MS Abstracts 9a-2g1

BiExponential T₂ relaxation of the localized water signal can be used for segmentation of Spectroscopic volumes.

To assess the specificity of the components an iterative relaxation measurement of the localized water signal.

(STEAM, 12 Echo times, geometric spacing from 30 ms to 2000 ms) was combined with Magnetization Transfer (MT) saturation.

40 single Lobe pulses, 12 ms duration, 1440 degrees nominal flip angle, 1 kHz offset, repeated every 30 ms.

Voxels (volume elements) including CSF were examined in Parietal Cortex and PeriVentricular Parietal White Matter (10 each).

As well as 13 Voxels in Central White Matter and 16 T₁-HypoIntense Non-Enhancing Multiple Sclerosis lesions without CSF inclusion.

BiExponential models (excluding Myelin water) were fitted to the relaxation data.

In PeriVentricular VOIs the component of long T₂ (1736 +/- 168 ms) that is attributed to CSF was not affected by MT.

In Cortical VOIs this component had markedly shorter T₂'s (961 +/- 239 ms) and showed both attenuation and prolongation with MT. Indicating contributions from tissue.

MS lesions and central WM showed a second tissue component of intermediate T₂ (160-410 ms).

In White Matter similar MT attenuation indicated strong exchange between the two tissue components, prohibiting segmentation.

In MS lesions, however, markedly less MT of the intermediate component was found, which is consistent with decreased cellularity and exchange in a region that is large compared to Diffusion motion.

In patients suspected of Multiple Sclerosis (MS), even in the case of MRI positive changes, CerebroSpinal Fluid (CSF) analysis is of great practical value.

Until now, however, only IgG index, restricted OligoClonal IgG Bands and the examination of IgG subclasses have been of practical value.

The determination of other Immunological markers is too expensive to be introduced into laboratory diagnostic standards in MS.

However, the clinical trials should be monitored by a large set of markers of MS activity, including the estimation of anti MBP AntiBodies, presence of MBP material in the CSF and concentration of sVCAM-1, ICAM-1, PECAM-1 in CSF and Serum.

We studied in vitro production of Interferon- and Interferon- by peripheral blood Leukocytes from 15 patients with Multiple Sclerosis.

The priming effects of Interferon preparations weakly correlated with Interferon- production by Leukocytes from patients with Multiple Sclerosis, but negatively correlated with Interferon- production.

The effects of Interferon inducers in most cases positively correlated with its spontaneous production.

We found a weak positive correlation between the priming effect of natural Interferon- and the effect of recombinant Interferons.

There were positive or strong positive correlations between the effects of recombinant Interferons on Leukocytes from patients with Multiple Sclerosis.

The relationship between the effects of medicinal Interferon inducers and Interferon preparation varied from negative to strong positive correlations.

These data suggest that correlation analysis can be used for dynamic control and elaboration of methods for combined ImmunoTherapy of Multiple Sclerosis with various Interferon preparations or Interferon and its inducers.

Twenty-three plaques obtained at early autopsy from 2 patients with Secondary/Progressive Multiple Sclerosis were examined ImmunoHistoChemically for Microglia/Macrophages, and for ImmunoGlobulins and components of activated Complement.

Most of the lesions examined in both cases exhibited evidence of low-grade active DeMyelination of an unusual type (frustrated Phagocytosis) in Periplaque White Matter.

This included linear groups of Microglia engaging short segments of disrupted Myelin that were associated with deposits of C3d, an Opsonin formed during Complement activation.

Similar Microglia/C3d/Myelin profiles were not observed in newly forming lesions in cases of acute Multiple Sclerosis or Other Central White Matter Diseases.

As C3d coupling is known to increase the Immunogenicity of potential Antigens enormously, present findings point to disrupted Myelin close to plaques as a possible source of the putative Multiple Sclerosis Antigen.

Ongoing Myelin destruction found in a high proportion of old, established plaques was surprising.

It suggests that slowly expanding lesions (progressive plaques), in which ongoing Myelin breakdown occurs in the absence of florid PeriVascular Cell cuffing.

Or other Histological signs of acute inflammation, contribute to disease progression in cases of Secondary/Progressive Multiple Sclerosis.

Background
As a parameter for the evaluation of the Vestibular Evoked Myogenic Potential (VEMP), amplitude has been used clinically. However, the significance of latency has not been considered.

Objective & Design & Setting
To clarify the diagnostic value of latencies of the VEMP.

Data were collected from patients in an outpatient clinic of a tertiary care center and healthy volunteers.

Subjects & Intervention
Diagnostic Clinical records of 134 patients (61 men and 73 women, aged 20-75 years) were reviewed.

Diagnoses were Meniere Disease in 43 patients, Acoustic Neuroma in 62 patients, Vestibular Neuritis in 23 patients, and Multiple Sclerosis in 6 patients.

Also, 18 healthy volunteers (13 men and 5 women, aged 25-38 years) were enrolled.

