MS Abstracts 7d-2g1

Recently evidence has been provided for a Genetic control of T-Cell dependent Cytokine production by HLA-Class II.

Candidate Genes in Multiple Sclerosis, a T-Cell mediated AutoImmune Disease, are the disease-associated DR2, DQ6, Dw2 haplotype.

Previous observations by us and others imply a HLA-DR2 dependent propensity of Antigen-specific T-Cell lines to produce increased amounts of TNF-/ß.

Here, we tested a possible association between HLA or disease status with Cytokine production employing the simple and widely used method of bulk cultures.

Peripheral blood cells of 48 patients and 68 healthy individuals were analyzed.

We observed no significant differences of the Cytokine production in relation to disease status or any HLA polymorphism.

Our data indicate that, in contrast to MonoClonal T-Cell cultures, bulk cultures are not suitable to detect ImmunoGenetic control of T-Cell function.

Background
Although the exact Etiology of MS remains elusive, there is good evidence that Genetic factors play an important role.

These factors are likely to be polygenic, exerting both independent and interactive effects on the expression of MS. They may determine susceptibility and/or shape the clinical course.

Methods
The authors studied clinical phenotype in 245 concordant parent-child pairs recruited from a national register of familial disease over a 10-year period.

Data were examined in order to determine the effect of parental sex on expression of disease in the offspring.

Results
Allowing for the observed sex ratio of 2.6 F:1 M in this group of patients, sex pairings of parents and offspring were close to those expected.

When assessed independently there was no evidence that either the sex of the affected offspring or the line of inheritance influenced disability, age at onset, or disease course.

However, trends were observed toward greater disability and an increased frequency of Primary/Progressive Disease in offspring of affected Fathers and an earlier age at onset in offspring of affected Mothers.

The highest mean Expanded Disability Status Scale score was observed in male offspring of affected Fathers (5.64) and this group was also more likely to have Primary/Progressive Disease (OR 1.92).

Thirty-one percent of families had an additionally affected offspring with no preferential Maternal or Paternal transmission.

Conclusions
In offspring of concordant parent-child families with MS who are at high risk of inheriting increased numbers of susceptibility Genes.

There is no evidence for a parent of origin effect distorting sex ratios in affected offspring.

But parent of origin may influence disability and disease course as well as increasing the risk to additional offspring within the same family.

The mechanism of these effects is not clear but may result from interactions between Genes encoded at different Loci (Epistasis), which each independently influencing susceptibility and phenotype.

Introduction
Magnetic Resonance Imaging (MRI) lesion load is widely used in the clinical evaluation of patients with Multiple Sclerosis (MS), but little is known about the associated changes in Cortical activation.

For this purpose, we studied the association between the CorticoCerebral metabolic rate of Glucose (CMRglc) and the MRI T₂-weighted total lesion area (TLA).

In addition, we investigated the correlation between Cognitive and Neurological disability and CMRglc.

Methods
Twenty-three patients with Clinically Definite MS underwent measurements of the CMRglc, TLA, Motor-Evoked Potentials (MEP), and Cognitive and Neurological disability.

CMRglc was calculated with Positron Emission Tomography (PET) and 18-F-deoxyglucose (FDG) and compared to that of nine healthy controls.

Results
A reduction in CMRglc (p < 0.01) was found in Cortical global and regional lobar measurements.

Furthermore, regional CMRglc (rCMRglc) was reduced in the DorsoLateral PreFrontal Cortex, OrbitoFrontal Cortex, Caudate, Putamen, Thalamus, and Hippocampus.

Global Cortical CMRglc correlated with TLA (rho = -0.66; p = 0.001), and rCMRglc correlated with the regional lesion load in all Cerebral Lobes (p < or = 0.05).

Global Cortical CMRglc and Cognitive disability were also correlated (rho = 0.58; p = 0.015), and stepwise regression analysis showed a significant association between rCMRglc of the Right Thalamus and Cognitive performance, as well as the TLA.

There was no correlation between CMRglc and Neurological disability (Expanded Disability Status Scale [EDSS]) or MEP.

Conclusion
Global and regional Cortical CMRglc is significantly reduced in patients with MS compared to healthy controls.

The reductions in CMRglc furthermore correlate with the TLA, as well as with Cognitive Dysfunction, which indicates that MRI White Matter lesion burden has a deteriorating effect on CorticoCerebral Neural function.

We report a 62-year-old woman with a past history of painful Central Visual loss who developed progressive QuadriParesis and Bulbar Palsy.

Neurological examination revealed widespread Upper and Lower Motor Neuron signs in the Bulbar region and extremities.

ElectroMyography demonstrated widespread active and chronic Motor Axon loss.

Magnetic Resonance NeuroImaging studies revealed enhancing Callosal and PeriVentricular White Matter lesions and Cervical and Thoracic Cord HyperIntensities.

