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Long-term S-antigen (S-Ag)-specific T lymphocyte lines were derived from the lymph nodes of immunized Lewis rats that had been pretreated with low-dose cyclophosphamide. The protocol consisted of functional selection by alternating cycles of stimulation with S-Ag presented on syngeneic accessory cells and proliferation in IL 2-containing spleen-conditioned medium, coupled with early phenotypic selection for cells bearing the helper/inducer membrane marker (W3/25), by panning on antibody-coated plastic dishes. This protocol consistently resulted in the rapid generation of in vivo functional cell lines capable of mediating experimental autoimmune uveoretinitis (EAU) when transferred into naive rats at 5 to 10 X 10(6) cells/rat systemically or 1 to 2 X 10(6) cells/rat intravitreally. The disease appeared within 6 to 8 days, usually with minimal anterior chamber involvement, and was often unilateral. Pathologic changes resembled those seen in EAU induced by active immunization. The disease could be transferred without concomitant formation of serum antibodies. The uveitogenic line cells were negative for Ia antigen and positive for the W3/25 membrane marker, which appeared to be stable in long-term culture. They proliferated vigorously in vitro and produced IL 2 in response to S-Ag or Con A, but not to unrelated antigens. The establishment of uveitogenic T helper lymphocyte lines will permit the analysis of the cellular mechanisms involved in EAU in a more defined system than has been available.
Caspi et al. (Sat,) studied this question.