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Results for “Extracellular Traps”

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At least 847 records · Page 47Linked to original sources

PubMed1994

Macrophages loaded with doxorubicin by ATP-mediated permeabilization: potential carriers for antitumor therapy.

In many cell types extracellular ATP (ATPe) has been shown to cause reversible plasma membrane permeabilization to low molecular weight (< 900 Da) water-soluble compounds. In the present report we have exploited this technique to incorporate the anticancer drug doxorubicin (DXR), molecular mass 543 Da, into the cytoplasm of two mouse cell lines that had previously been shown to express the ATPe-gated pore, J774 macrophages and tumor necrosis factor (TNF)-resistant L929 fibroblasts. Compared to passively loaded cells, ATPe-mediated reversible permeabilization allowed an at least 4-fold increase in DXR intracellular trapping (0.5 pg/cell versus 2 pg/cell). Analysis of the release kinetics at 37 degrees C showed that about 40% of total intracellular DXR was discharged during the first hour from both ATPe-permeabilized and passively loaded cells; about 15% further release was observed upon incubation up to 4 h. DXR release profiles were similar in ATPe-permeabilized and passively loaded cells. ATPe-permeabilized, DXR-loaded (ATPe-DXR) cells strongly inhibited the proliferation of K562 tumor cells. Taken together these results indicate that ATPe-mediated reversible plasma membrane permeabilization can be effectively used to load cells of different histotypes with high concentrations of DXR. This approach could permit to vehicle high doses of anticancer agents by using living cells while reducing systemic toxic effects.

Adenosine Triphosphate↗

PubMed1997

The beta-amyloid epitope masking activity in human brain is identified as albumin.

Human brain homogenate proteins were analyzed for binding and processing activity in relation to brain beta-amyloid precursor protein (APP). The homogenate was purified by arginine-Sepharose 4B affinity chromatography, which traps proteins with affinity to certain groups of arginine residue, such as serine proteases and zymogens. A 69 kDa protein that masks epitope(s) of brain APP was found in a weakly bound fraction. The nature of the 69 kDa brain protein was identified as albumin by N-terminal amino acid sequencing and Western blot analysis using anti-human albumin antibody. Western blot analysis with domain-specific anti-APP antibodies revealed that the masking activity is complete for beta-amyloid epitope(s), but incomplete for cytoplasmic and extracellular domain epitopes, suggesting that the interaction site of the albumin is beta-amyloid itself. Therefore, it seems that brain albumin is not merely a carrier protein for beta-amyloid in cerebrospinal fluid, but also a modulator which interferes with processing of beta-amyloid precursor protein and its peptides.

Aged↗

PubMed1998

Characterization of two Mg2+ transporters in sealed plasma membrane vesicles from rat liver.

The plasma membrane of mammalian cells possesses rapid Mg2+ transport mechanisms. The identity of Mg2+ transporters is unknown, and so are their properties. In this study, Mg2+ transporters were characterized using a biochemically and morphologically standardized preparation of sealed rat liver plasma membranes (LPM) whose intravesicular content could be set and controlled. The system has the advantages that it is not regulated by intracellular signaling machinery and that the intravesicular ion milieu can be designed. The results indicate that 1) LPM retain trapped intravesicular total Mg2+ with negligible leak; 2) the addition of Na+ or Ca2+ induces a concentration- and temperature-dependent efflux corresponding to 30-50% of the intravesicular Mg2+; 3) the rate of flux is very rapid (137.6 and 86.8 nmol total Mg2+ . micrometer -2 . min-1 after Na+ and Ca2+ addition, respectively); 4) coaddition of maximal concentrations of Na+ and Ca2+ induces an additive Mg2+ efflux; 5) both Na+- and Ca2+-stimulated Mg2+ effluxes are inhibited by amiloride, imipramine, or quinidine but not by vanadate or Ca2+ channel blockers; 6) extracellular Na+ or Ca2+ can stimulate Mg2+ efflux in the absence of Mg2+ gradients; and 7) Mg2+ uptake occurs in LPM loaded with Na+ but not with Ca2+, thus indicating that Na+/Mg2+ but not Ca2+/Mg2+ exchange is reversible. These data are consistent with the operation of two distinct Mg2+ transport mechanisms and provide new information on rates of Mg2+ transport, specificity of the cotransported ions, and reversibility of the transport.

