Determination of the four natural Ra isotopes in thermal waters by gamma-ray spectrometry.

Our method for the simultaneous determination of the four natural Ra isotopes ((226)Ra, (228)Ra, (224)Ra and (223)Ra) in thermal waters involves a separation of Ra on a selective filter (3M EMPORE Radium Rad disk), and a single counting using a broad-energy HPGe detector (BE Ge manufactured by CANBERRA). The calculation of (223)Ra and (228)Ra activities requires interference and cascade summing corrections. The (226)Ra activities in CO(2)-rich thermal waters of the Lodève Basin (South of France) range from 530 to 2240mBq/l. The low ((228)Ra/(226)Ra) activity ratios (0.19-0.29) suggest that Ra is mostly derived from the aquifer carbonates. The short-lived (224)Ra and (223)Ra are probably added to the water through recoil or desorption processes from Th-enriched coatings on the fracture walls.

A simplified method to estimate kinetic and thermodynamic parameters on the solid-liquid separation of pollutants.

The aim of the present study was to propose a simplified experimental-theoretical method for estimating the kinetic and thermodynamic parameters for the solid-liquid separation of pollutants by using kinetic studies with batch reactors, i.e., the removed quantity of dissolved ion as a function of time at different initial concentration. This method was applied to the removal of uranyl ion (UO(2+)(2)) from aqueous solutions onto synthetic manganese oxide (birnessite). The pseudo-second-order kinetics and one-site saturation models were proposed to fit the experimental and calculated data, the fitting parameters being estimated by nonlinear regression, using the least-squares method. For initial concentration range 0.2-11.8 microM, the results showed that the uranyl removal process in dispersed batch reactors can be efficiently modeled by the proposed models. Then, several kinetic and thermodynamic parameters were calculated, such as maximal removed quantity of uranyl, q(r,max), half-removal time, t(1/2), initial rate of uranyl-ion removal, v(0), initial uranyl-removal coefficient, K, maximal rate of uranyl removal, v(0,max), mass transfer coefficient, D(transfer), equilibrium Langmuir constant, K(L), and constant separation factor, K(s). These parameters make it possible to demonstrate that the removal of U onto birnessite is favorable, and that the maximum surface coverage of the uranyl ions represents about 3% of vacant sites in the Mn layer.

Electrical Double-Layer Structure at the Rutile-Water Interface as Observed in Situ with Small-Period X-Ray Standing Waves.

X-Ray standing wave (XSW) measurements were made of Rb and Sr adsorbed from aqueous solutions at the rutile (110)-water interface. These experiments were performed to address the extent to which direct measurements of electrical double-layer structure are possible. The experimental results show that the Bragg XSW technique, using small-period standing waves generated by Bragg diffraction from the substrate, can precisely measure ion locations within the condensed layer and the in situ partitioning of ions between the condensed and diffuse layers. Differences in condensed layer ion positions were observed for Sr ions (measured in situ) as compared with Rb ions (in situ) and also for Sr ions (ex situ). An additional constraint on the ex situ Sr site geometry was provided by polarization-dependent surface EXAFS measurements. Such measurements can provide important constraints for the development and verification of electrical double-layer theory especially as applied to ion adsorption at the solid-water interface. Copyright 2000 Academic Press.

Regulation of cytokine expression and leukotriene formation in human basophils by growth factors, chemokines and chemotactic agonists.

Basophils stimulated with IL-3 plus C5a selectively express IL-4 and IL-13 and continuously produce leukotrienes (LT) for hours. C5a combined with IL-5 or granulocyte-macrophage colony-stimulated factor was, however, much less effective in promoting cytokine expression and a late continuous phase of LTC4 production, possibly due to lower expression levels of their receptor alpha chains. Basophils also express several chemoattractant receptors, including high levels of C5a receptors, macrophage chemotactic protein (MCP) receptors (CCR2) and eotaxin receptors (CCR3), intermediate levels of CXCR1, CXCR2 and platelet-activating factor receptors, and lower levels of N-formyl-Met-Leu-Phe (fMLP) receptors. However, among the corresponding agonists, only C5a, fMLP and much more weakly MCP1, were found to induce cytokine expression and continuous LTC4 release, and only when combined with IL-3. CCR3, which is highly expressed on basophils and has been shown to mediate strong migratory but weak release responses, does not regulate cytokine expression. The weakly expressed fMLP receptor is an efficient activator of several cell functions including LTC4 formation, while CXCR2 hardly affects basophil function despite considerable expression. Thus, chemoattractant-receptors mediate different cellular responses unrelated to their expression levels.

