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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.