New approach to study of in vitro toxicity of multiple sclerosis and other sera

Bradbury, K.; Aparicio, S. R.; Sumner, David; Bird, C. C.

doi:10.1016/0022-510x(84)90080-7

Cited by 10 publications

(8 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In both of these the cellular degradative activity was high ( Table 2), suggesting that the in vitro demyelinating factor may be secreted by the circulating leucocytes, Grundke-Iqbal & Bornstein (1980) speculated that the demyelinating factor found in MS sera may be a proteolytic enzyme secreted by activated leucocytes in the blood or locally in the perivenous cuffs in the brain. However, cellular demyelinating activity was not usually accompanied by serum myelin degradative activity (Table 2), Our data also show that apart from one case of peripheral neuropathy in vitro myelin degradative activity was absent from the sera of patients with other neurological diseases including motor neurone disease, Guillain-Barre syndrome, inflammatory conditions and from all healthy controls, A possible explanation of these contradictory results is that demyelinating factors previously detected in serum may be acting on the cultured oligodendrocyte and not primarily on myelin, Bradbury et al (1984b) have developed a new technique of assessing sera-induced myelinotoxicity and cellular toxicity by using a combination of visual assessment of myelin damage and measuring radiolabel release from damaged glia and myelin in CNS tissue culture. In their studies (Bradbury et al, 1985) the sera from the MS group induced the greatest radiolabel release, which they suggested may be associated with cytotoxicity towards the oligodendrocyte, in addition to a direct effect on myelin.…”

Section: Serum Demyelinating Factorsmentioning

confidence: 50%

Degradation of human myelin in vitro by leucocytes from patients with multiple sclerosis

Owen¹,

Watson²,

Davison³

1990

Clinical and Experimental Immunology

View full text Add to dashboard Cite

SUMMARYIn order to study the possible autoimmune basis of multiple sclerosis (MS) a quantitative method has been used to investigate breakdown of human myelin in vitro. We found that serum from MS patients and controls was generally devoid of any myelin degradative activity. However, isolated peripheral blood mononuclear cells from 43% of MS patients showed significant myelin degradative activity as did those from 61-5% of patients with rheumatoid arthritis (RA). Myelin degradation by cells was found in only 13% of patients with other neurological diseases and in no healthy controls. It is proposed that this non-specific peripheral cellular immune degradative activity originates from cells activated within the central nervous system of MS patients or the joints of individuals with RA. As a result, activity in the blood only indirectly reflects the ongoing inflammatory response at the primary site, accounting for the lack of correlation between changes in the blood and the clinical status of the MS patient. We further propose that the lack of m vitro myelin degradative activity in cells recovered from the cerebrospinal fluid is due to autoaggressive cells being sequestered to the brain.

show abstract

Section: Serum Demyelinating Factorsmentioning

confidence: 50%

Degradation of human myelin in vitro by leucocytes from patients with multiple sclerosis

Owen¹,

Watson²,

Davison³

1990

Clinical and Experimental Immunology

View full text Add to dashboard Cite

show abstract

“…Sera from MS patients have been reported to induce demyelination in cultures of mammalian CNS (1)(2)(3)(4). Most often these studies have been carried out on CNS explants cultures or on cerebral slices.…”

mentioning

confidence: 99%

Inhibition of glial proliferationin vitroby serum from patients with multiple sclerosis

Ferraro

Salemi

Cestelli

et al. 1987

Acta Neurologica Scandinavica

View full text Add to dashboard Cite

Primary cell cultures from fetal rat CNS have been employed to evaluate the effects caused by the addition of serum from patients affected by multiple sclerosis (MS). MS-serum supplemented media caused a decrease in [3H]-thymidine incorporation into the cultures, thus indicating an inhibitory effect on proliferating glial cells. Sera from patients in remission stage of the disease showed an inhibitory effect not significatively lower than those from patients in acute stage. These results suggest that glial cells may be a target of circulating factors present in MS.

show abstract

“…The earliest of these studies go back over 50 years, when it was reported that sera from MS patients with active disease caused demyelination of in vitro cultured slices of rat cerebellum (Bornstein and Appel, 1965). However, subsequent studies in the 1970s and 1980s either reported that the toxicity observed in the earlier studies was an indicator of chronic disease, but not linked specifically to MS (Ulrich and Lardi, 1978), or else that both control and patient sera caused demyelination in culture (Wolfgram and Duquette, 1976;Lumsden, 1971;Bradbury et al, 1984;Suzumura et al, 1986;Silberberg et al, 1984), even after heat inactivation (Ruijs et al, 1990). Few follow up studies were done until quite recently.…”

