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PubMed1996

Comparison of basophil histamine release, eosinophil cationic protein and non-specific airway responsiveness between mite-sensitive asthmatic and non-asthmatic children and non-allergic controls.

To understand the relevance of allergy to the development of asthma in children, we examined basophil histamine release (HR) with Df antigen, blood eosinophil counts, serum eosinophil cationic protein (ECP) levels, and bronchial responsiveness to methacholine (PC20) in three groups of children, including 36 asthmatics with high RAST titre for Df (group 1), 36 non-asthmatics with similarly high RAST titre for Df (group 2) and 21 non-asthmatics with negative RAST titre for Df (group 3). The amount of Df antigen inducing 50% HR from basophils did not vary significantly between group 1 and 2 (P > 0.05), while none of the cells responded to higher concentrations of Df in group 3. The mean number of blood eosinophils and level of serum ECP were highest in group 1, and lowest in group 3, with group 2 being intermediate, and the differences were significant between all three groups (P < 0.01). The mean PC20 value was the lowest in group 1, intermediate in group 2, and the highest in group 3, and the differences were significant between all three groups (P < 0.01). While correlation studies showed that PC20 values of group 2 subjects significantly correlated with their eosinophil numbers (r = -0.48, P < 0.01) and ECP levels (r = -0.49, P < 0.01), such correlations were not found in group 1 subjects. These results suggest that the degree of the eosinophilic inflammation caused by the allergic reaction to mites is an important factor in determining the clinical expression of asthma in atopic subjects.

Adolescent↗
PubMed1996

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.

Antibodies, Monoclonal↗
PubMed1996

Human monocyte chemoattractant protein (MCP)-4 is a novel CC chemokine with activities on monocytes, eosinophils, and basophils induced in allergic and nonallergic inflammation that signals through the CC chemokine receptors (CCR)-2 and -3.

The chemokines are a large family of cytokines that regulate the complex and precise recruitment of immune cells into inflammatory foci. To fully appreciate their role in the pathogenesis of human diseases, the entire spectrum of chemokines, their receptors, their cellular targets, and mechanisms of regulation need to be delineated. Using eotaxin as a probe, we isolated a cDNA for a novel human beta (or CC) chemokine that, based on its biological and structural features, we have named monocyte chemoattractant protein (MCP)-4. Purified recombinant MCP-4 protein was a potent chemoattractant for monocytes and eosinophils and stimulated histamine release from basophils. MCP-4 induced a calcium flux in HEK-293 cells transfected with the monocyte selective MCP-1 receptor (CCR-2B) and the eosinophil selective eotaxin receptor (CCR-3), but not in the more widely expressed CCR-1 or CCR-5. This novel chemokine is expressed in TNF-alpha and IL-1 activated epithelial and endothelial cells in vitro, and in the epithelial mucosa of patients with both Th2-type allergic and Th1-type nonallergic sinusitis. Furthermore, both IFN-gamma and IL-4, products of Th1 and Th2 cells, respectively, synergized with TNF-alpha and IL-1 in inducing MCP-4 mRNA accumulation. These properties of MCP-4 offer a molecular explanation for the observed accumulation of monocytes, eosinophils and basophils in both Th1- and Th2-type immune responses.

Base Sequence↗
PubMed1979

Adenosine receptor on human basophils: modulation of histamine release.

Adenosine, at physiologic concentrations, inhibits in vitro IgE-mediated human basophil histamine release in a dose-dependent fashion. The inhibition dose-response curve is paralleled by an adenosine-induced increase in cAMP levels of human leukocyte preparations. Further evidence that the adenosine effect is related to changes in cAMP levels is that the nucleoside inhibits only in the first stage of antigen-induced histamine release and fails to inhibit the release caused by ionophore A23187. A poorly metabolized derivative of adenosine, 2-chloroadenosine inhibits as effectively as adenosine; dipyridamole, which blocks adenosine uptake, does not impair the inhibition caused by adenosine. Finally, theophylline, which is a competitive antagonist of adenosine in human lymphocytes also blocks the inhibition of release caused by adenosine. These data suggest that adenosine acts via a specific cell-surface receptor linked to adenylate cyclase. It appears that the human basophil has a specific receptor for adenosine and that this nucleoside may modulate the in vivo release of the mediators of immediate hypersensitivity reactions.

