Low-level laser therapy stimulates the oxidative burst in human neutrophils and increases their fungicidal capacity.

Low-level laser therapy (LLLT) is known to enhance mitochondrial electron transfer and ATP production; thus, this study asked whether LLLT could stimulate the oxidative burst in human neutrophils (PMN) and improve their ability to kill microorganisms. Blood from healthy human subjects was collected and PMN were isolated from the samples. PMN were treated in vitro with 660 nm or 780 nm CW laser light at 40 mW power and increasing energies up to 19.2 J and were subsequently incubated with Candida albicans cells. Generation of hydroxyl radicals, hypochlorite anions and superoxide anions by PMN were checked using fluorescent probes and chemiluminescence assays; a microbicidal activity assay against C. albicans was also performed. LLLT excited PMN to a higher functional profile, which was translated as superior production of reactive oxygen species (ROS) and increased fungicidal capacity. The most efficacious energy was 19.2 J and, interestingly, the 660 nm light was even more efficacious than 780 nm at increasing the respiratory burst of PMN and the fungicidal capacity. Human neutrophils (PMN) were stimulated in vitro with 660 nm or 780 nm CW laser light at 40 mW of power and a total energy of 19.2 J. Low-level laser therapy (LLLT) excited PMN to a higher functional profile, which was translated as a superior production of reactive oxygen species (ROS) such as hydroxyl radicals (HO• ) and hypochlorite anions (ClO- ) (Figure) and increased fungicidal capacity against Candida albicans cells.

Aminoguanidine treatment increased NOX2 response in diabetic rats: Improved phagocytosis and killing of Candida albicans by neutrophils.

In this study, we show that aminoguanidine (AMG), an inhibitor of protein glycation, increases the NOX2 (phagocyte NADPH oxidase) response and microbicidal activity by neutrophils, regardless of diabetic status. The non-enzymatic glycation of proteins, yielding irreversible advanced glycation end products (AGEs), is involved in the development of diabetes complications, including alterations of signaling pathways and the generation of reactive oxygen species by phagocytes. The phagocytes produce ROS (reactive oxygen species) through activation of the NOX2 complex, which generates superoxide. The purpose of this study was to evaluate the effect of hyperglycemia and the glycation of proteins on the NOX2 activity of neutrophils and its implications for cellular physiology, with a focus on the microbicidal activity of these cells. We treated diabetic rats with AMG and evaluated neutrophil ROS generation and Candida albicans killing ability. We observed a large increase in the microbicidal activity of peritoneal neutrophils from AMG-treated rats. The increase was independent of diabetic status and myeloperoxidase activity. Collectively, our results suggest that AMG has an immunomodulator role that triggers an increase in the microbicidal response of neutrophils mainly related to reactive oxygen species production by NOX2.

Neutrophil dysfunction induced by hyperglycemia: modulation of myeloperoxidase activity.

Our data suggest that impaired activity of myeloperoxidase (MPO) may play an important role in the dysfunction of neutrophils from hyperglycemic rats. Neutrophil biochemical pathways include the NADPH oxidase system and the MPO enzyme. They both play important role in the killing function of neutrophils. The effect of hyperglycemia on the activity of these enzymes and the consequences with regard to Candida albicans phagocytosis and the microbicidal property of rat peritoneal neutrophils is evaluated here. The NADPH oxidase system activity was measured using chemiluminescence and cytochrome C reduction assays. MPO activity was measured by monitoring HOCl production, and MPO protein expression was analysed using Western blot and immunofluorescence. C. albicans phagocytosis and death were evaluated by optical microscopy using the May-Grunwald-Giemsa staining method. ROS generation kinetic was slightly delayed in the diabetic group. MPO expression levels were higher in diabetic neutrophils; however, MPO activity was decreased in these same neutrophils compared with the controls. C. albicans phagocytosis and killing were lower in the diabetic neutrophils. Based on our experimental model, the phagocytic and killing functions of neutrophil phagocytosis are impaired in diabetic rats because of the decreased production of HOCl, highlighting the importance of MPO in the microbicidal function of neutrophils.