MS Abstracts 05e-2g1

Objective
To assess Endothelial Dysfunction in patients with MS and to investigate whether Plasma from patients with MS induces Endothelial Cell Dysfunction in vitro.

Background
Endothelial Cell dysfunction may contribute to the PathoGenesis of MS.

Elevations of Soluble Adhesion Molecules, IntraCellular Adhesion Molecule, Vascular Cell Adhesion Molecule, and Platelet-Endothelial Cell Adhesion Molecule-1 (CD31), have been reported as markers of Blood-Brain Barrier (BBB) damage in MS.

But direct assay of Endothelium has been difficult. Endothelial Cells release MicroParticles < approximately 1.5 &mgr;m (EMP) during activation or Apoptosis.

The authors developed a flow cytometric assay of EMP and studied EMP as markers of Endothelial damage in MS.

Methods
Platelet-Poor Plasma (PPP) from 50 patients with MS (30 in exacerbation and 20 in remission) and 48 controls were labeled with Fluorescein IsoThioCyanate (FITC)-conjugated anti-CD31 and anti-CD51 (vitronectin receptor) AntiBodies.

And two classes of EMP (CD31⁺ and CD51⁺) were assayed by flow cytometry.

For in vitro studies, patients' Plasma was added to the MicroVascular Endothelial Cell (MVEC) culture and release of CD31⁺ and CD51⁺ EMP were measured in the supernatant.

Results
Plasma from patients in Exacerbation had 2.85-fold elevation of CD31⁺ EMP as compared with healthy controls, returning to near control value during remission.

The CD31⁺ EMP concentration showed a positive association with Gadolinium enhancement in patients with MS. In contrast, CD51⁺+ EMP remained elevated in both exacerbation and remission.

This suggests that CD31⁺ EMP is a marker of acute injury, whereas CD51⁺+ EMP reflects chronic injury of Endothelium.

MS Plasma induced release of both CD31⁺ and CD51⁺+ EMP from MVEC culture in vitro.

Conclusion
Endothelial Dysfunction is evident during exacerbation of MS, evidenced by shedding of EMP expressing PECAM-1 (CD31). The in vitro data indicate contribution of one or more Plasma factors in Endothelial Dysfunction of MS.

The present review of the recent literature focuses on Antigen-specific Immune reactions in Multiple Sclerosis. New techniques have allowed precise quantitative analysis of the Antigen-receptor repertoire of infiltrating T-Cells in the Multiple Sclerosis Brain.

Novel candidate AutoAntigens, including B-Cell AutoAntigens, have been identified. 'Humanized' animal models allow the functional characterization of human Immune molecules in vivo.

Finally, several therapeutic trials have recently assessed the clinical benefit of selective ImmunoTherapies.

DeMyelination and inflammation both contribute to the Neurological deficits characteristic of Multiple Sclerosis and Guillain-Barre Syndrome.

Conduction deficits attributable to DeMyelination are well known, but it is becoming clear that factors such as Nitric Oxide, Endocaine, Cytokines, and AntiGanglioside AntiBodies also play significant roles.

DeMyelination directly affects Conduction and also causes changes in both the distribution and repertoire of expressed Axolemmal Ion Channels, which in turn affect Impulse propagation and can promote HyperExcitability.

In conducting Axons, sustained trains of Impulses can produce intermittent Conduction Failure, and, in the presence of Nitric Oxide exposure, can also cause Axonal Degeneration.

Other factors impairing Impulse transmission include Nodal widening, Glutamate toxicity, and disturbances of both the Blood-Brain Barrier and Synaptic transmission.

Magnetic Resonance Imaging methods allow observation of pathological changes in vivo.

Magnetic Resonance-based studies have provided a number of important insights into the spatio-temporal evolution of the pathology of Multiple Sclerosis in vivo, particularly with respect to the relation between pathology and progression of disability.

Magnetic Resonance techniques have shown that this pathology is not restricted to the plaques that are evident at autopsy, but also involve the so-called Normal-Appearing White Matter.

NonConventional Magnetic Resonance Imaging strategies such as Magnetization Transfer Imaging and Spectroscopic Imaging provide measures with higher pathological specificity for Myelin and Axonal injury.

These and other advanced Magnetic Resonance techniques (such as the measurement of Atrophy, lesion relaxation spectra, and lesion dynamics) are affording opportunities to use observations of patients to test biologically specific hypotheses.

This should help us to better define new targets for drug therapy and to assess responses to new therapeutic agents.

The association between susceptibility to Multiple Sclerosis (MS) and the Class II MHC allele HLA-DRB1*15 is well established although a possible relationship between this allele and outcome in MS is less clear.

