Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “BASOPHILS”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 559 records · Page 31Linked to original sources
PubMed1993

IL-3 alters free arachidonic acid generation in C5a-stimulated human basophils.

IL-3 is known to enhance the secretion of several mediators from human basophils activated by receptor-mediated stimuli. IL-3 can cause a qualitative change in the mediator release pattern for C5a-mediated stimulation; without IL-3, C5a causes no leukotriene release whereas in the presence of IL-3, significant leukotriene occurs. This study examines the influence of a 15-min pretreatment of basophils with IL-3 (10 ng/ml) on several signal transduction events. Basophils stimulated with C5a typically displayed only a transient cytosolic Ca2+ response [Ca2+]i, attributed to the release of intracellular calcium stores. IL-3 had sporadic, statistically nonsignificant, effects on the peak of this initial response as well as inconsistent effects on the generation of a second phase in the [Ca2+]i response (i.e., that which is caused by the influx of extracellular Ca2+). IL-3 also had no effect on resting [Ca2+]i levels. Challenge of basophils in the presence of EGTA had little effect on the amount of leukotrienes generated. This maneuver did not influence the initial transient elevation of [Ca2+]i. Although basophil leukotriene release is usually slow, after exposure to IL-3 it was found that leukotriene release occurred rapidly, during the brief window of time (0 to 45 s) in which the [Ca2+]i transient occurred. These results suggested that the generation of AA was accelerated by pretreatment with IL-3. Subsequent mass measurements of free AA by gas chromatography-mass spectroscopy showed: 1) IL-3 itself caused no free AA generation; 2) without IL-3, C5a stimulated little or no free AA generation; and 3) in the presence of IL-3, C5a generated substantial levels of free AA at an accelerated rate. A similar acceleration in the rate of free AA generation, without any apparent increase in the amount, occurred in IL-3-primed basophils stimulated with FMLP. Additional studies showed that IL-3 had no effects on whole cell ATP levels and that the kinase inhibitor, staurosporine, did not inhibit the ability of IL-3 to cause enhanced responses to C5a. We conclude that the primary effect of a short period of pretreatment with IL-3 is to couple the generation of free AA to C5a-mediated stimulation in particular and to accelerate its generation after other stimuli. Enhancements of the [Ca2+]i response, while sporadic and at best modest, may have some influence on enhanced mediator release but did not clearly explain the functional effects of IL-3.

Arachidonic Acid↗
PubMed1977

Histamine-induced inhibition of normal human basophil chemotaxis to C5a.

Histamine in concentrations as low as 10(-8) M inhibited the chemotactic response of normal human basophils to C5a. Histamine had no effect on basophil chemotaxis to lymphocyte-derived chemotactic factor. Histamine inhibition of basophil chemotaxis to C5a was prevented by metiamide, a drug which blocks H2 receptors for histamine. Since the accumulation of basophils in delayed cutaneous basophilic hypersensitivity (CBH) reactions may occur in part because of chemotaxis to C5a, and since C5a can induce histamine release, histamine inhibition of chemotaxis may limit basophil infiltration in CBH lesions.

Basophils↗
PubMed1977

Isolation of the cytoplasmic granules of guinea pig basophilic leukocytes: identification of esterase and protease activities.

Procedures were developed for isolating highly purified cytoplasmic granules of basophilic leukocytes from guinea pig peripheral blood. The methods involved disruption of cells in 0.34 M sucrose followed by a series of membrane filtrations and fractionation on sucrose density gradients. These preparations, up to 95% pure basophil granules by electron microscopy, contained a mixture of neutral esterases-proteases including caseinolytic activity; both trypsin- and chymotrypsin-like serine hydrolases were identified by means of appropriate inhibitors. Localization of at least one such activity to the basophil granule was confirmed by a cytochemical method; this activity was absent in contaminating lymphocytes and eosinophils. By contrast, several lysosomal enzymes, lactic dehydrogenase, and plasminogen activator activity, present in cell homogenates, were absent from purified granules. The granule matrix of guinea pig basophils, unlike the cytoplasmic granules of other granulocytes or mast cells, was little altered by high or low salt concentration but was disrupted into insoluble fragments by 0.01 N HCl and by Triton X-100. Granules were solubilized by papain and by urea-SDS but enzyme activity was destroyed. Triton X-100 incubation with freeze-thawing proved to be the optimal method for extracting esterase activities. Esterase activities were not released from basophils under conditions of anaphylactic degranulation that liberated the great majority of basophil granule histamine.

