Hydroxyl radical formation in phagocytic cells of the rat

1979 ◽  
Vol 46 (1) ◽  
pp. 136-140 ◽  
Author(s):  
D. B. Drath ◽  
M. L. Karnovsky ◽  
G. L. Huber
Keyword(s):  
Hydroxyl Radical ◽  
Polymorphonuclear Leukocytes ◽  
Cell Concentration ◽  
Intact Cell ◽  
Peritoneal Macrophages ◽  
Cell Type ◽  
Phagocytic Cells ◽  
Fourfold Increase ◽  
Highly Active ◽  
Effective Particle

Polymorphonuclear leukocytes (PMN) and macrophages, harvested from the peritoneum and lung, release superoxide (O-.2) and hydrogen peroxide (H2O2) during phagocytosis. These two agents are thought to react with each other to produce a highly active oxidative substance known as hydroxyl radical (OH.). We present evidence suggesting that these radicals are generated by phagocytic cells of the rat. Our findings are based upon an assay where ethylene gas is generated from methional by the action of this radical. Ethylene generation was shown to be inhibited by superoxide dismutase, catalase, and scavengers of OH.. Of the cells examined, PMN generated the most ethylene from methional, exhibiting a fourfold increase during phagocytosis. Pulmonary and peritoneal macrophages caused smaller amounts of this gas to be formed. Regardless of cell type, an intact cell was required for ethylene generation. Zymosan appeared to be the most effective particle for all cells in ethylene formation from methional, although opsonization was critical only for PMN. Ethylene generation was dependent on cell concentration to an extent and increased with time.

Interaction of pseudomonas exoproducts with phagocytic cells

1982 ◽  
Vol 28 (6) ◽  
pp. 679-685 ◽  
Author(s):  
Bruce Weber ◽  
Michele M. Nickol ◽  
Kathleen S. Jagger ◽  
Catharine B. Saelinger
Keyword(s):  
Protein Synthesis ◽  
Alkaline Protease ◽  
Polymorphonuclear Leukocytes ◽  
Mononuclear Cells ◽  
Proteolytic Enzymes ◽  
Membrane Integrity ◽  
Peritoneal Macrophages ◽  
Phagocytic Cells ◽  
Complement Components ◽  
Mouse Peritoneal Macrophages

Polymorphonuclear leukocytes play the major role in host defense against infections with Pseudomonas aeruginosa; however, mononuclear cells also may contribute to defense against pulmonary infections with P. aeruginosa. Therefore, we examined the effects of three extracellular products of P. aeruginosa, exotoxin A, alkaline protease, and elastase, on the function of phagocytic cells. Phagocytosis or killing, protein synthesis, and membrane integrity were used as assays of cellular function. Pseudomonas toxin readily inhibited protein synthesis in mouse peritoneal macrophages; in contrast, proteolytic enzymes did not alter protein synthesis, but transiently decreased the sensitivity of macrophages to toxin. High levels of toxin reduced protein synthesis in human peripheral polymorphonuclear leukocytes but did not alter the ability of these cells to kill P. aeruginosa. Elastase and alkaline protease did not cause release of marker enzymes and did not directly inhibit the bactericidal activity of polymorphonuclear leukocytes; killing was reduced due to inactivation of complement components. In conclusion, these potential virulence products do not modify phagocyte function directly and thus do not directly interfere with host response in pseudomonas infections.

scholarly journals Phagolysosome formation in normal and colchicine-treated macrophages.

1975 ◽  
Vol 142 (4) ◽  
pp. 903-913 ◽  
Author(s):  
E L Pesanti ◽  
S G Axline
Keyword(s):  
Acid Phosphatase ◽  
Lysosomal Enzyme ◽  
Peritoneal Macrophages ◽  
Cell Type ◽  
Inert Particles ◽  
Time Periods ◽  
Mouse Peritoneal Macrophages ◽  
Over Time

Intracellular lysosomal fusion has been evaluated in cultivated mouse peritoneal macrophages by measurement of transfer of acid phosphatase to polyvinyltoluene (PVT)-containing phagolysosomes. Enzyme transfer was found to be directly and significantly related to the uptake of PVT and to be independent of time allowed for phagolysosome formation over time periods of 15 min to 18 h. In addition, the extent of transfer of lysosomal enzyme to phagolysosomes was unaffected by treatment of the cells with 10(-6) M colchicine, a dose which eradicates morphologically identifiable microtubules in this cell type within 2 h. The data indicate that intracellular fusion of lysosomes with phagosomes in the macrophage does not require formed microtubules and suggest that fusion occurs promptly after interiorization of inert particles.

scholarly journals Role of Toll-Like Receptor 4 in Macrophage Activation and Tolerance during Salmonella enterica Serovar Typhimurium Infection

2003 ◽  
Vol 71 (9) ◽  
pp. 4873-4882 ◽  
Author(s):  
Qian Li ◽  
Bobby J. Cherayil
Keyword(s):  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Peritoneal Macrophages ◽  
Phagocytic Cells ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Serovar Typhimurium ◽  
Tlr4 Gene ◽  
Factor Alpha

