P2X7 from J774 murine macrophages acts as a scavenger receptor for bacteria but not yeast
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We studied the effects of extracellular ATP and Ca2 on uptake of bacteria (Staphylococcus aureus or Escherichia coli) and live yeast (Candida glabrata) by J774 macrophages to determine the role of endogenous P2X7 receptors in phagocytosis. Our findings show that phagocytosis of bio-particles coated with S. aureus or E. coli was blocked by ATP and the P2X7 receptor agonist BzATP, while yeast phagocytosis was not. A438079, an antagonist of P2X7 receptors, partially reverted the effects of ATP on bacterial phagocytosis. To determine if P2X7-mediated Ca2 entry into macrophages was blocking the engulfment of bacteria, we measured phagocytic activity in the absence or presence of 2 mM extracellular Ca2 with or without ATP. Ca2 , in the absence of ATP, was required for engulfment of E. coli and C. glabrata but not S. aureus. Adding ATP inhibited phagocytosis of S. aureus and E. coli regardless of Ca2 , suggesting that Ca2 entry was not important for inhibiting phagocytosis. On the other hand, phagocytosis of normal or hyper-adherent C. glabrata mutants had an absolute requirement for extracellular Ca2 due to yeast adhesion to macrophages mediated by Ca2 -dependent adhesion proteins. We conclude that unstimulated P2X7 from J774 cells act as scavenger receptor for the uptake of S. aureus and E. coli but not of yeast; Ca2 entry via P2X7 receptors play no role in phagocytosis of S. aureus and E. coli; while the effect of Ca2 on C. glabrata phagocytosis was mediated by the adhesins Epa1, Epa6 and Epa7. © 2016 Elsevier Inc.
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We studied the effects of extracellular ATP and Ca2%2b on uptake of bacteria (Staphylococcus aureus or Escherichia coli) and live yeast (Candida glabrata) by J774 macrophages to determine the role of endogenous P2X7 receptors in phagocytosis. Our findings show that phagocytosis of bio-particles coated with S. aureus or E. coli was blocked by ATP and the P2X7 receptor agonist BzATP, while yeast phagocytosis was not. A438079, an antagonist of P2X7 receptors, partially reverted the effects of ATP on bacterial phagocytosis. To determine if P2X7-mediated Ca2%2b entry into macrophages was blocking the engulfment of bacteria, we measured phagocytic activity in the absence or presence of 2 mM extracellular Ca2%2b with or without ATP. Ca2%2b, in the absence of ATP, was required for engulfment of E. coli and C. glabrata but not S. aureus. Adding ATP inhibited phagocytosis of S. aureus and E. coli regardless of Ca2%2b, suggesting that Ca2%2b entry was not important for inhibiting phagocytosis. On the other hand, phagocytosis of normal or hyper-adherent C. glabrata mutants had an absolute requirement for extracellular Ca2%2b due to yeast adhesion to macrophages mediated by Ca2%2b-dependent adhesion proteins. We conclude that unstimulated P2X7 from J774 cells act as scavenger receptor for the uptake of S. aureus and E. coli but not of yeast; Ca2%2b entry via P2X7 receptors play no role in phagocytosis of S. aureus and E. coli; while the effect of Ca2%2b on C. glabrata phagocytosis was mediated by the adhesins Epa1, Epa6 and Epa7. © 2016 Elsevier Inc.
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ATP; Bacteria; Ca2 ; P2X7 receptor; Phagocytosis; Yeast 3 [[5 (2,3 dichlorophenyl) 1h tetrazol 1 yl]methyl]pyridine; 3' o (4 benzoylbenzoyl)adenosine triphosphate; adenosine triphosphate; adhesin; calcium ion; epa1 protein; epa6 protein; epa7 protein; purinergic P2X7 receptor; scavenger receptor; unclassified drug; adhesin; purinergic P2X7 receptor; scavenger receptor; animal cell; Article; bacterial cell; bacterial strain; bacterium adherence; calcium transport; Candida albicans; cell adhesion; controlled study; drug effect; Escherichia coli; extracellular calcium; fungal strain; fungus mutant; in vitro study; macrophage; mouse; nonhuman; phagocytosis; priority journal; Staphylococcus aureus; yeast cell; animal; bacterial phenomena and functions; calcium signaling; Candida glabrata; cell culture; cell line; macrophage; metabolism; microbiology; phagocytosis; physiology; Adhesins, Bacterial; Animals; Bacterial Physiological Phenomena; Calcium Signaling; Candida glabrata; Cell Line; Cells, Cultured; Macrophages; Mice; Phagocytosis; Receptors, Purinergic P2X7; Receptors, Scavenger
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ATP; Bacteria; Ca2+; P2X7 receptor; Phagocytosis; Yeast 3 [[5 (2,3 dichlorophenyl) 1h tetrazol 1 yl]methyl]pyridine; 3' o (4 benzoylbenzoyl)adenosine triphosphate; adenosine triphosphate; adhesin; calcium ion; epa1 protein; epa6 protein; epa7 protein; purinergic P2X7 receptor; scavenger receptor; unclassified drug; adhesin; purinergic P2X7 receptor; scavenger receptor; animal cell; Article; bacterial cell; bacterial strain; bacterium adherence; calcium transport; Candida albicans; cell adhesion; controlled study; drug effect; Escherichia coli; extracellular calcium; fungal strain; fungus mutant; in vitro study; macrophage; mouse; nonhuman; phagocytosis; priority journal; Staphylococcus aureus; yeast cell; animal; bacterial phenomena and functions; calcium signaling; Candida glabrata; cell culture; cell line; macrophage; metabolism; microbiology; phagocytosis; physiology; Adhesins, Bacterial; Animals; Bacterial Physiological Phenomena; Calcium Signaling; Candida glabrata; Cell Line; Cells, Cultured; Macrophages; Mice; Phagocytosis; Receptors, Purinergic P2X7; Receptors, Scavenger
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