Respiratory Burst

It is well documented that the polysaccharide glucomannan (GM), an abundant constituent of the fungal cell wall, in the form of particulate induces strong activation of phagocytes, however, the effects of soluble GM are not known. Activation of phagocyte anti-microbial mechanisms is a crucial part of the innate host defense against invading pathogens. However, under uncontrolled inflammatory conditions they contribute to damage of surrounding tissues. Thus, to prevent these deleterious effects, the activation of phagocytes is a tightly regulated process. Therefore, in this study we analyzed the effect of soluble GM on some neutrophil functions such as reactive oxygen species production, degranulation, and receptor mobilization at the plasma membrane. Soluble GM at the tested concentrations did not stimulate oxidative burst of phagocytes directly but significantly potentiated oxidative burst in response to opsonized zymosan particles. GM induced significant phosphorylation of p47phox subunit of NADPH oxidase on Ser345. This priming effect of GM was accompanied by time and concentration dependent degranulation characterized by increased surface expression of receptors stored in neutrophil granules (CD10, CD11b, CD14, CD35, and CD66b). Degranulation was further confirmed by increase of elastase activity in media. Thus, it could be suggested that soluble GM induces priming of phagocytes connected with their degranulation, the increase of surface receptor expression, and potentiation of oxidative burst response to opsonized particles through the activation of NADPH oxidase.

After transmission by infected mosquitoes, malaria sporozoites rapidly travel to the liver. To infect hepatocytes, sporozoites traverse Kupffer cells, but surprisingly, the parasites are not killed by these resident macrophages of the liver. Here we show that Plasmodium sporozoites and recombinant circumsporozoite protein (CSP) suppress the respiratory burst in Kupffer cells. Sporozoites and CSP increased the intracellular concentration of cyclic adenosyl mono-phosphate (cAMP) and inositol 1,4,5-triphosphate in Kupffer cells, but not in hepatocytes or liver endothelia. Preincubation with cAMP analogues or inhibition of phosphodiesterase also inhibited the respiratory burst. By contrast, adenylyl cyclase inhibition abrogated the suppressive effect of sporozoites. Selective protein kinase A (PKA) inhibitors failed to reverse the CSP-mediated blockage and stimulation of the exchange protein directly activated by cAMP (EPAC), but not PKA inhibited the respiratory burst. Both blockage of the low-density lipoprotein receptor-related protein (LRP-1) with receptor-associated protein and elimination of cell surface proteoglycans inhibited the cAMP increase in Kupffer cells. We propose that by binding of CSP to LRP-1 and cell surface proteoglycans, malaria sporozoites induce a cAMP/EPAC-dependent, but PKA-independent signal transduction pathway that suppresses defence mechanisms in Kupffer cells. This allows the sporozoites to safely pass through these professional phagocytes and to develop inside neighbouring hepatocytes.

Rodríguez AB, Nogales G, Ortega E, Barriga C. Melatonin controls superoxide anion level: Modulation of superoxide dismutase activity in ring dove heterophils. J. Pineal Res. 1998; 24:9–14. © Munksgaard, CopenhagenRodríguez AB, Nogales G, Ortega E, Barriga C. Melatonin controls superoxide anion level: Modulation of superoxide dismutase activity in ring dove heterophils. J. Pineal Res. 1998; 24:9–14. © Munksgaard, CopenhagenAbstractThe pineal gland and the immune system in mammals and birds are functionally connected, with some recent findings showing that melatonin, the major secretory product of the pineal gland in vertebrates, is a free-radical scavenger and general antioxidant. In birds, heterophils, with their ability to ingest and kill different antigens, play a central role in the host defense mechanism. This in vitro study investigated the possible antioxidant action of melatonin in heterophils isolated from adult male and female ring dove)Streptopelia risoria). The superoxide anion levels (O2−) and the superoxide dismutase (SOD) activity were evaluated as indicators of the metabolic burst of heterophils after ingestion of inert particles (latex beads) at the basal time (0 min) and 15, 30,45, and 60 min of incubation with 100 μM melatonin. The heterophil O2− levels, measured by the nitroblue tetrazolium reduction test, were lower after incubation in the presence of melatonin at all the times studied, a time-dependent effect. Latex beads produced a significant increase in SOD activity, whereas there was a significant decrease in the presence of melatonin. In addition, when heterophils were incubated in the presence of both latex beads and melatonin, the indole appeared to annul the enhancement of SOD activity (U-525nm) produced by latex beads. In conclusion, our findings show that the neurohormone melatonin could be a useful antioxidant.The pineal gland and the immune system in mammals and birds are functionally connected, with some recent findings showing that melatonin, the major secretory product of the pineal gland in vertebrates, is a free-radical scavenger and general antioxidant. In birds, heterophils, with their ability to ingest and kill different antigens, play a central role in the host defense mechanism. This in vitro study investigated the possible antioxidant action of melatonin in heterophils isolated from adult male and female ring dove)Streptopelia risoria). The superoxide anion levels (O2−) and the superoxide dismutase (SOD) activity were evaluated as indicators of the metabolic burst of heterophils after ingestion of inert particles (latex beads) at the basal time (0 min) and 15, 30,45, and 60 min of incubation with 100 μM melatonin. The heterophil O2− levels, measured by the nitroblue tetrazolium reduction test, were lower after incubation in the presence of melatonin at all the times studied, a time-dependent effect. Latex beads produced a significant increase in SOD activity, whereas there was a significant decrease in the presence of melatonin. In addition, when heterophils were incubated in the presence of both latex beads and melatonin, the indole appeared to annul the enhancement of SOD activity (U-525nm) produced by latex beads. In conclusion, our findings show that the neurohormone melatonin could be a useful antioxidant.

The immune response of European sea bass, Dicentrarchus labrax, to a natural infection by the copepod parasite Lernanthropus kroyeri was evaluated for the first time in vivo. The results clearly demonstrated the triggering of the fish immune system by the parasite. Lysozyme activity and alternative complement pathway were involved in the early action against the parasitical infection, whilst classical complement and respiratory burst (RB) activity took over in the later stages of infection. It was hypothesized that the levels of alternative and classical complement and RB stimulation indexes may determine the resistance capacity of the fish to the parasite. It is not clear how parasites can survive despite the strong immunological arsenal deployed by the fish. The continual increase of prevalence and severity of parasite infection suggested that the parasite’s mechanism of evasion of the immune system was extremely successful. The contrasting decrease in the negative effects of parasites on the fish health (such as gills anaemia) suggested that an equilibrium between the parasites and their hosts was reached in chronic infection. These dynamic interactions between parasites and fish hosts were probably the main determinant of host specificity.