Immunocytochemical Determination of the Role of Alveolar Macrophages in Endotoxin Processing in vitro and in vivo

1991 ◽  
Vol 96 (2) ◽  
pp. 149-155
Author(s):  
George E. Keller, III ◽  
Richard D. Dey ◽  
Robert Burrell
Keyword(s):  
Alveolar Macrophages

A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

Factors affecting in vitro and in vivo antioxidant effects. Experimental conditions and nature of oxidants determine antioxidant efficacy

2021 ◽  
pp. 1-9
Author(s):  
Etsuo Niki
Keyword(s):  
Biological Molecules ◽  
Experimental Conditions ◽  
Factors Affecting ◽  
Beneficial Effects ◽  
Multiple Oxidants ◽  
Antioxidant Effects

Reactive oxygen and nitrogen species have been implicated in the onset and progression of various diseases and the role of antioxidants in the maintenance of health and prevention of diseases has received much attention. The action and effect of antioxidants have been studied extensively under different reaction conditions in multiple media. The antioxidant effects are determined by many factors. This review aims to discuss several important issues that should be considered for determination of experimental conditions and interpretation of experimental results in order to understand the beneficial effects and limit of antioxidants against detrimental oxidation of biological molecules. Emphasis was laid on cell culture experiments and effects of diversity of multiple oxidants on antioxidant efficacy.

Multinucleated rat alveolar macrophages express functional receptors for calcitonin

1991 ◽  
Vol 261 (6) ◽  
pp. F1026-F1032 ◽  
Author(s):  
A. Vignery ◽  
M. J. Raymond ◽  
H. Y. Qian ◽  
F. Wang ◽  
S. A. Rosenzweig
Keyword(s):  
Plasma Membrane ◽  
Alveolar Macrophages ◽  
Giant Cells ◽  
Mononuclear Phagocytes ◽  
Inflammatory Reactions ◽  
Calcitonin Gene ◽  
Novel Model

The fusion of mononuclear phagocytes occurs spontaneously in vivo and leads to the differentiation of either multinucleated giant cells or osteoclasts in chronic inflammatory sites or in bone, respectively. Although osteoclasts are responsible for resorbing bone, the functional role of giant cells in chronic inflammatory reactions and tumors remains poorly understood. We recently reported that the plasma membrane of multinucleated macrophages is, like that of osteoclasts, enriched in Na-K-adenosinetriphosphatases (ATPases). We also observed that the localization of their Na-K-ATPases is restricted to the nonadherent domain of the plasma membrane of cells both in vivo and in vitro, thus imposing a functional polarity on their organization. By following this observation, we wished to investigate whether these cells also expressed, like osteoclasts, functional receptors for calcitonin (CT). To this end, alveolar macrophages were fused in vitro, and both their structural and functional association with CT was analyzed and compared with those of mononucleated peritoneal and alveolar macrophages. Evidence is presented that multinucleated alveolar macrophages express a high copy number of functional receptors for CT. Our results also indicate that alveolar macrophages, much like peritoneal, express functional receptors for calcitonin gene-related peptide. It is suggested that multinucleated rat alveolar macrophages offer a novel model system to study CT receptors and that calcitonin may control local immune reactions where giant cells differentiate.

Fructose Phosphorylation and Dephosphorylation by the Intestinal Mucosa of the Rat During Fructose Absorption

1957 ◽  
Vol 189 (2) ◽  
pp. 301-306 ◽  
Author(s):  
Nicholas M. Papadopoulos ◽  
Joseph H. Roe
Keyword(s):  
Intestinal Mucosa ◽  
Phosphate Esters ◽  
Reduction Techniques ◽  
Mucosal Cells ◽  
Normal Metabolism ◽  
Fasted Rats

