Macrophage metabolism: activation of NADPH oxidation by phagocytosis

Rabbit and guinea pig peritoneal and alveolar macrophages and rabbit polymorphonuclear leucocytes (PMN) have been tested for their capacity to oxidize NADPH and NADH. In all these cells granule-bound NADPH oxidase is much more active than NADH oxidase, thus confirming our previous observations on human blood and guinea pig PMN. If the phagocytes are challenged with bacteria, the activity of NADPH oxidase is considerably stimulated. The enhancement of the oxidase activity is due to an increase of its V_max and, in the case of the PMN, also to a decrease of the K_m. We conclude that NADPH oxidase might play a relevant role in the metabolic stimulation of both PMN and macrophages by phagocytosis.     

Macrophage cell cycling: influence on Fc receptors and antibody-dependent phagocytosis

The influence of cell cycling on the density and binding properties of IgG2a Fc receptors and their associated antibody-dependent phagocytic activity was investigated with the P388D1 murine macrophage cell line. Unseparated macrophages and subpopulations of elutriated macrophages, enriched for cells in G1, S, and G2 + M phases were compared to detect possible differences in IgG2a-dependent phagocytosis. Suspensions of G2 + M phase cells were appreciably enhanced in phagocytic activity over G1-phase cells, which were less phagocytic than unseparated macrophage populations. An analysis of the binding of 125I-IgG2a myeloma protein disclosed that the IgG2a Fc receptor avidity remained essentially unchanged during cell cycle traverse, whereas the number of IgG2a Fc receptors more than doubled as cells cycled from G1 to G2 + M (1.5 X 10(5) vs 3.4 X 10(5) receptors per cell). With their increased size relative to G1 cells, and the resultant increase in receptor number, G2 phase cells should have more productive collisions with the antibody-coated target cells and greater phagocytic capacity.     

Macrophage metabolism during phagocytosis and digestion of normal and IgG antibody-coated sheep erythrocytes.

The respiration rate, aerobic glycolysis and glycolytic enzyme activities of purified mouse peritoneal macrophages increase during phagocytosis of either free or antibody-coated SRBC. These changes, however, seem to be roughly parallel to the amount of phagocytized antigen. In the digestive phase, Ag-Ab complexes induce more marked changes in macrophage metabolism than a comparable amount of free antigen. Lactate production, activity of several glycolytic enzymes and alanine-aminotransferase as well as glycogen content are greatly affected. Since in cell transfer experiments macrophage-associated SRBC can prime normal recipients for antibody response, whereas cell-associated SRBC-antibody complexes cannot, it is concluded that the changed macrophage metabolism drives antibody-coated antigen into metabolic channels allowing its rapid and total degradation.     

Macrophage recognition of externalized phosphatidylserine and phagocytosis of apoptotic Jurkat cells--existence of a threshold

Phosphatidylserine (PS) is predominantly confined to the inner leaflet of plasma membrane in cells, but it is externalized on the cell surface during apoptosis. This externalized PS is required for effective phagocytosis of apoptotic cells by macrophages. Because PS trans-bilayer asymmetry is not absolute in different types of nonapoptotic cells, we hypothesized that the amounts of externalized PS may be critical for macrophage discrimination between apoptotic and nonapoptotic cells. We developed a sensitive electron paramagnetic resonance method to quantify the amounts of externalized PS based on specific binding of paramagnetic annexin V-microbead conjugates with PS on cell surfaces. Using this technique, we found that nonapoptotic Jurkat cells externalize 0.9 pmol of endogenous PS/10(6) Jurkat cells. For cells with different amounts of integrated exogenous PS on their surface, no phagocytic response was observed at PS levels <5 pmol/10(6) Jurkat cells; at higher PS concentrations, phagocytosis increased in a concentration-dependent manner. Apoptosis in Jurkat cells caused externalization of approximately 240 pmol PS/10(6) Jurkat cells; these amounts of externalized PS are manyfold higher than the threshold amounts of PS required for phagocytosis. Thus, macrophages have a sensitivity threshold for PS externalized on the cell surface that provides for reliable recognition and distinction between normal cells with low contents of externalized PS and apoptotic cells with remarkably elevated PS levels.     