Regulation of cytokine expression by human blood basophils.

IL-4 and IL-13 are key immunoregulatory cytokines because of their ability to induce and amplify Th2-type immune responses and by promoting IgE formation. The basophil is a particularly prominent source of IL-4/IL-13, which are rapidly produced upon Fc epsilonRI cross-linking. Cytokine expression by basophils is unique and distinct from other cell types, since: (1) Basophils are the only cell type constitutively expressing IL-4 and IL-13 mRNA. IL-4/IL-13 message and protein are expressed in a very restricted manner, since neither the mRNAs nor the protein products of most proinflammatory Th1-type, and even Th2-type, cytokines or chemokines are expressed; (2) Basophils secrete IL-4/IL-13 also upon IgE-independent activation (IL-3 plus C5a), and they are thereby potentially capable of initiating a Th2 response. Furthermore, we identified an adjuvant from helminths capable of directly inducing IL-4 formation, and (3) IL-4 expression by basophils is resistant to counter-regulatory effectors inhibiting Th2 development. Studies about the regulation of IgE-dependent and -independent IL-4/IL-13 expression by different cytokines, growth factors and chemokines demonstrate that the different basophil effector functions such as chemotaxis, exocytosis, leukotriene C4 formation and cytokine expression are regulated separately. Thus, our study supports a key immunoregulatory role of basophils in the skewing of immune responses to Th2.

Basophil priming by neurotrophic factors. Activation through the trk receptor.

There is increasing evidence that nerve growth factor (NGF) acts on cells of the immune system, apart from its neurotrophic effects. In human basophils, NGF potentiates mediator release and primes the cells to produce leukotriene C4 in response to C5a. It is, however, unknown whether other homologous neurotrophins also act outside the nervous system, and whether activation of basophils by NGF requires interaction with trk tyrosine kinase receptors, the low affinity NGF receptor (LNGFR), or both. A triple mutant NGF designed to interrupt binding to the LNGFR was found to activate basophils with equal efficacy as wild-type NGF, demonstrating that the LNGFR is not necessary. Despite a 10 times lower potency of mutant NGF, no LNGFR expression was detected by FACS analysis. Brain-derived neurotrophic factor, which interacts with trkB, was inactive at concentrations up to 1000 ng/ml (> 30,000-fold lower potency than NGF), while neurotrophin-3, which is thought to interact with trkC, trkB, and more weakly with trk, induced a threshold effect at 300 ng/ml (approximately 10,000-fold lower potency), demonstrating that 1) the LNGFR cannot deliver a direct signal; and 2) basophils do not express functional trkB and trkC receptors. In agreement with the functional data, basophils (in contrast to other granulocyte types) expressed mRNA for trk, but not trkB or trkC, and no or minimal mRNA for LNGFR. These data demonstrate that human blood basophils express functional trk receptors that do not require the participation of LNGFR, and that, among the neurotrophin family, NGF is unique in priming basophils.

Differential expression of alpha E beta 7 integrins on bronchoalveolar lavage T lymphocyte subsets: regulation by alpha 4 beta 1-integrin crosslinking and TGF-beta.