CerebroSpinal Fluid analysis was consistent with Multiple Sclerosis. The patient died of respiratory failure two years after presentation.

And autopsy revealed multifocal DeMyelination involving the Corpus Callosum, Cerebellum and Spinal Cord as well as pathologic findings typical of Amyotrophic Lateral Sclerosis.

A review of the literature confirms the exceedingly unusual combination of Amyotrophic Lateral Sclerosis with Multiple Sclerosis.

Changes were assessed in Serum levels of soluble InterLeukin-2 Receptor (sIL-2R alpha) and soluble InterCellular Adhesion Molecule-1 (sICAM-1), indirect indices of activation of Immunological System, in the course of Multiple Sclerosis (MS).

12 patients (av. age 39.2 +/- 9.4 y.) with the first relapse that fulfilled criteria of clinical Probable MS according to Poser Committee were included into the study.

Blood samples were taken at the beginning of the relapse and then every 2-month periods. Simultaneously, Neurological Impairment (EDSS scale) was assessed.

When the next relapse occurred examination was repeated from the beginning. The total time of observation was between 12 and 18 months. The levels of both soluble molecules were examined with ELISA test.

In relapse mean Serum levels of sIL-2R alpha and sICAM-1 were significantly elevated in comparison to the results obtained in remission (respectively: p < 0.001 and p = 0.03).

Changes in Serum level of both soluble molecules during the first 2 months after relapse were significantly higher than in subsequent 2-month periods (p < 0.001).

Each relapse was accompanied by elevation of Serum levels of sIL-2R alpha and sICAM-1.

There was no obtainable correlation between improvement in EDSS scale and changes in sIL-2R alpha level during the whole time of observation.

Improvement in EDSS scale was correlated with lowering of sICAM-1 but only during the first two months after relapse.

Conclusions
Serum level of sIL-2R alpha and sICAM-1 in MS patients during relapse is significantly higher in comparison to the results obtained during remission.

Each relapse is accompanied by elevation of sIL-2R alpha and sICAM-1 in Serum, during remission Serum levels of both molecules do not changed significantly.

Purpose
To report the clinical features and outcome of a series of children with Optic Neuritis.

Methods
We reviewed the medical records of patients up to 16 years old with Optic Neuritis.

Group 1 comprised children seen up to two weeks after the onset of Visual loss.

Group 2 comprised patients already harboring Optic Atrophy.

Results
There were 15 boys and 12 girls. The mean age was 10.9 years.

Bilateral Optic Neuritis occurred in 10. Optic Disc pallor was found in 35%, Edema in 46%, and 19% had normal Fundus.

During follow-up Visual Acuity improved in all but one eye in Group 1, and in six of seven eyes in children in Group 2. Just one child converted to Multiple Sclerosis.

Conclusions
This study shows that the clinical features of childhood Optic Neuritis differ from those observed in adults.

In children it has a better Visual outcome and a lower conversion rate to Multiple Sclerosis than in adults.

In this study, we demonstrate that disruption of NeuroEndocrine signaling is a major factor driving disease progression in Myelin Oligodendrocyte Glycoprotein-induced Chronic Relapsing Experimental AutoImmune EncephaloMyelitis, an animal model of Multiple Sclerosis.

Although the initial episode of Chronic Relapsing Experimental AutoImmune EncephaloMyelitis is associated with a robust HypoThalamic-Pituitary-AdrenoCortical Axis response.

We show that subsequent disease progression is associated with a selective desensitization of HypoThalamic-Pituitary-AdrenoCortical responsiveness to inflammatory mediators.

Inflammatory activity in the Central Nervous System during relapse is therefore unable to produce an endogenous ImmunoSuppressive CorticoSterone response, and disease progresses into an ultimately lethal phase.

However, disease progression is inhibited if the circulating CorticoSterone level is maintained at levels seen during the initial phase of disease.

The effect of HypoThalamic-Pituitary-AdrenoCortical Axis desensitization on the clinical course of Experimental AutoImmune EncephaloMyelitis is aggravated by a marked reduction in ProInflammatory Cytokine synthesis in the CNS.

In the later stages of disease, reflecting an increasing involvement of AntiBody, rather than T-Cell-dependent Effector mechanisms, in disease PathoGenesis, with time.

Thus, our data indicate that distinct Immune-Endocrine effects play a decisive role in determining disease progression in Multiple Sclerosis.

A concept supported by reports that a subpopulation of Multiple Sclerosis patients shows evidence of HypoThalamic-Pituitary-AdrenoCortical Axis desensitization.

Macroscopic sampling of Multiple Sclerosis lesions in the Brain tends to find chronic lesions.

For a better understanding of the dynamics of the Multiple Sclerosis disease process, research into new and developing lesions is of great interest.

As MRI in vivo effectively demonstrates lesions in Multiple Sclerosis patients, we have applied it to unfixed post-mortem Brain slices to identify abnormalities, in order to obtain a higher yield of active lesions.

