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Toll-Like Receptor 2- and MyD88-Dependent Phosphatidylinositol 3-Kinase and Rac1 Activation Facilitates the Phagocytosis of Listeria monocytogenes by Murine Macrophages

Shen, Yanna, Kawamura, Ikuo, Nomura, Takamasa, Tsuchiya, Kohsuke, Hara, Hideki, Dewamitta, Sita R., Sakai, Shunsuke, Qu, Huixin, Daim, Sylvia, Yamamoto, Takeshi, Mitsuyama, Masao
Infection and immunity 2010 v.78 no.6 pp. 2857-2867
Gram-positive bacteria, Listeria monocytogenes, cycloheximide, cytokines, guanosinetriphosphatase, intravenous injection, lipopolysaccharides, macrophages, mice, models, phagocytosis, phosphatidylinositol 3-kinase, receptors
Toll-like receptors (TLRs) play a key role in the innate immune response by sensing bacterial ligands. The mechanisms involved in the TLR-mediated cytokine response are well established; however, the possible contribution of TLR-dependent recognition of bacteria to macrophage phagocytosis remains unclear. Listeria monocytogenes is an intracellular, parasitic, Gram-positive bacterium recognized mainly by TLR2. In this study, we investigated whether TLR2-dependent signaling is involved in the phagocytosis of L. monocytogenes by macrophages. We found no difference in the number of L. monocytogenes cells associating with wild-type (WT) and TLR2⁻/⁻ macrophages 1 h after infection. However, the number of L. monocytogenes cells phagocytosed in TLR2⁻/⁻ and MyD88⁻/⁻ macrophages was significantly lower than that of WT macrophages. In addition, lipopolysaccharide (LPS) treatment restored impaired phagocytic activity of TLR2⁻/⁻ macrophages but did not enhance the activity of MyD88⁻/⁻ macrophages. The efficiency of phagocytosis was suppressed by inhibitors of phosphatidylinositol 3-kinase (PI3K) and the small Rho GTPases but not by cycloheximide. Moreover, functional activation of PI3K and Rac1 was impaired in TLR2⁻/⁻ and MyD88⁻/⁻ macrophages. In an in vivo infection model, we found significantly lower numbers of L. monocytogenes cells phagocytosed in peritoneal macrophages of TLR2⁻/⁻ and MyD88⁻/⁻ mice after intraperitoneal infection. Moreover, a lower number of bacteria were detected in the spleens of TLR2⁻/⁻ mice 1 day after intravenous infection than in WT mice. These results clearly indicated that TLR2-MyD88-dependent signaling enhances the basal level of phagocytosis of L. monocytogenes by macrophages through activation of PI3K and Rac1, not by synthesis of proinflammatory cytokines or expression of phagocytic receptors.