The reported effects of different families of fatty acids (FA; SFA, MUFA, n-3 and n-6 PUFA) on human health and the importance of macrophage respiratory burst and cytokine release to immune defence led us to examine the influence of palmitic acid (PA), oleic acid (OA), linoleic acid, arachidonic acid, EPA and DHA on macrophage function. We determined fungicidal activity, reactive oxygen species (ROS) and cytokine production after the treatment of J774 cells with non-toxic concentrations of the FA. PA had a late and discrete stimulating effect on ROS production, which may be associated with the reduced fungicidal activity of the cells after treatment with this FA. OA presented a sustained stimulatory effect on ROS production and increased fungicidal activity of the cells, suggesting that enrichment of diets with OA may be beneficial for pathogen elimination. The effects of PUFA on ROS production were time- and dose-dependently regulated, with no evident differences between n-3 and n-6 PUFA. It was worth noting that most changes induced after stimulation of the cells with lipopolysaccharide were suppressed by the FA. The present results suggest that supplementation of the diet with specific FA, not classes of FA, might enable an improvement in host defence mechanisms or a reduction in adverse immunological reactions.

Dietary long-chain n-3 PUFA (n-3 LCPUFA) are thought to have immune-modulating effects, but the specific effects and mechanisms are not fully elucidated. The aim of this study was to determine whether dietary n-3 LCPUFA could affect ex vivo oxidative burst in healthy young men. The study had a randomised 2 × 2-factorial design in which subjects were randomly assigned to 8-week supplementation with capsules containing fish oil (about 2·9 g n-3 LCPUFA/d) or olive oil (control). Subjects were also randomly assigned to household use of oils and fat spreads with a high or a low 18 : 2n-6 content. At baseline and at the end of the intervention, the fatty acid composition of peripheral blood mononuclear cells (PBMC) was analysed by GLC and oxidative burst was studied in whole blood stimulated with zymosan using luminol-enhanced chemiluminescence. The PBMC content of n-3 LCPUFA was markedly increased by the fish oil-supplementation (P < 0·001, compared to the olive oil groups). No effect of the intervention was observed on neutrophil count, but one measure of the zymosan-induced oxidative burst was higher in the fish oil groups (P = 0·03) compared to the olive oil groups. The fat intervention did not in itself affect oxidative burst neither did it change the effect of the fish-oil intervention. The measures of oxidative burst at the end of the intervention period were found to be associated with the DHA content of PBMC (r 0·44, P = 0·016), suggesting a dose–response relationship. These results indicate that n-3 LCPUFA may have immuno-stimulating effects.

We investigated population differences in immunological adaptation of three-spined sticklebacks (Gasterosteus aculeatus) to one of their most abundant macroparasites, the eye fluke Diplostomum pseudospathaceum. We compared infection success in lab-bred fish of 2 populations in northern Germany, from a lake, where eye flukes are prevalent, and a river, where these parasites do not occur. In order to discriminate between protection through innate and acquired immunity, we exposed fish either only once or repeatedly. Lake fish were significantly less susceptible than river sticklebacks already after a single exposure, indicating that in sympatric hosts innate immunity plays the major role in the defence against this helminth infection. In both habitat types, previous exposures only marginally decreased infection rates within 12 weeks. Lake fish showed higher immunocompentence by means of respiratory burst activity and spleen size, regardless of the infection status. Furthermore, they were in a better energy status than river fish, as indicated by a higher hepatosomatic index and haematocrit value. Interestingly, F1 hybrid fish of both populations ranged between the pure habitat types in parasite susceptibility as well as in immunological and condition parameters. Our results suggest that sticklebacks from lakes are better adapted to cope with higher parasite abundance in this habitat.

To determine the effects of EPA, stearidonic acid (STA) or γ-linolenic acid (GLA) on immune outcomes, healthy male subjects consumed one of seven oil blends for 12 weeks. EPA consumption increased the EPA content of peripheral blood mononuclear cells (PBMC). Consumption of GLA (2·0 g/d) in the absence of STA or EPA increased di-homo-GLA content in PBMC. Neither STA nor its derivative 20:4n-3 appeared in PBMC when STA (<1·0 g/d) was consumed. However, STA (1·0 g/d), in combination with GLA (0·9 g/d), increased the proportion of EPA in PBMC. None of the treatments altered neutrophil or monocyte phagocytosis or respiratory burst, production of inflammatory cytokines by monocytes, T lymphocyte proliferation or the delayed-type hypersensitivity response. Production of cytokines by T lymphocytes increased in all groups, with no differences among them. The proportion of lymphocytes that were natural killer cells decreased significantly in subjects receiving 2·0 g EPA or GLA/d. There were no other effects on lymphocyte sub-populations. Plasma IgE concentration decreased in most groups, but not in the control group. Plasma IgG2 concentration increased in the EPA group. Thus, EPA or GLA at a dose of 2·0 g/d have little effect on key functions of neutrophils, monocytes and T lymphocytes, although at this dose these fatty acids decrease the number of natural killer cells. At this dose EPA increases IgG2 concentrations. STA can increase immune cell EPA status, but at 1·0 g/d does not affect human immune function.

