Macrophage Spreading Research Articles

Phosphatidylinositol 3-kinase is involved in the colony-stimulating factor-1-induced spreading of macrophages and osteoclasts

Phosphatidylinositol 3-kinase is involved in the colony-stimulating factor-1-induced spreading of macrophages and osteoclasts

Role of Ca2+ and calmodulin in ehrlichial infection in macrophages.

Replication of Ehrlichia risticii was inhibited in P388D1 cells and murine peritoneal macrophages when a calmodulin antagonist (W-7, chlorpromazine, or trifluoperazine); a Ca2+ channel blocker (verapamil, diltiazem, nifedipine, or flunarizine); an extracellular Ca2+ chelator, EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid]; an inhibitor of intracellular Ca2+ mobilization, TMB-8; or Ca2+ ionophore A23187 was added after internalization of the organism at 3 h postincubation. When intracellular ehrlichiae at their logarithmic stage of growth were treated with these reagents, not only was further proliferation prevented but also there was significant reduction in numbers of intracellular ehrlichiae. These reagents prevented spreading of E. risticii from P388D1 cells to THP-1 cells. None of these reagents prevented binding of [35S]methionine-labeled E. risticii to P388D1 cells, but all of these reagents prevented internalization of [35S]methionine-labeled E. risticii. Protein kinase C inhibitors, H-7 and staurosporin, had no effect. 14CO2 production from L-[14C]glutamine in Percoll-density-gradient-purified E. risticii was inhibited by A23187 but not by W-7 or verapamil, suggesting that Ca2+ but not calmodulin directly regulates ehrlichials glutamine oxidation. Pretreatment of E. risticii with W-7 or verapamil did not reduce its infectivity. These results indicate that calmodulin and Ca2+ are essential for ehrlichial internalization, replication, and spreading in macrophages but are not essential for binding.

Localization of cyclic GMP-dependent protein kinase in human mononuclear phagocytes

The presence and physiological role of cGMP-dependent protein kinase (G-kinase) was investigated in human mononuclear phagocytes. Western blots of monocyte extracts revealed a single polypeptide band that comigrated with purified bovine lung G-kinase. G-kinase was localized by immunofluorescence microscopy in freshly isolated adherent human monocytes, monocyte-derived macrophages cultured from 4 to 14 days, and alveolar macrophages. In monocytes, G-kinase was localized in granules or vesicles in the cytoplasm, at the microtubule organizing center, on filaments, and in the nucleus. In monocyte-derived macrophages, intense staining for G-kinase was found in the vicinity of the Golgi, in vesicles throughout the cytoplasm, and diffusely in the nucleus. Dual-label confocal laser scanning microscopy demonstrated that G-kinase was colocalized with the endoplasmic reticulum. For comparison, G-kinase was localized in alveolar macrophages that were adhered from 3 to 30 min. In these cells, G-kinase was prominent within the organelle-rich area pericortical to the nucleus. However, a well-defined area of intense staining was also observed at the cell periphery at early time points during adherence and spreading. Rhodamine-labeled phalloidin showed that this peripheral area was rich in F-actin. Cytochalasin D, but not nocodazole, inhibited G-kinase targeting to the cell margin. Furthermore, the guanylate cyclase inhibitor LY83583 inhibited alveolar macrophage spreading and staining for G-kinase at the cell periphery. These data suggest that G-kinase may play an important role in cGMP-mediated regulation involved in protein processing and cell motility.

The Effect of Space and Parabolic Flight on Macrophage Hematopoiesis and Function

We used weak electric fields to monitor macrophage spreading in microgravity. Using this technique, we demonstrated that bone marrow-derived macrophages responded to microgravity within 8 s. We also showed that microgravity differentially altered two processes associated with bone marrow-derived macrophage development. Spaceflight enhanced cellular proliferation and inhibited differentiation. These data indicate that the space/microgravity environment significantly affects macrophages.

Diagnosis and epidemiological association of Listeria monocytogenes strains in two outbreaks of listerial encephalitis in small ruminants.