Adenosine↗
PubMed1977

Basophil-derived platelet-activating factor (PAF) as an in vivo mediator of acute allergic reactions: demonstration of specific desensitization of platelets to PAF during IgE-induced anaphylaxis in the rabbit.

The anaphylactic response to an i.v. injection of antigen into rabbits making only IgE antibody against the antigen was shown to be preceded by a disappearance of the metachromatic staining properties of the circulating basophils and was accompanied by marked but transient thrombocytopenia. The platelets which returned to the circulation 1 hr after the anaphylaxis were shown to be unresponsive to the secretion-inducing activity of basophil-derived platelet-activating factor (PAF) when compared with platelets examined before antigen challenge. By contrast, platelet responsiveness to other stimuli such as collagen, thrombin, and C3b was unchanged. The specific desensitization to PAF provides strong evidence for the action of this mediator on platelets in vivo during IgE-mediated anaphylaxis and provides a useful tool for detecting the effects of particular cell activators in inflammatory reactions.

Acute Disease↗
PubMed1997

Effect of an ectokinase inhibitor, K252b, on degranulation and Ca2+ signals of RBL-2H3 cells and human basophils.

We examined the effects of K252b, an ectoprotein kinase inhibitor of microbial origin, on the activation process of RBL-2H3 cells by cross-linking of IgE receptors by the endoplasmic reticulum Ca2+-ATPase inhibitor 2,5-di(tert-butyl)-1,4-hydroquinone or by the Ca2+ ionophore A23187. Analysis of phosphorylation of ectoproteins following IgE receptor cross-linking revealed that K252b mainly inhibited the phosphorylation of a 130-kDa protein. The inhibitor simultaneously inhibited degranulation and the sustained increase in the cytosolic calcium ion concentration even after addition of Ag. In contrast, K252b did not inhibit the increase in degranulation and cytosolic calcium ion concentration caused by stimulation with 2,5-di(tert-butyl)-1,4-hydroquinone and A23187. Permeation of K252b into RBL-2H3 cells, assessed by fluorescence intensity, was very low. K252b also inhibited degranulation caused by IgE receptor cross-linking in human basophils, but did not inhibit the degranulation caused by A23187. Thus, our findings suggest that the effects of K252b may be mediated by outer surface-bound or -anchored K252b-sensitive molecules on RBL-2H3 cells and human basophils, and that the phosphorylation of ectoprotein may involve a transmembrane influx of Ca2+ by IgE receptor cross-linking.

Basophils↗
PubMedDate not supplied

[Mast cells and basophilic leukocytes in allergic inflammation].

The human mast-cell population is composed of a heterogenous group of cells with respect to structure and function. Mast-cells can no longer be regarded simply as cells that initiate acute allergic reactions through the release of rapidly metabolized mediators, such as histamine and products of arachidonic acid oxidation. The production of a wide range of cytokines by mast-cells in the centre of the allergic inflammation. These cytokines influence migration and activation of other different cells including basophils and eosinophils and also lymphocytes, thus perpetuating allergic inflammation. Mediators release from human basophils are considered to contribute to the late phase of allergic response.

Animals↗
PubMed1976

Cell cycle and morphological changes during growth and differentiation of a rat basophilic leukemia cell line.

Cells of the rat basophilic leukemia cell line RBL-1 differentiated maximally when permitted to achieve growth arrest in a high-density stationary phase, in which the cell number is constant, and the cells are arrested in a G phase of the cycle. Features of differentiation are the accumulation of large basophilic granules and increases in membrane receptors for immunoglobulin E. However, changes in histamine content did not parallel granule development or changes in immunoglobulin receptor concentration. During rapid "forced exponential" growth, the cell number doubles every 8 hr, 50% of the cells are in S phase, and differentiation is minimal.

