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  • 11
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Allergy 55 (2000), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background: The study aimed to investigate whether CD69 expression on granulocytes is subject to specific regulation by inflammatory mediators, and, if so, to identify these factors in relation to eosinophil activity markers such as the EG2 epitope and ECP release. Methods: Peripheral blood leukocytes from healthy donors were used. The surface and intracellular distribution of CD69 was investigated with a whole-blood cell-membrane permeabilization technique, the FOG method, and flow cytometry. In vitro stimulation was performed with GM-CSF, IL-5, IL-5 plus eotaxin, LPS, and fMLP. Results: A preformed intracellular pool of CD69 was demonstrated in both eosinophils and neutrophils, but not in monocytes. Almost no resting eosinophils, neutrophils. or monocytes expressed CD69 on the cell surface. However, in vitro stimulation with selected stimuli increased the proportion of CD69-positive eosinophils to various extents, with GM-CSF being the most and fMLP the least efficient stimulus. The neutrophils did not respond under these conditions. Increased expression of the EG2 epitope and initiation of degranulation preceded CD69 upregulation. Conclusions: Eosinophils and neutrophils from healthy donors have a preformed intracellular pool of CD69, which is mobilized on the cell surface on eosinophils, but not on neutrophils, to various extents by selected stimuli. Monocytes, however, do not have a preformed intracellular pool of CD69. Our data indicate that a kinetic order exists among the EG2 expression, the degranulation process, and CD69 upregulation. Due to a quantitative, rather then a qualitative, upregulation of CD69 by stimuli associated with both allergic and bacterial inflammation, CD69 may be a potential activity marker of clinical value.
    Type of Medium: Electronic Resource
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  • 12
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Allergy 48 (1993), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The FOG method is a new cell membrane permeabilization technique which makes it possible to identify the eosinophils in unseparated peripheral blood and allows analyses of both surface and intracellular antigens by flow cytometry. This technique has been adapted to analyse eosinophil activity in peripheral blood from asthmatic patients by measuring the expression of the EG2-epitope on intracellular ECP.
    Type of Medium: Electronic Resource
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  • 13
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 14
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Allergy 54 (1999), S. 0 
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We here report a girl, now 3 years old, who has suffered from severe food allergy since her first year of life. She was strongly allergic to cow's milk, and had high levels of IgE antibody (AB) to casein (210 kU/l), β-lactoglobulin (43 kU/l), and α-lactalbumin (23 kU/l) at 12 months of age. In addition, at the same age, she showed positive (2–4+) skin prick reactions to both unboiled and boiled formulas (Profylac, Nutramigen, and Neocate), besides being positive in RAST to Nutramigen (0.6 kU/l). During the first 3 years, IgE Ab levels against casein and Nutramigen increased to 310 and 1.6 kU/l, respectively. Furthermore, at 3 years of age, she had positive RAST to 14 of 15 tested food allergens, being negative only to codfish. Assessment of eosinophil-related markers revealed high total eosinophil count, increased eosinophil activity, and a low ratio of interferon (IFN)-γ:IL-5, indicating enhanced IL-5 production. The food allergy was correlated to poor weight gain and increasing problems with atopic allergy in the airways.
    Type of Medium: Electronic Resource
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  • 15
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    Archives of Biochemistry and Biophysics 247 (1986), S. 20-27 
    ISSN: 0003-9861
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 16
    Electronic Resource
    Electronic Resource
    Amsterdam : Elsevier
    FEBS Letters 174 (1984), S. 289-293 
    ISSN: 0014-5793
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology , Chemistry and Pharmacology , Physics
    Type of Medium: Electronic Resource
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  • 17
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 28 (1988), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Complement receptors, CR1 and CR3, on neutrophils increase their cellular spontaneously at 37°Cor after mechanical stress during the cell preparation. We have established a cell preparation procedure and a cytofluorometric immunoassay method to evaluate the receptor expression in vivo in this study. The expression of CRI and CR3 was studied after haemolysis in NH4C1 at different temperatures and incubation intervals. It was shown that cell preparation and receptor analysis must be performed at 15°C or lower to avoid up-regulation of the receptor structures. Two minutes' Incubation at 20°C was sufficient to modulate the cells in this regard. Granulocytes from healthy blood donors were analysed and the mean fluorescence intensity (MF1), which reflects the number of receptors on (he ceil surface, showed a normal distribution for the CRI (n=158) and CR3 (n=76) expression in the healthy population. The MFI of the two receptors showed a correlation (r=0.71). Granulocytes from all donors could be modulated at 37°C to a similar degree for both receptors (r=0.76), despite the fact that they are supposed to be mobilized from different intracellular pools. A group of patients with localized inflammatory processes, such as sinusitis, differed from the healthy controls. In this group, the CR1 expression was not normally distributed, but IS out of 26 patients (58%) had granulocytes with a CR1 expression exceeding the mean±SD of the normal population.