HLA-DRB1 typing was performed on 375 unrelated white patients with Clinically Definite MS and on 367 healthy controls.

Putative associations of the Gene with outcome were examined by dividing patients into two groups: those with an EDSS of 0-5.5 (mild/moderate disease) and those with an EDSS of 6-10 (severe disease).

In order to minimise the effects of disease variability patients with a disease duration of at least 10 years or 15 years were examined.

As subsidiary HLA-DRB1*03 and HLA-DRB1*04 associations have been previously reported, the effect of these alleles was also examined. As expected, HLA-DRB1*15 was found more frequently in patients than in controls (P < 0.000001).

HLA-DRB1*15 positive patients had a significantly earlier age at onset than HLA-DRB1*15 negative patients.

No significant associations were noted between HLA-DRB1*15 and outcome in the total patient group or in patients with a disease duration of 10 years or longer.

In patients with a disease duration of at least 15 years HLA-DRB1*15 negative status was associated with a worse prognosis, although this did not remain significant after correction for multiple testing.

It is thus likely that the contribution of HLA in MS is primarily towards onset and initial triggering mechanisms rather than influencing disease progression, chronicity and severity.

Autonomic Dysfunction has rarely been studied in patients suffering from Multiple Sclerosis (MS). Some hypotheses have concerned the PathoPhysiology, especially with regard to a possible Spinal Cord origin.

However, there have been no previous studies on Autonomic Dysfunction in MS and Spinal Cord Lesions.

This study assessed the frequency of Autonomic Dysfunction (AD) in MS and the correlation to Spinal Cord Magnetic Resonance Imaging (MRI) findings.

We prospectively studied 75 MS patients:

1. 25 with Relapsing/Remitting forms
2. 25 with Secondary/Progressive forms
3. 25 with Primary/Progressive forms

We performed Sympathetic Skin Response, R/R interval variability and Orthostatic Hypotension testing. Spinal Cord MRI was performed to detect DeMyelinating lesions (sagittal and axial plane) or Spinal Cord Atrophy .

Clinical and laboratory evidence of AD was found in 84% and 56% of MS patients, respectively. The correlation of the latter with Disability was evaluated using the Expanded Disability Status Scale.

AD was more frequent in Primary/Progressive MS than in the other two forms. AD was correlated with Spinal Cord Cross-Sectional Area reduction but not with Spinal Cord HyperIntensities.

This study confirms that the frequency of AD in MS, especially in Primary/Progressive forms, has until now been underestimated. Furthermore, AD appears to be more closely related to Axonal loss, as demonstrated by Spinal Cord Atrophy, than to DeMyelinating lesions.

Inflammatory Optic Neuropathies are common in clinical practice. MonoSymptomatic Optic Neuritis has important implications for the development of Multiple Sclerosis.

In the patient presenting with MonoSymptomatic Optic Neuritis, MR imaging provides critical prognostic information concerning the development of MS.

The ONTT provided valuable information regarding the natural history and therapy of Optic Neuritis.

Oral Prednisone alone is contraindicated in the treatment of Optic Neuritis because of its association with increased recurrence rate of Optic Neuritis.

Intravenous MethylPrednisolone remains a viable treatment option to slightly increase the rate of recovery and provide a degree of short-term protection against the subsequent development of MS.

Other Inflammatory Optic Neuropathies include Sarcoidosis, NeuroRetinitis, and Devic's Disease with each possessing distinct clinical characteristics and treatment approaches.

Optic Neuritis is a clinical syndrome resulting from inflammation, DeMyelination, or infection of the Optic Nerve. Its diagnosis and treatment are complicated.

In 1884, Nettleship first reported 28 cases of Optic Neuritis whose clinical symptoms have been accepted up to the present without any change.

On the other hand, the development of diagnostic procedures and Steroid therapy have also altered the clinical features of Optic Neuritis.

Among several developed diagnostic procedures, the Visually Evoked Cortical Potential (VECP) has become a good tool to prove the impairment of the Optic Nerve.

In 1971, we reported a decrease of threshold intensity required to evoke VECPs in Optic Neuritis patients whose Visual Acuity was relatively well preserved.

In the same year, Halliday et al reported that Pattern VECP (PVECP) was delayed in 93% of patients with multiple sclerotis (MS) without Optic Neuritis.

Stimulated by this report, a great number of studies appeared to show the usefulness of PVECP in the diagnosis of MS.

However, few of these studies gave descriptions of Ophthalmic findings. PVECP later become known to be closely related with Ophthalmic conditions.

In the Ophthalmological field, we reported the influence of Pupillary size, accommodation power, refractive powers, eccentricity of stimulated Retinal area, Retinal luminance, contrast, wavelengths, spatial and temporal frequencies, stimulus field, etc.