T lymphocytes expressing the alpha E beta 7 integrin are localized and selectively retained in mucosal tissues. To investigate a potential relationship between alpha E beta 7 expression and pulmonary inflammation, the distribution of alpha E beta 7-bearing CD4+ and CD8+ T cells in peripheral blood and bronchoalveolar lavage (BAL) fluids obtained from patients with allergic asthma, sarcoidosis, hypersensitivity pneumonitis, and idiopathic pulmonary fibrosis (IPF) was determined. In contrast to the distribution in peripheral blood, BAL fluid from these patients contained high number of cells expressing alpha E beta 7 with markedly different expression patterns on CD4 or CD8 cells as well as among the various diseases. Despite similar numbers of activated CD4 cells, alpha E beta 7+CD4+ T cells ranged from 15% in asthmatics to 70% in IPF. In contrast, even in normal individuals, 60% to 90% of BAL fluid CD8+ T cells express alpha E beta 7, suggesting differential induction mechanisms on CD4 and CD8 cells. In vitro experiments revealed that a substantial proportion of peripheral blood CD+ T cells express alpha E beta 7 after stimulation with anti-CD3 antibodies, and up to 80% positive cells were found after the addition of TGF-beta. In contrast, less than 10% of CD4 cells express this particular integrin after in vitro stimulation, and the presence of TGF-beta only increased the number to 30%. Supernatants from in vitro-activated BAL cells as well as concentrated BAL fluid from patients with high alpha E beta 7 expression had no further enhancing effect. However, crosslinking of alpha 4 beta 1-, but not beta 2-integrins, significantly increased the number of alpha E beta 7 expressing CD4+ and CD8+ T cells, even in the absence of TGF-beta. These data indicate that in addition to TGF-beta, the interaction of particular T-cell subsets with specific endothelial cell and extracellular matrix proteins may upregulate alpha E beta 7 integrin expression and thereby contribute to the selective accumulation of these cells in inflammatory lung diseases.

Human blood basophils produce interleukin-13 in response to IgE-receptor-dependent and -independent activation.

Interleukin-13 (IL-13) is a recently discovered immunoregulatory cytokine. The cellular sources of IL-13 and the regulation of its expression are largely unknown. Here we show that human basophils produce IL-13 in response to IgE-receptor (IgER) crosslinking, IL-3, IL-3 plus C5a, but not C5a alone. Human basophils express IL-13 in a restricted manner since, apart from IL-4, no other cytokines encoded on the cytokine gene cluster (IL-3, IL-5, and granulocyte macrophage-colony-stimulating factor [GM-CSF]), are induced. Highest levels of IL-13 are formed after IgE-independent activation leading to a prolonged secretion of IL-13. The response to IgER-cross-linking is more transient preferentially inducing IL-4, IL-3 is a unique cytokine regulating IL-13 production by human basophils: Among a large number of cytokines tested, only IL-3 is capable of directly inducing IL-13 expression. Furthermore, although some IL-13 is produced in response to C5a in the presence of IL-5, GM-CSF, IGF-1 or IL-1 beta, IL-3 is by far the most effective. IL-13 production was blocked by actinomycin D and cycloheximide and conditions leading to IL-13 release also lead to the induction of IL-13 mRNA. This study supports an important immunoregulatory role of human blood basophils, owing to their capacity to simultaneously express IL-13 and IL-4 in a restricted manner.

Influence of bee venom immunotherapy on degranulation and leukotriene generation in human blood basophils.

BACKGROUND: Rapid clinical tolerance can be induced over several hours by very fast bee venom immunotherapy (VIT) protocols. OBJECTIVE: To investigate the mechanisms underlying VIT we examined the changes of blood basophil responsiveness during VIT. METHODS: Seven bee venom allergic patients with a history of severe systemic reactions after a bee sting were investigated. A cumulative dose of 111.1 micrograms bee venom (BV) was administered sc over 3.5 h under intensive care conditions according to an ultra-rush protocol. The release of histamine and the formation of leukotrienes in response to BV, major BV allergen Phospholipase A2 (PLA), IgE receptor cross-linking with the use of monoclonal antibodies against IgE and IgE receptor, as well as IgE independent activation in response to C5a were determined in vitro before and after ultra-rush VIT. RESULTS: We demonstrated a decrease of total histamine in peripheral blood leucocytes just after VIT. Histamine release in response to all the stimuli used is not affected by ultra-rush VIT, if expressed as per cent release of total histamine. However, the absolute amount product released in response to stimulation was decreased, particularly with allergen (BV, PLA). We also found a significant reduction of LTC4 formation after VIT in samples stimulated with specific allergen (BV, PLA). CONCLUSION: Blood basophils are a target for VIT, which induces impaired release of both preformed and newly generated mediators. However, we believe the basic mechanisms of rapid clinical tolerance induced by ultra-rush VIT remain to be investigated.