Two outbreaks of epizootic listerial encephalitis, one in sheep and one in goats, were investigated through pathology, microbiology, and DNA amplification-based techniques. Efforts were made to survey the diversity of Listeria monocytogenes strains in the silage consumed by affected animals and to verify the causal relationship between silage and disease outbreak. In both outbreaks, L. monocytogenes was isolated from silage and brain tissue samples. Random amplified polymorphic DNA patterns revealed two distinct L. monocytogenes strains, one of which was identical to the sheep brain isolate, in the silage associated with the outbreak in sheep. Three brain isolates and one silage isolate, all of which had different random amplified polymorphic DNA patterns, were found in the outbreak involving goats. All isolates from both outbreaks were indistinguishable in an in vitro assay for cell-to-cell spread and growth in macrophages. All brain isolates from the goat outbreak had identical intracellular ActA patterns, which were different from the pattern for the silage isolate. While the sheep brain isolate had an ActA pattern different from that of the corresponding silage isolate, the patterns for the brain isolates from the two outbreaks were not identical. This survey demonstrates the diversity of L. monocytogenes in silage and suggests the existence of one or more selective processes by which certain strains are more prone to give rise to disease.

Biocompatibility evaluation of laser-induced AAm and HEMA grafted EPR. Part 1: In-vitro study

Samples based on ethylene-propylene rubber (EPR) have been surface grafted with acrylamide (AAm) and 2-hydroxyethyl methacrylate (HEMA) using CO 2-pulsed laser as a stimulation source. Scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDXA) and attenuated total reflectance infrared (ATR-IR) spectra were performed on the modified samples. These techniques revealed the formation of grafted poly(AAm) and poly(HEMA) on the surface of EPR. The surface grafted poly(AAm) and poly(HEMA) were found to have a fractal type of morphology. EDXA showed insignificant grafted AAm and HEMA in regions where fractals were absent. Fractal patterned surfaces provide hydrophilic and hydrophobic sites, making EPR suitable as a biomaterial. In-vitro adhesion and spreading of alveolar macrophages (AMs) cultured on the surface of modified samples have been evaluated by hemocytometry and SEM, respectively, and compared with unmodified controls. Relationships between AM adhesion and their spreading, with surface morphology, graft level and water compatibility are also discussed. Generally, more AMs attach onto unmodified surfaces with a greater degree of spreading, than on the modified EPR. Samples grafted between 0·7mg/cm 2 and Img/cm 2 showed fairly low AM density compared with both unmodified EPR and lightly modified samples (less than 0·2 mg/cm 2). AMs cultured on the unmodified EPR were larger and displayed pronounced ruffling of the plasma membrane, an increased capacity for adherence and spreading on the surface, and an increased number of extensive filopodia. Moreover, AMs attached onto the surface of modified samples appeared rounded, with minimal cytoplasmic spreading and ruffling.

Macrophage recognition of polymers: effect of carboxylate groups

The effect of anionic carboxylate groups in copolymers of 2-hydroxyethyl methacrylate with sodium methacrylate on the adsorption of albumin, adhesion, spreading and fusion of macrophages has been demonstrated. The results show an inhibitory effect of increasing concentration of carboxylate groups of polymers on all studied parameters.

Suppression of the Delayed Type Hypersensitivity Response by Tumor Facilitating Factor of B16 Melanoma

Tumor facilitating factor is a cell surface glycoprotein produced by B16 melanoma that has been found to reduce the lethal inoculum for B16. Tumor facilitating factor induces macrophage spreading in vitro, reduces macrophage chemotaxis in vivo, and depresses lymphocyte mitogenesis in vitro. It is assumed that the immune modifying effects are responsible for tumor facilitation. As tumors may be poor immunogens or inducers of inflammation, studies were conducted to determine whether tumor facilitating factor alters the inflammatory cascade of cells found in infiltrates of delayed type hypersensitivity. Freeze-thawed B16 cells, used as the source of TFF, caused a suppression of delayed type hypersensitivity measured as ear swelling in the mouse. When culture supernatant was substituted for freeze-thawed cells as a source of TFF and injected at different time points of the delayed type hypersensitivity response, the greater suppression was with tumor facilitating factor injections at 24 hours pre-elicitation only (82%), and 24 hours both presensitization and pre-elicitation (89%). Immunohistological staining demonstrated that tumor facilitating factor decreases ear thickness and cellular infiltrates, specifically Mac-1 staining cells, to a site of delayed hypersensitivity. Peritoneal cell analysis confirmed these findings. These data are consistent with the hypothesis that tumor facilitating factor alters immune functions including macrophage and lymphocyte mobility and recruitment to a target site, thereby allowing for facilitation of tumor growth.

Are hepatocyte growth factor‐like protein and macrophage stimulating protein the same protein?

