MS Abstracts 05b-2g5

Multiple Sclerosis (MS) is known to accumulate within families. The magnitude of the familial risk, however, remains uncertain.

Using a nationwide MS register and other national registers, the authors estimated relative and absolute risks of MS in a population-based cohort that included 19,615 first-degree relatives of 8,205 Danish MS patients followed from 1968 to 1997.

The ratio of observed to expected numbers of MS cases served as the measure of the relative risk of MS. Lifetime risks of MS in first-degree relatives were estimated as the product of the relative risk and the national lifetime risk of MS.

Overall, first-degree relatives had a sevenfold increased risk of MS (relative risk = 7.1, 95% confidence interval: 5.8, 8.8) (n = 90) compared with the background population.

By modeling the individual incidence rate of MS as the sum of a familial component and a sporadic risk component, the familial excess lifetime risk was found to be 2.5% (95% confidence interval: 2.0, 3.2) among first-degree relatives of MS patients, irrespective of the gender of the proband and the relative.

This percentage should be added to a sporadic absolute risk in the general population of 0.5% in women and 0.3% for men. Spouses of MS patients did not experience an increased risk of MS, suggesting no major role for environmental factors acting in adulthood.

Objectives
To determine whether a low fat diet supplemented with Omega-3 positively affects quality of life (QOL) in Relapsing/Remitting MS (RRMS) patients.

In this 1-year long double-blind, randomized trial, patients were randomized to two dietary interventions: the "Fish Oil" (FO) group received a low fat diet (15% fat) with omega-3 FOs and the "Olive Oil" (OO) group received the AHA Step I diet (fat 30%) with OO supplements.

The primary outcome measure was the Physical Components Summary Scale (PCS) of the Short Health Status Questionnaire (SF-36).

Additional measures using MS specific QOL questionnaires, Neurological status and relapse rate were obtained.

Results
31 RRMS patients were enrolled, with mean follow up over 11+/-SD 2.9 months.

Clinical benefits favoring the FO group were observed on PCS/SF-36 (P=0.050) and MHI (P=0.050) at 6 months. Reduced Fatigue was seen on the OO diet at 6 months (P=0.035).

The relapse rate decreased in both groups relative to the rates during the 1 year preceding the study: mean change in relapse rate in the FO group: -0.79+/-SD 1.12 relapses/year (P=0.021) vs. -0.69+/-SD 1.11 (P=0.044) in the OO group.

This study suggests that a low fat diet supplemented with omega-3 PUFA can have moderate benefits in RRMS patients on concurrent disease modifying therapies.

AutoAntiBodies to Myelin Oligodendrocyte Glycoprotein (MOG) can induce DeMyelination and Oligodendrocyte loss in models of Multiple Sclerosis (MS).

Whether Anti-MOG Abs play a similar role in patients with MS or inflammatory CNS diseases by Epitope Spreading is unclear.

We have therefore examined whether AutoAntibodies that bind properly folded MOG protein are present in the CNS Parenchyma of MS patients.

IgG was purified from CNS tissue of 14 postmortem cases of MS and 8 control cases, including cases of Encephalitis. Binding was assessed using two independent assays, a fluorescence-based solid-phase assay and a solution-phase RIA.

MOG AutoAntiBodies were identified in IgG purified from CNS tissue by solid-phase immunoassay in 7 of 14 cases with MS and 1 case of Subacute Sclerosing Panencephalitis, but not in IgG from noninflamed control tissue.

This finding was confirmed with a solution-phase RIA, which measures higher affinity AutoAntiBodies.

These data demonstrate that AutoAntiBodies recognizing MOG are present in substantially higher concentrations in the CNS Parenchyma compared with CerebroSpinal Fluid and Serum in subjects with MS.

Indicating that local production/accumulation is an important aspect of AutoAntiBody-mediated pathology in DeMyelinating CNS Diseases. Moreover, chronic inflammatory CNS disease may induce AutoAntiBodies by virtue of Epitope spreading.