The role of phosphorylation in the absorption of fructose from the intestinal tract of the fasted rat by in vitro and in vivo techniques was studied. The authors' method for the determination of fructose phosphate esters was used and these esters were identified by paper chromatography and copper reduction techniques. Buffered homogenate of intestinal mucosa of a fasted rat, mixed with ATP, MgCl2, KF and fructose, when incubated at 30°, showed the formation of fructose-6-phosphate and fructose-1, 6-diphosphate at a rate that corresponded to the decrease in free fructose. The same homogenate, mixed with fructose-1, 6-diphosphate and incubated at 37°, showed the formation of fructose-6-phosphate and free fructose at a rate that corresponded to the decrease in the concentration of the diphosphate ester. Following intraduodenal injection of fructose solution into anesthetized fasted rats, homogenates of the intestinal mucosa showed the presence of fructose-6-phosphate and fructose-1, 6-diphosphate in average concentrations 14 and 5 times, respectively, those found in control muocsa, also concentrations of free fructose in the blood of the portal vein up to 24.6 mg % were observed. The large increase in fructose phosphate esters in the intestinal mucosa, observed after fructose administration, suggests that phosphorylation of sugars in absorption serves a more extensive function than to initiate glycolysis for the normal metabolism of the mucosal cells. The data obtained suggest that phosphorylation and dephosphorylation are functional steps in the absorption of fructose from the alimentary tract of the rat.

THE ROLE OF A 3-0 SULFO GROUP IN THE GLUCOSAMINE RESIDUE OF A SYNTHETIC OLIGOSACCHARIDE FOR THE DETERMINATION OF ANTITHROMBOTIC ACTIONS

1987 ◽  
Author(s):  
J M Walenga ◽  
J Fareed ◽  
M Petitou ◽  
J C Lormeau ◽  
M Samama ◽  
...  
Keyword(s):  
Sulfo Group ◽  
Factor Xa ◽  
Antithrombotic Effect ◽  
Antithrombotic Activity ◽  
Thrombosis Model ◽  
Factor Xa Inhibition

We have previously reported on the antithromboticaction of a chemically synthesized heparin pentasaccharide which exhibits high affinity to anti thrombinIII and sole anti-factor Xa activity. In order to investigate the relative importance of the 3-0 sulfo group of this pentasaccharide, we evaluated the in vitro and in vivo antithrombotic activity of a synthetic pentasccharide devoid of the sulfo group at the third position of the glucosamine residue. In amidolytic and clot-based assays the 3-0 de- sulfated pentasaccharide (3-0-DP) failed to exhibit any antifactor Xa actions at concentrations <100 ug/ml in humanor rabbit plasmas, whereas pentasaccharide showed strong factor Xa inhibition at 1.0 ug/ml IK-=3.2x10 M)and at 10.0 ug/ml in rabbit plasma (K.=9.0×10™7 M). Using a rabbit stasis thrombosis model in which thrombosis was induce by human serum or an activated pro-thrombin complex concentrate, 3-0-DP failed to produce any antithrombotic action in acute intravenous regimens at dosages up to 200 ug/kg. In these two models, pentasaccharide produced >80% inhibition of induced thrombosis. These studies demonstrate the critical importance of the 3-0 sulfo group in this heparin pentasaccharide for the determination of antithrombotic activity, and that in this type of oligosaccharide, anti-factor Xa activity is responsible for producing the antithrombotic effect.

scholarly journals Trehalose Alleviates Crystalline Silica-Induced Pulmonary Fibrosis via Activation of the TFEB-Mediated Autophagy-Lysosomal System in Alveolar Macrophages

Cells ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 122 ◽  
Author(s):  
Xiu He ◽  
Shi Chen ◽  
Chao Li ◽  
Jiaqi Ban ◽  
Yungeng Wei ◽  
...  
Keyword(s):  
Pulmonary Fibrosis ◽  
Alveolar Macrophages ◽  
Nuclear Translocation ◽  
Protective Role ◽  
Crystalline Silica ◽  
Autophagic Flux ◽  
Lysosomal System