Animals↗
PubMed1976

Metabolic studies of guinea pig basophilic leukocytes in short-term tissue culture. I. Measurement of histaminesynthesizing capacity by using an isotopic-thin layer chromatographic assay.

Mature circulating guinea pig basophils, purified to comprise 25% or more of leukocytes, have been successfully maintained in short-term tissue culture for up to 72 hr. These cells were found to retain the ability to synthesize histamine, as assayed by a new isotopic-thin layer chromatographic assay which can reliably detect as little as 0.5 pg of 3H-histamine. Cell-associated, newly synthesized histamine was detectable as early as 1 hr of culture, was substantially increased at 6 hr, and reached maximal levels at 24 hr, when it accounted for approximately 6.5% of total cell histamine. Newly synthesized histamine was still detectable at 48 and 72 hr of culture. Histamine synthesis was decreased by lwoering the concentration of histidine in the culture medium, and was markedly reduced by all the specific histidine dedarboxylase (HDC)3 inhibitors tested, but not by alpha-methyl-DOPA, pyrilamine maleate, or metiamide. Increasing the concentration of pyridoxal phosphate, the HDC coenzyme, above that normally present in culture medium resulted in only an equivocal increase in the amount of newly synthesized histamine, whereas aminoguanidine, an inhibitor of histaminase, had no detectable effect. Uptake of exogenous histamine by cultured basophils was trivial compared to histamine synthesis. Both newly synthesized and previously manufactured, nonisotopic, histamine seemed to be stored in the same pool, as the same proportion of both was released by concanavalin A (Con A). Cellular histamine was largely conserved, with little or no spontaneous release into the medium of detectable isotopic or nonisotopic histamine. These techniques provide a model for studying granulocyte metabolic processes in vitro, and should assist in the direct investigation of a variety of their physiologic functions.

Animals↗
PubMed1975

Microspectrophotometric detection of heparin in mast cells and basophilic granulocytes stained metachromatically with Toluidine Blue O.

A qualitative microspectrophotometric detection method for heparin in situ has been developed, using data obtained previously with a model system of polyacrylamide films containing pure glycosaminoglycans (Tas, 1975). This technique, based on the unique metachromatic properties of heparin with Toluidine Blue O in glycerol, has been worked out with rat peritoneal and mesenteric mast cells. After the smears containing the stained cells had been mounted in glycerol, a change with time of the recorded metachromatic peaks to lower wavelengths was found, leading to an equilibrium phase after some days. The metachromatic peaks recorded in this phase appeared to resemble closely the peak obtained for the heparin-Toluidine Blue O complex under similar conditions in the model experiments. With rat mast cells it was found that nucleic acids, basic proteins, histamine and lipids had no appreciable influence on the position of the final recorded peaks, nor did they influence the slope of the time course very much. This observed decrease with time in the wavelengths of the metachromatic peaks can be explained by the time necessary for equilibration of the cells in glycerol and by the possible influence of lower sulphated glycosaminoglycans on the peak of the heparin-Toluidine Blue O complex. It was found that the method can be used to detect unequivocally the presence of heparin in cells, even if they also contain up to 75% (mole/mole) of other, lower sulphated glycosaminoglycan. Only a limited number of cells is necessary with this method - in contrast to biochemical determinations. For the first time the presence of heparin in normal human basophilic granulocytes and mast cells has been proved directly. The experiments indicate the occurrence of virtually similar sulphated heparins in human mast cells and basophilic granulocytes, as well as in pig mast cells. A higher sulphated heparin, however, might be present in rat mast cells.

Adolescent↗
PubMed1987

Boophilus microplus (ixodid tick): fine structure of the gut basophilic cell in relation to water and ion transport.

Basophilic cells in the guts of female ticks are derived from the basal remnants of type 2 secretory cells. As viewed by electron microscopy, these cells have microvilli uniformly distributed on the luminal surface, but they lack the abundant pinocytotic vesicles and lysosomes characteristic of digest cells. The cytoplasm is filled with well organized rough endoplasmic reticulum, Golgi complexes and secretory granules. Infoldings of a basal labyrinth extend the contact of the cell with the underlying haemolymph, and there are many mitochondria in the cell processes between folds. This morphology appears to fit the cell for functioning in active water transport across the gut wall. Subsequent to a final rapid phase of engorgement, the basophilic cell reorganizes its cisternae of rough endoplasmic reticulum into whorls and parallel arrays and resumes a secretory role.