Glutamine (GLN) and arginine (ARG) are recognized for their ability to modulate immune cell function. However, the metabolic pathways involved in their action remain unclear. It was recently shown that GLN- or ARG-enriched diets increased radical oxygen species (ROS) production by neutrophils from stressed rats. Since these two amino acids have a tied metabolism, we hypothesized that conversion between GLN and ARG (and its active metabolites NO• and polyamines) might be involved. To test this hypothesis male Sprague–Dawley rats (n 117) were randomized into thirteen groups: rats in eleven groups were rendered catabolic by dexamethasone injection (1·5 mg/kg per d for 5 d) and 6·8 mmol either GLN, ARG or non-essential amino acids (NEAA; glycine, alanine and histidine)/kg per d were given by the enteral route; one group was pair-fed to the treated groups. The regimens of all the groups were rendered isonitrogenous by the addition of NEAA. The last group was not treated and was fed ad libitum. For each supplementation three subgroups were formed, each of which received a specific inhibitor: methionine sulfoximine (inhibitor of GLN synthase; 100 mg/kg per d), S-methylthiourea (inhibitor of inducible NO• synthase (iNOS); 50 mg/kg per d) and difluoromethylornithine (inhibitor of ornithine decarboxylase (ODC); 50 mg/kg per d). Oxidative metabolism, intracellular H2O2, and extracellular O2•- production were measured in unstimulated and phorbol myristate acetate-stimulated polymorphonuclear neutrophils. GLN- and ARG-enriched diets increased respiratory burst by neutrophils (oxidative metabolism of 152 (SEM 24) AND 138 (sem 45) v. 57 (sem 18) mV for GLN-, ARG- and NEAA-enriched diets respectively, P<0·05). In vivo inhibition of iNOS or ODC decreased ROS production induced by GLN and ARG. In vivo inhibition of GLN synthase did not modify the effect of ARG on ROS production. In conclusion, GLN and ARG modulate ROS production in neutrophils from stressed rats by the same pathway involving polyamine and NO• synthesis.

The effect of regular intake of low doses of an effervescent multivitamin preparation on the free-radical-producing activity of murine peritoneal macrophages under conditions resembling a possible infection was studied in vitro. Initially, several groups of mice were fed a basal diet and given, for 2 weeks, water without or with supplementation of either α -tocopherol, ascorbic acid, riboflavin or a multivitamin preparation. The supplementation period was followed by a 2-week wash-out time interval during which control and multivitamin groups received deionized water. Macrophage stimulation tests using chemiluminescent spectroscopy were performed at the end of the supplementation and wash-out periods to determine cell counts and their capacity to produce free radicals. Multivitamin supplementation increased the number, and the reactive oxygen species-producing activity, of macrophages. This effect persisted for 2 weeks after higher doses of supplementation were stopped. Multivitamin supplementation lowered the steady-state free radical concentrations of liver and spleen as measured by electron paramagnetic resonance spectroscopy. It also increased the antioxidant reactivity of the same organs, while there was no effect on the free radical concentration and antioxidant capacity of the kidney and brain. When taken regularly, low doses of multivitamin supplementation may have a beneficial effect on the defence mechanisms of the organism.

The non-stimulated and phorbol 12-myristate 13-acetate (PMA)-stimulated luminol-augmented cellular chemiluminescence (CL) response and viability of milk and blood polymorphonuclear leukocytes (PMN) were determined in lactating dairy cows during different stages of lactation. In the first study, ten healthy cows each in early, mid and late lactation were compared. In a second study, the same measurements as in the first study were evaluated longitudinally in 12 cows during 1 month following parturition. The CL activity and myeloperoxidase (MPO) content of milk PMN and macrophages (M) were also compared. Milk M did not possess MPO activity and were devoid of any luminol-enhanced CL. The CL activity of milk and blood PMN was significantly lower in early lactation than in mid and late lactation (P < 0·001). Whereas little changes were observed in viability of blood PMN, the viability of milk PMN was lower in early lactation than in mid and late lactation (P < 0·001). The percentage of PMN in isolated milk cells was also lower during early lactation than during mid and late lactation (P < 0·001). The CL activity in response to PMA during early, mid and late lactation increased 13, 59 and 42-fold in blood PMN and 1·7, 2·6 and 2·4-fold in milk PMN, respectively, in comparison with non-stimulated PMN. The CL activity, both in milk and blood PMN, the milk PMN viability and the percentage of milk PMN were lowest between 3 d and 11 d post partum. These observed changes immediately after calving could contribute to a higher susceptibility to mastitis in that period.