Silicosis is an occupational lung disease characterized by persistent inflammation and irreversible fibrosis. Crystalline silica (CS) particles are mainly phagocytized by alveolar macrophages (AMs), which trigger apoptosis, inflammation, and pulmonary fibrosis. Previously, we found that autophagy-lysosomal system dysfunction in AMs was involved in CS-induced inflammation and fibrosis. Induction of autophagy and lysosomal biogenesis by transcription factor EB (TFEB) nuclear translocation can rescue fibrotic diseases. However, the role of TFEB in silicosis is unknown. In this study, we found that CS induced TFEB nuclear localization and increased TFEB expression in macrophages both in vivo and in vitro. However, TFEB overexpression or treatment with the TFEB activator trehalose (Tre) alleviated lysosomal dysfunction and enhanced autophagic flux. It also reduced apoptosis, inflammatory cytokine levels, and fibrosis. Both pharmacologically inhibition of autophagy and TFEB knockdown in macrophages significantly abolished the antiapoptotic and anti-inflammatory effects elicited by either TFEB overexpression or Tre treatment. In conclusion, these results uncover a protective role of TFEB-mediated autophagy in silicosis. Our study suggests that restoration of autophagy-lysosomal function by Tre-induced TFEB activation may be a novel strategy for the treatment of silicosis.

In vivo and in vitro uptake of surfactant lipids by alveolar type II cells and macrophages

2002 ◽  
Vol 283 (3) ◽  
pp. L648-L654 ◽  
Author(s):  
D. L. H. Poelma ◽  
L. J. I. Zimmermann ◽  
H. H. Scholten ◽  
B. Lachmann ◽  
J. F. van Iwaarden
Keyword(s):  
Alveolar Macrophages ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Alveolar Cells ◽  
Alveolar Type Ii Cells ◽  
Small Subpopulation

The uptake of fluorescent-labeled liposomes (with a surfactant-like composition) by alveolar macrophages and alveolar type II cells was studied using flow cytometry, in vivo by instillation of the labeled liposomes in the trachea of ventilated rats followed by isolation of the alveolar cells and determination of the cell-associated fluorescence, and in vitro by incubation of isolated alveolar cells with the fluorescent liposomes. The results show that the uptake of liposomes by the alveolar cells is time and concentration dependent. In vivo alveolar macrophages internalize more than three times as many liposomes as alveolar type II cells, whereas in vitro, the amount of internalized liposomes by these cells is approximately the same. In vitro, practically all the cells (70–75%) internalize liposomes, whereas in vivo only 30% of the alveolar type II cells ingest liposomes vs. 70% of the alveolar macrophages. These results indicate that in vivo, only a small subpopulation of alveolar type II cells is able to internalize surfactant liposomes.

scholarly journals IFNγ and TNFα mediate CCL22/MDC production in alveolar macrophages after hemorrhage and resuscitation

2020 ◽  
Vol 318 (5) ◽  
pp. L864-L872
Author(s):  
Nadine Beckmann ◽  
Jeffrey M. Sutton ◽  
Richard S. Hoehn ◽  
Peter L. Jernigan ◽  
Lou Ann Friend ◽  
...  
Keyword(s):  
Alveolar Macrophages ◽  
Inflammatory Cell ◽  
Pulmonary Inflammation ◽  
Major Complication ◽  
Signaling Mechanism ◽  
Precise Role ◽  
Autocrine Mechanism

Acute lung injury is a major complication of hemorrhagic shock and the required resuscitation with large volumes of crystalloid fluids and blood products. We previously identified a role of macrophage-derived chemokine (CCL22/MDC) pulmonary inflammation following hemorrhage and resuscitation. However, further details regarding the induction of CCL22/MDC and its precise role in pulmonary inflammation after trauma remain unknown. In the current study we used in vitro experiments with a murine alveolar macrophage cell line, as well as an in vivo mouse model of hemorrhage and resuscitation, to identify key regulators in CCL22/MDC production. We show that trauma induces expression of IFNγ, which leads to production of CCL22/MDC through a signaling mechanism involving p38 MAPK, NF-κB, JAK, and STAT-1. IFNγ also activates TNFα production by alveolar macrophages, potentiating CCL22/MDC production via an autocrine mechanism. Neutralization of IFNγ or TNFα with specific antibodies reduced histological signs of pulmonary injury after hemorrhage and reduced inflammatory cell infiltration into the lungs.

