scholarly journals Analysis of the nonfunctional respiratory burst in murine Kupffer cells.

1988 ◽  
Vol 167 (3) ◽  
pp. 1154-1170 ◽  
Author(s):  
A Ding ◽  
C Nathan
Keyword(s):  
Kupffer Cells ◽  
Respiratory Burst ◽  
Peritoneal Macrophages ◽  
Complete Suppression ◽  
Hexose Monophosphate ◽  
Cytochrome B559 ◽  
Prolonged Incubation ◽  
Hexose Monophosphate Shunt ◽  
Spectral Technique

Murine Kupffer cells (KCs), which constitute one of the largest populations of tissue macrophages, differ from most other cells of the myelomonocytic lineage in lacking the capacity for a respiratory burst. A collagenase perfusion technique followed by adherence to plastic at low temperature yielded pure cultures of KCs uniformly expressing receptors for Fc and C3bi, and containing virtually no morphologically detectable intracytoplasmic debris. Such KCs took up and oxidized glucose via the hexose monophosphate shunt about the same as peritoneal macrophages (PCs). Respiratory burst stimuli failed to enhance the hexose monophosphate shunt in KCs, probably because no H2O2 was produced. Detergent-permeabilized KCs generated no O2- in the presence of 1 mM NADPH, in striking contrast to all PC populations studied. Yet, KCs contained at least one component of the O2(-)-producing oxidase, cytochrome b559, in the same quantities as PCs and neutrophils. Cytochrome b559 was demonstrated by a novel double-reduction spectral technique that eliminated interference from hemoglobin and mitochondrial cytochromes. Consistent with the presence of the oxidase, KCs acquired normal respiratory burst capacity after prolonged incubation in vitro. The defect in triggering the respiratory burst in KCs was selective for the reduction of O2 by NADPH, in that reduction of O2 by endogenous arachidonate was readily demonstrate in response to zymosan. The percent of arachidonate released, the percent oxygenated, and the suppression of prostacyclin and leukotriene C production, as well as the pattern of LFA-1 expression, all resembled the pattern reported with PCs several days after exposure to bacteria. Indeed, exposure of PCs to low numbers of zymosan particles led gradually to complete suppression of respiratory burst capacity and refractoriness to its enhancement by rIFN-gamma, as evident in KCs both before and after their explanation. Thus, the modulation of oxidative metabolism that characterizes KCs probably arises from frequent endocytic encounters. This phenomenon may permit macrophages to act as scavengers without oxidative damage to bystander cells.

scholarly journals Dexamethasone inhibits the hexose monophosphate shunt in activated rat peritoneal macrophages by reducing hexokinase-dependent sugar uptake

1991 ◽  
Vol 278 (1) ◽  
pp. 129-135 ◽  
Author(s):  
R J Rist ◽  
R J Naftalin
Keyword(s):  
Sugar Transport ◽  
Peritoneal Macrophages ◽  
Free Sugar ◽  
Sugar Uptake ◽  
Sugar Accumulation ◽  
Dependent Manner ◽  
Hexose Monophosphate ◽  
Simultaneous Exposure ◽  
Hexose Monophosphate Shunt