IgE-independent interleukin-4 expression and induction of a late phase of leukotriene C4 formation in human blood basophils.

T-helper cells can differentiate into at least two subtypes secreting distinct profiles of cytokines, Th1 and Th2, regulating immunoprotection and different immunopathologies. Interleukin-4 (IL-4) is both the product and the inducer of Th2 cells, raising the question whether IL-4 can be produced in response to antigen-independent stimuli. Here we show that human basophils produce IL-4 on stimulation with IL-3 and C5a or C5adesarg in similar amounts as induced by IgE-receptor-cross-linking. C5a-induced IL-4 production requires the presence of IL-3, with little effect of the sequence of stimuli addition. No "Th1-cytokines" (interferon-gamma and IL-2) and even no "Th2-cytokines" (IL-3, IL-5, IL-10, and granulocyte-macrophage colony-stimulating factor) are produced by basophils in response to either IgE-dependent or IgE-independent activation. The generation of leukotriene C4 (LTC4) is regulated in a similar manner. However, C5a induces a rapid, transient burst of leukotriene formation only if added after IL-3. Interestingly, upon prolonged culture, a late phase of continuous LTC4 production is observed, which also requires two signals (IL-3 and C5a), but rather depends on their continuous presence than on their sequence of action. These data describe an antigen-independent pathway of very restricted IL-4 expression. Thus, basophils must be considered as central immunoregulatory cells of the innate immune system. Furthermore, the results show that LTC4 can also be generated more continuously for many hours, a phenomenon that may be of particular importance in chornic allergic inflammation, such as asthma.

Nerve growth factor from the venom of the Chinese cobra Naja naja atra: purification and description of non-neuronal activities.

Nerve growth factor (NGF) was separated from crude Naja naja atra venom by using weak cation-exchange chromatography, followed by reversed-phase liquid chromatography. The yield of the purification was 0.2-0.5% (w/w). The mol. wt was determined to be 13,600 and the protein still induced the typical fibre outgrowth of cultured PC-12 cells in a concentration range of 5-10 ng/ml. Beside this neuronal effect we demonstrated non-neuronal effects of cobra venom NGF, such as induction of plasma extravasation and histamine release from whole blood cells. With human leucocyte preparations, including enriched basophils, there was an increase in C5a-induced histamine release, whereas NGF alone was inactive. Cobra NGF was one-tenth as potent as human recombinant NGF, with a half-maximal stimulation occurring at 10 ng/ml. Cobra NGF and human recombinant NGF showed a modulatory effect on histamine release comparable to the haematopoietic growth factor IL-3. Thus, the non-neuronal effects of cobra NGF may account for immunomodulatory activities during inflammatory events.

Immunolocalization of intracellular interleukin-4 in normal human peripheral blood basophils.

The question as to whether other cell types apart from helper T lymphocytes are capable of producing interleukin-4 (IL-4) has gained much interest over the last years. Recent studies indicate that human basophils also produce IL-4, although direct proof is missing so far. In this study we demonstrate the presence of IL-4 in the cytoplasm of in vitro activated human peripheral blood basophils derived from normal donors. Cytokine-producing cells were revealed at the single-cell level by intracellular immunofluorescence staining using IL-4-specific monoclonal antibodies. Basophils showed a characteristic, apparently granular staining pattern easily discerned from the eccentric dot-shaped staining pattern in activated T cells used in control experiments. Cell counts following priming with IL-3 and stimulation with polyclonal sheep anti-IgE antibody or the anaphylatoxin C5a revealed a significant increase in IL-4-positive basophils to about 19% as compared with unprimed, unstimulated control cells (6%). The amount of IL-4 in the supernatant of these cell preparations paralleled these observations with an at least five- to sevenfold increase following stimulation as compared with control cells (< 5 ng/ml). Using confocal scanning laser microscopy, the intracellular presence of IL-4 was confirmed, and the cells were identified as being basophils on terms of their characteristic multilobed nucleus. This observation was supported by double labeling studies using antibodies to IL-4 and to the high-affinity IgE receptor (Fc epsilon R1). Interestingly, stimulation of cells led to a decrease in the number of Fc epsilon R1-positive cells. The above results show direct evidence that IL-4 is produced by activated human basophils.

