Rationale: Mycolic acid (MA) constitutes a major and distinguishing cell wall biolipid from Mycobacterium tuberculosis. MA interferes with the lipid homeostasis of alveolar macrophages, inducing differentiation into foamy macrophages exhibiting increased proinflammatory function. Objectives: We verified the interference of this altered macrophage function with inhaled antigen-triggered allergic airway inflammation and underlying Th2 lymphocyte reactivity. Methods: Using ovalbumin (OVA) as model allergen, C57BL/6 or BALB/C mice were sensitized by OVA-alum immunization. Experimental asthma, triggered subsequently by repetitive nebulized OVA inhalation, was assessed using as readout parameters eosinophilia, peribronchial inflammation and Th2 cytokine function. Measurements and Main Results: A single intratracheal treatment of sensitized mice with MA, inserted into liposomes as carriers, prevented the onset of OVA-triggered allergic airway inflammation and promoted unresponsiveness to a secondary set of allergen exposures. The development of this tolerant condition required an 8-day lapse following MA-instillation, coinciding with the appearance of foamy alveolar macrophages. MA-conditioned CD11b⁺F4/80⁺ macrophages, transferred to the airways, mimicked the tolerogenic function of instilled MA, however without the 8-day lapse requirement. Indicative for a macrophage-mediated tolerogenic antigen-presenting function, MHC-mismatched donor macrophages failed to promote tolerance. Furthermore, Treg markers were strongly increased and established tolerance was lost following in situ depletion of CD25⁺ Treg cells. Contrarily to the IL-10-dependence of tolerogenic dendritic cells, IFN- deficiency but not IL-10 deficiency abrogated the tolerogenic capacity of MA-conditioned macrophages. Conclusions: These results document an innate-driven, Mycobacterium tuberculosis MA-triggered immune regulatory mechanism in control of pulmonary allergic responses by converting macrophages into IFN--dependent tolerogenic antigen presenting cells.