Induction Of Eam In Mice By Adoptive Transfer Of Cardiac Myosinstimulated T Cells

It may be experimentally useful to induce EAM in mice in the absence of endogenous cell-mediated immunity or exogenous adjuvants. EAM can be induced by the transfer of cardiac myosin-stimulated T cells from immunized congenic C.B-17 donors into immu-nodeficient SCID (Severe Combined Immunodeficient) recipients. The SCID mice have no functional T or B cells and are an excellent model system to study the role of specific cellular effectors on the induction and pathogenesis of EAM; moreover, although recipient irradiation is often required for successful adoptive transfer of autoimmune diseases in mice, use of SCID recipients makes this unnecessary in this procedure. EAM has been successfully induced by adoptive transfer in BALB/c mice without recipient irradiation, using lipopolysaccharide to stimulate the recipients prior to transfer of stimulated T cells (see Bachmaier, 1999). The donor C.B-17 mice are immunized with purified cardiac myosin, as in Basic Protocol 1, and splenic T cells are isolated by nonadherence to nylon wool followed by cytotoxic elimination of B cells and accessory cells using anti-MHC class II antibodies and complement. After a 3-day in vitro stimulation with concanavalin A (Con A), T cells are injected intravenously into the SCID recipients. Successful transfer of myocarditis can be determined histologically within 14 to 21 days of T cell transfer.

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