Increased expression of CD11b and functional changes in eosinophils after migration across endothelial cell monolayers.

Eosinophils from sputum, nasal polyps, and bronchoalveolar lavages of asthmatics demonstrated a considerably increased CD11b expression, compared with blood eosinophils. Furthermore, the tissue eosinophils expressed ICAM-1 (CD54) and HLA-DR, whereas peripheral blood eosinophils did not. In vitro migration of peripheral blood eosinophils across IL-1-activated human umbilical vein endothelial cell monolayers caused a considerable up-regulation of CD11b and CD35 expression, no induction of ICAM-1 or HLA-DR, and a small but significant decrease in CD11a, CD29, and CD32 expression. These changes were only partially inducible with supernatants from nonactivated or IL-1-activated endothelial cells, platelet-activating factor, or a variety of recombinant cytokines. Thus, cell-cell interactions mediated by receptor-ligand binding or endothelial cell membrane-bound mediators, rather than soluble factors, are responsible for the altered eosinophil surface marker expression. Indeed, preparations of membrane fragments from IL-1-stimulated endothelial cells were able to induce up-regulation of CD11b, which was not inhibitable with the platelet-activating factor antagonist WEB 2086 or antibodies against ELAM-1, VCAM-1, or ICAM-1. Investigation of the functional significance of the increased CD11b expression on eosinophils revealed only minimal changes in the adherence or transmigration capacity. Nevertheless, increased CD11b expression was related to an increased capacity to generate superoxide after stimulation with opsonized zymosan. Thus, cell-cell interactions between eosinophils and endothelial cells induce a considerable up-regulation of CD11b and CD35 on eosinophils and an increased capacity to generate an oxidative burst.

Human peripheral blood eosinophils produce and release interleukin-8 on stimulation with calcium ionophore.

We present evidence that human blood eosinophils produce interleukin (IL)-8 when stimulated with calcium ionophore. Following in vitro culture of 99% pure eosinophils with calcium ionophore, released IL-8 was detectable by enzyme-linked immunosorbent assay in supernatants. Eosinophil IL-8 production was considerably greater than that of IL-3 or granulocyte macrophage colony-stimulating factor. Furthermore, eosinophil production of IL-8 in the presence of calcium ionophore could be inhibited with the immunomodulating agent cyclosporin A and the protein synthesis inhibitor cycloheximide. In addition, following stimulation of highly purified blood eosinophils with calcium ionophore, IL-8 mRNA was detectable after polymerase chain reaction amplification. In comparison with other cells on stimulation with calcium ionophore, eosinophils produce about half as much IL-8 as neutrophils but significantly more than purified T cells. In contrast to monocytes and neutrophils, IL-8 production was not inducible with IL-1 or tumor necrosis factor. Finally, following calcium ionophore stimulation blood eosinophils were shown to contain cytoplasmic IL-8 by employing a monoclonal antibody against IL-8 in conjunction with immunohistochemistry. These observations demonstrate that eosinophils are capable of IL-8 production and release, which may contribute to defense against parasites and to the pathophysiology of allergic and asthmatic disease.

Lack of increased numbers of low-density eosinophils in the circulation of asthmatic individuals.