On the basis of our results, we developed a television display system in 1975 and applied it clinically. In the present study, we reviewed the medical records of a total of 272 cases of Optic Neuritis who presented in our clinic between 1978 and 1999.

In the diagnostic, therapeutic point of view in relation with the data of other countries, the study was important regarding the racial differences and recent conceptions of Optic Neuritis.

The results showed that there were no racial difference in Optic Neuritis as had been thought. The development from Optic Neuritis to Multiple Sclerosis was not less than in Caucasian patients.

Regarding Steroid therapy, we found that the most effective method was sub-Tenon injection. For cases which recur and progress to Optic Atrophy, Optic Nerve transplantation will be needed.

Therefore, we have been studying the reconstruction of the Optic Nerve in Wister rats. We experimentally damaged the Ganglion Cells by causing Ischemic Retina with ligation of the Ophthalmic Artery and cutting the Optic Nerve just behind the eyeball.

To prevent the Apoptosis of Ganglion Cells, we injected various NeuroTrophic Factors such as BDNF, GDNF, and HSP 27 into the vitreous.

For effective injection of DNA, electropolation was applied and the best condition for avoiding Apoptosis was chosen. Further, in Mx-c-fox transgenic mice, we found that regeneration of Ganglion Cells was inhibited.

Based on the rescue study of the Ganglion Cells, Optic Nerve transplantation was performed using an artificial graft in which cultured Schwann Cells from the Ischiatic Nerve, BDNF, CNTF, Insulin, and Forscolin were compound and bridged to the Superior Colliculus.

The results showed a regeneration rate of the Optic Nerve Axon of 15%. This rate was much higher than in other reports.

Keratoplasty and IntraOcular Lens implantation had a relatively long history of research before achieving clinical success. We believe that Optic Nerve transplantation will one day be successful in clinical treatment in the same way.

Persons with Multiple Sclerosis (MS) commonly experience restrictions in mobility and everyday functional activities. A wide range of factors including Physical, Psychological, Environmental and Economic issues may contribute to these difficulties.

This is particularly the case as the disease evolves, and the Impairments and Disabilities become more numerous, inter-related and hence more complex.

Effective management of these complex problems requires assessment and intervention from a variety of different perspectives by using a coordinated, goal-oriented, multi-disciplinary management approach.

Crucially, it requires management to be considered from a long-term perspective rather than as a fragmented series of isolated "quick-fixes".

Interferon-ß is a remarkably Pleiotropic molecule. AntiViral, Pro- and AntiProliferative, Pro- and AntiApoptotic, and complex ImmunoRegulatory activities have all been described.

The precise mechanism(s) that underlie the beneficial effects of Interferon-ß in Multiple Sclerosis remain poorly understood; this has hindered progress in the search for more effective therapies.

An increasing body of literature supports the hypothesis that Interferon-ß-mediated changes in the production and activities of the ImmunoRegulatory Cytokines IL-12 and IL-10 are important to the therapeutic benefits of Interferon-ß in Multiple Sclerosis. These data are reviewed here.

ProteoLipid Protein (PLP) is the most abundant Protein of CNS Myelin, and is PostTranslationally Acylated by covalent attachment of Long Chain Fatty Acids to Cysteine residues via a Thioester linkage.

Two of the Acylation sites are within Epitopes of PLP that are Encephalitogenic in SJL/J mice (PLP(104-117) and PLP(139-151)) and against which increased Immune Responses have been detected in some Multiple Sclerosis patients.

It is known that attachment of certain types of Lipid side chains to Peptides can result in their enhanced ImmunoGenicity.

The aim of this study was to determine whether ThioAcylated PLP peptides, as occur in the native protein, are more ImmunoGenic than their NonAcylated counterparts.

And whether ThioAcylation influences the development of AutoReactivity and Experimental AutoImmune EncephaloMyelitis.

The results show that in comparison with NonAcylated Peptides, ThioAcylated PLP LipoPeptides can induce greater T-Cell and Ab responses to both the Acylated and NonAcylated Peptides.

They also enhanced the development and chronicity of Experimental AutoImmune EncephaloMyelitis.

Synthetic Peptides in which the Fatty Acid was attached via an amide linkage at the N terminus were not Encephalitogenic, and they induced greater proportions of CD8⁺ Cells in initial in vitro stimulation.

Therefore, the lability and the site of the linkage between the Peptide and Fatty Acid may be important for induction of Encephalitogenic CD4⁺ T-Cells.

These results suggest that Immune Responses induced by endogenous ThioAcylated LipoPeptides may contribute to the ImmunoPathogenesis of Chronic Experimental DeMyelinating Diseases and Multiple Sclerosis.