scholarly journals The Role of Antibodies Against the Crude Capsular Extract in the Immune Response of Porcine Alveolar Macrophages to In Vitro Infection of Various Serovars of Glaesserella (Haemophilus) parasuis

2021 ◽  
Vol 12 ◽  
Author(s):  
Katarína Matiašková ◽  
Lenka Kavanová ◽  
Pavel Kulich ◽  
Jan Gebauer ◽  
Kateřina Nedbalcová ◽  
...  
Keyword(s):  
Immune Response ◽  
Alveolar Macrophages ◽  
Specific Binding ◽  
Outer Membrane Proteins ◽  
Disease Outbreaks ◽  
Haemophilus Parasuis ◽  
Associated Proteins

In Glässer’s disease outbreaks, Glaesserella (Haemophilus) parasuis has to overcome the non-specific immune system in the lower respiratory tract, the alveolar macrophages. Here we showed that porcine alveolar macrophages (PAMs) were able to recognize and phagocyte G. parasuis with strain-to-strain variability despite the presence of the capsule in virulent (serovar 1, 5, 12) as well in avirulent strains (serovar 6 and 9). The capsule, outer membrane proteins, virulence-associated autotransporters, cytolethal distending toxins and many other proteins have been identified as virulence factors of this bacterium. Therefore, we immunized pigs with the crude capsular extract (cCE) from the virulent G. parasuis CAPM 6475 strain (serovar 5) and evaluated the role of the anti-cCE/post-vaccinal IgG in the immune response of PAMs to in vitro infection with various G. parasuis strains. We demonstrated the specific binding of the antibodies to the cCE by Western-blotting assay and immunoprecipitation as well as the specific binding to the strain CAPM 6475 in transmission electron microscopy. In the cCE, we identified several virulence-associated proteins that were immunoreactive with IgG isolated from sera of immunized pigs. Opsonization of G. parasuis strains by post-vaccinal IgG led to enhanced phagocytosis of G. parasuis by PAMs at the first two hours of infection. Moreover, opsonization increased the oxidative burst and expression/production of both pro- and anti-inflammatory cytokines. The neutralizing effects of these antibodies on the antioxidant mechanisms of G. parasuis may lead to attenuation of its virulence and pathogenicity in vivo. Together with opsonization of bacteria by these antibodies, the host may eliminate G. parasuis in the infection site more efficiently. Based on these results, the crude capsular extract is a vaccine candidate with immunogenic properties.

scholarly journals Changes in erythrocyte deformability during day and possible role of melatonin

2018 ◽  
Vol 52 (1) ◽  
pp. 17-20 ◽  
Author(s):  
Rastislav Vazan ◽  
Katarina Plauterova ◽  
Gabriela Porubska ◽  
Jana Radosinska
Keyword(s):  
Erythrocyte Deformability ◽  
Diurnal Changes ◽  
Sample Collection ◽  
Pass Through ◽  
In Vivo Effects ◽  
Vitro Effect

Abstract Objectives. The deformability of erythrocytes is their ability to change shape in order to pass through the capillaries. Th is is necessary for quality of microcirculation and sufficient delivery of oxygen to the tissues. Th e aim of our study was to investigate the possible spontaneous changes in the erythrocyte deformability during day and evaluation of the possible direct effects of melatonin (hormone involved in regulation of biorhythms) on the erythrocyte deformability. Methods. Samples of capillary blood were taken from 12 healthy volunteers in the morning (8:00) and early in the evening (16:30). Determination of erythrocyte deformability was done based on the measurement of their filtrability. It was measured immediately aft er the sample collection and 2-hour lasting incubation without or with melatonin (2000 μmol/L). Results. Erythrocyte deformability was significantly lower in the morning (filtrability index: 0.68±0.01 morning vs. 0.71±0.01 early evening, p<0.05). Th e incubation of blood samples with melatonin did not have impact on deformability. Conclusions. We suggest the presence of diurnal changes in erythrocyte deformability with worse values in the morning that may contribute to higher risk of ischemic attacks in the morning hours. Direct in vitro effect of melatonin on deformability was not observed, but possible in vivo effects cannot be excluded.

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