Protein ScienceVolume 2, Issue 4 p. 666-668 For the RecordFree Access Are hepatocyte growth factor-like protein and macrophage stimulating protein the same protein? Jorge A. Bezerra, Jorge A. Bezerra Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorSu Han, Su Han Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorMary Jo S. Danton, Mary Jo S. Danton Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorSandra J. Friezner Degen, Corresponding Author Sandra J. Friezner Degen Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Division of Basic Science Research, Children's Hospital Research Foundation, Elland and Bethesda Avenues, Cincinnati, Ohio 45229–2899Search for more papers by this author Jorge A. Bezerra, Jorge A. Bezerra Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorSu Han, Su Han Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorMary Jo S. Danton, Mary Jo S. Danton Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Search for more papers by this authorSandra J. Friezner Degen, Corresponding Author Sandra J. Friezner Degen Division of Basic Science Research, Children's Hospital Research Foundation and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio 45229Division of Basic Science Research, Children's Hospital Research Foundation, Elland and Bethesda Avenues, Cincinnati, Ohio 45229–2899Search for more papers by this author First published: April 1993 https://doi.org/10.1002/pro.5560020416Citations: 20AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat References 1 Bhargava, M., Joseph, A., Knesel, J., Halaban, R., Li, Y., Pang, S., Goldberg, I., Setter, E., Donovan, M.A., Zarnegar, R., Michalopoulos, G.A., Nakamura, T., Faletto, D., & Rosen, E.M. (1992). Scatter factor and hepatocyte growth factor: Activities, properties and mechanism. Cell Growth Differ. 3, 11– 20. 2 Bottaro, D.P., Rubin, J.S., Faletto, D.L., Chan, A.M.-L., Kmiecik, T.E., Vande Woude, G.F., & Aaronson, S.A. (1991). Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. Science 251, 802– 804. 3 Han, S., Stuart, L.A., & Degen, S.J.F. (1991). Characterization of the DNF15S2 locus on human chromosome 3: Identification of a gene coding for four kringle domains with homology to hepatocyte growth factor. Biochemistry 30, 9768– 9780. 4 Leonard, E.J. & Skeel, A. (1976). A serum protein that stimulates macrophage movement, chemotaxis and spreading. Exp. Cell Res. 102, 434– 438. 5 Michalopoulos, G.K. & Zarnegar, R. (1992). Hepatocyte growth factor [editorial]. Hepatology 15, 149– 155. 6 Nakamura, T., Nishizawa, T., Hagiya, M., Seki, T., Shimonishi, M., Sugimura, A., Tashiro, K., & Shimizu, S. (1989). Molecular cloning and expression of human hepatocyte growth factor. Nature 342, 440– 443. 7 Naldini, L., Vigna, E., Narsimhan, R.P., Gaudino, G., Zarnegar, R., Michalopoulos, G.K., & Comoglio, P.M. (1991). Hepatocyte growth factor (HGF) stimulates the tyrosine kinase activity of the receptor encoded by the proto-oncogene c-MET. Oncogene 6, 501– 504. 8 Skeel, A., Yoshimura, T., Showalter, S.D., Tanaka, S., Appella, E., & Leonard, E.J. (1991). Macrophage stimulating protein: Purification, partial amino acid sequence, and cellular activity. J. Exp. Med. 173, 1227– 1234. Citing Literature Volume2, Issue4April 1993Pages 666-668 ReferencesRelatedInformation

Macrophages in the urine in acute bacterial cystitis.

In the previous study in this series of studies concerning the role of macrophages in urinary tract infection, we attempted to detect macrophages in the urine of acute bacterial cystitis patients by nonspecific esterase staining of urinary sediment, however none of the leukocytes stained, probably because of cell damage caused by the urine and by centrifugation. In the present study, detection of macrophages in urine was again attempted, this time by prompt transfer of urinary leukocytes to culture medium after minimum centrifugation, 1 hr culture in a glass bottom chamber and non-specific staining of leukocytes adhering to glass. Macrophages in urine were detected by this method, and they comprised 5.9% of the adherent leukocytes, although macrophage spreading, which implies macrophage activation and is often seen in the early stage of nonbacterial prostatitis, was hardly ever observed. The percentages of adherent leukocytes were not correlated with urine osmolarity, probably because the effect of urine was minimized by prompt transfer of urinary leukocytes to culture medium after the urine samples had been collected. There have been quite few studies involving culture of urinary leukocytes in the past. Our simple techniques, such as prompt transfer of urinary leukocytes to culture medium after centrifuging with minimum gravity and for a minimum period of time, appear to be useful in the study of urinary leukocytes using other cells which appear in urinary tract infection, as well as cytokines and antibiotics, to clarify cellular mechanisms of defenses in urinary tract infection.

