Establishment of an antitoxoplasma state by stable expression of mouse indoleamine 2,3-dioxygenase

Abstract

Indoleamine 2,3-dioxygenase (IDO), a tryptophan-degrading enzyme, is inducible by various interferons (IFNs). IDO-mediated tryptophan degradation, but not the formation of IDO-catalyzed tryptophan metabolites, has been suggested as a mechanism for the antiparasitic action of IFN-gamma. To determine whether the IFN-gamma-induced IDO alone is sufficient for establishing the antiparasitic state, we constructed a mouse IDO expression plasmid containing a heavy metal-responsive metallothionein promoter and obtained a stable transformant (C6) by transfection of this plasmid into mouse rectal cancer (CMT-93) cells. In the presence of 100 microM ZnSO4, C6 cells yielded a high level of IDO; and after a 2-day culture period, the enzyme induction resulted in complete depletion of tryptophan from the culture medium. Under these conditions, the growth of Toxoplasma gondii in C6 cells infected with the organisms on day 3 after enzyme induction was completely blocked. In the absence of ZnSO4, however, IDO induction was negligible in C6 cells, and T. gondii continued to grow. Furthermore, in a transformant (CC10) carrying an antisense mouse IDO plasmid or in parental CMT-93 cells, IDO was not induced at all even in the presence of 100 microM ZnSO4, and T. gondii continued to grow in these cells as well. These results taken together indicate that complete depletion of tryptophan from the culture by IDO alone is sufficient to establish the antitoxoplasma state in mouse cells.