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C Capo

Publications and source records attributed to C Capo.

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Non-specific binding by macrophages: existence of different adhesive mechanisms and modulation by metabolic inhibitors.

Rat peritoneal cells were made to bind test particles of five different species: immunoglobulin-coated sheep red cells (IGSRC), glutaraldehyde-treated sheep red cells (GSRC), leishmania, latex beads and tumour cells. The dependence of binding on various physicochemical parameters was studied: the binding of latex or leishmania resisted cold (4 degrees), azide, cytochalasin B, ethyleneglycol and dimethylsulphoxide (DMSO). The binding of IGSRC resisted cold and azide, but it was inhibited by cytochalasin B, ethyleneglycol and DMSO. The binding of GSRC was inhibited by cold, azide and ethyleneglycol, but not by cytochalasin B and DMSO. The binding of tumour cells was inhibited by azide, cytochalasin B, ethyleneglycol and DMSO. An attempt was made to saturate selectively some adhesive sites of macrophage membranes: glutaraldehyde-treated bovine albumin (GBSA) inhibited the ingestion of latex and GSRC, not leishmania and IGSRC, whereas a crude leishmania extract (CLE) inhibited the ingestion of all tested particles except latex. Antigen-antibody complexes inhibited the ingestion of IGSRC and leishmania, not latex and GSRC. Rat macrophages were made to bind radio-iodinated GBSA (GIBSA). The uptake of glutaraldehyde-treated albumin was proportional to the amount of substrate the cells were incubated with over a wide concentration range. This uptake was not a result of endocytosis, and it was far more efficient than that of peroxidase. Macrophage-particle interaction was studied with electron microscopy. The binding of IGSRC and leishmania to macrophages involved large areas where the interacting membranes were separated by a low density gap of constant width. The interaction of GSRC and latex with macrophages was much more patchy and irregular. Further, no redistribution of the macrophage surface polysaccharides seemed associated with the binding of GSRC. It was concluded that several different mechanisms and different membrane adhesive structures are involved in non-specific recognition by macrophages. Further, non-specific binding sometimes requires an active cell participation. Several testable hypotheses are described, which suggest further experiments in order to obtain a fuller insight into the mechanism of rosette formation.

Animals↗

[Existence of several non-specific recognition systems in macrophages].

Rat peritoneal cells were made to bind five particle species: immunoglobulin-coated Sheep red cells, glutaraldehyde-treated Sheep red cells, latex beads, leishmania and tumor cells. The dependence of binding on various physico-chemical parameters was studied. The binding of latex beads or Leishmania was not inhibited by cold (4 degrees C), sodium azide, cytochalasin B and ethyleneglycol or dimethylsulphoxide. The binding of immunoglobulin-coated Sheep red cells was unaffected by cold and azide, but it was inhibited by cytochalasin B, ethyleneglycol and dimethylsulphoxide. The binding of glutaraldehyde-treated Sheep red cells was inhibited by cold, azide and ethyleneglycol, but it resisted cytochalasin B and dimethylsulphoxide. The binding of tumor cells was inhibited by azide, cytochalasin B, ethyleneglycol and dimethylsulphoxide. It is concluded that: (a) macrophages are endowed several sets of non-specific binding structures that are differently affected by physico-chemical parameters, which provides a simple way of characterizing them; (b) the expression of a given binding structure on the macrophage membrane is modulated by metabolic inhibitors; (c) some lymphocytes were able to bind tumor cells or Leishmania. Thus, lymphocytes and macrophages might share some non-specific adhesive structures.

Agglutination Tests↗

Evaluation of intercellular adhesion with a very simple technique.

