scholarly journals Dynamics of leukotriene C production by macrophages.

1980 ◽  
Vol 152 (5) ◽  
pp. 1236-1247 ◽  
Author(s):  
C A Rouzer ◽  
W A Scott ◽  
A L Hamill ◽  
Z A Cohn
Keyword(s):  
Time Course ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Experimental Conditions ◽  
Silicic Acid Chromatography ◽  
Pg Release ◽  
Antigen Antibody ◽  
Mouse Peritoneal Macrophages ◽  
Hydroxyeicosatetraenoic Acids

A method for the radiochemical assay of LTC production by mouse peritoneal macrophages in vitro is presented. The method involves labeling macrophages in culture with [5,6,8,9,11,12,14,15-3H]20:4 followed by stimulation of arachidonic acid (20:4) release under the experimental conditions desired. Radiolabeled leukotriene C (LTC) is recovered from the culture medium by extraction and silicic acid chromatography in 40% yield with full retention of biological activity. Because this LTC is radiochemically pure, the quantity of LTC release may be estimated from the amount of radioactivity in the sample. Use of the radioassay to study parameters affecting LTC synthesis by macrophages indicated that the time course of LTC synthesis and its relationship to the dose of a phagocytic stimulus (zymosan) were very similar to those of prostaglandin (PG) release. LTC release was also similar to that of PG in that lower levels of both metabolites were produced by Corynebacterium parvum-elicited macrophages than by resident cells. Finally, LTC release was stimulated in response to a challenge with antigen-antibody complexes, but lower maximal levels were attained than those with zymosan. The data presented here are consistent with the hypothesis that challenge of macrophages with a phagocytic stimulus leads to the release of 20:4 by an inducible phospholipase. Cyclooxygenase and lipoxygenase then compete for the released 20:4, leading to the production of PG, hydroxyeicosatetraenoic acids, and LTC.

scholarly journals IN VITRO STUDIES ON THE INTERACTION BETWEEN MOUSE PERITONEAL MACROPHAGES AND STRAINS OF SALMONELLA AND ESCHERICHIA COLI

1960 ◽  
Vol 112 (2) ◽  
pp. 403-417 ◽  
Author(s):  
Charles Jenkin ◽  
Baruj Benacerraf
Keyword(s):  
Escherichia Coli ◽  
Normal Mouse ◽  
Peritoneal Macrophages ◽  
In Vitro Studies ◽  
Experimental Conditions ◽  
Mouse Plasma ◽  
Virulent Strains ◽  
Mouse Macrophages ◽  
Mouse Peritoneal Macrophages

Virulent strains of Salmonella opsonized with normal mouse plasma are never phagocytosed as well as avirulent strains. The virulent strains of Salmonella phagocytosed after opsonization with normal mouse plasma are able to multiply within normal mouse peritoneal macrophages, whereas under similar experimental conditions the avirulent strains are killed. When virulent strains of Salmonella are opsonized with specific antiserum or plasma from BCG-infected mice, they are treated by normal mouse macrophages as if they were avirulent. Virulent bacteria opsonized with BCG plasma are phagocytosed and killed better by peritoneal macrophages from BCG-infected mice, than peritoneal macrophages from normal mice.

scholarly journals THE INTERACTION OF SOLUBLE HORSERADISH PEROXIDASE WITH MOUSE PERITONEAL MACROPHAGES IN VITRO

1972 ◽  
Vol 55 (1) ◽  
pp. 186-204 ◽  
Author(s):  
Ralph M. Steinman ◽  
Zanvil A. Cohn
Keyword(s):  
Horseradish Peroxidase ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Wide Range ◽  
Mouse Macrophages ◽  
Secondary Lysosomes ◽  
Endocytic Vesicles ◽  
In Vitro Interaction ◽  
Mouse Peritoneal Macrophages