Animals↗

PubMed2006

Protein profile of osteoarthritic human articular cartilage using tandem mass spectrometry.

Articular cartilage contains both chondrocyte cells and extracellular matrix (ECM) components. Currently, comprehensive information concerning the protein composition of human articular cartilage tissue is somewhat lacking. In this report we detail the use of tandem mass spectrometry (MS/MS) for a preliminary global identification of proteins from human articular knee cartilage tissue from patients diagnosed with osteoarthritis. Knee cartilage supernatant was fractionated using one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE), in-gel digested and peptide sequences were then determined by performing on-line nano-liquid chromatography (LC)/MS/MS experiments using an ion trap mass spectrometer. Altogether, over 100 different proteins from nearly 700 unique peptide sequences were detected by MS/MS. The majority of the proteins identified are involved in ECM organization (35%), signal transduction and cell communication (14%), immune response (11%) and metabolism and energy pathways (11%). Proteins observed included several well-known cartilage components as well as lower abundant lesser known ECM proteins. Possible degradation products in the cartilage sample, such as from cartilage link protein, could also be detected by our mass spectrometry methods. We show here that mass spectrometry can be utilized as a tool for a fast, accurate and sensitive analysis of a complex mixture of cartilage proteins. It is believed that this type of proteomic analysis will aid future work centered on investigating the pathology of this and other related joint diseases.

Adult↗

PubMed2003

Cocaine treatment increases expression of a 40 kDa catecholamine-regulated protein in discrete brain regions.

Previous reports from our laboratory have described brain-specific catecholamine-regulated proteins, which bind dopamine and related catecholamines. Evidence from the molecular cloning of a 40 kDa catecholamine-regulated protein (CRP40) revealed that CRP40 is dopamine-inducible and has properties similar to those of the 70 kDa heat shock protein (HSP70) family. The present study investigates the effects of acute and chronic cocaine treatment on CRP40 expression in the striatum, nucleus accumbens, prefrontal cortex, and medulla. Acute treatment with cocaine increased CRP40 expression in the nucleus accumbens and striatum, whereas chronic treatment with cocaine increased CRP40 expression in the nucleus accumbens only. Neither of these treatments affected CRP40 levels in the prefrontal cortex or medulla. In addition, pretreatment with the spin-trapping agent alpha-phenyl-tert-butylnitrone did not attenuate cocaine-induced expression of CRP40, suggesting that the observed increases in CRP40 levels were not caused by free radicals. On the other hand, pretreatment with anisomycin, a protein synthesis inhibitor, blocked the cocaine-induced expression of CRP40. Thus, protein synthesis may be involved in the observed CRP40 level increases. Furthermore, neither acute nor chronic cocaine treatment affected levels of inducible or constitutively expressed HSP70, which indicates a specificity of cocaine's effects on CRP40. Since cocaine has been shown to increase extracellular dopamine levels, these findings suggest that increased expression of CRP40 is associated with high extracellular levels of dopamine (or its metabolites). Elevated levels of CRP40 could play a protective role for dopamine neurons in response to increased oxidative stress that has been shown to be induced by cocaine and that can lead to apoptosis and neurodegeneration.

Animals↗

PubMed2002

Hemocyanin from the keyhole limpet Megathura crenulata (KLH) carries a novel type of N-glycans with Gal(beta1-6)Man-motifs.

Keyhole limpet (Megathura crenulata) hemocyanin (KLH), an extracellular respiratory protein, is widely used as hapten carrier and immune stimulant. Although it is generally accepted that the sugar constituents of this glycoprotein are likely to be implicated in the antigenicity and biomedical properties of KLH, knowledge of its carbohydrate structure is still limited. Therefore, we have investigated the N-linked oligosaccharides of KLH. Glycan chains were enzymatically liberated from tryptic glycopeptides, pyridylaminated and separated by two-dimensional HPLC. Only neutral oligosaccharides were obtained and characterized by carbohydrate constituent and methylation analyses, MALDI-TOF-MS, ESI-ion trap-MS and sequential exoglycosidase digestion. The results revealed that KLH is carrying high mannose-type glycans and truncated sugar chains derived thereof. As a characteristic feature, a number of the studied N-glycans contained a Gal(beta1-6)Man-unit which has not been found in glycoprotein-N-glycans so far. Hence, our studies demonstrate that this marine mollusk glycoprotein is characterized by a unique oligosaccharide pattern comprising, in part, novel structural elements.