ABSTRACT Toll-like receptors (TLRs) play an important role in the innate immune response, particularly in the initial interaction between the infecting microorganism and phagocytic cells, such as macrophages. We investigated the role of TLR4 during infection of primary murine peritoneal macrophages with Salmonella enterica serovar Typhimurium. We found that macrophages from the C3H/HeJ mouse strain, which carries a functionally inactive Tlr4 gene, exhibit marked impairment of tumor necrosis factor alpha (TNF-α) secretion in response to S. enterica serovar Typhimurium infection. However, activation of extracellular growth factor-regulated kinase and NF-κB signaling pathways was relatively unaffected, as was increased expression of TNF-α mRNA. Furthermore, macrophage tolerance, which is associated with increased expression of the NF-κB p50 and p52 subunits, was induced by S. enterica serovar Typhimurium even in the absence of functional TLR4. These results indicate that during infection of macrophages by S. enterica serovar Typhimurium, TLR4 signals are required at a posttranscriptional step to maximize secretion of TNF-α. Signals delivered by pattern recognition receptors other than TLR4 are sufficient for the increased expression of the TNF-α transcript and at least some genes associated with macrophage tolerance.

Functional characteristics of PMJ2-R macrophages

2020 ◽  
Vol 18 (4) ◽  
pp. 133-137
Author(s):  
P.M. Kozhin ◽  
◽  
A.L. Rusanov ◽  
O.O. Shoshina ◽  
N.G. Luzgina ◽  
...  
Keyword(s):  
Cell Biology ◽  
Alternative Pathway ◽  
Peritoneal Macrophages ◽  
Alternative Activation ◽  
M2 Macrophages ◽  
Phagocytic Cells ◽  
Phenotypic Properties ◽  
M1 Macrophages ◽  
Oxidized Dextran ◽  
Tnf Α

Objective. To evaluate the ability of PMJ2-R cells for classical and alternative activation and to assess the effect of oxidized dextran on the functional status of polarized cells of this line. OD is a promising lysosomotropic agent used for targeted drug delivery to phagocytic cells. Materials and methods. We analyzed ability of immortalized murine peritoneal macrophages PMJ2R (ATCC CRL2458) to classical and alternative polarization, including that upon exposure to OD. We used real-time polymerase chain reaction to assess gene expression of competing arginine pathways. The capacity of phagocytes to engulf zymosan granules was evaluated using fluorescence microscopy. Results. We observed metabolic changes in PMJ2-R cells following their classical and alternative activation; these changes were typical of M1 and M2 macrophages, respectively. M1 macrophages demonstrated most active phagocytosis, while the activity of phagocytosis in M2 macrophages increased dose-dependently upon AD exposure. OD upregulates production of proinflammatory cytokine TNF-α in intact PMJ2-R cells and M1 macrophages. Conclusion. PMJ2-R cells have the capacity to phagocytose particles, can be polarized via the classical and alternative pathway, can modulate their functional activity in response to OD (a macrophagotropic substance), and exhibit the main phenotypic properties typical of peritoneal macrophages from C57Bl/6J mice. Therefore, cells of this line can be used as model cells in the investigation of phagocytic cell biology and pathology. Key words: alternative activation, classical activation, oxidized dextran, peritoneal macrophages, phagocytosis, C57Bl/6J, PMJ2-R

scholarly journals Guanylate Binding Proteins Restrict Leishmania donovani Growth in Nonphagocytic Cells Independent of Parasitophorous Vacuolar Targeting

mBio ◽  
2020 ◽  
Vol 11 (4) ◽  
Author(s):  
Arun Kumar Haldar ◽  
Utsav Nigam ◽  
Masahiro Yamamoto ◽  
Jörn Coers ◽  
Neena Goyal
Keyword(s):  
Host Defense ◽  
Leishmania Donovani ◽  
Binding Proteins ◽  
Critical Role ◽  
Intrinsic Property ◽  
Epithelial Cell Line ◽  
Cell Type ◽  
Phagocytic Cells ◽  
Defense Proteins ◽  
Content Type