Section: 1mentioning

confidence: 99%

“…While these results will need to be replicated in additional studies, the findings within the cultures have been consistent, and, since the cultures were analysed in a blinded fashion, the results appear to be relatively robust. As discussed in Chapter 6, early studies that used very high concentrations of human serum on rat cerebellar slices concluded that almost any effects of human serum were a reflection of non-immunoglobulin cytotoxic components of serum, and did not reflect any antibody-mediated effects (Lumsden, 1971;Wolfgram and Duquette, 1976;Ulrich and Lardi, 1978;Bradbury et al, 1984;Suzumura et al, 1986;Silberberg et al, 1984;Ruijs et al, 1990). …”

Section: Principal Findings Of the Thesismentioning

confidence: 99%

“…Early studies that found that serum from patients with any sort of chronic disease (and even from healthy controls) could bind to brain slices and cause demyelination in cerebellar slice cultures typically used serum dilutions of 1/2 or 1/4 in their experiments (Bradbury et al, 1984, Lumsden, 1971, Ruijs et al, 1990, Suzumura et al, 1986, Ulrich and Lardi, 1978, Wolfgram and Duquette, 1976. In the work presented in this chapter, high concentrations of serum (1/10 dilution) did have a tendency to bind non-specifically to all cells, irrespective of whether the sera came from healthy individuals or MS patients, so it is not surprising that the earlier studies reported non-specific effects with such high serum concentration.…”

Section: 4mentioning

confidence: 99%

See 1 more Smart Citation

Development of a simple in vitro mature mixed glial model to investigate differential expression of cell markers on glial cells and their potential as targets in multiple sclerosis

Beasley¹

View full text Add to dashboard Cite

Autoantibodies are present in many patients with demyelinating diseases and have been shown to have pathogenic potential in the case of people with neuromyelitis optica (NMO). However, it is still debated whether antibodies that are present in people with multiple sclerosis (MS) are pathogenic. There are multiple different mechanisms by which autoantibodies could exert their effects in MS, and they could potentially target any cell/structure within the central nervous system (CNS), although the focus of antibody studies in MS has been almost solely on myelin antigens. Recently, however, some evidence has suggested that astrocytes could also be targeted in MS. Therefore, there is a need for a simple but robust in vitro model to allow the testing of antibodies against oligodendrocytes and astrocytes. The hypothesis tested in this thesis is that a cell culture model containing a mixture of mature glial cells could be a useful model by which to test the pathogenicity of the antibodies from people with MS, and the antigens that they are targeting. In order to test this hypothesis, this thesis set out to develop a mature mixed glial cell culture model, to test whether one cell type could be killed without impacting on the health of the other cells, and to then use this model to test antibodies from people with MS.Chapter 2 describes the development of a rat neonatal-derived mature mixed glial culture. A method was developed that allowed cultures with a mixture of protoplasmic (type 1) and fibrous (type 2) astrocytes to grow alongside mature oligodendrocytes. This method did not rely on addition of extra growth factors, presumably due to the production of all factors required for growth of each cell type by the other cells in the mixed culture. Because of this co-dependence on the other cells in the mixed culture for growth and development, the next step was to test whether specific targeting (killing) of one cell would result in the death of the other cells in culture, as that could potentially interfere with the interpretation of results in later chapters investigating the effects of antibodies in patient sera. In Chapter 3, it was found that oligodendrocytes could be selectively targeted (using a commercially-available antibody against sulfatide) and the astrocytes remained unaffected. It proved to be more difficult to find a cell surface-expressed molecule that could target only astrocytes and not have ii adverse effects on oligodendrocytes. In the literature, one molecule that is generally considered to be expressed on astrocytes, but not oligodendrocytes, is GLAST, a glutamate transporter. Initial studies using an antibody against the intracellular N-terminus of GLAST confirmed that only astrocytes in culture were stained by this antibody. An antibody was therefore generated that was specific for an epitope of GLAST ) that is expressed on the extracellular side of the membrane. However, when this antibody was tested for its ability to kill astrocytes in culture, both oligodendrocytes and astrocytes were ...

show abstract

New approach to study of in vitro toxicity of multiple sclerosis and other sera

Cited by 10 publications

References 14 publications

Degradation of human myelin in vitro by leucocytes from patients with multiple sclerosis

Degradation of human myelin in vitro by leucocytes from patients with multiple sclerosis

Inhibition of glial proliferationin vitroby serum from patients with multiple sclerosis

Development of a simple in vitro mature mixed glial model to investigate differential expression of cell markers on glial cells and their potential as targets in multiple sclerosis

Contact Info

Product

Resources

About