Macrophage colony-stimulating factor enhances phagocytosis and oxidative burst of mononuclear phagocytes against Penicillium marneffei conidia

The responses of rabbit pulmonary alveolar macrophages (PAMs) and elutriated human monocytes (EHMs) to Penicillium marneffei, an emerging dimorphic fungus that may cause fatal disease in human immunodeficiency virus-infected patients, were studied. PAMs and EHMs comparably phagocytosed conidia of two P. marneffei strains in the presence of serum. Electron microscopy showed intraphagosomal destruction of conidia after 12 h. Serum-opsonized conidia elicited significantly more superoxide anion (O(2)(-)) release from EHMs compared to non-opsonized conidia, but equivalent O(2)(-) amounts to that elicited by serum-opsonized Aspergillus fumigatus conidia. Macrophage colony-stimulating factor (M-CSF) significantly enhanced phagocytosis of P. marneffei conidia by PAMs and EHMs, as shown by light microscopy. Moreover, M-CSF enhanced O(2)(-) production by EHMs in response to both serum-opsonized (P<0.001) and non-opsonized (P=0.03) conidia of A. fumigatus as well as conidia of the P. marneffei isolates (P<0.001 and 0.03). We conclude that M-CSF enhances phagocytosis and oxidative metabolism of mononuclear phagocytes suggesting a potential role for this cytokine in host defense against pulmonary and disseminated P. marneffei infection.     

Macrophage heterogeneity in prostaglandins and thromboxane synthesis: differential activation by Fc- and C3b-dependent bacterial phagocytosis

Resident peritoneal macrophages, obtained from rats, were separated into subpopulations by centrifugation on a Percoll discontinuous density gradient. Nine fractions of pure macrophages were isolated. Each subpopulation was studied for Fc- and C3b-dependent bacterial phagocytosis and assayed for the related synthesis of PGE2, TxA2 and PGI2, measured by their stable metabolites TxB2 and 6-Keto-PGF1. The results show that with decreasing density, which corresponds to a greater maturity, the production of PGE2 increases and that of TxB2 and 6-Keto-PGF1 decreases. The cells of low density were mostly stimulated by IgG-opsonized bacteria, whereas those of high density responded preferentially to C3b- opsonized bacteria. This pattern is roughly similar to the one characterizing the phagocytosis via these two receptors although the correlation is not absolute. It can be concluded that enzymes involved in the metabolism of arachidonic acid, as well as receptors for C3b and IgG, are differentially expressed among resident macrophage subpopulations and thus during macrophage maturation.     

Macrophage phagocytosis of apoptotic neutrophils is compromised by matrix proteins modified by cigarette smoke and lipid peroxidation products

Clearance of apoptotic cells by phagocytosis plays an important role in the resolution of an inflammatory response. Macrophages interacting with extracellular matrix (ECM) proteins upregulate their phagocytic capacity. Cigarette smoke contains highly reactive carbonyls that modify proteins which directly/indirectly affects cellular function. We observed, in vitro, that human macrophages interacting with carbonyl or cigarette smoke modified ECM proteins dramatically down regulated their ability to phagocytose apoptotic neutrophils. We also show that this interaction with carbonyl-adduct modified ECM proteins led to increased macrophage adhesion in vitro. We hypothesise that changes in the ECM environment as a result of cigarette smoking affect the ability of macrophages to remove apoptotic cells. Moreover, we postulate that this decreased phagocytic activity was as a result of sequestration of receptors involved in the uptake of apoptotic cells towards that of recognition of carbonyl adducts on the modified ECM proteins leading to increased macrophage adhesion.     

Macrophage hydrogen peroxide production and phagocytic function are decreased following phagocytosis mediated by Fc receptors but not complement receptors

Previous in vivo and in vitro studies have shown that the phagocytosis of IgG-coated erythrocytes results in a depression of macrophage function. The present study compared the effect of phagocytosis mediated by Fc receptors with that mediated by complement receptors. The phagocytosis of IgG-coated erythrocytes by elicited peritoneal macrophages depressed their capacity to produce hydrogen peroxide as well as phagocytic function. Phagocytosis of erythrocytes coated with IgM and complement had neither of these effects. These results implicate the intracellular signaling that results from Fc receptor mediated phagocytosis in the depression of macrophage function that is caused by phagocytosis.     

