Can we pass from the experimental to the clinical phase in MS stem cell research?

A crucial point in development of new treatments is the step from the experimental level to the first clinical trials. For stem cell treatments in general, but for stem cell treatment in Multiple Sclerosis specifically, this is the question for the moment. To answer this question a rational analysis of the hypotheses and the suppositions behind the application of stem cells is necessary, as well as a review of the present knowledge, the risks and the gains to be expected. This is a personal analysis of 32 oral presentation and discussions of the European Charcot Foundation Symposium, Taormina 2006. It is the application of the Kenter and Cohen [Kenter MJH, Cohen AF. Establishing risk of human experimentation with drugs: lessons from TGN 1412. Lancet 2006; 368:1387-91] approach, adapted for stem cell treatment in MS. About half of the pertinent issues plead for the start of clinical experiments now. However, the absence of knowledge on deleterious effects and their predictability heavily weights against it. Organisational and funding aspects were discussed to prevent uncritical, uncontrolled clinical approaches.

The measurement of fatigue in patients with multiple sclerosis. A multidimensional comparison with patients with chronic fatigue syndrome and healthy subjects.

OBJECTIVE: To provide a multidimensional characterization of fatigue in patients with multiple sclerosis (MS). DESIGN: Cross-sectional design. Fifty patients with clinically definite MS were compared on the dimensions of fatigue with 51 patients with chronic fatigue syndrome (CFS) and 53 healthy subjects. RESULTS: Fourty-six percent of the patients with MS reported fatigue to be present at least once a week. Patients with MS and patients with CFS had significantly higher subjective fatigue severity scores than healthy subjects. Patients with MS and patients with CFS had significantly higher scores on measures of psychological well-being than healthy subjects. Patients with MS had scores similar to those of patients with CFS, except that patients with CFS had significantly higher somatization scores. High somatization scores reflect strong focusing on bodily sensations. Both groups of patients were significantly less active than the healthy subjects. The Kurtzke Expanded Disability Status Scale (EDSS) and the Beck Depression Inventory scores were not related to subjective fatigue severity. In patients with MS and in patients with CFS, subjective fatigue severity was related to impairment in daily life, low sense of control over symptoms, and strong focusing on bodily sensations. In CFS, but not in MS, evidence was found for a relationship between low levels of physical activity and attributing symptoms to a physical cause and between subjective fatigue severity and physical activity. CONCLUSIONS: Patients with MS experienced significant fatigue, which had a significant impact on daily functioning and was not related to depression on Expanded Disability Status Scale score. Psychological factors, such as focusing on bodily sensations and low sense of control play a role in the experience of fatigue in MS and CFS.

Quantitative MRI changes in gadolinium-DTPA enhancement after high-dose intravenous methylprednisolone in multiple sclerosis.

In 12 patients with definite multiple sclerosis who received a total of 21 courses of high-dose (1 gram daily for 10 consecutive days) intravenous methylprednisolone, we performed MRI of the brain with and without gadolinium-DTPA before and after treatment. On the initial MRI, there was a total of 98 enhancing lesions, 93 of which were also represented on the unenhanced images. After treatment, 13 patients improved clinically, and 78 of the lesions lost enhancement but remained visible on the unenhanced images. There were six new enhancing lesions on the second MRI. Thus, the blood-brain-barrier integrity improved after high-dose IV methylprednisolone, which correlated well with the clinical improvement. The lesions remaining visible on the unenhanced images indicate an incomplete histologic recovery at the time of the second scan, and also demonstrate that unenhanced MRI alone is not sufficient to monitor disease activity in the short term in multiple sclerosis.

A correlative triad of gadolinium-DTPA MRI, EDSS, and CSF-MBP in relapsing multiple sclerosis patients treated with high-dose intravenous methylprednisolone.