The density distribution pattern of eosinophils over discontinuous isotonic Percoll gradients from the blood of normal, asymptomatic allergic and non-allergic asthmatic individuals was investigated. There was a completely identical distribution pattern between the investigated groups. Analysis of the expression of surface markers for complement receptors CR1 and CR3 and immunoglobulin G receptor on eosinophils derived from the density bands 1.080, 1.085 and 1.090 g/ml supported this finding since they did not reveal differences in expression between the bands within one group but also not between the three groups. Eosinophils of the various density bands were further purified and stimulated in vitro to produce leukotriene C4 (LTC4) by the calcium ionophore A23187 or serum treated zymosan. Equal amounts of LTC4 were synthesized by the eosinophils of the various density bands within one group. However, it appeared that the eosinophils of all density bands of allergic and non-allergic asthmatics synthesized significantly more LTC4 than the eosinophils from normal individuals (five- to tenfold). Probably this indicates in vivo priming of the eosinophils in asthmatic individuals which is not reflected by a change in density. Control experiments, dealing with possible artifacts due to the isolation procedure or the patient selection, to find differences in distribution patterns over discontinuous Percoll density gradients of the eosinophils of asthmatic compared to normal individuals failed to show such a difference. Therefore, the density distribution pattern of eosinophils over these gradients does not reflect cell activation, whereas LTC4 formation clearly does. This could mean that LTC4 formation is a more sensitive parameter for cell activation than density distribution or cell surface marker expression.

Eosinophil migration in atopic dermatitis. I: Increased migratory responses to N-formyl-methionyl-leucyl-phenylalanine, neutrophil-activating factor, platelet-activating factor, and platelet factor 4.

Eosinophil granular protein deposits have been demonstrated in lesional atopic dermatitis skin. This suggests active tissue infiltration of eosinophils. To find an explanation for the tissue influx of eosinophils, eosinophil migration was studied in vitro by means of a microchemotaxis assay. Eosinophils from the circulation of patients with atopic dermatitis showed an altered capacity to respond to chemotactic stimuli in vitro compared with eosinophils from healthy donors. Eosinophils from patients with atopic dermatitis had significantly increased migratory responses toward dose ranges of N-formyl-methionyl-leucyl-phenylalanine, neutrophil-activating factor, platelet-activating factor, and platelet factor 4. Eosinophils from normal individuals did not respond to N-formyl-methionyl-leucyl-phenylalanine and neutrophil-activating factor and responded only slightly to platelet factor 4. The migratory responses toward tumor necrosis factor-alpha and complement factor C5a were identical in both groups. Interleukin-5, an eosinophil-selective cytokine, is a strong modulator of the migratory responses to these chemotaxins in eosinophils from normal donors. A migratory response toward N-formyl-methionyl-leucyl-phenylalanine and neutrophil-activating factor was induced by interleukin-5, whereas the migratory response toward platelet-activating factor and platelet factor 4 was markedly potentiated. In contrast, the response to complement fragment C5a was only slightly influenced. Our findings indicate that the increased migratory responsiveness of eosinophils from patients with atopic dermatitis to various chemotaxins reflects in vivo "priming" of eosinophils, presumably by circulating cytokines such as interleukin-5. This in vivo "priming" is not optimal because it can be further potentiated by renewed contact with interleukin-5.

Eosinophils and cytokines.

Cytokines act on eosinophils to regulate eosinophil function, with IL-5 recognised to be especially important in control of eosinopoiesis, eosinophil survival and eosinophil priming. In addition, eosinophils have the capacity to produce cytokines involved in acute and chronic inflammatory and repair processes, as well as to produce cytokines that stimulate eosinophils within an autocrine loop. This paper describes (A) an immunomagnetic selection technique for the purification of human blood eosinophils, and (B) a method that employs immunofluorescence with flow cytometry for measurement of blood and sputum eosinophil surface markers. Having demonstrated that sputum eosinophils express HLA-DR, highly purified blood eosinophils were used to analyse (C) the induction and function of eosinophil HLA-DR. Cytokines have the capacity to induce eosinophil HLA-DR, and are produced by eosinophils as an accessory function during antigen presentation. Finally, preliminary data on (D) eosinophil production of IL-8 is presented. Hence, eosinophils have the capacity to act as immunomodulatory cells within cells networks in allergic and asthmatic inflammation.

Induction and function of eosinophil intercellular adhesion molecule-1 and HLA-DR.