Main Outcome Measures
Click-evoked myogenic potentials were recorded with surface electrodes over each SternocleidoMastoid muscle. Latencies and amplitudes of responses were measured.

Results
Vestibular Evoked Myogenic Potentials were absent or decreased in 51% of patients with Meniere Disease (n = 22), 39% with Vestibular Neuritis (n = 9), 77% with Acoustic Neuroma (n = 48), and 25% with Multiple Sclerosis (3 of 12 sides of 6 patients).

Concerning latency, patients with Meniere Disease or Vestibular Neuritis hardly showed any latency prolongation. Four patients with Acoustic Neuroma showed prolonged p13; all had large tumors.

All patients with Multiple Sclerosis showed prolonged p13.

Conclusion
Prolonged latencies of the VEMP suggest lesions in the RetroLabyrinthine, especially in the VestibuloSpinal Tract.

In Japanese, susceptibility to the conventional form of Multiple Sclerosis (C-MS) is associated with the HLA-DRB1*1501-DRB5*0101 haplotype.

While susceptibility to the OpticoSpinal form of MS (OS-MS) is associated with HLA-DPA1*0202-DPB1*0501.

To clarify the characteristics of T-Cells AutoReactive to Myelin proteins in each MS subtype, we established T-Cell lines reactive to such Myelin Antigens as Myelin Basic Protein (MBP), ProteoLipid Protein (PLP) and Myelin Oligodendrocyte Glycoprotein (MOG).

From 5 of 10 OS-MS patients, 6 of 11 C-MS patients and 7 of 13 healthy controls (HCs), and T-Cell Epitopes and their restriction molecules were determined.

We found that:
1. Intermolecular Epitope spreading was found to be significantly more frequent in MS patients than in HCs (P=0.0128),

2. Intramolecular Epitope spreading also tended to occur more frequently in MS patients than in HCs (P=0.0584)

3. In OS-MS, HLA-DR-restricted and MOG-AutoReactive T-Cells were more frequently established as compared with those reactive to MBP or PLP Epitopes

4. In C-MS, HLA-DQ-restricted and PLP-AutoReactive T-Cells dominated those AutoReactive to MBP or MOG Epitopes

A DPB1*0501-restricted MBP-reactive T-Cell clone from a patient with OS-MS provided evidence that the first HLA Class II anchor Amino Acid of Peptide bound to disease-susceptible DP5 molecule was distinct from that for the DR2 molecule.

Taken together, these differences in specificities of Myelin-AutoReactive T-Cells between C-MS and OS-MS as well as the difference in the anchor motif of the binding Peptides.

Between each MS subtype-susceptible HLA Class II molecule may contribute to the development of distinct clinical phenotypes.

Multiple Sclerosis (MS) is considered an AutoImmune disease that is mediated by proinflammatory T-Helper-1 Lymphocytes.

The putative mechanism of Interferon-beta (IFN-ß), includes the inhibition of T-Cell proliferation, blocking of Blood-Brain Barrier opening and T-Cell transmigration into the Brain.

Via interference with Cell Adhesion, and the upregulation of anti-inflammatory Cytokines.

In the present study, a Gene expression analysis of IFN-ß-treated peripheral blood MonoNuclear Cells by cDNA microarray documents the broad effects of IFN-ß that are not purely anti-inflammatory.

Specifically, we addressed the effect of IFN-ß on T-Helper-1 differentiation- or lineage markers such as the IL-12 Receptor ß2 chain and the Chemokine Receptor CCR5 that have been implicated in the PathoGenesis of MS.

Both markers were significantly upregulated in vitro and in vivo under IFN-ß therapy, supporting that this Cytokine exerts complex effects on the Immune System.

The combination of cDNA microarray and quantitative PCR will expand our knowledge of the Immunological effects of such PleioTropic agents as IFN-ß.

May provide a key to why certain patients fail to respond, and eventually may influence our view of the disease PathoGenesis.

Ocular flutter is a rare horizontal eye movement disorder characterized by rapid Saccadic oscillations.

It has been hypothesized that it is caused by loss of "pause" Neuronal Inhibition of "burst" Neuron function in the ParaMedian Pontine Reticular Formation (PPRF).

However, there have been no imaging studies confirming such anatomical localization.

We report the case of a woman with an acute attack of Multiple Sclerosis associated both with Ocular flutter and a circumscribed Pontine lesion, mainly involving the PPRF on Magnetic Resonance Imaging.

As she recovered from the attack, both the MidLine Pontine lesion and the Ocular flutter dramatically improved.

This case is the first clear evidence that at least some cases of Ocular flutter are due to lesions involving the PPRF.

The presence of OligoClonal Bands (OCB) of ImmunoGlobulin G (IgG) is in our days the most useful finding in the study of the CSF for the diagnosis of Multiple Sclerosis (MS).

The most sensitive method for the detection of OCB is the IsoElectric focusing followed by ImmunoBloting. The prevalence of OCB changes in different populations with a rank of results from 60 to 95 97%.

