Ingibjorg Hardardottir

Effects of endotoxin and cytokines on lipid metabolism.

Endotoxin, via cytokines, induces marked changes in lipid metabolism: serum VLDL increases, whereas the effect on LDL levels varies among species. The increase in VLDL is caused by stimulation of hepatic VLDL secretion or inhibition of clearance, or both. These alterations can be deleterious or beneficial effects.

Endotoxin and Cytokines Decrease Serum Levels and Extra Hepatic Protein and mRNA Levels of Cholesteryl Ester Transfer Protein in Syrian Hamsters

Endotoxin alters the metabolism of lipoproteins, including that of high density lipoprotein (HDL). Cholesteryl ester transfer protein (CETP) facilitates exchange of HDL cholesterol for very low density lipoprotein (VLDL) triglyceride, leading to catabolism of HDL. We investigated the effects of endotoxin and cytokines on CETP in Syrian hamsters. Endotoxin induced a rapid and progressive decrease in serum CETP levels, by 48 h CETP had decreased to < 20% of control levels. Endotoxin also decreased CETP mRNA and protein levels in adipose tissue, heart, and muscle, the tissues with highest levels of CETP mRNA, providing a plausible mechanism for the endotoxin-induced decrease in circulating CETP. Dexamethasone did not mimic the effects of endotoxin on CETP, but the combination of tumor necrosis factor and interleukin-1 did, indicating that these cytokines may in part mediate the effects of endotoxin on CETP. The endotoxin-induced decrease in CETP may help maintain HDL cholesterol levels during infection and inflammation when increased triglyceride levels could drive the exchange of HDL cholesteryl ester for VLDL triglyceride. Maintaining circulating HDL may be important because HDL protects against the toxic effects of endotoxin and provides cholesterol for peripheral cells involved in the immune response and tissue repair.

Tumor necrosis factor production by murine resident peritoneal macrophages is enhanced by dietary n−3 polyunsaturated fatty acids

Tumor necrosis factor (TNF) is a macrophage derived peptide that has an antitumor action and modulates immune and inflammatory reactions. Dietary fatty acids may modulate TNF production as dietary n-3 polyunsaturated fatty acids suppress human monocyte TNF production, but enhance its secretion by murine peritoneal macrophages. Mice were maintained for 5 weeks on diets containing different amounts of n-3 and n-6 fatty acids. TNF, PGE2 and 6-keto PGF1 alpha production was monitored following in vitro stimulation of resident peritoneal macrophages with lipopolysaccharide. Macrophages from mice fed the high n-3 diet produced 8-fold more TNF and half the PGE2 produced by macrophages from mice on the other diets. Indomethacin caused an increase in the TNF production by macrophages from mice on all diets but macrophages from mice on the high n-3 diet produced more TNF than macrophages from mice on the other diets. Exogenous PGE2 (100 nM) greatly decreased TNF production by macrophages from mice on all diets, but macrophages from mice on the high n-3 diet secreted 70% more TNF than macrophages from mice fed the other diets, indicating that PGE2 is only partly responsible for the effects observed. The results show that feeding n-3 polyunsaturated fatty acids may cause enhanced TNF production by resident peritoneal macrophages and that PGE2 is partly responsible for the effect.

Endotoxin and cytokines increase hepatic messenger RNA levels and serum concentrations of apolipoprotein J (clusterin) in Syrian hamsters.

Infection and inflammation induce alterations in hepatic synthesis and plasma concentrations of the acute phase proteins. Our results show that apolipoprotein (apo) J is a positive acute phase protein. Endotoxin (LPS), tumor necrosis factor (TNF), and interleukin (IL)-1 increased hepatic mRNA and serum protein levels of apo J in Syrian hamsters. Hepatic apo J mRNA levels increased 10- to 15-fold with doses of LPS from 0.1 to 100 micrograms/100 g body weight within 4 h and were elevated for > or = 24 h. Serum apo J concentrations were significantly increased by 16 h and further elevated to 3.3 times that of control, 24 h after LPS administration. Serum apo J was associated with high density lipoprotein and increased fivefold in this fraction, after LPS administration. Hepatic apo J mRNA levels increased 3.5- and 4.6-fold, with TNF and IL-1, respectively, and 8.2-fold with a combination of TNF and IL-1. Serum apo J concentrations were increased 2.3-fold by TNF, 79% by IL-1, and 2.9-fold with a combination of TNF and IL-1. These results demonstrate that apo J is a positive acute phase protein.