    Type of Medium: Electronic Resource
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  • 18
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The objective of the present study was to investigate the potential of the long-acting p-aonist salmeterol as an inhibitor of various components of IgE-mediated inflammation in man. For this purpose, we measured gross skin reactivity (diameters of wheal and flare reaction [WFR] and late cutaneous reaction [LCR]) as well as inflammatory cells, mediators, and protein in cutaneous suction blister chambers in eight subjects with allergic rhinitis. Blisters were induced, two on each forearm, by gentle suction and heating, and were unroofed 12 h later, after which plastic chambers were placed over the denuded area. The chambers were challenged for 2 h with antihuman IgE (titer 1:10) in the presence and absence of salmeterol or terbutaline. Normal goat IgG served as negative control. Chamber fluids were removed hourly for the first 4 h, and this was followed by a 4-h incubation before final collection. Salmeterol (10-6M) and terbutaline (10-5 M) injected intradermally 30 min before, as well as together with anti-IgE (titer 1:100), inhibited the WFRs by up to 30%. The effect of salmeterol on the ensuing LCR (75% inhibition at 24 h) tended to be more pronounced than the corresponding inhibition by terbutaline. Both salmeterol and terbutaline very effectively inhibited the anti-IgE-induced extravasation of α2-macroglobulin into skin chambers, with a significantly more sustained effect by salmeterol. Interestingly, only terbutaline reduced the histamine release evoked by anti-IgE. With the present experimental design, where both drugs were washed out from the chambers after 2 h, neither drug inhibited recruitment of leukocytes (including eosinophils). Taken together, salmeterol had a more sustained inhibitory effect than terbutaline on indices of IgE-mediated edema formation (late induration and plasma protein extravasation). On the other hand, under the present experimental conditions, salmeterol failed to reduce the histamine release (in contrast to terbutaline), and neither salmeterol nor terbutaline affected the recruitment of leukocytes.
    Type of Medium: Electronic Resource
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  • 19
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We have previously found that CD9, CD11b, and intracellular ECP (EG2) may be used as activity markers for eosinophils in vitro. The main object of the present study was to determine whether these markers can reflect eosinophil activation in vivo in relation to allergen exposure. For this purpose, six patients with a history of allergic rhinitis and occasional asthma symptoms during the pollen season participated. Blood donors served as controls. Peripheral blood eosinophils were analyzed according to the FOG method and flow cytometry, before and during one birch pollen season with high pollen load (HPL) and one with low pollen load (LPL). The CD9 expression on peripheral eosinophils from the patients was significantly increased both before (P〈0.05) and during (P〈0.01) HPL season, and CD11b expression solely during HPL season (P = 0.01) as compared to controls. The intracellular expression of the EG2 epitope was increased before (P〈0.01) and during (P〈0.05) HPL season, and increased significantly (P〈0.05) during season as compared to before. No changes were observed before and during LPL season. The proportion of eosinophils was increased both before (P〈0.05) and during (P〈0.001) the HPL season as compared to controls. The markers CD9, EG2, and, to a lesser extent, CD11b seem to detect activated eosinophils in the circulation, whereas EG2 may also reflect increased antigen exposure during season.
    Type of Medium: Electronic Resource
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  • 20
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: To investigate allergic rhinitis induced by two experimental provocation models, we compared local inflammation with markers of eosinophil activity in peripheral blood. Patients with strictly seasonal allergic rhinitis were selected and investigated outside the pollen season. An acute challenge with increasing doses of allergen every 15 min until symptoms occurred was performed in nine patients. Nasal lavage and blood samples were taken before and 4 and 24 h after challenge. After a 6-week washout period, 10 patients were submitted to 7 days of repeated low allergen exposures. One small dose (≅/100 of the acute dose) was given each day. Blood and lavage samples were taken prior to and after the period. As control four patients were challenged with diluent only. The acute challenge resulted in sneezing and nasal discharge and blockage and was accompanied by a rise in histamine and eosinophil cationic protein (ECP) in lavage fluid after 4 h and continuing after 24 h, when there also was a rise in the number of eosinophils and ECP in peripheral blood. The repeated low-dose exposures caused very few symptoms but produced increased ECP in the lavage Auid and a trend toward increased histamine concentration. There were no changes in ECP, intracellular EG2 binding, or number of eosinophils in the blood. No changes were seen in the control group. Our findings show that changes in eosinophil mediator release in nasal lavage can be seen after very low, but repeated, allergen exposures despite no, or minimal, clinical symptoms.
    Type of Medium: Electronic Resource
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