Rosetting techniques are widely used to quantify or purify various lymphocytic subpopulations; however, these techniques cannot discriminate between different receptors of similar specificities and different binding strengths, further, they do not provide any information concerning the molecular mechanisms involved in cell-cell adhesion. This paper describes a very simple technique of assaying rosette stability: cell suspensions are driven with known pressure through a calibrated needle with a syringe. Adhesion is quantified before and after this treatment. This procedure did not damage rat peritoneal cells used in a model system. Further, this method yielded fairly reproducible results and allowed a crude estimate of the force involved in the binding of glutaraldehyde-treated sheep red cells (GSRC) or immunoglobulin-coated sheep red cells (IGSRC) by rat macrophages (an average force of 0.8 x 10(-7) Newton was needed to separate 50% of bound IGSRC from macrophages). Binding and binding strength were found to be independent parameters. Last, this method possibly provided a way of separating two distinct subpopulations of rat macrophages. It is suggested that this technique might be routinely used to refine rosette studies.

Animals↗

[Macrophage receptors].

Macrophages are involved in many immunological functions such as phagocytosis, cytotoxicity, antigen binding and cooperation with lymphocytes. The triggering of those functions involves membrane receptors. Several receptors species are well characterized, but some phenomena can be accounted for only if macrophages are endowed with nonspecific adhesive structures. A study of the effect of various physical or chemical factors on the binding of several particle species by rat peritoneal macrophages allowed us to classify nonspecific receptors and discuss the mechanisms involved in some types of cellular interaction.

Animals↗

Non-specific recognition in phagocytosis: ingestion of aldehyde-treated erythrocytes by rat peritoneal macrophages.

Particles were chemically modified with aldehydes and incubated with rat peritoneal cells for phagocytosis. All dialdehydes and lower monaldehydes tested (methanal, ethanal and propanal) made sheep erythrocytes phagocytosable. Failure of higher monaldehydes to induce phagocytosis of treated erythrocytes was not due to lack of reactivity with red cell membranes. All erythrocytes tested (bird and mammal red cells were used) and rat thymocytes were phagocytosed by rat macrophages after incubation with aldehyde. Treatment of Candida albicans did not induce phagocytosis: this failure was not due to lack of aldehyde binding (as demonstrated with [14C]-methanal) nor to anti-phagocytic properties of the parasite membrane. Sheep erythrocytes were submitted to enzymatic treatment (pronase, trypsin, neuraminidase) or incubated with succinic anhydride (to block free NH2 groups) or iodacetamide (to block free SH groups) before aldehyde treatment: phagocytosis was not decreased, which suggested that aldehydes did not act by altering some definite surface structure of the treated particles. Treatment of erythrocytes with cross-linking compounds such as tetraazotized o-dianisidine (coupling occurs mainly on tyrosine and histidine residues) or l-ethyl(3-dimethyl aminopropyl) carbodiimide (a bivalent reagent binding free COOH groups) did not induce any substantial phagocytosis of erythrocytes. Phagocytosis of aldehyde treated erythrocytes was partly correlated with hydrophobicity of these cells, as measured with a two-phase partition system. It is concluded that aldehyde-mediated phagocytosis of erythrocytes is mainly due to cross-linking of red cell membrane structures, probably involving free OH groups, which must increase local rigidity and thereby modify hydrophobicity of the red cell surface.

Aldehydes↗

Dependence of phagocytosis on strength of phagocyte-particle interaction.

Sheep erythrocytes were pretreated with concanavalin A (Con-A-SRC), or glutaraldehyde (G-SRC), or specific rabbit immunoglobulin G (IgG-SRC), or specific rabbit immunoglobulin M and complement (C-SRC). Each erythrocyte type was made to adhere to rat peritoneal cells and adhesion was measured; binding decreased as follow: conA-SRC greater than IgG-SRC greater than less than G-SRC greater than C-SRC Peritoneal cell-erythrocyte complexes were then submitted to a laminar shear flow, and resistance of binding was assayed. Binging strength decreased in the following order: G-SRC greater than C-SRC greater than IgG-SRC greater than ConA-SRC Cell suspensions were incubated at 37 degrees, and phagocytosis was measured. Ingestion decreased in the following order: G-SRC greater than IgG-SRC greater than C-SRC greater than ConA-SRC It is concluded that: Binding strength may be of importance in triggering phagocytosis; when immunocytoadherence is studied, two independent parameters should be considered: binding and binding strength. This report describes a new method that may allow discrimination between different cell subpopulations of similar binding specificities.