Cytokines stimulate lipolysis and decrease lipoprotein lipase activity in cultured fat cells by a prostaglandin independent mechanism.

We previously showed that indomethacin blocked the effect of tumor necrosis factor (TNF) and other cytokines on lipolysis. We now show that TNF stimulates prostaglandin (PG) production, enhances lipolysis and decreases lipoprotein lipase (LPL) activity in 3T3-F442A adipocytes and indomethacin blocks these activities, suggesting that the actions of TNF are mediated by PGs. However, exogenous PGE2 at the levels induced by TNF is not sufficient to affect lipolysis or LPL activity and low doses of indomethacin and flurbiprofen block PG production without affecting TNFs action. Interleukin-1 and interferon-alpha and gamma induce lipolysis and decrease LPL activity but do not stimulate much PG production. These results demonstrate that cytokines enhance lipolysis and decrease LPL activity in 3T3 adipocytes by a PG independent mechanism.

LPS and cytokines regulate extra hepatic mRNA levels of apolipoproteins during the acute phase response in Syrian hamsters.

Altered hepatic expression of apolipoproteins occurs during the acute phase response. Here we examined whether the acute phase response alters extra hepatic expression of apolipoproteins. Syrian hamsters were injected with endotoxin (LPS), tumor necrosis factor (TNF), interleukin (IL)-1, or the combination of TNF + IL-1 and mRNAs for serum amyloid A (apoSAA), apolipoprotein (apo) J, apo E. apo A-I, and apo D, were analyzed. LPS increased mRNA levels for apoSAA in all tissues examined. LPS and TNF + IL-1 increased mRNA levels for apo J in kidney, heart, stomach, intestine, and muscle. Individually, TNF and IL-1 were less potent than the combination of the two cytokines. LPS decreased mRNA levels for apo E in all tissues, except for mid and distal intestine. TNF and IL-1 were less effective than LPS. LPS, TNF + IL-1 and TNF decreased mRNA levels for apo A-I in duodenum. mRNA for apo D decreased in heart, were unchanged in brain and increased in muscle, following LPS. The widespread extra hepatic regulation of the apolipoproteins during the acute phase response may be important for the alterations in lipid metabolism that occur during infection and inflammation as well as the immune response.

Dietary Fish Oil Supplementation Increases Survival in Mice Following Klebsiella pneumoniae Infection

INTRODUCTION Epidemiological studies have shown that high intake of omega-3 fatty acids correlates with low incidence of various diseases such as cardiovascular diseases, asthma, diabetes mellitus and various auto-immune disorders. It may therefore be suggested that omega-3 fatty acids have substantial impact on the immune system. Studies of the effect of omega-3 fatty acids on survival in bacterial infections have however been contradicting. A Dutch study from 1991 showed increased survival in mice fed fish-oil following infection with Klebsiella pneumoniae. Because of the contradicting results the authors conducted a study with the hypothesis that fish-oil intake increases survival after severe Klebsiella pneumoniae infection. METHODS Thirty mice were fed fish-oil enriched diet (10%), olive-oil enriched diet (10%) or standard chow diet. After six weeks the mice were injected intramuscularly with l.óxlO2 cfu of Klebsiella pneumoniae. The survival was measured at regular time intervals for 120 hours. RESULTS After 56 hours, 93% of the mice fed fish-oil were alive and 68% and 40% of the mice fed olive-oil and standard chow respectively. The overall survival after 120 hours was 40% in the fish-oil group, 25% in the olive-oil group and 20% in the standard group. The survival after 120 hours of the mice fed the fish-oil enriched diet was significantly better when compared to the two other groups (p=0.0034). DISCUSSION We conclude that fish-oil enriched diet increases survival of NMRI mice following infection with Klebsiella pneumoniae when compared to olive-oil supplementation or standard chaw. We therefore conclude that the difference in survival is probably based on the effect of omega-3 fatty acid on the immune system. The immunological pathway is still unknown and our results encourage further studies.

