scholarly journals LOSS OF IRON FROM MOUSE PERITONEAL MACROPHAGES IN VITRO AFTER UPTAKE OF [55FE]FERRITIN AND [55FE]FERRITIN RABBIT ANTIFERRITIN COMPLEXES

1974 ◽  
Vol 62 (3) ◽  
pp. 802-814 ◽  
Author(s):  
Martha E. Fedorko

Mouse peritoneal macrophages in culture for 24 h were exposed to horse [55Fe]ferritin and rabbit antihorse [55Fe]ferritin antibody complex and the amount of 55Fe in the medium was assayed up to 2 days after the pulse uptake. Cell survival was assayed by photographing the same areas of the tissue culture Petri dish on successive days and by counting cell numbers per unit area. In experiments in which quantitative assay for cell death is negligible, about 10–20% of the iron ingested by pinocytosis or phagocytosis is released to iron-free medium containing either freshly dialyzed or deironized newborn calf serum (10%). Over the 2-day postpulse period, iron loss is linear. This loss of iron to the medium is significantly reduced by adding iron-saturated newborn calf serum in the postpulse recovery period. A significant portion of the iron released to the medium is bound to transferrin. When human serum is used in the tissue culture system, similar quantities (10–25%) of the ingested iron are lost to the medium 2 days after the pulse.

1971 ◽  
Vol 133 (2) ◽  
pp. 231-259 ◽  
Author(s):  
Thomas C. Jones ◽  
James G. Hirsch

Methods have been devised for establishing infection in vitro of mouse macrophages and fibroblasts with Mycoplasma pulmonis. The mycoplasmas attached to the cells and under appropriate cultural conditions grew into a lawn of microorganisms covering most of the cell surface. The mycoplasmas grew abundantly on fibroblasts cultured in minimal essential medium containing 20% fetal calf serum; supplementation of this medium with heart infusion broth was necessary to obtain similar growth on macrophages. The infection of these cells appeared to be essentially an extracellular process; only rarely were partially degraded mycoplasmas seen with phagocytic vacuoles. The addition to heavily infected macrophage cultures of low concentrations of anti-mycoplasma antibody stimulated rapid, massive phagocytosis of the surface microorganisms. In sharp contrast, the same antiserum had no discernable effect on the mycoplasma-fibroblast relationship. The antibody effect in the macrophage system was apparently a direct opsonic one rather than an indirect result of microbial killing, since the mycoplasmas in macrophage or fibroblast cultures incorporated labelled thymidine into DNA after the addition of antiserum to the medium. The phagocytic event and the subsequent fate of the mycoplasmas were studied in detail after the addition of antibody to the macrophage cultures. Phase-contrast cinemicrophotography revealed membrane ruffles surrounding the surface mycoplasmas and disappearance from view of the organisms; 10–30 min later translucent grapelike clusters were seen in large phagocytic vacuoles. On electronmicroscopic study the surface mycoplasmas were surrounded by pincers-like projections of the macrophage. Numerous mycoplasmas were seen in phagocytic vacuoles; in the early minutes after the addition of antibody the intracellular mycoplasmas appeared normal, but within 2 hr they appeared partially degraded with a central electron-lucent area and electron-opaque deposits at the microbial cell margin. 24 hr after the addition of antiserum, digestion of the mycoplasmas was nearly complete; the cells appeared normal except for large residual bodies composed of amorphous moderately dense material and increased lipid deposits. Degradation of mycoplasmas within macrophages was also studied using infected cultures in which the mycoplasmas, but not the macrophages, had incorporated tritiated thymidine into DNA. The appearance of large amounts of acid-soluble radiolabel after phagocytosis stimulated by antibody confirmed the degradation of the intracellular mycoplasmas.


1974 ◽  
Vol 140 (4) ◽  
pp. 1085-1096 ◽  
Author(s):  
Haakon Melsom ◽  
Gilla Kearny ◽  
Stanislav Gruca ◽  
Rolf Seljelid

Mouse peritoneal macrophages cultivated in vitro acquire a strong extracellular cytotoxic activity towards isotope labeled syngeneic erythrocytes as demonstrated by isotope release to the medium. This lytic process is mediated by an extremely labile macrophage cytolytic factor (MCF) which is not detected under ordinary tissue culture conditions with serum present in the medium. By the use of serum-free medium containing low doses of 2-mercaptoethanol MCF is stabilized and found to be an easily dialysable, low molecular substance which resists heating at 60°C for 30 min.


Parasitology ◽  
1973 ◽  
Vol 66 (2) ◽  
pp. 343-353 ◽  
Author(s):  
Kazem Behbehani

Mouse peritoneal macrophage cells suspended in a TC 199 calf serum medium and cultured in Leighton tubes, were infected with Trypanosoma (Schizotrypanum) cruzi culture forms in order to study the development of this parasite in vitro. Three cycles of development were thought to occur: 1. Epimastigotes or trypomastigotes were taken up by macrophages and transformed into amastigotes. These multiplied by binary fission and ruptured the infected cell after 4‐5 days. Released amastigotes were taken up by uninfected macrophages and the cycle was repeated. 2. A small proportion of intracellular amastigotes developed into ‘ovoid forms’ which transformed progressively into promastigotes, epimastigotes and trypomastigotes. 3. Some amastigotes became ‘round forms’ from which sphaeromastigotes developed. These transformed directly into trypomastigotes without the formation of epimastigotes.


2009 ◽  
Vol 6 (5) ◽  
pp. 387-392 ◽  
Author(s):  
Yinan Wang ◽  
Xueling Cui ◽  
Guixiang Tai ◽  
Jingyan Ge ◽  
Nan Li ◽  
...  

2014 ◽  
Vol 9 (6) ◽  
pp. 1934578X1400900 ◽  
Author(s):  
Evgeny A. Pislyagin ◽  
Dmitry L. Aminin ◽  
Alexandra S. Silchenko ◽  
Sergey A. Avilov ◽  
Pelageya V. Andryjashchenko ◽  
...  

Stimulation of lysosomal activity and ROS formation in mouse peritoneal macrophages by five triterpene glycosides, typicosides A1 (1), A2 (2), B1 (3), C1 (4) and C2 (5) has been studied and compared with their cytotoxic activities. Glycosides 1–3 possess moderate activities, but the most cytotoxic glycoside 5 is not active. Typicoside C1 (4), with low toxicity, was proved to be the most active concerning stimulation of ROS formation. This is the first example of a triterpene glycoside from sea cucumbers with low cytotoxicity, but which demonstrates a strong immunostimulatory effect on mouse peritoneal macrophages in vitro.


1975 ◽  
Vol 95 (2) ◽  
pp. 385-395 ◽  
Author(s):  
Martha E. Fedorko ◽  
Carmine Lanni

1980 ◽  
Vol 152 (5) ◽  
pp. 1236-1247 ◽  
Author(s):  
C A Rouzer ◽  
W A Scott ◽  
A L Hamill ◽  
Z A Cohn

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.


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