Dexamethasone decreases 2-D-deoxyglucose (2-dGlc) uptake and accumulation into rat peritoneal macrophages in vitro in a concentration- and time-dependent manner (Ki for 1 microM-dexamethasone after a 2 h exposure = 0.71 +/- 0.21 microM; Ki for 0.1 microM-dexamethasone after exposure for 4 h = 0.10 +/- 0.06 microM). The inhibition of 2-dGlc uptake is consistent with a decrease in the coupling between endofacial hexokinase activity and the sugar transporter. The evidence for this is: (1) the Km for zero-trans 2-dGlc uptake in quiescent macrophages was increased by dexamethasone, but there was no significant effect on the Vmax.; (2) dexamethasone increased the rate of exit of sugar from cells preloaded with 2-dGlc; (3). the free sugar accumulation within the cytosol of the cells above the external solution concentration was significantly decreased by dexamethasone. These effects of dexamethasone on 2-dGlc transport were antagonized by simultaneous exposure to the steroid RU 38486 (Ki = 0.04 +/- 0.01 microM; 4 h incubation). Although dexamethasone inhibited zero-trans uptake, the maximum rate of infinite-trans exchange uptake of 2-dGlc into cells preloaded with 3-O-methyl-D-glucose (40 mM) was unaltered by dexamethasone or RU 38486, indicating that the dexamethasone-dependent decrease in zero-trans uptake was not due to a change in the number of transporters in the plasma membrane. Dexamethasone also inhibited the phorbol myristate acetate-induced stimulation of hexose monophosphate shunt (HMPS) activity, and this was reversed by RU 38486. Cytochalasin B, the potent sugar-transport inhibitor, inhibited HMPS activity and 2-d[2,6-3H]Glc uptake equally, indicating a single site of action. By contrast, dexamethasone showed differential inhibition of HMPS activity and 2-d[2,6-3H]Glc uptake, suggesting that it not only acts by decreasing the coupling between hexokinase and sugar transport, but also at one or more additional points.

scholarly journals Upregulation of mouse CD14 expression in Kupffer cells by lipopolysaccharide.

1994 ◽  
Vol 179 (5) ◽  
pp. 1671-1676 ◽  
Author(s):  
K Matsuura ◽  
T Ishida ◽  
M Setoguchi ◽  
Y Higuchi ◽  
S Akizuki ◽  
...  
Keyword(s):  
Cell Line ◽  
Kupffer Cells ◽  
Intraperitoneal Injection ◽  
Specific Binding ◽  
Flow Cytometric Analysis ◽  
Peritoneal Macrophages ◽  
Macrophage Cell ◽  
Inflammatory Monocytes

Western blot analysis showed that a monoclonal antibody against recombinant mouse CD14 (mCD14), designated rmC5-3, specifically reacted with mouse macrophage cell line J774, but not myeloma cell line NS1. Fluorographic and immunocytochemical analysis demonstrated specific binding of rmC5-3 with mouse resident macrophages, inflammatory monocytes and neutrophils, and macrophage cell lines. Immunohistochemical staining using rmC5-3 showed that CD14-positive Kupffer cells (KC) were small in number in the liver in nonstimulated mice. The number of stained KC, which were rich in the midzonal and periportal regions, gradually increased with time after intraperitoneal injection of lipopolysaccharide (LPS), peaked 6 h after injection, and returned to normal by 20 h after injection. Staining intensity over time was proportional to the number of KC. A slight increase in mCD14 expression was observed in peritoneal macrophages 2 h after LPS administration in vivo using flow cytometric analysis. mCD14 mRNA became detectable at 1 h after the intraperitoneal injection of LPS (20 micrograms/mice), and the level dramatically increased with time, peaking at 3 h, and sharply dropped at 6 h. The resident peritoneal macrophages demonstrated a constitutively high mCD14 mRNA expression, which slightly increased 2 h after LPS (100 ng/ml) stimulation in vitro. The level of mCD14 expression in macrophages did not increase after intraperitoneal injection of LPS (20 micrograms/mice).

THE EFFECTS OF ADENINE AND GLUCOSE ON SYNTHESIS OF NUCLEOTIDES BY EHRLICH ASCITES CARCINOMA CELLS IN VITRO

1962 ◽  
Vol 40 (3) ◽  
pp. 343-352 ◽  
Author(s):  
D. B. Ellis ◽  
P. G. Scholefield
Keyword(s):  
Ehrlich Ascites Carcinoma ◽  
Pentose Phosphate ◽  
Carcinoma Cells ◽  
Hexose Monophosphate ◽  
Hexose Monophosphate Shunt ◽  
Nucleotide Content ◽  
Ehrlich Ascites

Incubation of Ehrlich ascites carcinoma cells with glucose and adenine in a phosphate-containing medium leads to an increase in the total nucleotide content of the cells. The use of adenine-8-C14, glucose-U-C14, glucose-1-C14, and P32 has shown that there is true synthesis of nucleotides, and the relative rates of incorporation suggest that a purine riboside triphosphate such as ATP is the predominant nucleotide synthesized.The relative rates of incorporation of radioactivity from glucose-1-C14 and glucose-U-C14 into the acid-soluble nucleotides cannot be explained in terms of the hexose monophosphate shunt or of the transketolase–transaldolase pathway for pentose phosphate formation.

