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PubMed1994

Enumeration of macrophage carbohydrates in protein-deficient rats: relation to cell surface properties.

In order to study the effect of protein malnutrition on macrophage glycoproteins the carbohydrate composition of peritoneal macrophages from protein-deficient rats has been studied by paper chromatography and HPLC. The results show that the carbohydrate content of resident cells recovered from protein-deficient group was significantly greater than control and decreased on prolonged incubation. In the protein-deficient samples there was a significant decrease in the content of galactose, fucose and galactosamine known to be binding to specific ligands and increase in glucose and mannosamine. In both control and deficient groups, thioglycollate (TG) elicitation resulted in higher total sialic acid content. Prolonged incubation (18 hr) caused an elevation of sialic acid levels in the resident cells, whereas a drastic reduction was observed in the TG elicited cells. In the protein-fed (20%) group, the cell surface sialic acid which contributes to the negative charge of the cells, reduced significantly on culturing the TG cells but not the resident cells. In the protein-deficient group, this effect was seen in the resident cells also; in the TG cells the cell surface sialic acid was significantly low at the isolation stage suggesting that these cells had become comparatively more positively charged in vivo itself. This observed reduction could be correlated to the enhanced sialidase levels in these cells. These protein deficiency related changes in the carbohydrate composition of macrophages could lead to modification of their receptor activity and charge related functions.

Animals↗
PubMed1982

Inducible macrophage cytotoxins. I. Biokinetics of activation and release in vitro.

Peritoneal exudate macrophage monolayers (PEMM) from C57BL/6 and DBA/2 mice inoculated ip with tumor allografts were induced to release in vitro labile cell toxin(s), herein called "macrophage cytotoxin(s)" (MCT). Macrophages released MCT spontaneously for a short interval when initially established as monolayers, and they were reinduced to secrete MCT by exposure to allogeneic and syngeneic tumor cells (but not to normal cells) and by exposure to polyinosinic-polycytidylic acid (poly I . poly C) and lipopolysaccharide (LPS). PEMM from normal mice treated ip 3 days previously with thioglycollate were also induced to release toxins in vitro. These cells did not release MCT spontaneously before or after treatment with neoplastic cells but were induced to release MCT by exposure to poly I . poly C or LPS. Resident peritoneal macrophages did not release MCT either spontaneously or after treatment with tumor cells, poly I . poly C, or LPS. MCT released from alloimmune mice stimulated with syngeneic or allogeneic tumor cells were resolved by molecular sieving into a major peak at 140,000--160,000 daltons, called "alpha-MCT," and into a minor peak at 60,000 daltons, called "beta-MCT." However, supernatants from thioglycollate-induced PEMM, stimulated with poly I . poly C or LPS, appeared to be composed entirely of the alpha-class. alpha-MCT from poly I . poly C-stimulated PEMM caused 31--56% lysis of syngeneic EL-4 and allogeneic L-929, NS-1, and YAC-1 tumor cells in vitro but was not cytotoxic for normal cells. Secretion of the MCT by PEMM derived from thioglycollate-treated animals stimulated with poly I . poly C was inhibited by colchicine, emetine, iodoacetic acid, trypan blue, and cytochalasin B.

Animals↗
PubMed1987

Neutral amino acid transport in mouse peritoneal macrophages.

Neutral amino acid transport has been characterized in the mouse peritoneal macrophages elicited by thioglycolate broth. Serine, alanine, cysteine, and leucine were transported mainly in a Na+-independent manner and, within the concentration ranges tested, the transport of each amino acid was mediated by a single, saturable system. The transport of these amino acids was strongly inhibited by a wide variety of neutral amino acids, 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid, a putative System L substrate, but not by glycine, proline, or 2-methylaminoisobutyrate. In this study the transport of serine has been characterized in detail. Inhibition of the transport of serine by 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid or by leucine was competitive. The transport of serine which exhibited trans-stimulation was slightly stimulated by lowering the pH and was little affected by N-ethylmaleimide. It was also shown that most of the neutral amino acids were concentrated only slightly (less than 2-fold) in the macrophages, unlike in the lymphocytes and granulocytes. From these results we have concluded that in the macrophages serine, cysteine, alanine, leucine, and presumably many other neutral amino acids are transported almost exclusively through a common transport system analogous to System L.