Animals↗
PubMed1996

Counter-receptors on human basophils for endothelial cell adhesion molecules.

Ligands on human basophils for the endothelial adhesion molecules intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), mucosal addressin cell adhesion molecule-1 (MAdCAM-1), and E-selectin were investigated. Adhesion of basophils to endothelial cells was inhibited by mAb recognizing CD18, CD11a, and/or CD11b, with the pattern and magnitude of inhibition dependent upon the activation state of the basophils and endothelium. Adhesion to recombinant VCAM-1 was completely inhibited by mAb recognizing alpha 4 integrin and partially by mAb to the beta 1 or beta 7 subunit; surface expression of these integrins was also detected. Adhesion to recombinant MAdCAM-1 expressed on Chinese hamster ovary cells was completely inhibited by mAb recognizing alpha 4 and/or beta 7 integrins. Adhesion to recombinant E-selectin was completely inhibited by basophil pretreatment with neuraminidase and partially inhibited by endo-beta-galactosidase. By flow cytometry, bimodal patterns of expression of sialyl-Lewis X- and sialyl-dimeric-Lewis X were observed, and adherent cells tended to be sialyl-dimeric-Lewis X positive. Thus, basophils express beta 1, beta 2, and beta 7 integrins along with sialylated surface ligands that may interact with the endothelium during basophil recruitment responses.

Animals↗
PubMed1997

IgE regulates mouse basophil Fc epsilon RI expression in vivo.

The binding of IgE to high-affinity IgE receptors (Fc epsilon RI) on the surface of mast cells and basophils primes these cells to secrete a panel of proinflammatory mediators upon subsequent exposure to specific Ag. We now find that the level of Fc epsilon RI expression on bone marrow basophils in mice infected with the nematode Strongyloides venezuelensis exhibits a strong positive correlation with the serum concentration of IgE, as was previously reported for human blood basophils. Moreover, the administration of IgE in vivo can significantly upregulate Fc epsilon RI expression on mouse basophils, and genetically IgE-deficient (IgE -/-) mice exhibit a dramatic (approximately 81%) reduction of basophil Fc epsilon RI expression compared with the corresponding normal (IgE +/+) mice. The finding that IgE can be a major regulator of mouse basophil Fc epsilon RI expression in vivo identifies a potentially important mechanism for enhancing the expression of effector cell function in IgE-dependent allergic reactions or immunologic responses to parasites.

Animals↗
PubMed1997

Regulation of IgE-dependent IL-4 generation by human basophils treated with glucocorticoids.

Glucocorticoids are widely used in the therapeutic intervention of allergic diseases, affecting the function of a variety of proinflammatory cell types that participate in these disorders. These drugs have been shown to inhibit the release of histamine by human basophils, but not by mast cells, in a reaction requiring 8 to 24 h. To address whether the generation of IL-4 by basophils is similarly affected, we investigated the actions of glucocorticoids on the in vitro release of this cytokine induced by anti-IgE Ab or by the human recombinant histamine-releasing factor. The ability of basophils to generate IL-4 was immediately affected, with secretion of this protein being inhibited >50% with <1-h preexposure to steroid. However, the release of histamine in these cultures was inhibited only after 24-h preincubation, suggesting that the mechanisms controlling the release of this mediator differ significantly from those regulating cytokine secretion. A rank order of potency of the steroids tested for inhibition of IL-4 protein was as follows: triamcinolone > dexamethasone > betamethasone > hydrocortisone. The sex steroids, testosterone and estrogen, showed no effect on basophil secretion. Experiments using reverse transcription-PCR indicate that glucocorticoids inhibit IL-4 generation in basophils on the level of transcription. These studies suggest that the success of glucocorticoids in the treatment of allergic conditions is due in part to their ability to inhibit both mediator release and cytokine secretion by basophils.

Antibodies, Monoclonal↗
PubMed1997

The in vitro and in vivo effect of corticosteroids on basophil releasability in patients with mild and severe bronchial asthma.