Support and deformability in insect wings

Coupled investigations of insect wing movements and detailed wing morphology are in progress, and some functional principles underlying wing design are emerging. High speed cine and still photography and stroboscopy indicate that most wings undergo orderly deformation in flight. Common patterns are described and their significance discussed in the light of recent aerodynamic studies.
Many aspects of wing morphology–venational features, relief, thickened areas, flexionlines and vein fractures–may be related to the control of three-dimensional shape while beating. It is usually possible to distinguish areas specialized for deformability, and for support and the limiting of deformation. Some structural adaptations for these roles are described and illustrated.     

Phosphoinositides and phagocytosis

Phosphoinositide 3 kinases (PI3Ks)*Abbreviation used in this paper: PI3K, phosphoinositide 3 kinase. are known as regulators of phagocytosis. Recent results demonstrate that class I and III PI3Ks act consecutively in phagosome formation and maturation, and that their respective products, phosphatidylinositol 3,4,5-trisphosphate (PI[3,4,5]P(3)) and phosphatidylinositol 3-phosphate (PI[3]P), accumulate transiently at different stages. Phagosomes containing Mycobacterium tuberculosis do not acquire the PI(3)P-binding protein EEA1, which is required for phagosome maturation. This suggests a possible mechanism of how this microorganism evades degradation in phagolysosomes.     

红细胞变形性测定

血液流变学给临床研究展现了新的前景,揭示出免疫学、生物精神病学、生物心理学之间、心血管疾病与癌瘤之间人们忽视了的联系。血液流变学的测定给内科和外科医生提供了新的工具。     

Superoxide radicals and phagocytosis

Escherichia coli B, grown in iron-rich media, were more resistant toward the aerobic bactericidal action of the formed elements of blood than were comparable iron-deficient cells. The iron replete cells contained 2.5 times more ferrisuperoxide dismutase, 12 times more peroxidase, and 1.5 times more catalase than did the iron-deficient cells. The iron-deficient cells were more susceptible to exogenous O₂^? and to H₂O₂ than were iron-replete cells. Cyanide permitted a differentiation between ferrisuperoxide dismutase and catalase or peroxidase since it inhibited the latter peroxide-consuming enzymes but had no effect on the superoxide-utilizing enzyme. In the presence of 2 mm cyanide, the iron-replete E. coli were much more resistant toward phagocytic kill than were the iron-deficient cells even though this level of cyanide completely inhibited catalase and peroxidase. It can be concluded that a large part of the enhanced resistance toward phagocytic kill, exhibited by iron-replete E. coli B, was due to their increased content of the periplasmic ferrisuperoxide dismutase. It follows that O₂^? is probably an important agent in the killing of phagocytized E. coli B.     

Decreased deformability in aging erythrocytes

Deformability of bovine erythrocytes separated according to density (and age) was estimated by a modified Teitel's filterability test, the centrifugational test of Sirs, and viscosity measurements of cell suspensions. Both youngest and oldest erythrocytes were found to be less deformable than middle-aged cells, a result speaking against any chief role for deformability in the recognition of senescent erythrocytes and their removal from the circulation.     

Erythrocyte deformability: physiological aspects

Analysis of published and author's data on the physiological role of deformability of erythrocytes, general mechanisms of modifications and disturbances, assessment methods, and the role of this blood feature for assessing the body status. This parameter is one of the most labile features of blood that shows highly sensitive reaction to practically any changes in metabolic process in erythrocytes and in a whole body. Degradation of deformability of erythrocytes under various types of oxygen deficit deteriorates functioning of the system of oxygen transportation at various levels: heart, vascular flow, oxygen-transporting blood function. Under hypoxia the parameters of oxygen-transporting blood function, of peroxide oxidation of lipids and of the antioxidation system correlate well with degradation of deformability of erythrocytes. Therefore, this parameter can be used an integrated criterion of disturbances in oxygen supply, and of prooxidation-antioxidation body status. Deformability of erythrocytes is a factor generating adequate the oxygen supply to tissue, and its degradation aggravates substitution of the oxidase pattern of oxygen utilization with the oxygenase pattern. This parameter is extremely important for the body functional status.