The ability of macrophages to control the growth of microorganisms is increased by macrophage activation. Previously, it was shown that epinephrine activated mouse macrophages to resist the growth of Mycobacterium avium via ␣ 2 -adrenergic stimulation. In the present study, we show that the ␣ 2 -adrenergic agonist (␣ 2 -agonist) clonidine induced resistance to M. avium growth in the RAW264.7 mouse macrophage cell line. The ability of catecholamines to induce resistance to mycobacteria was specific to ␣ 2 -adrenergic stimulation, as ␣ 1 -,  1 -, and  2 -agonists had no effect. Receptor signaling through Gi proteins was required. A G-protein antagonist specific for the ␣ subunits of the Go/Gi family blocked the increased resistance induced by clonidine, while a Gs-protein antagonist was without effect. Both nitric oxide (NO) production and superoxide (O 2 ؊ ) production were required for the increased resistance to M. avium growth induced by clonidine. Although NO production was required, clonidine did not increase the level of NO in M. avium-infected cells. Since NO and O 2 ؊ interact to produce peroxynitrite (ONOO ؊ ), we examined whether ONOO ؊ mediates the increased resistance to M. avium induced by clonidine. 5,10,15,20-Tetrakis(4-sulfonatophenyl)prophyrinato iron (III) chloride (FeTPPS), a specific scavenger of ONOO ؊ , inhibited the effect of clonidine on M. avium growth. Clonidine also increased the production of ONOO ؊ in M. avium-infected RAW264.7 cells, as measured by the oxidation of 123-dihydrorhodamine and the production of nitrated tyrosine residues. We therefore conclude that ␣ 2 -adrenergic stimulation activates macrophages to resist the growth of M. avium by enhancing the production of ONOO ؊ .Macrophages are among the first cells of the host to confront microbes and are important effector cells in innate resistance to intracellular microbial pathogens. The outcome of this initial encounter with an intracellular pathogen is that either the macrophage resists the growth of the microorganism or the microorganism adapts and replicates within the macrophage. The ability of the macrophage to resist the growth of the microorganism is dependent on the activation state of the macrophage. Cytokines, such as gamma interferon (IFN-␥), granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor alpha (TNF-␣), activate macrophages to resist the growth of intracellular pathogens by enhancing the production of the major antimicrobial effector molecules, including reactive oxygen species and nitric oxide (38,46,53).The sympathetic nervous system acts to maintain homeostasis during periods of stress by releasing norepinephrine at sympathetic nerve endings and epinephrine from the adrenal medulla (3,20,65). These catecholamine hormones modulate the activities of cells, including cells of the immune system (35). Macrophage function can be either activated (50,59,60) or suppressed (9,31,58,63,67) by catecholamines. A study by Boomershine et al. (9) showed that the addition of epinephrine to IFN-␥-...