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PubMed1998

Identification of the Fc epsilonRI-activated tyrosine kinases Lyn, Syk, and Zap-70 in human basophils.

BACKGROUND: In human blood basophils, cross-linking the high-affinity IgE receptor Fc epsilonRI with multivalent antigen activates a signaling pathway leading to Ca2+ mobilization, actin polymerization, shape changes, secretion, and cytokine production. METHODS AND RESULTS: The role of tyrosine kinases in human Fc epsilonRI signaling was explored by using human basophils isolated by Percoll gradient centrifugation followed by negative and/or positive selection with antibody-coated magnetic beads. Fc epsilonRI cross-linking of more than 95% pure basophil preparations activates the protein-tyrosine kinases Lyn and Syk, previously linked to Fc epsilonRI-coupled rodent mast cell activation, as well as Zap-70, previously implicated in T-cell receptor signaling, and causes the tyrosine phosphorylation of multiple proteins. The presence of Lyn, Syk, and Zap-70 in basophils was confirmed by Western blotting in lysates of highly purified basophils and independently by confocal fluorescence microscopy in cells labeled simultaneously with kinase-specific antibodies and with the basophil-specific antibody 2D7. Comparable amounts of Lyn and Syk were found in basophils and B cells, whereas T cells appear to have greater amounts of Zap-70 than basophils. The tyrosine kinase inhibitor piceatannol spares IgE-mediated Lyn activation but inhibits IgE-induced Syk and Zap-70 activation as well as overall protein tyrosine phosphorylation and secretion. Overall protein-tyrosine phosphorylation increases steadily over a range of anti-IgE concentrations that are low to optimal for secretion. However, tyrosine phosphorylation continues to increase at high anti-IgE concentrations that elicit very little secretion (the characteristic high-dose inhibition of secretion). CONCLUSIONS: Our data demonstrate the association of anti-IgE-stimulated, protein-tyrosine phosphorylation by a cascade of tyrosine kinases, including Zap-70 as well as Lyn and Syk, with the initiation of Fc epsilonRI-mediated signaling in human basophils.

Basophil Degranulation Test↗
PubMed1998

Degranulation of human basophils by picomolar concentrations of IL-3, IL-5, or granulocyte-macrophage colony-stimulating factor.

In most secretory cells, an increase in the cytosolic free Ca2+ concentration ([Ca2+]i) is associated with the exocytosis response. In this study we have evaluated the effect of thapsigargin on histamine release from purified (70% to 97% pure) human basophils of nonallergic donors. Thapsigargin (2 micromol/L), by inhibiting the uptake of Ca2+ in the stores of the endoplasmic reticulum, leads within 1 minute to a gradual increase in [Ca2+]i in human basophils. Incubation of basophils with thapsigargin by itself induced only a very small release of histamine (5.6% +/- 1.8%). However, under suboptimal conditions of stimulation with other agonists, preincubation of basophils with thapsigargin significantly enhanced histamine release. Most strikingly, addition of thapsigargin made basophils extremely sensitive for histamine release induced by IL-3 (maximum histamine release, 71% +/- 7%), IL-5 (maximum histamine release, 43% +/- 8%), or granulocyte-macrophage colony-stimulating factor (GM-CSF) (maximum histamine release, 57% +/- 10%). These cytokines by themselves did not induce histamine release in purified basophils. The effect of thapsigargin was mimicked to a limited extent by addition of platelet-activating factor. We conclude that depletion of the Ca2+ stores may be a critical event in the activation of receptor-mediated histamine release in human basophils.

Basophil Degranulation Test↗
PubMed1986

Bronchospasm following the use of vecuronium.

A case history is presented which records bronchospasm due to vecuronium. Immunological investigations, including basophil degranulation tests, indicated that the bronchospasm was not caused by direct histamine release and was not IgE mediated. It was of interest that intradermal testing gave a positive wheal response against neuromuscular agents other than those involved in the anaesthetic procedure under investigation. A positive reaction to vecuronium was only obtained when given in a high concentration and accords with the general belief that vecuronium has extremely low potential for histamine release.

Aged↗
PubMed1993

Comparison of mechanisms of IL-3 induced histamine release and IL-3 priming effect on human basophils.

We have investigated the mechanisms by which interleukin-3 (IL-3) induces histamine release and primes basophils for increased histamine release in response to anti-IgE- and N-formyl-methionyl-leucyl-phenylalanine (fMLP). The responsiveness of basophils from atopic donors was variable, only 5/11 subjects showing release of > 10%, to IL-3 in the range 0.1-100 ng/ml. IL-3-induced histamine release required both extracellular Ca2+ and cell membrane IgE, removal of membrane IgE by lactate stripping or desensitization of basophils by incubation with anti-IgE in a Ca2+-free medium blocking IL-3-induced histamine release. IL-3 also primed basophils for histamine release by anti-IgE and fMLP in the same concentration range as it evoked histamine release. When IL-3 and either anti-IgE or fMLP were combined, the result was additive or supra-additive depending on the basophil donor. Unlike IL-3-evoked histamine release, IL-3 priming of basophils for fMLP-induced histamine release was shown to be independent of the presence of both cell surface IgE and of extracellular calcium. The protein kinase C (PKC) inhibitor, staurosporine (10 nM), inhibited anti-IgE induced histamine release, but neither IL-3 induced histamine release nor IL-3 priming of IgE- and fMLP-induced histamine release. Pertussis toxin (1.0 microg/ml) inhibited fMLP-induced histamine release but not anti-IgE-induced histamine release, IL-3-evoked histamine release or IL-3 priming. These results indicate that IL-3 modulates mediator release from human basophils by two mechanisms; a direct release of histamine which involves cell surface IgE and the influx of extracellular calcium but which is unlikely to proceed by the same mechanism as cross-linkage of IgE, and a priming effect which is independent of IgE and extracellular Ca2+ and which enhances the secretory effects of a wide range of unrelated secretagogues.