Amino Acid Motifs↗

PubMed1998

Cloning and characterization of developmental endothelial locus-1: an embryonic endothelial cell protein that binds the alphavbeta3 integrin receptor.

We have taken advantage of an enhancer trap event in a line of transgenic mice to identify a unique developmentally regulated endothelial cell locus (Del1). The protein encoded in this locus contains three EGF-like repeats homologous to those in Notch and related proteins, including an EGF-like repeat that contains an RGD motif, and two discoidin I-like domains. Del1 is shown to be a matrix protein and to promote adhesion of endothelial cells through interaction with the alphavbeta3 integrin receptor. Embryonic endothelial-like yolk sac cells expressing recombinant Del1 protein, or grown on an extracellular matrix containing Del1 protein, are inhibited from forming vascular-like structures. Expression of Del1 protein in the chick chorioallantoic membrane leads to loss of vascular integrity and promotes vessel remodeling. Del1 is thus a new ligand for the alphavbeta3 integrin receptor and may function to regulate vascular morphogenesis or remodeling in embryonic development.

Amino Acid Sequence↗

PubMed2000

Extracellular targeting of choline-binding proteins in Streptococcus pneumoniae by a zinc metalloprotease.

A genetic-based search for surface proteins of Streptococcus pneumoniae involved in adhesion identified a putative zinc metalloprotease (ZmpB). ZmpB shared high amino acid sequence similarities with IgA1 proteases of Gram-positive bacteria, but ZmpB had neither IgA1 nor IgA2 protease activity. Analysis of a family of surface-expressed proteins, the choline-binding proteins (Cbp's), in a zmpB-deficient mutant demonstrated a global loss of surface expression of CbpA, CbpE, CbpF and CbpJ. CbpA was detected within the cytoplasm. The zmpB-deficient mutant also failed to lyse with penicillin, a sign of lack of function of the Cbp LytA. Immunodetection studies revealed that the autolysin (LytA), normally located on the cell wall, was trapped in the cytoplasm colocalized with DNA and the transformation protein CinA. Trafficking of CinA and RecA to the cell membrane during genetic competence was also not observed in the zmpB-deficient mutant. These results suggest a protease dependent regulatory mechanism governing the translocation of CinA and the Cbp's LytA and CbpA of S. pneumoniae.

Alleles↗

PubMed1993

The diagnostic significance of autoantibodies against cartilage components or their counterparts in destructive articular diseases.

In this work four different types of antibodies to collagens, membrane protein of chondrocytes (MP), cartilage matrix proteins (CMP) and heat shock proteins (HSP-s) were tested in order to find an available clinical laboratory method for diagnostic and monitoring purposes in patients suffering from osteoarthritis. From the point of view of diagnostic efficiency, the estimation of antibody level to MP seemed to be the best and as a supplementary method the determination of rheuma factor was recommended. The anticollagen antibody estimation is less sensitive, anti CMP antibodies are not detectable. In spite of immunological crossreaction between HSP and cartilage matrix component the antibody level against HSP has no correlation with osteoarthritis. The appearance of humoral reaction, antibodies against different cartilage specific collagens and chondrocyte membrane proteins, is an epiphenomenon, however as the supposed immune complexes, trapped in a cartilage, play an important role in the damage of cartilage which may explain the self-perpetuating and chronic nature of cartilage degradation on osteoarthritis.

Adolescent↗

PubMed1982

Kinetic studies on the effects of ouabain on Na+ fluxes in frog skin.

Among 48 pieces of paired frog skins of Rana pipiens in Ringer's solution, 10 pieces showed a strictly monotone decrease in the short circuit current (SCC) following ouabain treatment (10(-4) M). In 9 cases a transient attenuation, and in 27 cases a distinct wave in the ebb of the SCC, was seen. In 2 instances, two waves were seen. Associated with the not-monotone events was a transient rise in electrical skin conductance. The reasons for these mixed skin responses are unknown. One possible reason is considered here: Early during the ouabain action, some of the Na+ entering from the mucosal side is trapped in the skin by electroneutral processes, in keeping with the already known fact that ultimately cellular KCl is partly replaced by NaCl. Computer assisted model studies show how monotone, and not-monotone "transepithelial" net Na+ flux curves can be generated. Essential conditions for the generation of not-monotone Na+ flux curves are: 1. Presence of two distinct "cellular", active Na+ pools in the model. 2. Presence of a loop pathway in which a principal "transepithelial Na+ transport compartment", and a constituent "Na+/K+ maintenance compartment", are connected to each other and to the "extracellular" compartment. The model, then, predicts under which kinetic conditions monotone and not-monotone transepithelial Na+ flux curves will be seen.