The in vitro interaction of soluble horseradish peroxidase (HRP) with homogeneous mono layers of mouse macrophages has been studied using sensitive biochemical and cytochemical techniques. The compartmentalization of HRP in extracellular and intracellular sites has been quantitatively evaluated. A significant fraction is bound to a serum-derived layer, which coats the surface of culture vessels and may be removed by appropriate washes. Macrophages interiorize HRP as a solute in pinocytic vesicles without appreciable binding of the glycoprotein to the plasma membrane. Uptake is directly proportional to the concentration of HRP in the culture medium. 1 x 106 cells ingest 0.0025% of the administered load per hr over a wide range of concentrations. Cytochemically, all demonstrable HRP is sequestered within the endocytic vesicles and secondary lysosomes of the vacuolar apparatus. After uptake, the enzymatic activity of HRP is inactivated exponentially with a half-life of 7–9 hr, until enzyme is no longer detectable. When macrophages have pinocytosed trace-labeled HRP-125I, cell-associated isotope disappears with a t ½ of 20–30 hr and they release monoiodotyrosine-125I into the culture medium. We were unable to obtain evidence that significant amounts of HRP (>2%) can be exocytosed after uptake, can exist intact on the cell surface, or can be digested extracellularly. It is difficult to reconcile these observations with several of the postulated mechanisms whereby macrophages are thought to play a prominent role in the induction of an immune response.

scholarly journals The ingestion and digestion of human lactoferrin by mouse peritoneal macrophages and the transfer of its iron into ferritin.

1977 ◽  
Vol 146 (3) ◽  
pp. 817-827 ◽  
Author(s):  
J L van Snick ◽  
B Markowetz ◽  
P L Masson
Keyword(s):  
Iron Content ◽  
Culture Medium ◽  
Reticuloendothelial System ◽  
Peritoneal Macrophages ◽  
Human Lactoferrin ◽  
Saturation Point ◽  
Intracellular Digestion ◽  
Additional Support ◽  
Mouse Peritoneal Macrophages

Human lactoferrin (Lf) labeled with 125I and/or 59Fe was found to be ingested in vitro by mouse peritoneal macrophages (MPM). The uptake measured after 15 h incubation reached a saturation point at a concentration of 200 microgram/ml in the culture medium, whatever was the iron content of Lf. In such conditions, the uptake of transferrin (Tf) used as a control was 10 times lower. At a concentration of 80 microgram/ml in the medium, one cell picked up about 0.7 X 10(6) molecules of Lf per hour, and 0.13 X 10(6) molecules of Tf per hour. Iron-saturated Lf disappeared from MPM with a half life of 14.5 h, whereas the halflife of iron-free Lf was 4.2 h. Concomitant with the intracellular digestion of Lf, the iron was transmitted to ferritin. These data provide additional support for the hypothesis that Lf plays a key role in iron turnover, especially at the level of the reticuloendothelial system where iron is recovered from the catabolism of erythrocytes.

Active entry of bloodstream forms of Trypanosoma cruzi into macrophages

Parasitology ◽  
1979 ◽  
Vol 78 (1) ◽  
pp. 89-98 ◽  
Author(s):  
T. L. Kipnis ◽  
V. L. G. Calich ◽  
W. Dias da Silva
Keyword(s):  
Trypanosoma Cruzi ◽  
Cytochalasin B ◽  
Previous Treatment ◽  
Peritoneal Macrophages ◽  
Experimental Conditions ◽  
Specific Antibodies ◽  
Previously Treated ◽  
Mouse Peritoneal Macrophages ◽  
Intracellular Amastigote

