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Streptococcus pyogenes CAMP factor promotes calcium ion uptake in RAW264.7 cells.

Microbiology and Immunology 2018 September 14
Streptococcus pyogenes is a bacterium that causes systemic diseases such as pharyngitis and toxic shock syndrome. S. pyogenes produces molecules that inhibit the function of the human immune system, thus allowing to grow and spread of the pathogen in tissues. It is known that S. pyogenes Christie Atkins Munch-Petersen (CAMP) factor induces vacuolation in macrophages, but the mechanism remains unclear. In the current study, we investigated the mechanism of vacuolation induced by CAMP factor in macrophages. CAMP factor induced calcium ion uptake in murine macrophage RAW264.7 cells. In addition, ethylenediaminetetraacetic acid (EDTA) inhibited calcium ion uptake and vacuolation in the cells. L-type voltage-dependent calcium ion channel blockers nifedipine and verapamil reduced vacuolation. Furthermore, phosphoinositide 3-kinase inhibitors LY294002 and wortmannin also inhibited the vacuolation induced by CAMP factor. Fluorescent microscopy analysis revealed that clathrin localized to the vacuoles. These results suggest that the vacuolation was related to the calcium ion uptake in RAW264.7 cells via L-type voltage-dependent calcium ion channels. Therefore, we conclude that the vacuoles induced by S. pyogenes CAMP factor in macrophages were clathrin-dependent endosomes induced by activation of the phosphoinositide 3-kinase signaling pathway through calcium ion uptake.

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