Arginine metabolism during macrophage autocrine activation and infection with mouse hepatitis virus 3

In contrast to BALB/c mouse macrophages (Mphi), Mphi from the A/J mouse strain, upon activation by exogenous interferon gamma (IFNgamma), develop an anti-mouse hepatitis virus 3 (MHV3) state which correlates with resistance to virus infection. To investigate the autocrine activation of BALB/c and A/J Mphi, we activated them with interleukin-12 (IL-12) and/or IL-18, and quantified IFNgamma production, the anti-MHV3 state and arginine metabolism. Synergistic activation by IL-12/IL-18 induced the expression of the IFNgamma gene in Mphi from both mouse strains. In bone marrow (BM) or peritoneal (P) Mphi of specific pathogen-free (spf) mice of both strains, IFNgamma synthesis occurred only with a synergistic IL-12/IL-18 activation and showed increasing levels from 24 to 72 h of activation. In contrast, when non-spf mice were used in the assay, their PMphi synthesized higher IFNgamma levels upon activation with only IL-12 or only IL-18 or both. The BALB/c Mphi were always capable of synthesizing higher amounts of IFNgamma than the A/J Mphi. An anti-MHV3 state was observed only in A/J Mphi upon activation with IL-12/IL-18 or IFNgamma regardless of their origin from the peritoneum or bone marrow. Arginine metabolism in activated and/or virus infected BMMphi was investigated through nitric oxide (NO) and arginase induction as well as the consumption of arginine and synthesis of citrulline, ornithine and spermine. The results showed that both BALB/c and A/J BMMphi populations released NO only after activation with IL-12/IL-18 or IFNgamma. Arginase was not induced in BMMphi from both strains by IL-12/IL-18 or IFNgamma but only by IL-4/IL-10. Higher arginine consumption was observed in BMMphi from both strains upon activation with IL-4 or IFNgamma which further increased, in this case, when the cells were infected with MHV3. As a consequence of nitric oxide synthase synthesis and arginine consumption in IFNgamma activated BMMphi, we observed a higher synthesis of citrulline. High levels of ornithine were induced only upon IL-4 activation. Polyamine synthesis was higher in A/J BMMphi than in BALB/c ones, which correlated with the slightly lower levels of ornithine observed. Upon infection with MHV3, we observed a higher synthesis of spermine. IL-12/IL-18 or IFNgamma activation, mainly in MHV3 infected cells, led to a decreased synthesis of polyamines, notably spermine, only in A/J BMMphi. Difluoromethylornithine treatment, which leads to inhibition of polyamine synthesis, induced a decreased MHV3 multiplication in both BALB/c and A/J BMMphi. Altogether these data show the relevance of IFNgamma, from the autocrine or paracrine pathway, and arginine metabolism for the control of MHV3 replication in Mphi of a resistant mouse strain.