenic mouse strains identified H-2(q) mice as highly susceptible to MOG-induced EAE with minimal adjuvant requirement.

We identified H-2(q) as a susceptible genotype for MOG-induced EAE by systematic screening of a series of H-2 congenic B10 mouse strains. A series of H-2(q)-bearing strains with divergent gene backgrounds were subsequently investigated. DBA/1 mice were highly susceptible to MOG(1-125)- and MOG(79-96)-induced EAE in the absence of pertussis toxin. Immunisation with MOG(1-125) and MOG(79-96) induced an autoreactive T-cell response in DBA/1 mice. Brain histopathology revealed T-cell and macrophage-infiltrated lesions with associated demyelination. The important features which make this an appropriate model of human disease are high sensitivity to MOG and dependence of an immunodominant peptide region homologous to that implicated in multiple sclerosis.

Genetics of rat neuroinflammation.

The definition of genes regulating the pathogenetic pathways of autoimmune neuroinflammation, may provide targets for new therapeutic strategies. This is not easily accomplished in human disease. Such genetic dissection can more readily be done by the use of inbred rodent strains. With these, genetic heterogeneity is avoided and variation in the environmental influences is minimized. Close mimicking of the human disease characteristics is desirable in such endeavors. Chronic relapsing experimental autoimmune encephalomyelitis (EAE) with MS-like histopathology is achieved after immunization of certain rat strains with myelin oligodendrocyte glycoprotein (MOG) or spinal cord homogenate. The major histocompatibility complex (MHC) regulate the ease by which the environmental trigger in the form of immunisation induces disease. Use of intra-MHC recombinant strains demonstrated major influences from the MHC class II genome region, but additional influences from both the MHC class I and III regions. These findings now provide a basis for studies of the mechanisms for MHC-controlled autoimmune pathogenicity leading to MS-like disease. Gene mapping of F2 crosses between susceptible and resistant rat strains demonstrated nine genome regions outside the MHC which regulate different phenotypes of rat EAE. Many of these co-localize with genome regions regulating other organ-specific disease such experimental arthritis, suggesting a sharing of disease pathways. Further finemapping can lead to the exact identification of disease regulating genes. Interestingly, we have also demonstrated a non-MHC gene control of the inflammatory response, in the form of glial cell activation, and neuronal degeneration, subsequent to anterior nerve root avulsion in rats. The genetic dissection of these influences may unravel pathways controlling CNS vulnerability.

Increased numbers of mononuclear cells from blood and CSF expressing interferon-gamma mRNA in multiple sclerosis are from both the CD4+ and the CD8+ subsets.

Activated, cytokine-producing lymphocytes may regulate central nervous system (CNS) inflammation in multiple sclerosis (MS). We utilize a novel combination of in situ hybridization (ISH) and immunocytochemical staining of peripheral blood lymphocytes (PBLs) to identify spontaneously interferon-gamma (IFNgamma) mRNA expressing cells as CD4+ or CD8+. A major proportion of the IFNgamma mRNA expressing lymphocytes belonged to the CD4+ lineage, which concords with the cellular composition of MS brain lesions, findings in experimental models and the HLA class II haplotype association in MS. There were also significantly more CD8+ IFNgamma mRNA expressing lymphocytes in the MS patients compared with healthy controls, further suggesting the contribution of activated cells from this lineage in the inflammatory response in MS. Both CD4+ and CD8+ IFNgamma mRNA expressing cells were enriched in the cerebrospinal fluid (CSF) as compared with the peripheral blood of the MS patients. Combined with emerging genetic data on HLA class I influences, our data argues for a joint role of activated CD8+ and CD4+ cells in the pathogenesis of MS.

Reduction of both pro- and anti-inflammatory cytokines after 6 months of interferon beta-1a treatment of multiple sclerosis.

Treatment of multiple sclerosis (MS) with interferon beta (IFNbeta) reduces relapse rate, magnetic resonance imaging (MRI) activity and progression of disability. It has been suggested that this beneficial effect is paralleled by an inhibition of proinflammatory cytokines such as interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) and an induction of anti-inflammatory cytokines such as interleukin-4 (IL-4) and interleukin-10 (IL-10). In this study, we record a reduced number of spontaneously IFNgamma mRNA-expressing cerebrospinal fluid mononuclear cells (CSF-MC) and IFNgamma, TNFalpha and IL-10 mRNA-expressing peripheral blood mononuclear cells (PBMC) after 6 months of IFNbeta-1a treatment, paralleled by a decreased purified protein derivate (PPD)-stimulated and unstimulated IFNgamma secretion by PBMC. These effects were not apparent after 2 weeks of treatment, and IFNbeta-1a induced IFNgamma production by naive PBMC in vitro. We did not record increased numbers of IL-4 mRNA-expressing CSF-MC or PBMC, increased plasma IL-10 levels, increased numbers of IgG, A or M secreting plasma cells or in vitro induction of IL-10 production by IFNbeta-1a. We conclude that long-term cytokine modulation by IFNbeta-1a differs from acute effects and that downregulation of both pro- and anti-inflammatory cytokines, rather than a shift in the cytokine profile, is apparent after 6 months of IFNbeta-1a treatment of MS patients.