ABSTRACT Interferon (IFN)-inducible guanylate binding proteins (GBPs) play important roles in host defense against many intracellular pathogens that reside within pathogen-containing vacuoles (PVs). For instance, members of the GBP family translocate to PVs occupied by the protozoan pathogen Toxoplasma and facilitate PV disruption and lytic parasite killing. While the GBP defense program targeting Toxoplasma has been studied in some detail, the role of GBPs in host defense to other protozoan pathogens is poorly characterized. Here, we report a critical role for both mouse and human GBPs in the cell-autonomous immune response against the vector-borne parasite Leishmania donovani. Although L. donovani can infect both phagocytic and nonphagocytic cells, it predominantly replicates inside professional phagocytes. The underlying basis for this cell type tropism is unclear. Here, we demonstrate that GBPs restrict growth of L. donovani in both mouse and human nonphagocytic cells. GBP-mediated restriction of L. donovani replication occurs via a noncanonical pathway that operates independent of detectable translocation of GBPs to L. donovan-containing vacuoles (LCVs). Instead of promoting the lytic destruction of PVs, as reported for GBP-mediated killing of Toxoplasma in phagocytic cells, GBPs facilitate the delivery of L. donovani into autolysosomal-marker-positive compartments in mouse embryonic fibroblasts as well as the human epithelial cell line A549. Together our results show that GBPs control a novel cell-autonomous host defense program, which renders nonphagocytic cells nonpermissible for efficient Leishmania replication. IMPORTANCE The obligate intracellular parasite Leishmania causes the disease leishmaniasis, which is transmitted to mammalian hosts, including humans, via the sandfly vector. Following the bite-induced breach of the skin barrier, Leishmania is known to live and replicate predominantly inside professional phagocytes. Although Leishmania is also able to infect nonphagocytic cells, nonphagocytic cells support limited parasitic replication for unknown reasons. In this study, we show that nonphagocytic cells possess an intrinsic property to restrict Leishmania growth. Our study defines a novel role for a family of host defense proteins, the guanylate binding proteins (GBPs), in antileishmanial immunity. Mechanistically, our data indicate that GBPs facilitate the delivery of Leishmania into antimicrobial autolysosomes, thereby enhancing parasite clearance in nonphagocytic cells. We propose that this GBP-dependent host defense program makes nonphagocytic cells an inhospitable host cell type for Leishmania growth.

scholarly journals Characterization of deoxyribonuclease activities derived from control and inflammation-associated mouse peritoneal macrophages

1984 ◽  
Vol 220 (2) ◽  
pp. 561-568
Author(s):  
G E Brown ◽  
T P Karpetsky ◽  
K Rictor ◽  
A Rahman
Keyword(s):  
Peritoneal Cavity ◽  
Peripheral Blood ◽  
Peritoneal Macrophages ◽  
Apparent Size ◽  
Blood Components ◽  
Cell Type ◽  
Specific Subset ◽  
Mouse Peritoneal Macrophages ◽  
Increased Activity

Native DNAase (deoxyribonuclease) activities derived from mouse peritoneal cavity and peripheral blood components were separated, detected, and characterized by electrophoresis into polyacrylamide gels containing DNA, followed by incubation of the gels, and staining of the substrate to reveal only the DNAase activities. Resident peritoneal macrophages contained 12 DNAase-II-like activities that were characteristic of that cell type, whereas lymphocytes and granulocytes each contained five DNAases. Induction of inflammation by peritoneal injection of thioglycollate resulted in changes in macrophage DNAase expression, including: increased total DNAase activity, a decrease in the number of activities from 12 to 11, increased activity of a specific subset of the enzymes, and a change in the apparent size of a specific subset of the enzymes. Electrophoretic and enzymic properties and sensitivity to endo-beta-N-acetylglucosaminidase H indicated that the macrophage activities probably represented charge variants of one or two parent peptide chains.

Metal ions and oxygen radical reactions in human inflammatory joint disease

1985 ◽  
Vol 311 (1152) ◽  
pp. 659-671 ◽  
Keyword(s):  
Hydroxyl Radical ◽  
Metal Ion ◽  
Tissue Injury ◽  
Copper Ion ◽  
Chelating Agent ◽  
Joint Disease ◽  
Radical Reactions ◽  
Phagocytic Cells ◽  
Bacterial Killing

Activated phagocytic cells produce superoxide (O 2 - ) and hydrogen peroxide (H 2 O 2 ) ; their production is important in bacterial killing by neutrophils and has been implicated in tissue damage by activated phagocytes. H 2 O 2 and O 2 are poorly reactive in aqueous solution and their damaging actions may be related to formation of more reactive species from them. One such species is hydroxyl radical (OH . ), formed from H 2 O 2 in the presence of iron- or copper-ion catalysts. A major determinant of the cytotoxicity of O - 2 and H 2 O 2 is thus the availability and location of metal-ion catalysts of OH* formation. Hydroxyl radical is an initiator of lipid peroxidation. Iron promoters of OH* production present in vivo include ferritin, and loosely bound iron complexes detectable by the ‘bleomycin assay’. The chelating agent Desferal (desferrioxamine B methanesulphonate) prevents iron-dependent formation of OH* and protects against phagocyte-dependent tissue injury in several animal models of human disease. The use of Desferal for human treatment should be approached with caution, because preliminary results upon human rheumatoid patients have revealed side effects. It is proposed that OH* radical is a major damaging agent in the inflamed rheumatoid joint and that its formation is facilitated by the release of iron from transferrin, which can be achieved at the low pH present in the micro-environment created by adherent activated phagocytic cells. It is further proposed that one function of lactoferrin is to protect against iron-dependent radical reactions rather than to act as a catalyst of OH· production.

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