The Application of Tissue Culture to the Study of Experimental Allergic Encephalomyelitis

Abstract

Myelinated cultures of rat and mouse cerebellum undergo a specific and characteristic pattern of demyelination in the presence of sera from animals with experimental allergic encephalomyelitis (EAE) (1, 2). The neuroglia swell; myelin sheaths become disfigured b y fusiform swellings and break away as small fragments; and finally, remaining intact sheaths and myelin fragments are reduced to small fat droplets. During this process neuron somas and axons appear little affected (Figs. 1 to 4).This communication will present evidence for an immunological mechanism responsible for the in vitro demyelination. MethodsThe techniques of rat cerebellar tissue culture, induction of EAE in animals, and the preparation of sera have been previously described (1). An additional technique was employed to develop a series of animals with varying incidence of clinical and pathological lesions and varying levels of circulating antibodies. Each of 10 rabbits received 3 intraperitoneal inoculations of bovine cord (250 mg each) over a 3 month period: 5 animals with cord suspension in Freund's adjuvant; and the remaining 5, with cord in saline. Four of the 5 animals inoculated with cord in adjuvant developed hindleg weakness following the third inoculation, which progressed over 2 to 4 weeks to complete paraplegia, urinary and fecal incontinence, and death. At autopsy, 4 months after the first inoculation, characteristic lesions of EAE were found. Two of the remaining animals, i.e. without adjuvant, developed hindleg weakness and ataxia following the third inoculation. However, at autopsy, 4 months after the first inoculation and 2 weeks after serum examination and the appearance of symptoms, no microscopic nervous system changes were noted.

Copper accumulation and lipid oxidation precede inflammation and myelin lesions in N,N-diethyldithiocarbamate peripheral myelinopathy

“…In particular IgG2a demonstrated increases greater than six fold. The observation that the IgG fraction of serum can produce complement-dependent demyelination in experimental models (Appel and Bornstein, 1961;Halstead et al, 2005;Saida et al, 1979) raises the question as to whether the elevation in IgGs could represent an autoimmune component in DEDC myelinopathy. Autoantibodies could possibly be produced from the generation of non-self oxidatively damaged proteins or from release of normally sequestered nerve proteins into the general circulation.…”

EAE cerebrospinal fluid augments in vitro phagocytosis and metabolism of CNS myelin by macrophages

“…In vitro studies have supported evidence for an anti-myelin antibody role in MS and EAE demyelination (Appel and Bornstein, 1964;Seil et al, 1968;Kim et al, 1970;Raine et al, 1973). Serum from animals with EAE as a result of immunization with whole myelin causes myelin breakdown through complement-and antibodydependent mechanisms (reviewed by Raine, 1984b).…”

The Membrane Attack Complex of Complement Causes Severe Demyelination Associated with Acute Axonal Injury

“…Early in vitro models of demyelination using cerebellar cultures showed that the demyelinating component of sera from animals with EAE was heat labile, a classical characteristic of the complement system (8). In rat EAE, complement depletion ameliorates disease (9), and in a demyelinating model, Ab-mediated demyelinating EAE (ADEAE), induced by injection of Abs to myelin oligodendrocyte glycoprotein (MOG) at the onset of clinical signs in EAE (10), complement is essential for demyelination (11,12).…”