BACKGROUND: While inhalation of corticosteroids (CST) is considered very effective in most asthmatic patients, some require a high dose of oral prednisone to control the disease. Basophils, which participate in inflammation, are responsive to corticosteroids by suppressing histamine release. OBJECTIVES: We investigated the in vivo (oral prednisone) and in vitro (dexamethasone, DEX) effect on basophil histamine release in mild and steroid-dependent asthmatics. METHODS: Histamine release from basophils to anti-IgE and anti-IgE + IL3 was evaluated following five days of prednisone given 20 mg twice daily in eight subjects with mild disease and 2 h following their daily prednisone ingestion in eight subjects with severe disease, as well as after in vitro DEX was added to the cells. RESULTS: Histamine release from basophils was seen following anti-IgE as well as anti-IgE + IL-3. The same amount of release was seen in the mild and severe asthmatics. In vivo prednisone suppressed histamine release to both stimuli and DEX added to the suppression in the mild asthmatics. In the severe ones, DEX showed no inhibitory effect on histamine release. CONCLUSION: Oral and in-vitro CST suppressed histamine release from basophils of mild but not severe CST-dependent asthmatics. Suppression of basophil releasability can be a reflection of asthma severity.

Administration, Oral↗
PubMed1976

Chemotactic responses of normal human basophils to C5a and to lymphocyte-derived chemotactic factor.

Peripheral blood basophils from normal human subjects responded chemotactically to the complement component C5a and also to the supernatant fluid from cultured human lymphocytes stimulated by mitogens or specific antigen. Basophil chemotactic factor (BCF) was detected in stimulated lymphocyte cultures within 24 hr and increased for the first 3 days. BCF was found after stimulation of lymphocytes by B or T cell mitogens as well as by specific antigen. The amount of BCF in culture fluids was independent of cell proliferation. Monocyte chemotactic factor (MCF) and BCF could not be separated from one another by Sephadex or DEAE chromatography. Both activities were eluted from Sephadex G-100 columns in a m.w. region of about 15,000 daltons and were found after ion exchange chromatography on DEAE in a peak eluted by 0.1 M NaCl. These findings suggest that mitogen or antigen-stimulated lymphocytes secrete a molecule that is chemotactic for both monocytes and basophils. Thus, in the absence of a basophil-specific chemotactic molecule, the preferential accumulation of basophils in certain types of delayed hypersensitivity skin reactions must be due to additional factors such as basophil chemotaxis augmentation factor.

Basophils↗
PubMed1998

In vitro regulation of FcepsilonRIalpha expression on human basophils by IgE antibody.

In vivo studies suggested the possibility of an IgE-dependent regulation of high-affinity (FcepsilonRI) IgE receptor expression on basophils. The current studies extend these observations to in vitro cultures of human basophils. Incubation of basophils for 3 to 4 weeks resulted in a slow dissociation of IgE antibody, during which time FcepsilonRI expression decreased, as measured by flow cytometry using the anti-FcepsilonRIalpha monoclonal antibody, 22E7, or by measuring FcRIalpha mass by Western blotting of whole-cell lysates. Culture of basophils with IgE resulted in upregulation of FcepsilonRIalpha expression by both flow cytometry and Western blotting of whole-cell lysates. Upregulation followed a linear time course during 2 weeks of culture. The relative increase in FcepsilonRIalpha density depended on the starting density; with starting densities of FcepsilonRIalpha of 10,000 to 170,000 per basophil, the upregulation varied 20- to 1.1-fold, respectively. Upregulation occurred in high-purity basophils, was not influenced by IgG at concentrations up to 1 mg/mL, and was inhibited by dimeric IgE. Heat-inactivated IgE was less effective and the monoclonal antibody CGP51901 that prevents IgE binding to FcepsilonRIalpha blocked the ability of IgE to induce upregulation. The dose-response curve for IgE-induced upregulation had an effective concentration50 of 230 ng/mL. Although the receptor through which IgE induces this upregulation is not yet known, several characteristics suggest that the upregulation is mediated by IgE interacting through FcepsilonRIalpha itself.

Antibodies, Monoclonal↗
PubMed1998

Characteristics of arachidonic acid generation in human basophils: relationship between the effects of inhibitors of secretory phospholipase A2 activity and leukotriene C4 release.