Myelin/Oligodendrocyte Glycoprotein (MOG) is found on the surface of Myelinating Oligodendrocytes and external lamellae of Myelin Sheaths in the Central Nervous System, and it is a target Antigen in Experimental Autoimmune Encephalomyelitis and Multiple Sclerosis.

We have isolated bovine, mouse, and rat MOG cDNA clones and shown that the developmental pattern of MOG expression in the rat Central Nervous System coincides with the late stages of Myelination.

The amino-terminal, extracellular domain of MOG has characteristics of an ImmunoGlobulin variable domain and is 46% and 41% identical with the amino terminus of bovine butyrophilin (expressed in the lactating mammary gland).

And B-G Antigens of the chicken Major Histocompatibility Complex (MHC), respectively; these proteins thus form a subset of the ImmunoGlobulin superfamily.

The homology between MOG and B-G extends beyond their structure and genetic mapping to their ability to induce strong antibody responses and has implications for the role of MOG in pathological, AutoImmune conditions.

We colocalized the MOG and BT genes to the human MHC on chromosome 6p21.3-p22. The mouse MOG gene was mapped to the homologous band C of chromosome 17, within the M region of the mouse MHC.

Three-millimeter-thick, fast Spin-Echo T₂-weighted MR images and Spin-Echo PostgGadolinium ₁-weighted images were obtained.

Computer software that which had been validated previously for quantitation of MS lesions was used to detect lesions on the T₂-weighted images.

The Expanded Disability Status Scale (EDSS), Ambulation Index (AI), and a battery of NeuroCognitive Tests were performed on each patient. Forty-two arcuate HyperIntensities along the U-fiber were detected by the software in 28 patients (53%).

Twenty-seven lesions (64.3%) were seen in the Frontal Lobe, eight (19.0%) in the Temporal Lobe, three (7.1%) in the Parietal Lobe, three (7.1%) in the Occipital Lobe, and one (2.4%) in both Frontal and Parietal Lobes. Four lesions (9.5%) showed Gadolinium enhancement.

Seventeen lesions (40%) were HypoIntense on the T₁-weighted images.

Scores of three of the 11 NeuroPsychological Tests reflecting performance in Executive control and Memory were significantly different at least at the p = 0.05 level between the eight patients with multiple, isolated U-fiber lesions and the 45 patients without any or with only a single U-fiber lesion.

No significant difference was noted for EDSS or AI. Isolated U-fiber involvement is an underappreciated MR finding in MS.

Our preliminary hypothesis is that U-fiber lesions may contribute to NeuroPsychological Impairment, although our observation requires confirmation.

Inflammation, DeMyelination, and Axonal damage of the Central Nervous System (CNS) are major pathological features of Multiple Sclerosis (MS).

ProteoLytic digestion of the Blood-Brain Barrier and Myelin protein by Serine Proteases is known to contribute to the development and progression of MS.

Neuropsin, a Serine protease, has a role in Neuronal plasticity, and its expression has been shown to be upregulated in response to injury to the CNS.

To determine the possible involvement of Neuropsin in DeMyelinating Diseases of the CNS, we examined its expression in Myelin Oligodendrocyte Glycoprotein (MOG)-induced Experimental Autoimmune Encephalomyelitis (EAE), a recognized animal model for MS.

Neuropsin mRNA expression was induced in the Spinal Cord White Matter of mice with EAE.

Combined in situ hybridization and ImmunoHistoChemistry demonstrated that most of the cells expressing Neuropsin mRNA showed ImmunoReactivity for CNPase, a cell-specific marker for Oligodendrocytes.

Mice lacking Neuropsin (Neuropsin(-/-)) exhibited an altered EAE progression characterized by delayed onset and progression of clinical symptoms as compared to wild-type mice.

Neuropsin(-/-) mice also showed attenuated DeMyelination and delayed Oligodendroglial death early during the course of EAE.

These observations suggest that Neuropsin is involved in the pathogenesis of EAE mediated by DeMyelination and Oligodendroglial death.

(c) 2005 Wiley-Liss, Inc.

Objective
With diagnostic criteria alterations, increased MRI availability, and awareness of therapies, temporal changes in incidence and prevalence rates may occur, with an increase in the proportion of mildly affected persons diagnosed with Multiple Sclerosis (MS).