Animals↗

PubMed1997

Osmotic 'detubulation' in frog muscle arises from a reversible vacuolation process.

Isolated Rana temporaria sartorius muscle fibres were subject to introduction and subsequent withdrawal of 400 mM extracellular glycerol, exposures to high divalent ion concentrations and then cooling. Tubular detachment was then assessed through changes in the action potential afterdepolarization. (1) The rapid (5-10 min) rather than slow cooling step (30 min) produced a gradual (30 min) development of detubulation arrested by the subsequent replacement of glycerol and reversed by addition of 350 mM sucrose. Such osmotic agents influenced neither resting potentials of intact or detubulated fibres nor action potentials in intact fibres. (2) Full tubular detachment was achieved by 40 min. Laser epifluorescence microscopy demonstrated an accompanying tubular vacuolation through its trapping of a Rhodamine dye. (3) Subsequent re-additions (at 10-80 min) of glycerol restored the afterdepolarization in 30% of detubulated fibres and correspondingly reduced vacuolation. Sustained (> 60 min) exposures to 350 mM sucrose, applied between 30-60 min, both reversed tubular isolation in 70% of detubulated fibres and abolished tubular vacuolation. Finally, results from transient (10-30 min) sucrose exposures resembled the consequences of sustained applications of glycerol, suggesting that detubulation and its reversal result from an osmotic mechanism. (4) Nevertheless, irreversible changes developed after 70-80 min in 70% of detubulated fibres, a process hastened by slow cooling steps in the initial osmotic stress. The present study thus correlates morphological and electrophysiological consequences of applying osmotic shock to skeletal muscle for the first time. It additionally differentiates reversible and irreversible components of detubulation. Finally, it suggests that detubulation results from the similarly reversible vacuolation observed under comparable osmotic conditions, and that such vacuolation can eventually lead to irreversible detubulation.

Action Potentials↗

PubMed2005

Er:YAG laser promotes gingival wound repair by photo-dissociating water molecules.

OBJECTIVE: The aim of the present study was to show that rapid wound repair following Er:YAG treatment and its bactericidal effect can also be related to reactive oxygen species (ROS) generation in irradiated tissue. BACKGROUND DATA: The Er:YAG laser with a wavelength of 2,940 nm (corresponding to the vibrational OH stretch frequency of water) is of great value in dental medicine, owing to its dual ability to ablate soft and hard tissues with minimal damage to surrounding structures. The relatively rapid postoperative healing time seen after ablation of the gingiva is attributed to the very narrow zone of thermal disruption. METHODS: Water was irradiated with an Er:YAG laser at an energy of 100-130 mJ/cm(2) and 10-30-Hz pulse repetition rate The concentration of OH radicals produced following irradiation was assessed by spin trapping coupled with electron paramagnetic resonance (EPR) spectroscopy. RESULTS: We found that the Er-YAG laser dissociates water and generates OH radicals. The concentration of radicals produced was strongly dependent on the pulse repetition rate and energy density per laser pulse. CONCLUSIONS: The dissociation of water needed to generate OH radicals is possibly due to intermolecular vibrational (V-V) energy transfer in water, competing with vibrational relaxation, thus leading to water dissociation. High amounts of oxygen radicals (e.g., hydroxyl groups) have a sterilization effect, whereas low concentrations of ROS stimulate fibroblasts, causing collagen and extracellular matrix formation. ROS formation may explain the wound healing effect of the Er-YAG laser in dentistry.

Laser Therapy↗

PubMed2003

Heteromeric MAPPIT: a novel strategy to study modification-dependent protein-protein interactions in mammalian cells.