We have previously established that eosinophils studied ex vivo from the sputum of asthmatics express intercellular adhesion molecule-1 (ICAM-1) and HLA-DR, whereas peripheral blood eosinophils do not express these surface proteins. On incubation of highly purified (greater than 99.5% pure) blood eosinophils from normal subjects with T cell supernatants, eosinophil ICAM-1 was induced in 24 h, whereas HLA-DR was maximally induced within 48 h. Recombinant cytokines that enable eosinophil survival (IL-5, IL-3, and granulocyte macrophage-CSF) were found to be unable to induce ICAM-1 or HLA-DR, even when pooled at concentrations individually required for eosinophil survival. However, synergy between these eosinophil survival factors and TNF (-alpha and -beta) was found mainly responsible for ICAM-1 induction, whereas synergy between IL-3 and IFN-gamma occurred for HLA-DR induction. Culture of eosinophils in the presence of cytokines and cycloheximide prevented expression of ICAM-1 and HLA-DR, showing that de novo eosinophil protein synthesis is occurring. At a functional level we demonstrate that ICAM-1-bearing eosinophils have increased adhesion capacity for autologous T cells. In contrast, HLA-DR-expressing eosinophils mediated Ag-specific proliferation of an autologous HLA-DR-restricted T cell clone that was inhibitable by anti-HLA-DR and anti-ICAM-1 mAb. Since eosinophil-mediated Ag presentation was inhibitable by treatment of eosinophils with glutaraldehyde or chloroquine, this suggests that eosinophils participate in Ag uptake, processing, and presentation and have accessory functions. Thus, through the induction of ICAM-1 and HLA-DR on tissue eosinophils, eosinophils have the capacity to interact with leukocytes and present Ag to T cells.

Early activation or "priming" of eosinophils in asthma.

Increased numbers of blood and tissue eosinophils are regularly observed in subjects suffering from bronchial asthma. The eosinophil number in the diseased organ is normally closely associated with the presence of clinical symptoms. Not only the cell number, but also the concentration of eosinophil-granule derived mediators is increased in the diseased organ. In particular toxic proteins released by the eosinophil may be responsible for the allergic inflammatory reaction and the concomitant tissue damage. Our recent investigations have shown that eosinophilic granulocytes from asthmatic individuals have the same phenotype as eosinophils from normal individuals (i.e. with respect to their density distribution pattern and surface receptor expression). In contrast, eosinophils from asthmatic individuals do possess increased metabolic activity (i.e. increased leukotriene C4 (LTC4) generating capacity and migration capacity). This increased metabolic activity is due to the presence of circulating factors, i.e. the cytokines interleukin 3 (IL-3), interleukin 5 (IL-5) and granulocyte-macrophage colony stimulating factor (GM-CSF). These cytokines are demonstrable in the circulation of asthmatic, but not normal individuals; they are synthetized by activated T-lymphocytes. This early activation, called "priming", should be the goal of future pharmacological endeavours in order to achieve more efficient treatment of asthma.

[Eosinophilic granulocytes and their significance in allergic diseases]. / Der eosinophile Granulozyt und seine Bedeutung bei allergischen Erkrankungen.

Eosinophils are found in the blood and tissues of patients with allergic diseases such as asthma, allergic rhinitis and also atopic dermatitis. The number of eosinophils in the diseased tissue generally correlates with the expression of clinical symptoms. Originally, the eosinophil was regarded as having an exclusively protective role, for example in host defense against parasites. More recently, the eosinophil is recognized as being a pro-inflammatory cell that can mediate allergic disease. The eosinophil is active in inflammation through the release of granule proteins and the synthesis and release of inflammatory mediators. The important eosinophil granule proteins are major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO). These proteins have toxic effects on the surrounding tissue. Of additional importance for the allergic inflammatory reactions are membrane-derived mediators such as leukotriene C4 (LTC4), platelet activating factor (PAF) and Charcot-Leyden crystals. These mediators are synthesized and released after eosinophil activation, and act toxic on surrounding cells. Eosinophils are active in asthma, and increased numbers and eosinophil-derived mediator concentrations have been documented in bronchial biopsies, BAL and sputum. In addition, eosinophil granule proteins and inflammatory mediators are found in nasal secretions in allergic rhinitis. In atopic dermatitis one finds activated eosinophils and depositions of eosinophil granule proteins in skin biopsies. Eosinophils are not only active in mediating allergic inflammation, but interact in cellular networks with antigen presenting cells (APC), mast cells, and T lymphocytes. These other cells influence eosinophil maturation, mobilization, tissue localization and activation.