The authors assessed temporal trends in the delay from symptom onset to diagnosis (DONDX), and determined whether the degree of disability at diagnosis differs by year of symptom onset (YONSET), using the NARCOMS Registry.

Methods
The authors selected US participants with an age at symptom onset of 10 to 60 years, and YONSET > or = 1980 (n = 16,581).

The authors divided YONSET into 5-year groups and compared DONDX between groups using multivariate Cox regression.

The authors classified participants enrolled within 2 years of diagnosis (n = 5,548) as having mild, moderate, or severe disability using Patient Determined Disease Steps, and assessed the association of disability with YONSET using polytomous logistic regression.

Results
DONDX decreased with later YONSET (r = -0.43, p < 0.0001). This association remained after adjustment for demographic factors in a multivariate Cox model.

Later YONSET was associated with increased odds of having mild disability at diagnosis as compared to severe disability (OR = 1.10 per year; 1.09 to 1.11).

Conclusion
The delay from symptom onset to diagnosis is steadily decreasing in MS. An increasing proportion of patients with MS have mild disability at diagnosis after accounting for confounders.

As the effectiveness of therapies is influenced by disease duration, this has implications for comparison of treatment effects in modern clinical trials to earlier study results.

Background
Bee Sting Therapy is increasingly used to treat patients with Multiple Sclerosis (MS) in the belief that it can stabilize or ameliorate the disease. However, there are no clinical studies to justify its use.

Methods
In a randomized, crossover study, we assigned 26 patients with Relapsing/Remitting or Relapsing Secondary/Progressive MS to 24 weeks of medically supervised Bee Sting Therapy or 24 weeks of no treatment.

Live bees (up to a maximum of 20) were used to administer Bee Venom three times per week. The primary outcome was the cumulative number of new Gadolinium-enhancing lesions on T₁-weighted MRI of the Brain.

Secondary outcomes were lesion load on T₂-weighted MRI, relapse rate, disability (Expanded Disability Status Scale, Multiple Sclerosis Functional Composite, Guy's Neurologic Disability Scale), Fatigue (Abbreviated Fatigue Questionnaire, Fatigue Impact Scale), and health-related quality of life (Medical Outcomes Study 36-Item Short Form General Health Survey).

Results
During Bee Sting Therapy, there was no significant reduction in the cumulative number of new Gadolinium-enhancing lesions.

The T₂-weighted lesion load further progressed, and there was no significant reduction in relapse rate.

There was no improvement of disability, Fatigue, and quality of life. Bee Sting Therapy was well tolerated, and there were no serious adverse events.

Conclusions
In this trial, treatment with Bee Venom in patients with Relapsing Multiple Sclerosis did not reduce disease activity, disability, or Fatigue and did not improve quality of life.

In established Multiple Sclerosis, Magnetization Transfer Ratio (MTR) Histograms reveal abnormalities of Normal-Appearing White Matter (NAWM) and Gray Matter (NAGM).

The aim of this study was to investigate for such abnormalities in a large cohort of patients presenting with Clinically Isolated Syndromes suggestive of Multiple Sclerosis.

Magnetization Transfer Imaging was performed on 100 patients (67 women, 33 men, median age 32 years) a mean of 19 weeks (SD 3.8, range 12-33 weeks) after symptom onset with a Clinically Isolated Syndrome.

And, in 50 healthy controls (34 women, 16 men, median age 32.5 years). SPM99 software was used to generate segmented NAWM and NAGM MTR maps.

The volumes of T₂ lesions, White Matter and Gray Matter were calculated. Eighty-one patients were followed up clinically and with conventional MRI after 3 years (n = 61) or until they developed Multiple Sclerosis if this occurred sooner (n = 20).

Multiple regression analysis was used to investigate differences between patients and controls with age, gender and volume measures as covariates to control for potential confounding effects.

The MTR Histograms for both NAWM and NAGM showed a reduction in the mean (NAWM, 38.14 versus 38.33, P = 0.001; NAGM 32.29 versus 32.50, P = 0.009; units in pu) and peak location, with a left shift in the histogram.