We recently reported a two-hybrid trap for detecting protein-protein interactions in intact mammalian cells (MAPPIT). The bait protein was fused to a STAT recruitment-deficient, homodimeric cytokine receptor and the prey protein to functional STAT recruitment sites. In such a configuration, STAT-dependent responses can be used to monitor a given bait-prey interaction. Using this system, we were able to demonstrate both modification-independent and tyrosine phosphorylation- dependent interactions. Protein modification in this approach is, however, strictly dependent on the receptor-associated JAK tyrosine kinases. We have now extended this concept by using extracellular domains of the heteromeric granulocyte/macrophage colony-stimulating factor receptor (GM-CSFR). Herein, the bait was fused to the (beta)c chain and its modifying enzyme to the GM-CSFRalpha chain (or vice versa). We demonstrate several serine phosphorylation-dependent interactions in the TGFbeta/Smad pathway using the catalytic domains of the ALK4 or ALK6 serine/threonine kinase receptors. In all cases tested, STAT-dependent signaling was completely abolished when mutant baits were used wherein critical serine residues were replaced by alanines. This approach operates both in transient and stable expression systems and may not be limited to serine phosphorylation but has the potential for studying various different types of protein modification-dependent interactions in intact cells.

Animals↗

PubMed2000

Fluvastatin, a new inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, resists hydroxyl radical generation in the rat myocardium.

The aim of this study was to determine whether fluvastatin, an inhibitor of low-density lipoprotein (LDL) oxidation, can resist Cu (II)-induced hydroxyl radical generation (*OH) in the extracellular fluid of rat myocardium. Rats were anaesthetized and sodium salicylate in Ringer's solution (0.5 nmol microL(-1) min(-1)) was infused through a microdialysis probe to detect the generation of *OH as reflected by the non-enzymatic formation of 2,3-dihydroxybenzoic acid (2,3-DHBA) in the myocardium. When Cu (II) (0, 10, 25 or 50 microM) was administered to 1 mM tyramine-pretreated rats, a marked elevation in the levels of 2,3-DHBA was found, indicating a positive linear correlation between Cu (II) and the increase in *OH formation trapped as 2,3-DHBA in the dialysate (r2 = 0.977). In the presence of fluvastatin (100 microM), a marked decrease in the levels of 2,3-DHBA was found. Corresponding experiments performed with iron (II) (0, 10, 25 or 50 microM), showed a marked elevation in the levels of 2,3-DHBA, indicating a positive linear correlation between iron (II) and the increase in *OH formation trapped as 2,3-DHBA in the dialysate (r2 = 0.986). However, in the presence of fluvastatin (100 microM) a small decrease in the level of 2,3-DHBA was found. The results show that iron (II) against LDL oxidation may be insensitive compared with Cu (II). Cu (II)-induced *OH formation may be reduced by inhibiting LDL with fluvastatin.

Animals↗

PubMed2003

Reaction of cysteine and glutathione (GSH) at the freshly fractured quartz surface: a possible role in silica-related diseases?

The reactivity of quartz dusts towards glutathione (GSH) and cysteine (Cys) has been investigated. Cys and GSH react, without being adsorbed (UV-Vis spectroscopy), with commercial quartz dusts in an exposed surface-dependent way, but not with amorphous silica. GSH and Cys have been contacted with freshly ground quartz (agate jar QZg-a and steel jar QZg-s) and quartz heated in air at 500 degrees C (QZs-500) and with a dust generated from a purified quartz (99.9999%) to detect the nature of the reacting surface sites. With both GSH and Cys, the highest reactivity was found on the particles ground in a steel jar, while pure quartz was fully inactive. Detection of the radical GS* (spin trapping) suggests a radical mechanism of oxidation to disulphide onto surface-bound iron traces, more abundant on QZg-s and absent on the pure quartz. Oxidation of thiol groups occurs at surface sites different from those involved in the homolytic rupture of a C-H bond. Both reactions are more pronounced on freshly ground samples, but the C-H rupture takes place at silicon-based surface radicals and Fe2+ centers, while oxidation of GSH and Cys requires Fe3+ centers. As all commercial quartz dusts contain surface iron as an impurity, depletion of extracellular or intracellular GSH may contribute to the oxidative damage caused by particle-derived and cell-derived reactive oxygen species.

Adsorption↗

PubMed1995

Detection of free radical activity during transient global ischemia and recirculation: effects of intraischemic brain temperature modulation.