Dietary n − 3 polyunsaturated fatty acids modify Syrian hamster platelet and macrophage phospholipid fatty acyl composition and eicosanoid synthesis: a controlled study

The aim of this study was to determine the effects of varying intakes of dietary n - 3 polyunsaturated fatty acids (PUFA) on the fatty acyl composition and arachidonic acid metabolite synthesis of platelets and macrophages in Syrian hamsters consuming diets that were strictly controlled for n - 6 PUFA content. Animals consumed highly controlled diets which were not supplemented with n - 3 PUFA (control) or supplemented with 0.4%, 0.8% or 2% (w/w) n - 3 fatty acids. The content of n - 3 PUFA in cellular phospholipids increased progressively with the intake of n - 3 PUFA, while n - 6 PUFA, including arachidonic acid, decreased despite the constant intake of 18:2(n - 6); this latter effect was more substantial in macrophages than in platelets. The synthesis by stimulated macrophages of prostaglandin E2, 6-keto-prostaglandin F1 alpha, thromboxane B2 and 11- and 15-hydroxyeicosatetraenoic acids decreased with the intake of 0.8% n - 3 PUFA to 30-50% of the control values. Little effect of diets on platelet aggregation and eicosanoid synthesis was observed reflecting the limited effect on platelet arachidonic acid content. The synthesis of 12-hydroxyeicosapentaenoic acid by stimulated platelets increased with n - 3 PUFA consumption in a dose-dependent fashion. Circulating triacylglycerols and HDL-cholesterol were decreased only in animals consuming 2% n - 3 PUFA. The strict control of n - 6 PUFA intake allows the determination of the effects of n - 3 PUFA intake on the measured parameters without confounding effects of other dietary lipids.

The Effects of Omega-3 Polyunsaturated Fatty Acids on TNF-α and IL-10 Secretion by Murine Peritoneal Cells In Vitro

Omega-3 polyunsaturated fatty acids (PUFA) affect immune response, partly by affecting cytokine secretion. Omega-3 PUFA decrease tumor necrosis factor (TNF)-α secretion by RAW 264.7 macrophages but increase TNF-α secretion by primary elicited peritoneal macrophages in vitro. In this study, the effects of omega-3 and omega-6 PUFA on lipopolysaccharide induced TNF-α and interleukin (IL)-10 secretion by murine primary resident and elicited peritoneal macrophages and by RAW 264.7 macrophages, were examined in vitro using an enzyme-linked immunosorbent assay. In addition, the effects of dietary omega-3 PUFA on the number of cells secreting these cytokines were examined with enzyme-linked immunospot assay. All cell types secreted more TNF-α but similar amounts of IL-10 when incubated with the omega-3 PUFA, eicosapentaenoic acid or docosahexaenoic acid, compared with that when incubated with the omega-6 PUFA, linoleic acid or arachidonic acid. Dietary fish oil did not affect the number of TNF-α secreting resident peritoneal macrophages but decreased the number of macrophages secreting IL-10 ex vivo. These results show that dietary omega-3 PUFA and omega-3 PUFA added to cells in vitro increase TNF-α secretion by resident peritoneal macrophages, probably by a direct effect on the cells. In contrast, omega-3 PUFA did not affect IL-10 secretion by the cells but decreased the number of cells secreting IL-10 ex vivo, possibly by affecting cell recruitment, maturation or proliferation.

Dietary fish oil decreases secretion of T helper (Th) 1-type cytokines by a direct effect on murine splenic T cells but enhances secretion of a Th2-type cytokine by an effect on accessory cells

Dietary fish oil is considered to have anti-inflammatory effects based primarily on its effects on T-cell proliferation and IL-2 secretion. Its effects on the secretion of T helper (Th) 1-type cytokines vary and few studies have examined its effects on the secretion of Th2-type cytokines. In the present study, we examined the effects of dietary fish oil on the secretion of Th1 and Th2-type cytokines by splenocytes and the mechanism by which dietary fish oil affects Th2-type cytokine secretion. Mice were fed diets supplemented with 18 % fish oil (w/w) +2 % maize oil or 20 % maize oil for 6 weeks. Spleen cells, isolated splenic T cells and accessory cells (splenocytes depleted of T cells) were stimulated with anti-CD3/anti-CD28. The secretion of interferon (IFN)-gamma, TNF-alpha, IL-4 and IL-10 was measured by ELISA. Dietary fish oil decreased the secretion of IFN-gamma and TNF-alpha by total splenocytes and isolated T cells. In contrast, dietary fish oil increased the secretion of IL-4 by total splenocytes but had no effect on IL-4 secretion by isolated T cells. When isolated T cells were cultured with CD11b+ cells (mainly macrophages), cells from mice fed the fish oil diet secreted more IL-4 than cells from mice fed the maize oil diet. These results demonstrate that dietary fish oil directs cytokine secretion by splenocytes towards a Th2 phenotype and that the effects of dietary fish oil on the secretion of a Th2-type cytokine are mediated by its effect on CD11b+ accessory cells.