The Netherlands Brain Bank organized the rapid autopsy of 29 Multiple Sclerosis patients. The Brain was cut in 1 cm Coronal slices.

One or two slices were subjected to T₁- and T₂-weighted MRI, and then cut at the plane of the MRI scan into 5 mm thick opposing sections.

Areas of interest were identified based on the MRI findings and excised. One half was fixed in 10% Formalin and paraffin-embedded, and the corresponding area in the adjacent half was snap-frozen in liquid Nitrogen.

In total, 136 out of 174 Brain tissue samples could be matched with the abnormalities seen on T₂-weighted MRIs. The stage of lesional development was determined ImmunoHistoChemically.

For 54 MRI-detectable samples, it was recorded whether they were macroscopically detectable, i.e. visible and/or palpable.

HistoPathological analysis revealed that 48% of the HyperIntense areas seen on T₂-weighted images represented active lesions, including lesions localized in the Normal-Appearing White Matter, without apparent loss of Myelin.

But nevertheless showing a variable degree of Edema, small clusters of Microglial Cells with enhanced Major Histocompatibility Complex Class II Antigen, CD45 and CD68 Antigen expression.

And a variable number of PeriVascular Lymphocytes around small blood vessels [designated as (p)reactive lesions].

From the macro-scopically not-visible/not-palpable MRI-detected abnormalities, 58% were (p)reactive lesions and 21% contained active DeMyelinating lesions.

In contrast, visible and/or palpable Brain tissue samples mainly contained chronic inactive lesions.

We conclude that MRI-guided sampling of Brain tissue increases the yield of active Multiple Sclerosis lesions, including active DeMyelinating and (p)reactive lesions.

Molecular mimicry is the process by which Virus infection activates T-Cells that are cross-reactive with self Antigens.

Infection of SJL/J mice with the NeuroTropic PicornaVirus Theiler's Murine EncephaloMyelitis Virus (TMEV) leads to a progressive CD4⁺ T-Cell-mediated DeMyelinating disease similar to Multiple Sclerosis.

To study the potential of Virus-induced molecular mimicry to initiate AutoImmune DeMyelination, a nonpathogenic TMEV variant was engineered to encode a 30-mer Peptide encompassing the ImmunoDominant Encephalitogenic Myelin ProteoLipid Protein (PLP139-151) Epitope.

Infection with the PLP139-151-encoding TMEV led within 10-14 days to a rapid-onset Paralytic DeMyelinating Disease characterized by PLP139-151-specific CD4⁺ Th1 responses; insertion of a non-self ovalbumin sequence led to restoration of the normal late-onset disease.

Early-onset disease was also observed in mice infected with a TMEV encoding PLP139-151 with an Amino Acid substitution at the secondary T-Cell receptor (TCR) contact residue (H147A).

But not in mice infected with TMEV encoding a PLP139-151 substitution at the primary TCR contact (W144A).

Most significantly, mice infected with TMEV encoding a Haemophilus influenzae mimic Peptide, sharing only 6 of 13 Amino Acids with PLP139-151, displayed rapid-onset disease and developed cross-reactive PLP139-151-specific CD4⁺ Th1 responses.

To our knowledge, this is the first study showing that a naturally infectious Virus encoding a Myelin Epitope mimic can directly initiate organ-specific T-Cell-mediated AutoImmunity.

The exclusive detrimental role of ProInflammatory Cytokines in DeMyelinating Diseases of the CNS, such as Multiple Sclerosis, is controversial.

Here we show that the Intrathecal delivery of an HSV-1-derived vector engineered with the mouse IFN- Gene leads to persistent (up to 4 wk) CNS production of IFN-.

And inhibits the course of a Chronic/Progressive form of Experimental AutoImmune EncephaloMyelitis (EAE) induced in C57BL/6 mice by Myelin Oligodendrocyte Glycoprotein (MOG)(35-55).

Mice treated with the IFN--containing vector before EAE onset, showed an earlier onset but a milder course of the disease compared with control mice treated with the empty vector.

In addition, 83% of IFN--treated mice completely recovered within 25 days post immunization, whereas control mice did not recover up to 60 days post immunization.

Mice treated with the IFN--containing vector within 1 wk after EAE onset partially recovered from the disease within 25 days after vector injection, whereas control mice worsened.

Recovery from EAE in mice treated with IFN- was associated with a significant increase of CNS-infiltrating Lymphocytes undergoing Apoptosis.

During the recovery phase, the mRNA level of TNFR1 was also significantly increased in CNS-infiltrating cells from IFN--treated mice compared with controls.

Our results further challenge the exclusive detrimental role of IFN- in the CNS during EAE/Multiple Sclerosis.

And indicate that CNS-confined Inflammation may induce protective Immunological counter-mechanisms leading to a faster clearance of Encephalitogenic T-Cells by Apoptosis, thus restoring the Immune privilege of the CNS.