SUMMARYThe uptake of bloodstream forms of Trypanosoma cruzi, Y and CL stocks, by mouse peritoneal macrophages and their intracellular differentiation and multiplication has been compared in vitro. After 48 h the number of macrophages showing intracellular amastigote forms was higher when the Y stock was used. The number of parasitized cells increased with the time of contact between parasites and macrophages. Prior treatment of the parasites with anti-T. cruzi antibodies and/or complement increased the number of infected macrophages, but did not interfere with their subsequent differentiation within the macrophages. The number of parasitized cells was greater when macrophages were obtained from mice previously treated with lipopolysaccharide, peptone or thioglycollate. Uptake was not appreciably affected when macrophages were pre-treated with trypsin or anti-macrophage serum, or when the parasites and macrophages were incubated in the presence of cytochalasin B. In the same experimental conditions, epimastigotes of T. cruzi were not able to differentiate into amastigotes. Their uptake was potentiated by previous treatment with specific antibodies and/or complement and was blocked by cytochalasin B. These results confirm that epimastigotes derived from T. cruzi cultures are phagocytosed and suggest that bloodstream forms penetrate actively into macrophages.

scholarly journals Induction of cystine transport activity in mouse peritoneal macrophages.

1987 ◽  
Vol 165 (3) ◽  
pp. 628-640 ◽  
Author(s):  
H Watanabe ◽  
S Bannai
Keyword(s):  
Amino Acids ◽  
Culture Medium ◽  
Glutamate Uptake ◽  
Human Fibroblasts ◽  
Peritoneal Macrophages ◽  
Transport Activity ◽  
Drastic Increase ◽  
Uptake Activity ◽  
Mouse Peritoneal Macrophages

Uptake of cystine was investigated in mouse peritoneal macrophages. The rates of the uptake of cystine in resident macrophages or macrophages elicited by some irritants were very low, but a drastic increase was observed when the cells were cultured in vitro. This increase was time-dependent and required protein synthesis. In macrophages elicited by thioglycollate broth, the rate of the uptake of cystine increased by about 40-fold after 16 h in culture. Contrary to the uptake of cystine, the rates of uptake of some neutral amino acids did not change markedly during culture. We characterized the induced activity of the cystine uptake in macrophages elicited by thioglycollate broth. Cystine was taken up in an Na+-independent and pH-sensitive manner, and the uptake was potently inhibited by extracellular glutamate and the analogous anionic amino acids, but not by aspartate. The activity of the glutamate uptake was also induced during the culture in a way similar to that of cystine uptake, and the uptake of glutamate was potently inhibited by cystine. From these results we concluded that the uptake of cystine and glutamate in macrophages was mostly mediated by a single transport system similar to the ones previously reported in human fibroblasts and some other cells. As a consequence of the induction of the activity of the cystine uptake, glutathione levels in macrophages doubled during culture, and a thiol compound, presumably cysteine, was released into the culture medium and accumulated there. When the macrophages were cultured hypoxically, the induction of the cystine uptake activity was markedly depressed, suggesting an involvement of oxygen in the induction.

scholarly journals Mouse Peritoneal Macrophages Contain Abundant ω-6 Lipoxygenase Activity That Is Independent of Interleukin-4

1996 ◽  
Vol 16 (12) ◽  
pp. 1488-1494 ◽  
Author(s):  
Joseph A. Cornicelli ◽  
Kathryn Welch ◽  
Bruce Auerbach ◽  
Steven J. Feinmark ◽  
Alan Daugherty
Keyword(s):  
Interferon Gamma ◽  
Time Course ◽  
Disease Process ◽  
Human Monocyte ◽  
Peritoneal Macrophages ◽  
Human Cells ◽  
Interleukin 4 ◽  
Mouse Peritoneal Macrophages