In a prospective study, we compared the number of gadolinium-DTPA (Gd-DTPA) enhancing lesions on MRI with the CSF and clinical findings before and after a total of 20 courses of high-dose intravenous methylprednisolone in relapsing multiple sclerosis patients. Before treatment, there was a significant correlation of Gd-DTPA enhancement (seen on 16 of 20 scans) and CSF myelin basic protein (MBP). If enhancement with Gd-DTPA represents inflammation and CSF-MBP indicates myelin breakdown, the amount of inflamed tissue should correlate with the amount of myelin being damaged. Clinical improvement occurred following 15 of 20 courses, and decrease of Gd-DTPA enhancement in 12 of 16 scans; the mean CSF-MBP level was the only CSF variable to return to reference values. There was a significant correlative triad of decrease in CSF-MBP, Gd-DTPA enhancement, and clinical disability. Thus, the clinical effect of methylprednisolone might be accompanied by a reduction of inflammation and myelin breakdown.

Accumulation of hypointense lesions ("black holes") on T1 spin-echo MRI correlates with disease progression in multiple sclerosis.

MRI findings are increasingly used as outcome measures in therapeutic trials in MS. The discrepancy between the extent of the lesions on conventional T2 images and the clinical condition of the patient is one of the problems encountered in such studies. This clinical-radiological paradox prevents the use of MRI data as surrogate markers of disability in MS. A recent pilot study suggested a relationship between hypointense lesions on T1 MRI and disability. To assess in more detail the correlation of changes in hypointense lesion load on T1-weighted spin-echo MR images ("black holes") with changes in disability in MS, we studied 46 patients with clinically definite MS at baseline and after a median follow-up of 40 months. There was a significant correlation between baseline disability and hypointense lesion load (Spearman rank correlation coefficient [SRCC] = 0.46, p = 0.001). In secondary progressive patients, the rate of accumulation of these "black holes" was significantly related to progression rate (SRCC = 0.81, p < 0.0001). We speculate that the appearance of hypointense lesions is the MRI equivalent of a failure of remission. Overall, T1 lesion load measurements correlated better with clinical assessments than T2 lesion load measurements. Quantification of hypointense lesion load on T1-weighted spin-echo MRI helps to resolve the clinical-radiological paradox between disability and MRI and has the potential to be a surrogate marker of disability in MS.

Magnetic resonance evaluation of disease activity during pregnancy in multiple sclerosis.

We followed two women with MS during pregnancy by means of serial unenhanced MR imaging. On each follow-up image, we assessed the number of new or enlarging lesions. Both women showed a decrease in MR disease activity during the second half of pregnancy and a return of MR disease activity to prepregnancy levels in the first months postpartum. These findings support the view that pregnancy reduces disease activity in MS.

Correlating MRI and clinical disease activity in multiple sclerosis: relevance of hypointense lesions on short-TR/short-TE (T1-weighted) spin-echo images.

Magnetic resonance imaging (MRI) is being used as an outcome criterion in therapeutic trials in multiple sclerosis (MS) on the assumption that it, as a sensitive marker of biologic disease activity, could serve as a surrogate marker of disability. We evaluated the relation between MRI findings and disability in a quantitative follow-up study of 48 MS patients. Median duration of follow-up was 24 months (range, 10 to 42 months). Computer-assisted volume measurements employing a seed-growing technique yielded a standard error of measurement of 0.275 cm2. The median total area of the hyperintense lesions on the initial T2-weighted images was 8.4 cm2. The median increase was 0.76 cm2/yr (9%). In a subgroup (n = 19) with short-TR/short-TE spin-echo (SE) images, we measured the hypointense lesion load. The median total area of the lesions at entry was 0.70 cm2, with a median increase of 0.28 cm2/yr (40%). The total area of the hyperintense lesions on the initial T2-weighted images showed a weak correlation with the Expanded Disability Status Scale score at entry (Spearman rank correlation coefficient [SRCC] = 0.30; 0.02 < p < 0.05). The increase in disability showed a positive correlation (SRCC = 0.19) with the increase in hyperintense lesion load on the T2-weighted SE images, but this correlation did not reach statistical significance (p = 0.09), probably because of lack of clinical progression.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of the development of experimental allergic encephalomyelitis by suramin.

Suramin was tested for its ability to suppress experimental allergic encephalomyelitis. Prophylactic administration caused significant reduction in the severity of the disease, incidence of paralysis and cellular infiltration in nervous tissue. Therapeutic treatment with suramin also caused a reduction in the severity of the disease, the incidence of paralysis and cellular infiltration, but to a lesser extent. Significantly fewer animals were paralysed for more than two days on therapeutic treatment.

