Dying neurons conduct repair processes in the injured brain through osteopontin expression in cooperation with infiltrated blood monocytes

As part of a larger study involving the co-infection of human monocyte cultures with HIV and protozoan parasites, electron microscopic observations were made on the course of HIV replication and infection in these cells. Although several ultrastructural studies of the cytopathology associated with HIV infection have appeared, few studies have shown the details of virus production in “normal,” human monocytes/macrophages, one of the natural targets of the virus, and suspected of being a locus of quiescent virus during its long latent period. In this report, we detail some of the interactions of developing virons with the membranes and organelles of the monocyte host.Peripheral blood monocytes were prepared from buffy coats (Portland Red Cross) by Percoll gradient centrifugation, followed by adherence to cover slips. 90-95% pure monocytes were cultured in RPMI with 5% non-activated human AB serum for four days and infected with 100 TCID50/ml of HIV-1 for four hours, washed and incubated in fresh medium for 14 days.

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Ultrastructural Changes in Tumor Cells During Human Peripheral Blood Monocyte-Mediated Cytotoxicity

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100098757 ◽

1981 ◽

Vol 39 ◽

pp. 452-453

Author(s):

K. E. Muse ◽

D. G. Fischer ◽

H. S. Koren

Keyword(s):

Target Cell ◽

Human Peripheral Blood ◽

Mononuclear Phagocytes ◽

Ultrastructural Changes ◽

Target Cells ◽

Limited Information ◽

Blood Monocytes ◽

Tumoricidal Activity ◽

Activated State

Mononuclear phagocytes, a pluripotential cell line, manifest an array of basic extracellular functions. Among these physiological regulatory functions is the expression of spontaneous cytolytic potential against tumor cell targets.The limited observations on human cells, almost exclusively blood monocytes, initially reported limited or a lack of tumoricidal activity in the absence of antibody. More recently, freshly obtained monocytes have been reported to spontaneously impair the biability of tumor target cells in vitro (Harowitz et al., 1979; Montavani et al., 1979; Hammerstrom, 1979). Although the mechanism by which effector cells express cytotoxicity is poorly understood, discrete steps can be distinguished in the process of cell mediated cytotoxicity: recognition and binding of effector to target cells,a lethal-hit stage, and subsequent lysis of the target cell. Other important parameters in monocyte-mediated cytotoxicity include, activated state of the monocyte, effector cell concentrations, and target cell suseptibility. However, limited information is available with regard to the ultrastructural changes accompanying monocyte-mediated cytotoxicity.

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Ultrastructure of rat alveolar macrophages activated in situ by poly:IC

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100128158 ◽

1987 ◽

Vol 45 ◽

pp. 768-769

Author(s):

A. M. Klinkner ◽

R. A. Weiss ◽

A. Kelley ◽

P. J. Bugelski

Keyword(s):

Venous Blood ◽

Intratracheal Instillation ◽

Buffy Coat ◽

Fischer 344 Rats ◽

Blood Monocytes ◽

Fischer 344 ◽

Polyinosinic:Polycytidylic Acid ◽

Macrophage Activator ◽

Endogenous Peroxidase

Polyinosinic:polycytidylic acid is an inducer of interferon and a macrophage activator. We have found that intratracheal instillation of polyI:C (IT-pI:C) activates rat bronchoalveolar lavage cells (BAL) for a variety of functions. Examination of Giemsa stained, cytocentrifuge preparations showed that IT-pI:C induced a population of BAL not seen in resident BAL. The morphology of these cells suggested that they might be derived from blood monocytes. To test this hypothesis we have examined several populations of macrophages that had been stained for endogenous peroxidase activity as a marker of cells derived from the monocyte-macrophage lineage.Macrophages were obtained from Fischer 344 rats. Peritoneal exudate cells (PEC) were collected by lavage 4 days after i.p. injection of 20 ml 3% thioglycolate. Buffy coat monocytes were separated from venous blood from naive rats.