Alanine↗
PubMed1971

Fatty Acid Requirements of the Kazan 5 and Reiter Strains of Treponema pallidum.

The fatty acid requirements of two avirulent treponemes were investigated by using a "lipid-poor" albumin-thioglycolate medium. The Kazan 5 and the Reiter stains of Treponema pallidum required a pair of fatty acids for growth. One member of the pair was saturated and the other was unsaturated. The saturated fatty acids could contain either an odd or even number of carbon atoms, but a chain length of at least 14 carbon atoms was necessary. Unsaturated fatty acids with one, two, or three double bonds were satisfactory if they contained 15 or more carbon atoms. The pair of fatty acids could be replaced with a single 18-carbon monounsaturated fatty acid if it was in the trans configuration rather than the naturally occurring cis form.

Journal Article↗
PubMed1980

Induction of cell infiltration and acid hydrolase release into the peritoneal cavity of mice.

An injection of thioglycollate into the peritoneal cavity of mice produced a peak of exudate at 4 h, a peak of total leukocytes at 24 h when the predominant cell was the polymorphonuclear neutrophil, and a secondard macrophage phase beginning 2--3 days after thioglycollate. The release of N-acetylglucosaminidase (EC 3.2.1.30) into the peritoneal fluid paralleled the macrophage phase. The amount of enzyme released was related to the dose of thioglycollate. Neither zymosan (400 micrograms) nor endotoxin (30 micrograms) produced a marked inflammatory response when injected into the peritoneal cavity.

Acetylglucosaminidase↗
PubMed1996

The relationship between altered membrane composition, eicosanoids and TNF-induced IL1 and IL6 production in macrophages of rats fed fats of different unsaturated fatty acid composition.

The study investigated the changes in individual molecular species in PE and the effects of a variety of dietary fats with varying proportions of saturated and unsaturated fatty acids on membrane composition, eicosanoid production and cytokine production in thioglycollate-elicited rat macrophages. The data obtained indicates that the greatest degree of modulation by dietary fats on cytokine production was observed after 8 weeks feeding and at this time, the total diacyl species containing linoleic acid (18:2 n-6) and arachidonic acid (20:4 n-6) at the sn-2 position related in a curvilinear fashion to total 18:2 n-6 intake and that IL1 and IL6 production related in a curvilinear fashion to the total diacyl species with 20:4 and 18:2 at the sn-2 position. After 4 weeks of feeding, fish and olive oils enhanced production of IL6 and LTB4, however, while IL1 production, after 8 weeks of dietary treatment, was greatest from macrophages of animals fed corn and olive oils, PGE2 production was greatest in the former group and LTB4 production in the latter. Thus an eicosanoid effect may explain the modulatory influence of olive oil and IL1 production but, cannot explain the effect of corn oil on production of the cytokine. The data from the present study provides some insight into how dietary fats could provide therapy for conditions in which inflammatory cytokines are implicated.

Animals↗
PubMed1986

Evidence for the presence of oxytocin in the corpus luteum of the goat.

Purified acetic acid extracts of corpora lutea (CL) of non-pregnant goats were found to contain substantial amounts of oxytocin (OT) as measured by radioimmunoassay. OT standard and the CL extracts released prostaglandin F2 alpha (PGF2 alpha) from rat isolated uteri in a quantitatively similar manner. Treatment of both OT standard and CL extract with sodium thioglycolate, oxytocin antiserum or oxytocin antagonist abolished this biological activity. Acid extracts of CL of pregnant goat were found to contain approximately 2% of levels during the cycle by day 21 after fertile mating and this had a reduced ability to release PGF2 alpha from rat uterus. It is concluded that both the immunoreactivity and the biological activity of the CL extracts are due to the presence of an oxytocin-like substance and that tissue levels of oxytocin are low in pregnant compared to non-pregnant goats.

Animals↗
PubMed1965

NITROGEN-CONTAINING AND CARBOHYDRATE-CONTAINING ANTIGEN FROM ACTINOMYCES BOVIS.