T-cell subsets in spinal fluid of multiple sclerosis patients.

CSF T-cells and T-cell subsets were characterized by monoclonal antibodies in 15 untreated multiple sclerosis (MS) patients, 17 immunosuppressed chronic progressive MS patients and 9 patients with other neurological diseases. A negative correlation was found between total cell numbers and T suppressor cell percentages in untreated and treated MS patients. A negative correlation (r = -0.71) was found between intrathecal IgG levels and T suppressor cell percentages in untreated MS patients. In peripheral blood, no correlation between T-cells and T-cell subsets with IgG levels was found. It is discussed that T-cell subsets and intrathecal IgG levels may be indicators of the activity of the inflammatory process in the brains of chronic progressive MS patients.

Modulation of experimental allergic encephalomyelitis in Lewis rats by monoclonal anti-T cell antibodies.

The effect of monoclonal antibodies to different T lymphocyte populations of the rat on the induction and the course of experimental allergic encephalomyelitis (EAE) was investigated. EAE was induced by injection of guinea pig spinal cord in adjuvant. Subcutaneous injections of monoclonal antibodies to all peripheral T lymphocytes (W3/13) abrogated or prevented the development of clinical EAE. Similar results were obtained in animals injected with monoclonal antibodies to T helper cells (W3/25) mixed with monoclonal antibodies to T nonhelper cells (OX8). Animals treated with either W3/25 or OX8 developed clinical EAE as the control rats (subcutaneous injected with normal mouse serum). Histological examination after the acute stage revealed no significant differences between rats treated prophylactically with W3/13, W3/25 or OX8 and rats injected with normal mouse serum. Animals treated prophylactically with a mixture of W3/25 and OX8 developed, on the whole, EAE with less histological severity compared to the control animals. Treatment of rats after the onset of the first clinical symptoms of EAE (tail flaccidity) with W3/13 resulted in a less fatal course of the disease. Compared to surviving rats injected with mouse serum (controls) the number of infiltrates were reduced in these rats treated therapeutically.

The effects of high-dose methylprednisolone on gadolinium-enhanced magnetic resonance imaging and cerebrospinal fluid measurements in multiple sclerosis.

Blood-brain barrier (BBB) disruption is probably the first event in the lesion development in multiple sclerosis (MS). This stage can be visualized by gadolinium-enhanced magnetic resonance (MR) imaging of the brain. Serial MR imaging studies have indicated a continuous spectrum of disease activity with waxing and waning of acute lesions, even in clinically stable MS patients. High-dose intravenous methylprednisolone (MP) has a beneficial clinical effect; reduces gadolinium enhancement, indicating improvement of BBB integrity; and, in MS patients, decreases intrathecal immunoglobulin synthesis with reduction of cerebrospinal fluid (CSF) myelin basic protein (MBP). A correlative triad is noted between gadolinium enhancement, clinical improvement, and decrease of CSF MBP following MP treatment, indicating a relationship between restoration of BBB integrity, clinical improvement and decrease of myelin breakdown. It is not clear whether MP interferes primarily with the process of demyelination or reacts non-specifically with its mediators.

Physical activity in chronic fatigue syndrome: assessment and its role in fatigue.

This paper describes the assessment of physical activity in chronic fatigue syndrome (CFS) and investigated the following questions: Do patients with CFS have low levels of physical activity; is there a relationship between actual level of physical activity and fatigue; can self-report measures adequately assess actual level of physical activity; what is the role of cognitions with respect to physical activity; and are results with respect to physical activity specific to CFS? Three different types of activity measures were used: self-report questionnaires, a 12-day self-observation list, and a motion-sensing device (Actometer) which was used as a reference for actual activity level. Fifty-one patients with CFS, 50 fatigued patients with multiple sclerosis (MS), and 53 healthy subjects participated in this study. Although none of the self-report questionnaires showed high correlations with the Actometer, questionnaires that require simple ratings of specified activities were related to the Actometer and can be used as acceptable substitutes, in contrast to instruments that require general subjective interpretations of activity that had low or non-significant correlations with the Actometer. Actometer results showed that CFS patients and MS patients had similar activity levels and both groups were significantly less active than healthy subjects. Compared to MS patients, CFS patients were more likely to indicate that they had been less active than other persons they knew. Activities which patients expected to result in higher fatigue levels were less frequently performed. Patients with CFS had significantly higher scores on this measure than MS patients and healthy subjects. Low levels of physical activity were related to severe fatigue in CFS but not in MS. In conclusion, although CFS patients have similar low activity levels than MS patients, there are also important differences between both groups: in CFS cognitive factors are more prominently involved in producing the low activity levels than in MS and in CFS patients activity level is related to fatigue but not in MS.