Macrophage response to microtextured silicone

Seven different silicone surface textures were tested for effect on macrophage spreading and metabolic activity in vitro. Variables of the textured arrays that could modify spreading were determined to be the size, spacing between, depth, density, and orientation of the individual surface events and the roughness of the surfaces. Cells were influenced by the size of the events and the roughness of the surfaces more than any other variables. Cell morphology data, surface area and perimeter, could be divided into discrete regions that correlated well with the size of the events. Cell dimensions on 5μm textures were smallest while those on smooth silicone and glass surfaces were the largest. Surface texture events may be modifying contact guidance of the cells or interacting with specific transmembrane proteins to alter cell shape and function. The mitochondrial activity of cells attached to the textured silicones was determined by measuring the amount of reduced MTT directly through live cells. Cells on polystyrene (PS), 5VP and 8VP textures were metabolically more active than cells on the other textures. PMA was used to stimulate cells on the various textures. PMA-stimulated cells, on the smaller textures, 2VP, 5VP and 5CP, were less active than test cells that were not stimulated. The inability of PMA to stimulate these cells may be due to a structural alteration of protein kinase C. An hypothesis is introduced that includes a possible mechanism of how a micrometre-sized surface texture could modify cell function.

Influence of circadian light-dark alternations on macrophages and lymphocytes of CBA mouse.

The influence of circadian 12 h light-12 h dark alternations on CBA mouse macrophages and lymphocytes was determined using tests for macrophage spreading and ingestion ability or flow cytometry immunophenotyping of blood, lymph node, and spleen lymphocytes. The animals were tested every 4 h around the clock. Collected macrophages were incubated in vitro for 3 or 18 h. Monoclonal antibodies permitted detection of T-lymphocytes, suppressor-cytotoxic T-lymphocytes, helper-inducer T-lymphocytes, or B-lymphocytes. Two types of analyses were performed: First, the difference between the same intervals of the 12 h light or dark period was determined. The macrophage ingestion was significantly lower at the beginning and higher at the end of the dark period. We have also found a significant increase in blood T-lymphocytes of helper-inducer T-lymphocyte percentages and of the T helper-inducer: T suppressor-cytotoxic ratio during the dark period. Second, the ultradian variation during the 12 h light or dark period was determined. The variability was significant both for macrophage spreading and ingestion. Multiple significant variations of lymph node, spleen, or blood lymphocyte percentages were also observed. All of these data indicate that daily alteration of the lighting regimen significantly influences mouse peritoneal macrophage functions and various lymphocyte subsets.

Suppression of human macrophage function in vitro by delta 9-tetrahydrocannabinol.

The ability of macrophages to function in the presence of delta 9-tetrahydrocannabinol (THC), the major psychoactive component in marijuana, was evaluated. THC added to macrophage cultures prepared from human peripheral blood inhibited macrophage spreading and phagocytosis of yeast. The effects of THC were concentration dependent, with inhibitory effects observed from 10 to 1 micrograms/ml or lower. These results suggest that macrophages are more sensitive to THC than are lymphocytes because macrophage functions were inhibited by THC at concentrations that did not affect lymphocyte function. Thus, inhibition of lymphocyte function(s) by THC could be attributed to a direct effect of the drug on macrophages which indirectly results in lowered lymphoid cell activity.

Single pulses of cytoplasmic calcium associated with phagocytosis of individual zymosan particles by macrophages

We have measured cytosolic free calcium levels ([Ca ++] i) in individual macrophages during the phagocytosis of single zymosan particles. We report here that the contact of a macrophage with zymosan results in a rapid transient elevation of [Ca ++] i. Each [Ca ++] i pulse is symmetrical lasting for up to 30 seconds. In contrast, macrophage spreading is associated with repetitive [Ca ++] i spiking occuring in salvos of up to four smaller spikes, each lasting for between 8 and 18 seconds. These qualitative and kinetic differences might suggest that the role of [Ca ++] i in phagocytosis is distinct from its role in spreading.

Suppression of T lymphocyte subpopulations by THC.

THC (delta-9-tetrahydrocannabinol) is considered the major psychoactive component of marijuana. It is widely acknowledged that THC can depress various parameters of the immune response. These parameters include lymphocyte proliferation, macrophage spreading, and antibody production (1–3). Studies have suggested that THC has different magnitudes of effect depending on the lymphoid organ analyzed (2) as well as the age of the animal (4). Different lymphoid organs of adult mice as well as identical organs from mice of various ages have unique proportions of lymphoid cell subpopulations. This study examines the effect of THC on lymphoid cell subpopulations using the lymphoblastogenic assay (LBT) as well as analysis of cell subpopulations by the fluorescent activated cell sorter (FACS).KeywordsSpleen CellFluorescent Activate Cell SorterLymphoid OrganIdentical OrganFluorescent Activate Cell Sorter AnalysisThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Essential fatty acid deficiency impairs macrophage spreading and adherence. Role of arachidonate in cell adhesion.

