Impairment of macrophage function by inhibitors of ornithine decarboxylase activity.

The effects of irreversible inhibition of ornithine decarboxylase on the capacity of murine macrophages to take up a protozoan organism (Trypanosoma cruzi) or inert particles were investigated. Incubation of macrophage cultures with four different ornithine decarboxylase inhibitors, namely, DL-alpha-difluoromethylornithine (DFMO, 0.5 to 20 mM), delta-methyl-acetylenic putrescine (1 to 5 mM), monofluoromethyldehydroornithine ethyl ester (1 to 5 mM), and monofluoromethyldehydroornithine methyl ester (1 to 5 mM), before the addition of the parasites significantly reduced the percentage of macrophages with parasites, indicating that some of the host cells were no longer capable of binding or ingesting the parasite. The average number of trypanosomes per 100 macrophages was also diminished, denoting a lesser phagocytic capacity as a consequence of the treatments. These effects were reversible within 2 h after removal of excess DFMO. No alteration in parasite-macrophage interaction was seen when the trypanosomes were treated with DFMO. That the effects of DFMO on the macrophages probably resulted from a reduction in polyamine levels caused by inhibition of ornithine decarboxylase was indicated by the fact that these effects were not seen when the macrophages were incubated with DFMO in the presence of putrescine, the product of ornithine decarboxylation by ornithine decarboxylase. DFMO treatment of macrophages also inhibited the capacity of these cells to ingest killed parasites or latex beads and thus appeared to generally affect phagocytosis. An effect of DFMO on the susceptibility of macrophages to penetration by the parasites seemed less likely because no significant alteration in cell-parasite association occurred when myoblasts--which, not being phagocytic, can be infected only by membrane penetration--were treated with DFMO. Taken together, these results emphasize a role of ornithine decarboxylase activity and polyamine biosynthesis in macrophage function.     

Arginine decarboxylase inhibitors reduce the capacity of Trypanosoma cruzi to infect and multiply in mammalian host cells.

The capacity of blood (trypomastigote) forms of Trypanosoma cruzi to infect mouse peritoneal macrophages or rat heart myoblasts in vitro was inhibited by treatment of the trypomastigotes with DL-alpha-difluoromethylarginine (F2Me Arg), monofluoromethylagmatine, or (E)-alpha-monofluoromethyl-3-4-dehydroarginine--all irreversible inhibitors of arginine decarboxylase. Similar results were obtained when F2MeArg-treated parasites were incubated with rat heart myoblasts. The inhibitory effects were characterized by marked reductions in both the proportion of infected cells and the number of parasites per 100 host cells. The concentrations of the arginine decarboxylase inhibitors that affected infectivity had no detectable effect on either the concentration or motility of the parasite and, therefore, could not have affected the collision frequency. F2MeArg appeared to inhibit the ability of T. cruzi to penetrate the host cells since the drug had no significant effect on the extent of parasite binding to the surface of the host cells. The inhibitory effect of F2MeArg was markedly reduced or abrogated in the presence of either agmatine or putrescine, as would have been expected if F2MeArg acted by inhibiting arginine decarboxylase. Addition of F2MeArg to macrophage or myoblast cultures immediately after infection or at a time when virtually all of the intracellular parasites had transformed into the multiplicative amastigote form, resulted in a markedly reduced parasite growth rate. This effect was also prevented by exogenous agmatine. These results indicate the importance of polyamines and polyamine biosynthesis in the following two important functions of T. cruzi: invasion of host cells and intracellular multiplication. Furthermore, concentrations of the inhibitors tested that affected the parasite did not alter the viability of the host cells, the cellular density of the cultures, or the ability of uninfected myoblasts to grow. Thus, arginine decarboxylase inhibitors may have a potential application in chemotherapy against T. cruzi infection.     

UNUSUAL EFFECT OF SOME α-MERCAPTOACRYLIC ACIDS ON METABOLISM OF ZINC IN THE RAT

1. Gavage of rats with β-aryl derivatives of α-mercaptoacrylic acid resulted in pronounced and sustained elevation of serum zinc concentration. Greater than ten-fold increases above normal zinc levels were attained 2–8 h after doses of 50 mg/kg of the phenyl and furan derivatives. 2. These compounds were rapidly absorbed from the rat gastrointestinal tract and could be detected in serum for several days after a single dose. The return of serum zinc concentration to the normal level paralleled clearance of the mercaptoacrylic acid from plasma. 3. Close to 100% of the zinc and of the α-mercapto-β-(2-furan)acrylic acid (MFA) in serum 4 h after administration of the compound were bound to serum macromolecules. 4. MFA decreased excretion of endogenous zinc; it altered neither the gastrointestinal absorption of zinc nor serum concentrations of copper, albumin and total protein. 5. These compounds appear to mobilize zinc from tissue stores and retard zinc clearance from plasma.