Experimental Autoimmune Encephalomyelitis in the

Studies of experimental autoimmune encephalomyelitis (EAE) in rats have provided important insight into mechanisms responsible for autoimmune tissue damage and the regulation of immune responses to "self" antigens. Rats of the inbred Lewis (LEW) and Dark Agouti (DA) strains are highly susceptible to EAE induced with guinea pig myelin basic protein (MBP) emulsified in a modified complete Freund's adjuvant (CFA). DA rats are also highly susceptible to EAE induced with proteolipid protein (PLP). EAE in rats is an acute paralytic disease mediated by CD4+ T cells. The earliest clinical signs begin 10 to 12 days after immunization, progressing from loss of tail tonicity to hind limb paralysis. Most rats recover by 18 to 20 days post immunization. A similar clinical picture is seen after adoptive transfer of activated T cells from MBP/CFA-immunized donors, with the exception that disease onset occurs 4 to 5 days post transfer. LEW rats have been used in these studies because the acute onset and spontaneous recovery bear resemblance to the exacerbations and remissions seen in multiple sclerosis (MS), thus providing a model to investigate the mechanisms underlying autoimmune demyelination.

This unit describes methods for induction of EAE in rats by active immunization with guinea pig MBP (see Basic Protocol 1) and for preparation of MBP (see Support Protocol 1). Alternatively, a crude guinea pig spinal cord homogenate can be used to actively induce EAE (see Alternate Protocol). MBP-specific T cells can be activated to establish T cell lines (see Support Protocol 2) for the adoptive transfer of EAE (see Basic Protocol 2). An in vitro proliferation assay (see Support Protocol 3) can be used to evaluate the T cell immune response to MBP. However, it should be stressed that an in vitro proliferative response to MBP does not imply that EAE would necessarily have developed had the T cells been adoptively transferred because T cell responses to nonencephalitogenic MBP epitopes may occur in the absence of disease. Thus, proliferation should be considered as a confirmatory assay to evaluate the immune response to MBP. It is important to use guinea pig spinal cord or brain tissue for the induction of EAE in rats, because MBP from other species (e.g., bovine, rabbit, or human MBP) is relatively inactive in the rat. Even rat central nervous system tissue is less encephalitogenic than guinea pig preparations.

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