THE EFFECTS OF ADENINE AND GLUCOSE ON SYNTHESIS OF NUCLEOTIDES BY EHRLICH ASCITES CARCINOMA CELLS IN VITRO

1962 ◽  
Vol 40 (1) ◽  
pp. 343-352 ◽  
Author(s):  
D. B. Ellis ◽  
P. G. Scholefield
Keyword(s):  
Ehrlich Ascites Carcinoma ◽  
Pentose Phosphate ◽  
Carcinoma Cells ◽  
Hexose Monophosphate ◽  
Hexose Monophosphate Shunt ◽  
Nucleotide Content ◽  
Ehrlich Ascites

Incubation of Ehrlich ascites carcinoma cells with glucose and adenine in a phosphate-containing medium leads to an increase in the total nucleotide content of the cells. The use of adenine-8-C14, glucose-U-C14, glucose-1-C14, and P32 has shown that there is true synthesis of nucleotides, and the relative rates of incorporation suggest that a purine riboside triphosphate such as ATP is the predominant nucleotide synthesized.The relative rates of incorporation of radioactivity from glucose-1-C14 and glucose-U-C14 into the acid-soluble nucleotides cannot be explained in terms of the hexose monophosphate shunt or of the transketolase–transaldolase pathway for pentose phosphate formation.

scholarly journals The origin, kinetics, and characteristics of the kupffer cells in the normal steady state

1978 ◽  
Vol 148 (1) ◽  
pp. 1-17 ◽  
Author(s):  
RW Crofton ◽  
MMC Diesselhoff-Den Dulk ◽  
RV Furth
Keyword(s):  
Bone Marrow ◽  
Steady State ◽  
Kupffer Cells ◽  
Peritoneal Macrophages ◽  
Turnover Time ◽  
Mononuclear Phagocytes ◽  
Labeling Index ◽  
Cell Labeling ◽  
Initial Suspension

Enzymatic digestion with pronase and DNAase was used to isolate Kupffer cells from mouse liver. The characteristics of these cells were found to be similar to those of peritoneal macrophages, except that in the initial suspension the percentage of Kupffer cells with Fc receptors was low, C receptors were absent and the ingestion of opsenized bacteria was very poor, because of the effect of pronase on the cell membrane. After 24 h incubation in vitro all these characteristics return. The in vitro and 1 h-pulse [(3)H]thymidine labeling of the Kupffer cells is low (0.8 and 1 percent, respectively) indicating that in essence these cells do not divide. It was also shown that the small percentage of in vitro labeled Kupffer cells was recently derived from the circulation. After an intravenous injection of zymosan the in vitro labeling index of the Kupffer cells increased 16-fold, but it was proven that these dividing cells were immature mononuclear phagocytes very recently recruited from the bone marrow. The labeling of Kupffer cells aider one or four injections of [(3)H]thymidine reached a peak of 10.4 percent at 48 h or 24.1 percent at 60 h, respectively, indicating that these cells are derived from labeled monocytes. Further evidence for this conclusion was obtained by the absence of an increase of labeled Kupffer cells during treatment with hydrocortisone, which causes a monocytopenia during which no circulating monocytes are available to migrate to the tissues. Labeling studies in animals X-irradiated with hind-limb shielding gave a Kupffer cell labeling index of 5-10 percent of the normal values, which confirms their bone marrow origin. A quantitative study on the production of labeled monocytes in the bone marrow and their transit through the circulation showed that in the normal steady state at least 56.4 percent of the monocytes leaving the circulation become Kupffer cells. Considering the Kupffer cells as kinetically homogeneous this gives a mean turnover time of the total population of Kupffer cells of 21 days.

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