Changes in carotid flow velocity induced by lowering cerebrospinal fluid pressure in normal pressure hydrocephalus.

This is a report of changes in blood flow velocity in the carotid system induced by lumbar puncture in five patients who had clinical and neuroradiological signs of normal pressure hydrocephalus. After lowering cerebrospinal fluid pressure an increase of carotid flow velocity was found on Doppler hematotachography. These changes of carotid blood flow velocity could not be demonstrated in a control group of four patients with normal cerebrospinal fluid dynamics. This method is easy to perform, does not overtax the patient and seems to be indicated for the diagnosis of NPH. It is presented as a new, simple, reproducible aid to the diagnosis of NPH by Doppler hematotachography.

Decreased vitamin B12 and folate levels in cerebrospinal fluid and serum of multiple sclerosis patients after high-dose intravenous methylprednisolone.

Twenty-one patients (15 women, 6 men) with definite multiple sclerosis (MS) were treated with 1000 mg intravenous methylprednisolone-succinate (MP) daily for 10 days. Before MP treatment there was a negative correlation (r = 0.59, P = 0.0084) between serum vitamin B12 and progression rate, defined as the ratio of the score on Kurtzke's Expanded Disability Status Scale and disease duration. A significant decrease was demonstrated in the cerebrospinal fluid (CSF) and serum levels of folate and in the CSF level of vitamin B12 after MP treatment. The decrease in serum B12 was not statistically significant. After MP treatment all median levels of vitamin B12 and folate were below the reference medians. We hypothesize that low or reduced vitamin B12/folate levels found in MS patients may be related to previous corticosteroid treatments. Otherwise a more causal relationship between low vitamin B12/folate and MS cannot be excluded. Further studies may be required to clarify the vitamin B12 and folate metabolism in patients with MS.

Functional correlates of callosal atrophy in relapsing-remitting multiple sclerosis patients. A preliminary MRI study.

In multiple sclerosis (MS), periventricular lesions produce atrophy of the corpus callosum (CC), as evidenced by magnetic resonance imaging (MRI). We investigated whether CC atrophy in relapsing-remitting MS patients is related to functional deficits. We compared 14 mildly disabled (mean Expanded Disability Status Scale score 2.7) relapsing-remitting MS patients with 14 age- und sex-matched controls. CC size was determined using sagittal T1-weighted MRI. The function of the CC was studied using a neuropsychological battery and neurophysiological evaluation based on visual stimulation using a divided visual field paradigm. The total area of the CC in patients (mean 5.3 cm2) was significantly (P = 0.002) smaller than in controls (mean 6.6 cm2). Patients showed left ear extinction using the dichotic listening test and impaired name learning, which was correlated with atrophy of the splenium. There were no differences in interhemispheric transfer time between patients and controls. Marked atrophy of the CC can be encountered in relapsing-remitting MS patients. The associated cerebral disconnection correlated with atrophy of expected regions of the CC, thus supporting topographical organization.

The preselection value of the Doppler LP test for shunt-therapy in patients with normal pressure hydrocephalus.

The results of this study confirm the hypothesis that the simple innocuous Doppler LP test may be useful in obtaining information about CSFP-CBF (cerebrospinal fluid pressure-cerebral blood flow) relationships. The value of this test in predicting the clinical outcome of CSF shunting in patients with normal pressure hydrocephalus (NPH) was studied. Fourteen patients with NPH were examined; eight patients showed a positive and six patients a negative Doppler LP test. Seven of the eight patients with a positive test improved following the shunting procedure, but none of the six patients improved after operation. This study suggests that the Doppler LP test is a useful preoperative selection test in patients with NPH. In addition this study supports the hypothesis that CBF autoregulation may be impaired with NPH.