Dietary polyunsaturated fatty acid manipulation exerts a strikingly protective effect in models of tissue inflammation and injury. A critical element of this effect appears to revolve around leukocyte trafficking but underlying mechanisms are ill understood. In the current study it was observed that essential fatty acid (EFA) deficiency markedly impaired the capacity of resident macrophages to spread and adhere. This effect was not a simple function of the alteration of membrane fatty acid composition. Elicited EFA-deficient macrophages were equally adherent to elicited control cells, despite the fact that they were equally EFA-deficient relative to resident EFA-deficient cells. With respect to the mechanism underlying defective macrophage adherence in EFA deficiency, no change in the expression of cell surface adherence molecules (Fc receptor, Mac-1, or LFA-1) was noted with the deficiency state. Also, an adherence defect could not be induced in normal cells pharmacologically with cyclooxygenase blockade, lipoxygenase blockade, or a platelet-activating factor receptor antagonist. In contrast, phospholipase inhibition was able to induce a spreading and adherence defect in resident macrophages similar to that seen with EFA deficiency. Using several phospholipase inhibitors, a correlation between phospholipase inhibition and impairment of adherence was observed. Adding back exogenous fatty acids to cells after phospholipase inhibition demonstrated that normal adherence was reconstituted with arachidonate. This alteration in macrophage spreading and adherence with EFA deficiency may be an important component of the anti-inflammatory effect of dietary polyunsaturated fatty acid manipulation. Additionally, these results suggest that arachidonate may be an intracellular mediator of leukocyte adherence.

Effects of saikosaponin-d on the functions and morphology of macrophages

The effects of saikosaponin-d, isolated from Bupleurum Radix, on phagocytosis and spreading of mouse peritoneal macrophages were investigated. Macrophages from saikosaponin-d-treated mice showed a significant increase in spreading activity followed by an increase in phagocytic activity. An intense distribution of microfilaments and microtubules was also observed in these macrophages by immunofluorescence microscopy.

Study of Macrophages in Prostatic Fluid from Nonbacterial Prostatitis Patients

Relationship between activation of macrophages in prostatic fluid and stage of nonbacterial prostatitis was studied by observing the rate of adherence of leukocytes, percentage of macrophages among adherent leukocytes and presence of spreading of macrophages. As a result, it was found that macrophages in the early stage of NBP were more activated than macrophages in the chronic stage.

Macrophage functions are regulated by the substratum of murine decidual stromal cells.

Because of their paternal antigens, the fetus and placenta may be considered an allograft in the maternal host. Local properties of the maternal-fetal interface, the placenta and decidua basalis, are important in preventing maternal immunologic rejection of the fetoplacental allograft. However, the exact nature of these local properties remains a fundamental unsolved problem in immunology. We now report that three macrophage functions were inhibited by the substratum formed by monolayers of decidual stromal cells via a novel pathway. Solid-phase inhibitors blocked macrophage adhesion, spreading, and lysis of tumor necrosis factor-alpha-resistant P815 mastocytoma tumor cells. Inhibition was not solely attributable to an inability of macrophages to adhere to decidual substratum because there were differences in macrophage functions on this surface versus polyhema where no adherence occurred. Because macrophages play a central role in cell-mediated immunity, including allograft rejection, inhibiting their function in the decidua basalis may help prevent maternal antifetal responses.

The influence of hydrogel functional groups on cell behavior

This study provides information on the behavior of macrophages on the surface of the subcutaneously implanted hydrogel strips. Hydrogel containing -OH, -CO-NH-, and (CH3)2N- groups induced a spreading of macrophages on the implants. The materials containing -SO3H groups slightly, and materials containing -COOH groups more intensively, inhibited spreading of the macrophages. The fusion of macrophages into multinucleate cells was inhibited on the surface of materials containing acidic groups (-SO3H, -COOH) and increased on the hydrogels containing 30 mol% of alkaline (CH3)2N- groups in comparison with hydrogels containing -OH or -CO -NH- groups. The differences of the behavior of macrophages on the surface of individual types of hydrogels are probably independent on the adsorption of plasma fibronectin onto the hydrogels. The correlation between the macrophages spreading and fusion and surface charge of the hydrogel implant can be hypothetically explained by electrostatic interaction between macrophages cell membrane and implant.