The action of an ω-6 lipoxygenase (LO) has been implicated in the development of atherosclerosis through a mechanism involving oxidation of LDL, and its regulation in macrophages may have important implications for the disease process. Human monocyte–derived macrophages (HMDMs) showed no demonstrable LO protein or activity unless they were incubated with interleukin-4 (IL-4). In contrast, mouse peritoneal macrophages (MPMs) possessed significant basal levels of LO activity and protein that were augmented by IL-4 treatment. Interferon gamma prevented the induction of LO in both HMDMs and MPMs. Whereas interferon gamma could completely block the IL-4 induction of LO in human cells, it did not suppress basal LO activity in MPMs. Both HMDMs and MPMs exhibited similar concentration-response relationships for stimulation of LO activity and protein, with maximal induction at 1 ng/mL IL-4. The time course of IL-4 induction of LO activity was markedly different in human and murine cells. IL-4 induced LO activity and protein in human cells by 48 hours that were maximal by 72 hours; there was a decline to a new baseline by 96 hours. MPMs have a significant amount of LO activity at baseline, which declined with time by nearly 10-fold in the absence of IL-4. IL-4 blunted the decline of LO activity with time and restored activity to that found at baseline by 48 hours. IL-4 was not responsible for the LO activity present in freshly isolated MPMs since both activity and protein content were similar in cells harvested from IL-4 +/+ and IL-4 −/− mice. Therefore, whereas IL-4 may be an important modulator of LO production in vitro, it is not essential for the in vivo expression of this protein. Further, these studies demonstrate that significant differences exist between monocyte-derived macrophages matured in vitro and tissue macrophages that have matured in vivo.

scholarly journals Mouse macrophage elastase. Purification and characterization as a metalloproteinase

1981 ◽  
Vol 193 (2) ◽  
pp. 589-605 ◽  
Author(s):  
M J Banda ◽  
Z Werb
Keyword(s):  
Culture Medium ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Serine Proteinase ◽  
Peritoneal Macrophages ◽  
Serine Proteinases ◽  
Endogenous Inhibitor ◽  
Purification And Characterization ◽  
Predominant Form ◽  
Mouse Peritoneal Macrophages

Macrophage elastase was purified from tissue-culture medium conditioned by inflammatory mouse peritoneal macrophages. Characterized as a secreted neutral metalloproteinase, this enzyme was shown to be catalytically and immunochemically distinct from the mouse pancreatic and mouse granulocyte elastases, both of which are serine proteinases. Inhibition profiles, production of nascent N-terminal leucine residues and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of degraded elastin indicated that macrophage elastase is an endopeptidase, with properties of a metalloproteinase, rather than a serine proteinase. Macrophage elastase was inhibited by alpha 2-macroglobulin, but not by alpha 1-proteinase inhibitor. Macrophage elastase was resolved into three chromatographically distinct forms. The predominant form had mol.wt. 22 000 and was purified 4100-fold. Purification of biosynthetically radiolabelled elastase indicated that this form represented less than 0.5% of the secreted protein of macrophages. Approx. 800% of the starting activity was recovered after purification. Evidence was obtained for an excess of an endogenous inhibitor masking more than 80% of the secreted activity.

Development of embryonic rat eyes in organ culture

Development ◽  
1968 ◽  
Vol 19 (3) ◽  
pp. 407-414
Author(s):  
R. Christy Armstrong ◽  
Joel J. Elias
Keyword(s):  
Organ Culture ◽  
Culture Medium ◽  
Folic Acid Deficiency ◽  
Experimental Conditions ◽  
Acid Deficiency ◽  
Series Of Experiments ◽  
Development In Vitro ◽  
Ocular System

Abnormalities of the ocular system which appear in organ culture in Waymouth's medium with freshly added glutamine (Armstrong & Elias, 1968) resemble those caused by transitory pteryolglutamic acid (PGA or folic acid) deficiency in vivo (Armstrong & Monie, 1966). The configurations of such malformations as lens herniations, retinal diverticula, and rosette-like formations of the retina are remarkably similar in both cases. The experiments reported in this paper were undertaken in an effort to understand the mechanisms involved in the production of similar abnormalities by two very different experimental conditions: the addition of glutamine in vitro and the transitory deficiency of PGA in vivo. One series of experiments involved the effects of manipulation of the PGA and glutamine content of the culture medium on eye development in vitro. Parallel studies on PGA-deficiency in vivo were undertaken in conjunction with organ-culture experiments in order to compare the effects on abnormal eye morphogenesis.

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