Mean NAWM and NAGM MTR were also reduced in the patients who developed Clinically Definite Multiple Sclerosis and Multiple Sclerosis according to the McDonald Criteria but not in the 24 patients with normal T₂-weighted Brain Magnetic Resonance Imaging.

MTR abnormalities occur in the NAWM and NAGM at the earliest clinical stages of Multiple Sclerosis

Hematopoietic Stem Cell (HSC) transplantation is a potential therapy that can offer Multiple Sclerosis patients a radical, potentially curative treatment.

Using Experimental Autoimmune Encephalomyelitis (EAE) as a model, we previously reported that retrovirally transduced B-Cells expressing Myelin Basic Protein (MBP), MBP Ac1-11, or Myelin Oligodendrocyte Glycoprotein p35-55 induced tolerance and reduced symptoms.

Here, we extend our tolerance approach using Bone Marrow (BM) cells expressing full-length PhosphoLipid Protein (PLP) in a model for Relapsing/Remitting EAE.

Using GFP expression as a marker, we found that up to 50% of cells were positive for transgene expression in peripheral blood after 900 rad irradiation and transduced BM transplantation, and expression was stable in hematopoietic lineages for over 10 weeks.

Upon challenge, T-Cell proliferation in response to PLP p139-151 was reduced and EAE was completely abolished in a pretreatment protocol. In addition, protection from EAE could be achieved with PLP-transduced BM cells given on day 12 after immunization, a potential therapeutic protocol.

Finally, the protective effect of PLP-expressing BM could also be observed using a NonMyeloAblative protocol, albeit with lower efficacy. Our results suggest that HSC may be useful to achieve long-lasting tolerance to protect mice from EAE and possibly to promote CNS repair in ongoing EAE.

Inflammatory processes play a key role in the pathogenesis of a number of common NeuroDegenerative Disorders such as Alzheimer's Disease (AD), Parkinson's Disease (PD), and Multiple Sclerosis (MS).

Abnormal Iron accumulation is frequently noted in these diseases and compelling evidence exists that Iron is involved in inflammatory reactions.

HistoChemical stains for Iron repeatedly demonstrate that Oligodendrocytes, under normal conditions, stain more prominently than any other cell type in the Brain.

Therefore, we examined the hypothesis that Cytokine toxicity to Oligodendrocytes is Iron mediated.

Oligodendrocytes in culture were exposed to Interferon-gamma (IFN-γ), InterLeukin-1beta (IL-1ß), and Tumor Necrosis Factor-alpha (TNF-).

Toxicity was observed in a dose-dependent manner for IFN-γ and TNF-.

IL-1ß was not toxic in the concentrations used in this study. The toxic concentration of IFN-γ, and TNF- was lower if the cells were Iron loaded, but Iron loading had no effect on the toxicity of IL-1ß.

These data provide insight into the controversy regarding the toxicity of Cytokines to Oligodendrocytes by revealing that Iron status of these cells will significantly impact the outcome of Cytokine treatment.

The exposure of Oligodendrocytes to Cytokines plus Iron decreased Mitochondrial membrane potential but activation of Caspase 3 is limited.

The antioxidant, TPPB, which targets Mitochondria, protected the Oligodendrocytes from the Iron-mediated CytoToxicity, providing further support that Mitochondrial dysfunction may underlie the Iron-mediated Cytokine toxicity.

Therapeutic strategies involving AntiInflammatory agents have met with limited success in the treatment of DeMyelinating Disorders.

A better understanding of these agents and the contribution of cellular Iron status to Cytokine toxicity may help develop a more consistent intervention strategy.

(c) 2005 Wiley-Liss, Inc.

Little is known about the temporal evolution of Gray Matter damage occurring early in the course of Multiple Sclerosis (MS). The authors investigated the evolution of Gray Matter volume loss in 117 patients with Relapsing/Remitting MS, scanned monthly for a 9-month period.

Time-trend analysis revealed a decrease of Gray Matter volumes over the study period (p < 0.001). This study shows that Gray Matter damage in Relapsing/Remitting MS evolves markedly over a short period of observation.