Pérez-Flores Gabriela, Hernández-Silva Cesar, Gutiérrez-Escobedo Guadalupe, De Las Peñas Alejandro, Castaño Irene, Arreola Jorge, Pérez-Cornejo Patricia
Unidad Académica Multidisciplinaria Zona Huasteca, Universidad Autónoma de San Luis Potosí, Cd. Valles, SLP 79060, Mexico.
Department of Physiology and Biophysics, School of Medicine, Universidad Autónoma de San Luis Potosí, San Luis Potosí, SLP 78210, Mexico.
Biochem Biophys Res Commun. 2016 Dec 2;481(1-2):19-24. doi: 10.1016/j.bbrc.2016.11.027. Epub 2016 Nov 8.
We studied the effects of extracellular ATP and Ca 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 Ca entry into macrophages was blocking the engulfment of bacteria, we measured phagocytic activity in the absence or presence of 2 mM extracellular Ca with or without ATP. Ca, 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 Ca, suggesting that Ca 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 Ca due to yeast adhesion to macrophages mediated by Ca-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; Ca entry via P2X7 receptors play no role in phagocytosis of S. aureus and E. coli; while the effect of Ca on C. glabrata phagocytosis was mediated by the adhesins Epa1, Epa6 and Epa7.
我们研究了细胞外ATP和Ca对J774巨噬细胞摄取细菌(金黄色葡萄球菌或大肠杆菌)和活酵母(光滑念珠菌)的影响,以确定内源性P2X7受体在吞噬作用中的作用。我们的研究结果表明,被金黄色葡萄球菌或大肠杆菌包被的生物颗粒的吞噬作用被ATP和P2X7受体激动剂BzATP阻断,而酵母吞噬作用未被阻断。P2X7受体拮抗剂A438079部分逆转了ATP对细菌吞噬作用的影响。为了确定P2X7介导的Ca进入巨噬细胞是否会阻断细菌的吞噬,我们在有无ATP的情况下,测量了存在或不存在2 mM细胞外Ca时的吞噬活性。在没有ATP的情况下,Ca是大肠杆菌和光滑念珠菌吞噬所必需的,但不是金黄色葡萄球菌吞噬所必需的。添加ATP会抑制金黄色葡萄球菌和大肠杆菌的吞噬作用,而与Ca无关,这表明Ca的进入对抑制吞噬作用并不重要。另一方面,正常或高粘附性光滑念珠菌突变体的吞噬作用绝对需要细胞外Ca,这是由于酵母通过Ca依赖性粘附蛋白介导与巨噬细胞粘附。我们得出结论,J774细胞未受刺激的P2X7作为摄取金黄色葡萄球菌和大肠杆菌而非酵母的清道夫受体;通过P2X7受体进入的Ca在金黄色葡萄球菌和大肠杆菌的吞噬作用中不起作用;而Ca对光滑念珠菌吞噬作用的影响是由粘附素Epa1、Epa6和Epa7介导的。