Pirtle, E. C. (National Animal Disease Laboratory, Ames, Iowa), P. A. Rebers, and W. W. Weigel. Nitrogen-containing and carbohydrate-containing antigen from Actinomyces bovis. J. Bacteriol. 89:880-888. 1965.-Water-soluble, heat-stable antigens have been isolated from the supernatant fluids of broth cultures of Actinomyces bovis ATCC 10048 after 8 days of growth in a broth medium composed of Casamino Acids, yeast extract, Tween 80 (polyoxyethylene sorbitan monooleate), sodium thioglycolate, dextrose, and sodium chloride. The antigens were precipitated from the culture supernatant fluid with alcohol and purified by fractional precipitation with alcohol in the presence of calcium or zinc and by chromatography on diethylaminoethyl Sephadex. Two serologically active substances which differed in chemical composition and size were characterized. The larger one contained 71% hexose and 2.8% nitrogen, and the smaller one contained 45% hexose and 5.4% nitrogen. Heterogeneity of these fractions could not be demonstrated by electrophoresis in free solution at pH 7.8 or 2, by ultracentrifugal analysis, by double diffusion in agar, or by immunoelectrophoresis. Despite their differences in chemical composition and size, they appeared identical in activity as antigens in complement fixation and in double-diffusion tests in agar. Each was found to contain mannose as its chief component after hydrolysis and paper chromatography with three solvent systems. A small percentage of the total nitrogen may be attributed to the presence of amino sugars but the remainder is as yet unidentified.

Acetates↗
PubMed1994

Evidence that prostaglandins modulate lipogenesis in cultured lymphocytes--a comparison with its effect on macrophages and tumour cells.

Lipogenesis is essential for the rapid proliferation of cells and it is established that the biosynthesis of selected lipids precedes S-phase, DNA synthesis and the initiation of cell division. Pyruvate was previously shown to be an important lipid precursor for lymphocytes, macrophages and tumour cells. This study now reports the role of prostaglandins (PG) on the regulation of lipogenesis from [3-14C] pyruvate in 24-h cultured lymphocytes. It is shown that indomethacin (10 microM) and ibuprofen (10 microM), both inhibitors of PG biosynthesis, increased [3-14C] pyruvate incorporations into phospholipid and cholesterol fractions in resting lymphocytes but reduced its incorporation into these fractions in concanavalin A (Con A)-stimulated lymphocytes and tumour cells. These two agents also affected [2-14C] thymidine incorporation into the DNA of these cells in the same manner. Lipids produced from [3-14C] pyruvate and exported into the cell culture medium were also measured. The PG biosynthesis inhibitions reduced the transfer to culture medium of arachidonic acid, phospholipids, cholesterol and fatty acids higher than C20 by lymphocytes and tumour cells. Although macrophages are not proliferative cells, the cytoplasmic export measurement is important because these cells have a high capacity for lipid secretion. The results show that the PG biosynthesis inhibitors do not affect the export of phospholipids and cholesterol in macrophages. They do however markedly change the export of arachidonic acid and fatty acids higher than C20 produced from [3-14C] pyruvate in macrophages. It is also reported that a cell-stimulating response diminished the above fatty acid outcome in resting cells and augmented it in thioglycollate-stimulated macrophages. The findings suggest that PG modulation of lipogenesis may depend on cell cycle phase as well as the intrinsic lipid metabolic diversity and capacity.

Acetates↗
PubMed2005

Ascorbate uptake and antioxidant function in peritoneal macrophages.

Since activated macrophages generate potentially deleterious reactive oxygen species, we studied whether ascorbic acid might function as an antioxidant in these cells. Thioglycollate-elicited murine peritoneal macrophages contained about 3 mM ascorbate that was halved by culture in ascorbate-free medium. However, the cells took up added ascorbate to concentrations of 6-8 mM by a high-affinity sodium-dependent transport mechanism. This likely reflected the activity of the SVCT2 ascorbate transporter, since its message and protein were present in the cells. Activation of the cells by phagocytosis of latex particles depleted intracellular ascorbate, although not below the basal levels present in the cells in culture. Glutathione (GSH) was unaffected by phagocytosis, suggesting that ascorbate was more sensitive to the oxidant stress of phagocytosis than GSH. Phagocytosis induced a modest increase in reactive oxygen species as well as a progressive loss of alpha-tocopherol, both of which were prevented in cells loaded with ascorbate. These results suggest that activated macrophages can use ascorbate to lessen self-generated oxidant stress and spare alpha-tocopherol, which may protect these long-lived cells from necrosis or apoptosis.