Animals↗
PubMed2001

Immunohistochemical and ultrastructural study of basophilic inclusions in adult-onset motor neuron disease.

We immunohistochemically and ultrastructurally studied basophilic inclusions (BI) in a patient with adult-onset sporadic motor neuron disease (MND). BI were frequently observed not only in degenerated anterior horn cells, such as central chromatolytic neurons, but also in normal-appearing large anterior horn neurons. They had various shapes, round, elliptical or irregular, and occasionally they had distinct basophilic rims. They also varied in size. There were no halos around them nor core in their centers. Immunohistochemically, some BI were immunostained for ubiquitin or SOD1, but BI were not immunoreactive with anti-phosphorylated neurofilament (SMI 31), phosphorylated tau, cystatin C or Golgi (MG-160) antibodies. Ubiquitin-positive skein-like inclusions (SI) were occasionally observed in the somata of anterior horn neurons. Ultrastructurally, BI consisted of filamentous structures associated with granules, which were attached to thick filaments. The thick filaments were straight without constriction or side arms and their diameter was twice that of the neurofilaments. BI occasionally contained tubular structures among the granule-associated filaments. The granulo-filamentous profiles varied from being compactly arranged to being more loosely packed. The structure of BI resembles that of the Lewy body-like hyaline inclusions (LBHI) observed in sporadic MND patients. Bundles of filaments resembling SI, which were composed of compactly packed filaments without fine granules running parallel to the longitudinal axis, were frequently observed inside or at the periphery of BI, and occasionally clustered in the perikarya. Each filament measured approximately 15-25 nm in diameter, and a bundle of these grouped filaments was sometimes surrounded by a unit membrane. We also occasionally observed in-between structures of BI and bundles of filaments resembling SI. These findings suggest a certain relationship between BI, SI and LBHI in the pathomechanism of BI development. Further studies are needed to elucidate whether sporadic adult-onset MND characterized by BI forms a different subtype of MND.

Anterior Horn Cells↗
PubMed1998

Trichinella spiralis: induction of the basophilic transformation of muscle cells by synchronous newborn larvae. II. Electron microscopy study.

The present ultrastructure observations apply to the same experimental groups that were investigated in the first part of the study. The results of electron microscopy investigations correspond to those obtained using light microscopy methods. The ultrastructure data demonstrated that 1-h-old sNBL (group I) penetrated into the sarcoplasm of the muscle cells and transformed it basophilically, finally creating the "nurse cell-muscle larva complex." These larvae also caused transformation of the same muscle cells without being present in the sarcoplasm. The larvae of group II (9-h-old sNBL) preserved transformation potential as well, but it was less intensive. Not all NBL settled in the muscle cells; some of them remained in the intercellular spaces. Group II larvae present in the muscle cells underwent early degeneration and necrosis more often than did group I larvae; the inflammatory cell reactions in the vicinity of the larvae were more intensive. The basophilic transformation of muscle cells that did not contain larvae in their sarcoplasm was not intensive and often stopped at the level of cell nuclei. The larvae of group III (6-day-old sNBL) neither settled in the muscle cells nor transformed the cell sarcoplasm.

Animals↗
PubMed1991

12-Lipoxygenase from rat basophilic leukemia cells: separation from 5-lipoxygenase and temperature-dependent inactivation by hydroperoxy fatty acid.

12-Lipoxygenase and 5-lipoxygenase from rat basophilic leukemia cells were separated by protein-HPLC in a single step. Upon incubation in the presence of Ca2+, 12-lipoxygenase converted arachidonic acid into 12(S)-hydroxyeicosatetraenoic acid and linoleic acid into 13(S)-hydro(pero)xyoctadecadienoic acid. The reaction products were analyzed by reversed-phase and chiral straight-phase HPLC with ultraviolet-detection. Using the cytosolic fraction of rat basophilic leukemia cells, optimal 12-lipoxygenase activity was observed at 10 degrees C. At 37 degrees C 12-lipoxygenase was very rapidly inactivated by its own product, hydroperoxy fatty acid, at low concentrations (10-100 nM).