To obtain direct evidence of oxygen radical activity in the course of cerebral ischemia under different intraischemic temperatures, we used a method based on the chemical trapping of hydroxyl radical in the form of the stable adducts 2,3- and 2,5-dihydroxybenzoic acid (DHBA) following salicylate administration. Wistar rats were subjected to 20 min of global forebrain ischemia by two-vessel occlusion plus systemic hypotension (50 mm Hg). Intraischemic striatal temperature was maintained as normothermic (37 degrees C), hypothermic (30 degrees C), or hyperthermic (39 degrees C) but was held at 37 degrees C before and following ischemia. Salicylate was administered either systemically (200 mg/kg, i.p.) or by continuous infusion (5 mM) through a microdialysis probe implanted in the striatum. Striatal extracellular fluid was sampled at regular intervals before, during, and after ischemia, and levels of 2,3- and 2,5-DHBA were assayed by HPLC with electrochemical detection. Following systemic administration of salicylate, stable baseline levels of 2,3- and 2,5-DHBA were observed before ischemia. During 20 min of normothermic ischemia, a 50% reduction in mean levels of both DHBAs was documented, suggesting a baseline level of hydroxyl radical that was diminished during ischemia, presumably owing to oxygen restriction to tissue at that time. During recirculation, 2,3- and 2,5-DHBA levels increased by 2.5- and 2.8-fold, respectively. Levels of 2,3-DHBA remained elevated during 1 h of reperfusion, whereas the increase in 2,5-DHBA persisted for 2 h. The increases in 2,3- and 2,5-DHBA levels observed following hyperthermic ischemia were significantly higher (3.8- and fivefold, respectively). In contrast, no significant changes in DHBA levels were observed following hypothermic ischemia.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Pseudomonas aeruginosa adhesion to normal and injured respiratory mucosa.

Human nasal polyps in outgrowth culture were used to study the adhesion of Pseudomonas aeruginosa to respiratory cells. By transmission electron microscopy, bacteria associated with ciliated cells were identified trapped at the extremities of cilia, usually as aggregates of several bacterial cells. They were never seen at the interciliary spaces or attached along cilia. Bacteria were also seen to adhere avidly to migrating cells of the periphery of the outgrowth culture. Using a model of repair of wounded respiratory epithelial cells in culture, we observed that the adhesion of P. aeruginosa to migrating cells of the edges of the repairing wounds was significantly higher than the adhesion to non-migrating cells and that adherent bacteria were surrounded by a fibronectin-containing fibrillar material. The secretion of extracellular matrix components is involved in the process of epithelium repair following injury. To investigate the molecular basis of P. aeruginosa adhesion to migrating cells, bacteria were treated with a fibronectin solution before their incubation with the respiratory cells. P. aeruginosa treatment by fibronectin significantly increased their adhesion to migrating cells. Accordingly, we hypothesize that during cell migration, fibronectin secreted by epithelial cells may favour P. aeruginosa adhesion by establishing a bridge between the bacteria and the epithelial cell receptors. Such a mechanism may represent a critical step for P. aeruginosa infection of healing injured epithelium.

Bacterial Adhesion↗

PubMed1997

Host-derived intracellular immunization against mouse hepatitis virus infection.

Mouse hepatitis virus (MHV) utilizes the murine biliary glycoprotein receptor (BgpA) for entry into susceptible cells. The Bgp1 gene was transfected into a murine DBT cell clone that expressed little if any BgpA receptor and resisted wild-type MHV infection. Clones which expressed low levels of receptor were efficient hosts for MHV-A59 replication. Clones which expressed 20- to 69-fold more BgpA receptor than controls were also susceptible to MHV-A59 infection, yet few infectious progeny virions were released. Pretreatment of clones with monoclonal antibody CC1, which binds to the N-terminus of BgpA, blocked MHV-A59 replication in DBT clones that expressed wild-type levels of receptor protein. In the overexpressing cell clones, however, CC1 pretreatment reversed the BgpA overexpression blockade and increased virus titers by 3-4 logs. BgpA overexpression inhibited the formation of infectious extracellular and intracellular virions, but levels of virus transcription were equivalent to those of controls. Ultrastructural studies revealed normal cell cytopathology in both the MHV-A59-infected controls and the overexpressing cell clones. Although equivalent numbers of virions were released compared with the control, peplomer spike glycoproteins were rarely evident in virions derived from the BgpA-overexpressing cell clones. Consonant with these findings, purified virions from BgpA-overexpressing cell clones demonstrated a 70% reduction in the amount of S glycoprotein, but not of N or M proteins. These data suggest that BgpA receptor overexpression establishes an intracellular trap which blocks MHV replication during the maturation and release of infectious progeny virions.

Animals↗