We have determinated the prevalence of OCB in our population and the sensitivity and the specificity of the technique used in our laboratory. We have included 391 patients in whom we analyzed the presence of OCB, subdivided in:
Group 0:
• Diagnosed of MS
Group 1:
• First episode of DeMyelinating process
Group 2:
• Neurological disorders considered NonInflammatory or NonAutoImmune (NINA)
Group 3:
• Neurological Disorders considered Inflammatory, Infectious or AutoImmune (IIA)

The presence of OCB was searched in CSF and Serum simultaneously using IsoElectric Focusing and ImmunoBlotting. In order to standardize the technique we achieved and internal and external validation.

Internal validation: sensitivity and specificity (using as a control group first the group NINA and after the group IA).

External validation: we choose 10 pairs of CSF/Serum from patients with different diagnostics and sent to a reference laboratory (Karolinska Institute Medical School) that was blind of our results and of the diagnostics.

The prevalence of OCB in each group has been:

1. Group 0 (MS): 87.7%
2. Group 1: 54.8%
3. Group 2 (NINA): 17.5%
4. Group 3(IIA): 52.7%

Sensitivity: 97.7%, specificity using group NINA as control 82.5% and using group IIA 45.7%.

Concordance with the reference laboratory in 9/10 determinations. We conclude that in our population the prevalence of OCB, in patients with MS, is lower than in Northern Europe.

The OCB appear in may Inflammatory, AutoImmune Diseases, their specificity for the diagnostic of MS is low.

IntraCerebral infection of mice with Mouse Hepatitis Virus (MHV) results in an acute EncephaloMyelitis.

Followed by a chronic DeMyelinating Disease with clinical and histological similarities with the human DeMyelinating Disease Multiple Sclerosis (MS).

Following MHV infection, Chemokines including CXC Chemokine Ligand (CXCL)10 (IFN inducible protein 10 kDa), CXCL9 (Monokine induced by IFN-), and CC Chemokine Ligand 5 (RANTES) are expressed during both acute and chronic stages of disease.

Suggesting a role for these molecules in disease exacerbation. Previous studies have shown that during the acute phase of infection, T-Lymphocytes are recruited into the CNS by the Chemokines CXCL10 and CXCL9.

In the present study, MHV-infected mice with established DeMyelination were treated with AntiSera against these two Chemokines, and disease severity was assessed.

Treatment with Anti-CXCL10 reduced CD4⁺ T-Lymphocyte and Macrophage invasion, diminished expression of IFN- and CC Chemokine Ligand 5, inhibited progression of DeMyelination, and increased ReMyelination.

Anti-CXCL10 treatment also resulted in an impediment of clinical disease progression that was characterized by a dramatic improvement in Neurological function.

Treatment with AntiSera against CXCL9 was without effect, demonstrating a critical role for CXCL10 in Inflammatory DeMyelination in this model.

These findings document a novel therapeutic strategy using Ab-mediated neutralization of a key Chemokine as a possible treatment for chronic human Inflammatory DeMyelinating Diseases such as MS.

CerebroSpinal Fluid (CSF) analysis aids in the diagnosis of Multiple Sclerosis. However, this examination is invasive. The aim of this study was to assess the potentials of a new method of tears IsoElectroPhoresis (IEF).

Silver staining of IEF was used to examined tears and CSF from 123 patients including 60 patients with Multiple Sclerosis (MS), 50 Other Neurological patients and 13 patients with Inflammatory Neurological Diseases.

Tears were collected on a Shirmer strip placed in one eye, avoiding reflex secretion.

This method of IEF with silver staining allowed the detection of OligoClonal Bands in tears that were truly ImmunoGlobulin G on ImmunoFixation.

The concordance rate between tears and CSF was 83%, meaning that CSF provided no more information than tears analysis in 83% of cases.

Sensitivity in tears (72%) and CSF (75%) was very close as was specificity (respectively 84 % and 86 %).

High concordance between tears and CSF is the first step in developing a non invasive test which could replace Lumbar Puncture, particularly when this procedure is not fearalele or is refused by the patient.

Background
Multiple Sclerosis is a chronic DeMyelinating Disease rare in children. Currently marketed disease modifying therapies are limited to adults.

Objective
To determine the tolerability of Interferon-beta-1a (IFN-ß-1a) 30 mcg injected intramuscularly once a week in children with Clinically Definite Relapsing/Remitting Multiple Sclerosis (RRMS).

Design/Methods
A standardized questionnaire was sent to Neurologists in the United States to determine the tolerability of IFN-ß-1 a in patients younger than 16 years.

Results
Tolerability data were available for 9 of 33 children who were reported to initiate IFN-ß-1 a.

Mean age on initiating treatment was 12.7 years (range 8 - 15) and mean duration of therapy was 17 months (range 5 - 36). No patient discontinued therapy due to an adverse event.

Conclusions
Preliminary data indicate that weekly intramuscular injections of IFN-ß-1a a are well tolerated.