Animals↗
PubMed1994

Thapsigargin-induced nuclear calcium signals in rat basophilic leukaemia cells.

By a confocal fluorescence microscope with an argon-ion laser (488 nm) and a He-Cd laser (325 nm) we have studied thapsigargin-induced calcium signals in individual rat basophilic leukaemia (RBL-2H3) cells. In the presence or absence of external calcium ions, thapsigargin-induced calcium signals were transferred to the nucleus as well as to the cytoplasm of RBL-2H3 cells. The calcium signals were generally much stronger in the nucleus than in the cytoplasm. However, some of the RBL-2H3 cells had apparently reduced nuclear calcium signals. They had a basophil-like bilobed (multilobed) nucleus, although most RBL-2H3 cells had a mast-cell-like monolobed nucleus. In the cells with a bilobed nucleus, IgE-receptor-mediated calcium signals were neither transferred to the nucleus nor to the cytoplasm. The results gave a new insight into the understanding of the mechanism of the nuclear calcium signals in RBL-2H3 cells.

Animals↗
PubMed2003

The protein tyrosine phosphatase PTP-Basophil/Basophil-like. Interacting proteins and molecular functions.

The protein tyrosine phosphatase PTP-Basophil (PTP-Bas) and its mouse homologue, PTP-Basophil-like (PTP-BL), are high molecular mass protein phosphatases consisting of a number of diverse protein-protein interaction modules. Several splicing variants of these phosphatases are known to exist thus demonstrating the complexity of these molecules. PTP-Bas/BL serves as a central scaffolding protein facilitating the assembly of a multiplicity of different proteins mainly via five different PDZ domains. Many of these interacting proteins are implicated in the regulation of the actin cytoskeleton. However, some proteins demonstrate a nuclear function of this protein tyrosine phosphatase. PTP-Bas is involved in the regulation of cell surface expression of the cell death receptor, Fas. Moreover, it is a negative regulator of ephrinB phosphorylation, a receptor playing an important role during development. The phosphorylation status of other proteins such as RIL, IkappaBalpha and beta-catenin can also be regulated by this phosphatase. Finally, PTP-BL has been shown to be involved in the regulation of cytokinesis, the last step in cell division. Although the precise molecular function of PTP-Bas/BL is still elusive, current data suggest clearly that PTP-Bas/BL belongs to the family of PDZ domain containing proteins involved in the regulation of the cytoskeleton and of intracellular vesicular transport processes.

Animals↗
PubMed1982

Increase in histidine decarboxylase activity in skin of genetically mast-cell-deficient W/Wv mice after application of phorbol 12-myristate 13-acetate: evidence for the presence of histamine-producing cells without basophilic granules.

Histidine decarboxylase (HisDCase, EC 4.1.1.22) activity in mouse skin increased by a factor of more than 10 after a single application of phorbol 12-myristate 13-acetate. The cell type that was responsible for the increase in HisDCase activity was examined by using (WB X C57BL/6)F1-W/Wv mice, which are genetically deficient in tissue mast cells. In contrast to a report that increase of ornithine decarboxylase (EC 4.1.1.17) activity occurs in the epidermis [O'Brien, T. G., Simisiman, R. C. & Boutwell, R. K. (1975) Cancer Res. 35, 2426-2433], the HisDCase activity was found to increase in the dermis. Although most of the histamine in the dermis was present in mast cells, the increase in HisDCase activity in the skin in W/Wv mice was comparable with that in congeneic +/+ mice. This increase of HisDCase activity in the skin of W/Wv mice was abolished by prior x-ray irradiation (800 rads; 1 rad = 0.01 gray) but was restored by subsequent bone marrow transplantation. Because mice, in general, are known to lack basophilic leukocytes, the present results suggest the existence of histamine-producing cells without basophilic granules that are derived from the bone marrow.

Animals↗