Animals↗

Phagocytosis.

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Animals↗

Analysis of the topological changes induced on cells exposed to adhesive or mechanical stimuli.

Fluorescent probes are widely used to study cell structure and function. However, few reports were devoted to a quantitative analysis of the intracellular distribution of fluorescent markers. In the present work, we describe the topographical changes of surface and cytoskeletal markers on individual cells subjected to adhesive or mechanical interaction. Conjugates were prepared with a cytotoxic T-lymphocyte clone and target cells. Specific antigens, membrane phospholipids, surface glycoconjugates, and polymerized actin were labeled with fluorescent antibodies or biochemical probes. The analysis of fluorescence distributions in conjugates demonstrated a selective reorganization of the plasma membrane with a gathering of some molecular species in the intercellular adhesion area. Furthermore, individual phagocytic cells were sucked into glass micropipets, then stained with fluorescent phallacidin to analyze the effect of mechanical efforts on the cytoskeleton organization. The concentration of polymerized actin was found to be similar in mechanically-induced protrusions and whole cells. It is concluded that adhesive interactions may result in marked cell polarization and formation of membrane zones with a particular biochemical composition. The submembranar cytoskeleton might play a role in this process.

Actins↗

Protein tyrosine kinases and TNF alpha secretion in human monocytes.

The role of protein tyrosine kinases (PTK) in TNF alpha secretion by human monocytes was investigated in this report. We showed that an immunomodulator such as Nocardia lysozyme digest (NLD) and a particulate agonist, zymosan, stimulated an increase in tyrosine phosphorylation of several endogenous substrates including 53-56 kDa protein which was the predominant phosphoprotein. In addition, NLD and zymosan induced TNF alpha secretion which was impaired by a PTK inhibitor, tyrphostin. We suggest that a cascade of kinases including PTK is involved in NLD and zymosan signalling.

Humans↗

Phagocytic cell function in aged subjects.

In order to study the activity of phagocytic cells in normal and pathological aging, we compared normal young and aged subjects and patients with Alzheimer's (AD) or Parkinson's (PD) disease. Blood granulocytes and monocytes were separately assayed for ingestion of three different particle species (opsonized zymosan, immunoglobulin-coated sheep red cells (IgG-SRC) and glutaraldehyde-treated sheep red cells (G-SRC]. The superoxide anion production induced by these particles was also measured. All granulocyte responses to zymosan and IgG-SRC were depressed in the three aged groups as compared to young controls. Hence, only functions involving a specific receptor (Fc or C3b receptor) seemed affected. Monocyte activity was slightly decreased in the same groups. No difference was found between AD or PD patients and normal aged subjects. Hence the phagocytic and oxidative defects we found were a consequence of aging.

Adult↗

Effect of botulinum D toxin on human neutrophilic leukocytes and localization of its substrates.

Botulinum D toxin has been shown to ADP-ribosylate 22-kD proteins in neutrophilic leukocytes, but the function of these GTP-binding proteins remains unknown. In analogy to small GTP-binding proteins like SEC4 to YPT1, it has been suggested that botulinum D toxin substrates might be involved in secretory process of myeloid cells. Three main findings lead to the opposite conclusion. First of all, in human neutrophils, botulinum D toxin does not modify the release of azurophilic and specific granules induced by a chemoattractant (a formylpeptide) or a phorbol ester. Second, botulinum D toxin ADP-ribosylates 24 to 26-kD proteins that are only present in plasma membranes of human neutrophils. The membrane location of these substrates differs largely from that of the GTP-binding proteins involved in exocytosis and located in granules. Finally, since the same quantity of the toxin substrates is present in neutrophils as in their precursors, HL60 cells (which are devoid of specific granules and characterized by immature azurophilic granules and NADPH oxidase), it is unlikely that endogenous botulinum D toxin substrates are directly involved in the secretory responses of neutrophils.

Botulinum Toxins↗