Background and objective Respiratory burst is an essential component of the neutrophil's biocidal function. In vitro, sodium thiopental, isoflurane and lidocaine each inhibit neutrophil respiratory burst. The objectives of this study were (a) to determine the effect of a standard clinical induction/tracheal intubation sequence on neutrophil respiratory burst and (b) to determine the effect of intravenous lidocaine administration during induction of anaesthesia on neutrophil respiratory burst.

Methods Twenty ASA I and II patients, aged 18–60 years, undergoing elective surgery were studied. After induction of anaesthesia [fentanyl (2 μg kg−1), thiopental (4–6 mg kg−1), isoflurane (end-tidal concentration 0.5–1.5%) in nitrous oxide (66%) and oxygen], patients randomly received either lidocaine 1.5 mg kg−1 (group L) or 0.9% saline (group S) prior to tracheal intubation. Neutrophil respiratory burst was measured immediately prior to induction of anaesthesia, immediately before and 1 and 5 min after lidocaine/saline.

Results Neutrophil respiratory burst decreased significantly after induction of anaesthesia in both groups [87.4 ± 8.2% (group L) and 88.5 ± 13.4% (group S) of preinduction level (P < 0.01 both groups)]. After intravenous lidocaine (but not saline) administration, neutrophil respiratory burst returned towards preinduction levels, both before (97.1 ± 23.6%) and after (94.4 ± 16.6%) tracheal intubation.

Conclusion Induction of anaesthesia and tracheal intubation using thiopentone and isoflurane, inhibit neutrophil respiratory burst. This effect may be diminished by the administration of lidocaine.

The activation of peritoneal macrophage effector functions after therapy with free PZQ and PZQ incorporated in liposomes (lip.PZQ) was studied in the Mesocestoides corti–mouse model system. Each drug formulation was administered to an infected group of mice in 6 daily doses from day 14 p.i. Phagocytic activity of macrophages increased significantly after the administration of both drug formulations, more after lip.PZQ with an earlier peak observed for PZQ (day 3) than for lip.PZQ (day 6). Empty liposomes had no significant effect. The average counts of ingested particles in phagocytosing cells were significantly higher only after lip.PZQ administration. The pattern of changes in phagocytic activity correlated with the reduction of parasite numbers in the peritoneal cavity, with the highest observed on day 6 after therapy with lip.PZQ. Phagocytosis of lip.PZQ in vivo stimulated significantly the respiratory burst in peritoneal macrophages, with the highest concentration of superoxide anions recorded on day 1 after the last dose, whereas therapy with PZQ itself did not increase this process significantly. The capacity for the respiratory burst declined in all groups with progressing infection. It is proposed that the phagocytic activity of peritoneal macrophages after therapy was stimulated indirectly as a consequence of activation of the specific immune response. The larvicidal effect of lip.PZQ on the tetrathyridia in the peritoneal cavity was synergistic with the phagocytic activity and might be the result of double action of drug and superoxide anions generated during the respiratory burst stimulated by this drug formulation.

The effects of different anaesthetics on the neutrophil oxidative response in vitro are compared. Neutrophils were stimulated with small amounts of the bacterial peptide N-formyl-L-methionyl-L-leucyl-phenylalanine as a physiological, receptor-dependent stimulus. A new flow cytometry-based method capable of detecting the small amounts of H2O2 generated by neutrophils in a heterogenous all-or-none response following submaximal stimulation with N-formyl-L-methionyl-L-leucyl-phenylalanine was used. Propofol and thiopentone suppressed the respiratory burst significantly (P < 0.05 and P < 0.01) in therapeutic concentrations, while midazolam and methohexitone inhibited significantly (P < 0.01) only at concentrations greater than the therapeutic range. Impairment of the neutrophil response was primarily because of a reduction in the number of neutrophils participating in the respiratory burst and not of a proportional decrease of the fluorescence of all neutrophils.

We have examined the effect of propofol on the neutrophil respiratory burst. Chemiluminescence was used as a measure of the respiratory burst following stimulation with 10−5m N-formyl-L-methionyl-L-leucyl-L-phenylalanine.Propofol 1.25, 2.5 and 5.0 × 10−5m inhibited neutrophil chemiluminescence by 29.6, 43.0 and 57.6%, respectively, in neutrophils prepared from healthy adult volunteers, and by 25.5, 37.4 and 54.7% in cells from patients with severe sepsis. We conclude that propofol interferes with the ability of human neutrophils to generate reactive oxygen species.