Antibodies, Anti-Idiotypic↗
PubMed1990

Enhancement of basophil histamine release by a human basophil-like cell promoting activity.

A basophil-like cell promoting activity (BaPA) is a lymphokine which has the ability to proliferate metachromatically staining cells in human bone marrow. We studied the effect of BaPA on histamine release from human peripheral basophils. BaPA did not directly induce histamine release from basophils. However BaPA ranging from 0.001 to 0.5 U/ml enhanced histamine release from basophils stimulated with anti-IgE, calcium ionophore A23187 or FMLP in a dose-dependent manner. On the basis of recent observations that in addition to their capacity for proliferating progenitor cells, colony-stimulating factors are capable of regulating functions of end-stage cells of the same lineage, this finding that BaPA is able to modulate a basophil function raises the possibility that BaPA also regulates progenitors of basophils.

Basophil Degranulation Test↗
PubMedDate not supplied

Inhibition of allergic reactions with antibodies to IgE.

Numerous clinical studies show that direct interference with the IgE response leads to a decrease or elimination of allergic symptoms. The aim of these studies was to design a therapy aimed at decreasing IgE levels in order to ameliorate atopic disease. To this end, a murine monoclonal antibody, MAE11, directed against IgE was identified, which had all the properties necessary to interfere with IgE responses, but lacked the harmful side effects of inducing receptor cross-linking. The antibody was selected on the basis of its ability to bind circulating IgE at the same site as the high-affinity receptor, thus blocking the binding of IgE to mast cells and basophils. To allow for possible chronic administration and to avoid the problems of antigenicity, MAE11 was humanized. The best of several humanized variants, version 25 (rhumAb-E25) was selected since it possessed binding affinity and biological activity comparable to MAE11. Clinical studies are underway to determine the safety and efficacy of this treatment for allergic rhinitis and asthma.

Allergens↗
PubMed1984

Unilateral central retinal vein occlusion in systemic lupus erythematosus.

A case of unilateral central vein occlusion occurring in a normotensive patient with systemic lupus erythematosus (SLE) is reported. This vasculitis is related to SLE. In spite of the observation of circulating immune complex and a positive response to the human basophil degranulation test in the presence of bovine soluble retinal antigen, the real pathogenesis of this unilateral vasculitis remains obscure.

Female↗
PubMed1984

Amiodarone-induced hypersensitivity pneumonitis. Evidence of an immunological cell-mediated mechanism.

Interstitial pneumonitis developed in a patient who had received a cumulative dose of 985 g of amiodarone in nine years. No other cause for pneumonitis was found. The following findings favor an immunologic mechanism of hypersensitivity due to amiodarone: positive skin and basophil degranulation tests with amiodarone; lymphocytosis and inverted ratio of helper/suppressor T lymphocytes in bronchoalveolar lavage fluid; secretion of leukocyte inhibitory factor, as shown by the inhibition of migration of peripheral blood leukocytes; and positive lymphoblastic transformation in the presence of amiodarone.

Aged↗
PubMedDate not supplied

[Influence of novodrin and obsidan on the secretory activity of tissue basophils of the thyroid and parathyroid glands].

The task was to investigate secretory activity of sets of tissue basophils of the thyroid and parathyroids during administration of isoproterenol (novodrin) and propranolol (obsidan); 32 rats, divided into 2 groups, were given a single intraperitoneal injection of 20 mg of isoproterenol (the 1 st group) and 2.5. mg of propranolol (the 2nd group) with their subsequent decapitation with a 12-minute interval for 3 h. Six rats were controls. Increased tissue activity of tissue basophils was noted and interpreted as a result of elevated secretion of calcium regulating hormones developing under the influence of beta-adrenergic drugs.

Animals↗
PubMed1979

[Two new cases of allergy to tartrazine (author's transl)].

Two patients developed allergy to tartrazine, and the symptoms, as in most such cases, were related to the skin and mucous membranes. The clinical diagnosis was confirmed by the basophil degranulation test. These observations underline the intrication of food substances and medications in the genesis of these accidents and demonstrate the notion of a quantitative contribution of food additives. The author discusses the frequency of these allergies to this colouring substance, and restates the position, which has just been taken by the Academy of Medicine, with the objective of prohibiting the use of tartrazine E 102 in food substances.

Azo Compounds↗
PubMed1991

[Post-vaccination reactions and complications of pertussis, diphtheria and tetanus vaccines].

Clinical manifestation of postvaccinal reactions and complications and the presence of sensibilizating antibodies to certain components of the applied vaccines was analyzed in 80 children immunized by Di-Te-Per, Di-Te or Te vaccine. Most children showed a mild reaction; concerning statistical significance it was most frequently limited to the site of the vaccine application if the immunization was performed by Di-Te and Te vaccines, as compared to the immunization by Di-Te-Per vaccine. Indirect basophil degranulation test revealed that 50% of the analyzed reactions were of allergic origin. Sensibilizating antibodies to pertussis antigen are statistically significantly more frequent than to other vaccinal antigens (diphtheria and tetanus toxoids) and additives contained in the vaccines (methiolate, sodium benzoate).

Child↗