Protective innate immunity against <i>Pneumocystis</i> does not require Stat6-dependent macrophage polarization

<i>Pneumocystis</i> species are respiratory fungal pathogens that cause life-threatening opportunistic infections in immunocompromised hosts. <i>Pneumocystis</i> typically evade pulmonary innate immunity but are efficiently eradicated by a functional adaptive immune response. FVB/NJ mice are unique in that they display protective alveolar macrophage-dependent innate immunity against <i>Pneumocystis</i>, and remain resistant to infection even in the absence of CD4⁺ T lymphocyte function. FVB/NJ alveolar macrophages (AMs) were found to display an M2-biased phenotype at baseline, which was potentiated after stimulation with <i>Pneumocystis</i>, suggesting that macrophage polarization may dictate the outcome of the <i>Pneumocystis</i>-macrophage interaction. To determine whether Stat6, a key global regulator of M2 polarization, was required for FVB/NJ innate immunity, FVB Stat6^-/- mice were generated. FVB Stat6-deficient AMs were markedly impaired in their ability to polarize to an M2 phenotype when stimulated with Th2 cytokines. However, FVB Stat6^-/- mice remained highly resistant to infection, indicating that Stat6 signaling is dispensable for innate FVB/NJ resistance. Despite the loss of Stat6 signaling, primary AMs from FVB Stat6^-/- mice maintained baseline expression of M2 markers, and also strongly upregulated M2-associated genes following direct stimulation with <i>Pneumocystis</i>. Additional FVB/NJ knockout strains were generated, but only FVB MerTK^-/- mice showed a marginally increased susceptibility to <i>Pneumocystis</i> infection. Together, these findings demonstrate that effective FVB/NJ innate immunity against <i>Pneumocystis</i> does not require Stat6 signaling and suggest that alternative pathways regulate M2 bias and macrophage-mediated innate resistance in FVB/NJ mice.