Gadopentetate dimeglumine enhancement of multiple sclerosis lesions on long TR spin-echo images at 0.6 T.

The authors sought to determine if Gd-DTPA enhancement of multiple sclerosis (MS) hampers lesion detection on long TR spin-echo images (TE 60 msec) at 0.6 T. They measured the signal intensity (SI) of 41 lesions (10 patients) and normal-appearing gray (NAGM) and white matter (NAWM) before and after administration of contrast. The change in SI of nonenhancing lesions and NAGM and NAWM was small (less than or equal to 1.5%), and of enhancing lesions (5.3%) moderate. The contrast of nonenhancing lesions to NAGM and NAWM changed insignificantly, but the contrast of enhancing lesions to NAGM and NAWM increased significantly. The authors conclude that long TR images can be obtained after Gd-DTPA without hampering lesion conspicuity in research MR protocols in multiple sclerosis.

The effect of gadolinium on the sensitivity and specificity of MR in the initial diagnosis of multiple sclerosis.

PURPOSE: To determine whether gadolinium can improve the sensitivity and specificity of MR imaging for the initial diagnosis of multiple sclerosis. METHODS: Patients (n = 57) with neurologic symptoms suggesting multiple sclerosis were studied prospectively. MR imaging consisted of T2-weighted and gadolinium-enhanced T1-weighted spin-echo images. Lumbar puncture was performed for cerebrospinal fluid analysis in 34 patients. RESULTS: After imaging, 17 patients (35%) had clinically definite multiple sclerosis. Cerebrospinal fluid examination had a sensitivity of 69% and specificity of 38%. Using liberal criteria, the sensitivity of T2-weighted MR imaging was 94% and the specificity 55%; using more strict criteria, the specificity increased to 65% with a sensitivity of 88%. Gadopentetate dimeglumine enhancement increased the specificity further to 80% with a loss of sensitivity (59%). CONCLUSION: Gadolinium enhancement increases the specificity of MR imaging in the early diagnosis of multiple sclerosis.

Comparison of two MR sequences for the detection of multiple sclerosis lesions in the spinal cord.

PURPOSE: To compare cardiac-triggered dual-echo spin-echo and magnetization transfer-prepared gradient-echo (MT-GE) MR imaging in the detection of multiple sclerosis (MS) lesions in the spinal cord. METHODS: The cervical spinal cord in 20 patients with MS and in nine healthy volunteers was examined with spin-echo and MT-GE MR imaging. Sagittal images were scored for number of lesions, certainty about lesions, image quality, and visual hindrance by artifacts in random order by two radiologists separately and in a blinded manner. RESULTS: In one healthy volunteer, a lesion was seen on images obtained with both images. Lesion/cord contrast-to-noise ratio was equal on both the MT-GE and T2-weighted spin-echo images. MT-GE images showed better image quality and fewer artifacts than the spin-echo images did. The readers found approximately the same number of lesions. However, the number of definite lesions was higher for the spin-echo sequence than for the MT-GE sequence. One reader found 45 definite lesions with spin-echo and 34 definite lesions with MT-GE. For the other reader, these numbers were 37 (spin-echo) and 31 (MT-GE). On the spin-echo images, 90% of the patients were considered to have definite lesions; on the MT-GE images, the readers found definite lesions in 65% (reader 1) and in 70% (reader 2) of the patients. CONCLUSION: Image quality was better with the MT-GE technique than with the spin-echo technique, and lesion/cord contrast-to-noise ratio on the MT-GE images was equal to that of T2-weighted spin-echo images. However, for detecting spinal cord MS lesions in the sagittal plane, the spin-echo images were preferred to the MT-GE images.

Clinical practice of immunosuppressive treatments in multiple sclerosis: results of a second international questionnaire.

Immunosuppressive treatment is a current practice in MS therapy. Mitoxantrone has gained largest acceptance. Differences in clinical indications of mitoxantrone, cyclophosphamide, azathioprine and methotrexate are described.