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Traumatic Brain Injury: A Trauma Surgeon's Perspective

Perspectives on Neurophysiology and Neurogenic Speech and Language Disorders ◽

10.1044/nnsld22.3.82 ◽

2012 ◽

Vol 22 (3) ◽

pp. 82-89

Author(s):

Oscar D. Guillamondegui

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

Multidisciplinary Approach ◽

Trauma Team ◽

The United States ◽

Long Term Outcomes ◽

Term Care ◽

Injured Brain ◽

The Moment

Traumatic brain injury (TBI) is a serious epidemic in the United States. It affects patients of all ages, race, and socioeconomic status (SES). The current care of these patients typically manifests after sequelae have been identified after discharge from the hospital, long after the inciting event. The purpose of this article is to introduce the concept of identification and management of the TBI patient from the moment of injury through long-term care as a multidisciplinary approach. By promoting an awareness of the issues that develop around the acutely injured brain and linking them to long-term outcomes, the trauma team can initiate care early to alter the effect on the patient, family, and community. Hopefully, by describing the care afforded at a trauma center and by a multidisciplinary team, we can bring a better understanding to the armamentarium of methods utilized to treat the difficult population of TBI patients.

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392: Osteopontin Expression in Metastatic Renal Cell Carcinoma

The Journal of Urology ◽

10.1016/s0022-5347(18)32648-x ◽

2006 ◽

Vol 175 (4S) ◽

pp. 128-128

Author(s):

Adrienne J.K. Carmack ◽

Daniel Saenz ◽

Merce Jorda ◽

Thomas Temple ◽

Balakrishna L. Lokeshwar

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Metastatic Renal Cell Carcinoma ◽

Renal Cell ◽

Osteopontin Expression

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Naphthalene as a model to study repair processes of the murine tracheal epithelium in vivo

Pneumologie ◽

10.1055/s-0032-1315553 ◽

2012 ◽

Vol 66 (06) ◽

Author(s):

K Döring ◽

G Knebel ◽

P König

Keyword(s):

Tracheal Epithelium ◽

Repair Processes

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Increased Expression of u-PA and u-PAR on Monocytes by LDL and Lp(a) Lipoproteins – Consequences for Plasmin Generation and Monocyte Adhesion

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614531 ◽

1999 ◽

Vol 81 (04) ◽

pp. 594-560 ◽

Cited By ~ 10

Author(s):

Florence Ganné ◽

Marc Vasse ◽

Jean-Louis Beaudeu ◽

Jacqueline Peynet ◽

Arnaud François ◽

...

Keyword(s):

Fold Increase ◽

Surface Expression ◽

Cell Surface Expression ◽

Atheromatous Plaque ◽

Monocyte Adhesion ◽

Blood Monocytes ◽

Proteolytic Cascade ◽

Ecm Degradation ◽

Dose Dependent Increase ◽

Dose Dependent

SummaryMonocyte-derived foam cells figure prominently in rupture-prone regions of atherosclerotic plaque. As urokinase/urokinase-receptor (u-PA/u-PAR) is the trigger of a proteolytic cascade responsible for ECM degradation, we have examined the effect of atherogenic lipoproteins on monocyte surface expression of u-PAR and u-PA. Peripheral blood monocytes, isolated from 10 healthy volunteers, were incubated with 10 to 200 µg/ml of native or oxidised (ox-) atherogenous lipoproteins for 18 h and cell surface expression of u-PA and u-PAR was analysed by flow cytometry. Both LDL and Lp(a) induced a dose-dependent increase in u-PA (1.6-fold increase with 200 μg/ml of ox-LDL) and u-PAR [1.7-fold increase with 200 μg/ml of ox-Lp(a)]. There is a great variability of the response among the donors, some of them remaining non-responders (absence of increase of u-PA or u-PAR) even at 200 μg/ml of lipoproteins. In positive responders, enhanced u-PA/u-PAR is associated with a significant increase of plasmin generation (1.9-fold increase with 200 μg/ml of ox-LDL), as determined by an amidolytic assay. Furthermore, monocyte adhesion to vitronectin and fibrinogen was significantly enhanced by the lipoproteins [respectively 2-fold and 1.7-fold increase with 200 μg/ml of ox-Lp(a)], due to the increase of u-PAR and ICAM-1, which are receptors for vitronectin and fibrinogen. These data suggest that atherogenous lipoproteins could contribute to the development of atheromatous plaque by increasing monocyte adhesion and trigger plaque weakening by inducing ECM degradation.