Animals↗
PubMed1989

Effects of diacerein on the quantity and phagocytic activity of thioglycollate-elicited mouse peritoneal macrophages.

Diacerein (DAR: 1,8-diacetoxy-9,10-dioxo-dihydroanthracene-3-carboxylic acid) is an anthraquinone drug which displays anti-inflammatory effects in experimental animals and antirheumatic activity in humans. The drug was administered for orally for four consecutive days to mice injected intraperitoneally with thioglycollate. The following dose levels of DAR were used: 2.5, 5 and 10 mg/kg/day. At the end of the experimental period the macrophage content of peritoneal exudate was dose-dependent and significantly lower in DAR-treated mice compared with animals that were given saline orally. The macrophages isolated from the peritoneal exudate of mice that received DAR displayed a dose-dependent reduced phagocytosis. The effects of DAR were found to be similar to those of indometacin and dexamethasone, which were used as reference drugs. The ability of DAR to interfere with macrophage functioning may contribute to its overall therapeutic activity.

Animals↗
PubMed2003

Effect of ascorbic acid on stimulatory status of activated mouse peritoneal phagocytes.

Mouse peritoneal macrophages (MPM) when elicited by the antioxidant ascorbic acid have been found to be significantly stimulatory, exhibiting marked alteration at the cellular and enzyme levels. Alterations recorded were as follows--cellular yield per mouse, their protein content, lysosomal acid hydrolase levels and capability to phagocyte, all were significantly enhanced. The new stimulant was observed to produce no synergistic action on MPM when thioglycollate, BCG or endotoxin along with the same stimulated the latter. Levels of antioxidants like ascorbic acid and glutathione were found to be enhanced in elicited macrophages whereas superoxide dismutase levels varied when the three above stimulators were administered. However, the ascorbic acid elicited cells showed an increase in glutathione levels and a decrease in SOD levels but no change in total intracellular ascorbic acid levels. Further, though ascorbic acid interaction enhanced the phagocytic capability of MPM as compared to resident cells, no significant boosting of phagocytic process could be observed when each of three stimulators coupled with ascorbic acid was used for macrophage elicitation.

Animals↗
PubMed1995

Influence of unsaturated fatty acids on the production of tumour necrosis factor and interleukin-6 by rat peritoneal macrophages.

The effect of individual unsaturated fatty acids on the release of tumour necrosis factor (TNF) and interleukin 6 (IL6) was investigated in thioglycollate-induced rat peritoneal macrophages. The intracellular mechanisms associated with the changes of cytokine production in response to fatty acids were also studied. Incubation of macrophages with 100 microM docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) increased TNF (21% and 15% respectively) and IL6 (69% and 40% respectively) production. Linoleic acid (LA) diminished TNF production by 16%. At 100 microM oleic acid (OA), LA and EPA concentration an increase in macrophage adenylate cyclase activity (110%, 72% and 39% respectively) and a decrease (14%) in the presence of DHA was observed. PGE2 production in the presence of 100 microM DHA was reduced by 36%, whereas in the presence of 100 microM LA an increase (75%) was observed. Phospholipase A2 (PLA2) activity was also found to be modified in the presence of EPA and DHA at 50 microM (20% and 60% respectively) and 100 microM (34% and 62% respectively) concentrations. The activities of both protein kinase A (PKA) and protein kinase C (PKC) were effected by the different fatty acids. At 50 microM all fatty acids suppressed PKA activity except OA which enhanced PKA activity by 14%. At 100 microM fatty acid concentration, EPA suppressed PKA activity by 40%. PKC activity was enhanced by LA and OA, by 18% and 21% respectively. However, at 100 microM EPA and DHA, PKC activity was suppressed by 37% and 17% respectively, whereas PKC activity was enhanced by 146% in the presence of 100 microM LA. These results show for the first time that unsaturated fatty acids have an effect on macrophage PLA2 activity and that PGE2 may be a potent modulator of IL6 production. From these studies it is tempting to speculate that macrophage TNF and IL6 release may, in part, occur via a PKC and PKA independent pathway and that PLA2 activity and PGE2 concentration are inversely related to production of TNF and IL6.

Adenylyl Cyclases↗