Peptidoglycan fromBacillus anthracisInhibits Human Macrophage Efferocytosis in Part by Reducing Cell Surface Expression of MERTK and TIM-3

Bacillus anthracispeptidoglycan (PGN) is a major component of the bacterial cell wall and a key pathogen-associated molecular pattern (PAMP) contributing to anthrax pathology, including organ dysfunction and coagulopathy. Increases in apoptotic lymphocytes are a late-stage feature of anthrax and sepsis, suggesting there is a defect in apoptotic clearance. Here, we tested the hypothesis thatB. anthracisPGN inhibits the capacity of human monocyte-derived, tissue-like macrophages (MΦ) to efferocytose apoptotic cells. Exposure of CD206+CD163+MΦ to PGN for 24h impaired efferocytosis in a manner dependent on human serum opsonins but independent of complement component C3. PGN treatment reduced cell surface expression of the pro-efferocytic signaling receptors MERTK, TYRO3, AXL, integrin αVβ5, CD36 and TIM-3, whereas TIM-1, αVβ5, CD300b, CD300f, STABILIN-1 and STABILIN-2 were unaffected. Soluble forms of MERTK, TYRO3, AXL, CD36, and TIM-3 were increased in PGN-treated supernatants, suggesting involvement of proteases. ADAM17 is a major membrane-bound protease implicated in mediating efferocytotic receptor cleavage. ADAM17 inhibitors TAPI-0 and Marimastat abolished TNF release, indicating effective protease inhibition, modestly increased cell-surface levels of MerTK and TIM-3 but only partially restored efferocytic capacity by PGN-treated MΦ. We conclude that human serum factors are required for optimal recognition of PGN by human MΦ and thatB. anthracisPGN inhibits efferocytosis in part by reducing cell surface expression of efferocytic receptors.