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Detection of both Type 1 and Type 2 Plasminogen Activator Inhibitors in Human Monocytes

Thrombosis and Haemostasis ◽

10.1055/s-0038-1645688 ◽

1990 ◽

Vol 63 (01) ◽

pp. 067-071 ◽

Cited By ~ 10

Author(s):

Joan C Castellote ◽

Enric Grau ◽

Maria A Linde ◽

Nuria Pujol-Moix ◽

Miquel LI Rutllant

Keyword(s):

Molecular Mass ◽

Plasminogen Activator ◽

Human Peripheral Blood ◽

Direct Interaction ◽

Apparent Molecular Weight ◽

Fibrinolytic System ◽

Human Monocytes ◽

Blood Monocytes ◽

Single Chain ◽

Sds Page

SummaryIncreasing evidence suggests the involvement of leukocytes in the fibrinolytic system. Monocytes secrete pro-urokinase (Grau, Thromb Res 1989; 53: 145) and it has been shown that these cells have specific receptors for urokinase and plasminogen (Miles, Thromb Haemostas 1987; 58: 936). The aim of this study was to analyse the presence of plasminogen activator inhibitor(s) in platelet-free suspensions of human peripheral blood monocytes and polymorphonuclear leukocytes (PMN). SDS-PAGE and reverse fibrin autography showed an inhibitory band of 50 kDa in the monocyte extracts (Triton X-100) but not in the PMN extracts. Urokinase (u-PA) was mixed with increasing amounts of monocyte extract for 10 min and the mixtures were added to 125Ifibrin coated wells containing plasminogen. A dose-dependent decrease in the u-PA fibrinolytic activity was observed. The amount of inhibition increased when the monocyte releasates were preincubated with u-PA (40% inhibition after 5 min preincubation and 80% after 15 min), indicating a direct interaction between this activator and an inhibitor(s). After SDS-PAGE of monocyte extracts, immunoblotting and peroxidase staining identified both PAI1 and PAI2, with an apparent molecular weight of 47-50 kDa. Monocyte-associated PAI1 formed complexes with single chain t-PA with a molecular mass 50 kDa higher than the molecular mass of the free PAI1. However, a significant amount of PAI remained unbound to t-PA. This inactive PAI1 could have come from a rapid inactivation of the primary active PAI1. These PAI1 and PAI2 detected in human monocytes may be transcendent in the regulation of the fibrinolytic system.

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Characterization of Monocyte-Associated Factor V

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649565 ◽

1993 ◽

Vol 70 (02) ◽

pp. 273-280 ◽

Cited By ~ 5

Author(s):

Janos Kappelmayer ◽

Satya P Kunapuli ◽

Edward G Wyshock ◽

Robert W Colman

Keyword(s):

Monoclonal Antibody ◽

Light Chain ◽

Peripheral Blood ◽

De Novo ◽

Factor V ◽

Blood Monocytes ◽

De Novo Synthesis ◽

Hep G2 ◽

Hep G2 Cells ◽

Peripheral Blood Monocytes

SummaryWe demonstrate that in addition to possessing binding sites for intact factor V (FV), unstimulated peripheral blood monocytes also express activated factor V (FVa) on their surfaces. FVa was identified on the monocyte surface by monoclonal antibody B38 recognizing FVa light chain and by human oligoclonal antibodies H1 (to FVa light chain) and H2 (to FVa heavy chain) using immunofluorescence microscopy and flow cytometry. On Western blots, partially cleaved FV could be identified as a 220 kDa band in lysates of monocytes. In addition to surface expression of FVa, monocytes also contain intracellular FV as detected only after permeabilization by Triton X-100 by monoclonal antibody B10 directed specifically to the Cl domain not present in FVa. We sought to determine whether the presence of FV in peripheral blood monocytes is a result of de novo synthesis.Using in situ hybridization, no FV mRNA could be detected in monocytes, while in parallel control studies, factor V mRNA was detectable in Hep G2 cells and CD18 mRNA in monocytes. In addition, using reverse transcriptase and the polymerase chain reaction, no FV mRNA was detected in mononuclear cells or in U937 cells, but mRNA for factor V was present in Hep G2 cells using the same techniques. These data suggest that FV is present in human monocytes, presumably acquired by binding of plasma FV, and that the presence of this critical coagulation factor is not due to de novo synthesis.

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