Mechanisms of Macrophage Stimulation Through CD8: Macrophage CD8α and CD8β Induce Nitric Oxide Production and Associated Killing of the ParasiteLeishmania major

Abstract

AbstractPrior studies demonstrated that rat macrophages express CD8, which differs from T lymphocyte CD8 within the ligand binding domain. We investigated whether stimulation of macrophage CD8 could induce mediator release and regulate host defense. Cross-linking either CD8α (OX8, 5 μg/ml) or CD8β (341, 10 μg/ml) stimulated nitric oxide (NO) production, which correlated with an up-regulation of inducible NO synthase protein. Cell signaling inhibitors were used to elucidate the pathways of CD8α and CD8β stimulation. Genistein (broad spectrum protein tyrosine kinase inhibitor, 10 μg/ml), PP1 (src family kinase inhibitor, 5 μg/ml), polymyxin B (protein kinase C (PKC) inhibitor, 100 μg/ml), and Ro 31-8220 (PKC inhibitor, 1 μM) significantly inhibited anti-CD8α- and anti-CD8β-stimulated NO production and inducible NO synthase up-regulation, suggesting that tyrosine kinase(s) (src family) and PKC are involved in CD8 signaling. In addition, cross-linking CD8α stimulated NO-dependent macrophage killing of the parasite Leishmania major. For the first time, this work demonstrates that the β-chain of macrophage CD8, in addition to the α-chain, can regulate mediator release. These results further illustrate the importance of this molecule and support our previous data demonstrating differences between macrophage and T lymphocyte CD8. Additional studies on the signaling mechanisms and possible ligand(s) for macrophage CD8 will lead to a greater understanding of inflammation and host defense.