In human basophils, degranulation stimulated by receptor activation or Ca++ ionophores is accompanied by an increase in free arachidonic acid (AA) as determined by gas chromatography negative ion chemical ionization mass spectrometry. Previous studies suggested that there was more than one pool of AA generated during stimulation and indirectly suggested that the leukotriene (LTC4) generated in these reactions was dependent on only one of these pools of AA. Our studies further examined these issues. Preliminary studies demonstrated discordance in the generation of free AA and LTC4 release. Treatment of basophils with triacsin C, a reacylation inhibitor, led to a marked increase in N-formyl-L-methionyl-L-leucyl-L-phenylalanine-(fMLP) stimulated free AA generation with no effect on LTC4 release. Similarly, incubation of basophils with recombinant human secretory phospholipase A2 (sPLA2), before and during fMLP stimulation, led to the generation of extremely high levels of free AA with no effect on LTC4 release. Pretreatment of basophils with anti-14 kDa phospholipase A2 monoclonal antibody (mAb 3F10) inhibited fMLP-induced synthesis of LTC4 but did not attenuate the mass of AA measured nor histamine release. Treating human basophils with zileuton (an inhibitor of 5-lipoxygenase) inhibited the stimulated synthesis of LTC4 and in combination with triacsin C increased the amount of observable AA by an amount approximately equal to the loss in LTC4 mass. Monoclonal antibody 3F10 blocked only the enhanced AA production caused by the combination of zileuton and triacsin C. Monoclonal antibody 3F10 did not inhibit the increases in free AA produced by pretreatment with triacsin C alone. These findings were supported by experiments using another relatively specific inhibitor of sPLA2, SB 203347. In all respects, SB 203347 mimicked the addition of mAb 3F10. Taken together, these data indicate that not all pools of AA are well used for LTC4 synthesis. These experiments also suggest that LTC4 synthesis in human basophils stimulated with fMLP depends on a SB 203347- and monoclonal antibody 3F10-inhibitable deacylation activity, presumably a sPLA2 acting at or near the cell surface. Furthermore, under normal conditions, this pool of AA is not observable because it is efficiently coupled to 5-lipoxygenase. Other deacylating enzymes, which do not supply AA for 5-lipoxygenase metabolism, also appear to be activated by fMLP and these other enzymes appear responsible for the net free AA normally observed after stimulation.

Arachidonate 5-Lipoxygenase↗
PubMed2005

Lysis with Saponin improves detection of the response through CD203c and CD63 in the basophil activation test after crosslinking of the high affinity IgE receptor FcepsilonRI.

BACKGROUND: The basophil activation test (BAT), in which translocation of markers to the surface of blood basophils is measured in response to allergen by flow cytometry, is a rapid assay that is gaining popularity. Two markers are currently being evaluated for the BAT; CD63 and the lineage-specific CD203c. In a recent report, detection of CD203c after lysis with Saponin was shown to be superior to detection of CD63 after lysis with formic acid. We wanted to compare a) lysis with formic acid and lysis with Saponin, b) the response through CD203c and CD63, and c) the definition 10% activated cells above background with the probability binning metric T(chi) > 4, on sets of data generated with blood basophils stimulated with varying concentrations of anti-FcepsilonRI antibody. METHODS: Blood from volunteers was incubated with serial logarithmic dilutions of anti-FcepsilonRI and subsequently with antibodies to CD203c PE and CD63 FITC. Sets of samples set up in parallel were lysed with either Saponin based Whole Blood Lysing reagent or with formic acid based Immunoprep/Q-prep. Samples were acquired on a FACS Calibur, but were compensated and analysed offline. Responders were defined as persons who had 10% or more activated basophils above background, or a T(chi) > 4, for two consecutive dilutions of anti-FcepsilonRI antibody. RESULTS: More basophils (median 1164 vs. median 397) and better discrimination of upregulated CD203c and CD63 amongst responders were obtained after lysis with Saponin than after lysis with formic acid. We suggest that CD203c may be a more sensitive marker for the BAT than CD63, as 6/11 responders were found with CD203c, compared with 3/11 with CD63. Most responders (7/11) were identified with probability binning. CONCLUSION: A combination of lysis with Saponin and the markers CD203c and CD63 computed by probability binning may be the most sensitive method of detecting activation of basophils after stimulation through FcepsilonRI.

Journal Article↗
PubMed1978

Cytological changes of the pituitary basophils in rats slowly infused with thyrotropin-releasing hormone (TRH).

Young male rats were slowly infused with synthetic TRH, 1 microgram/hr, for 1, 3, 24, 48 and 72 hr, respectively. In the control rats, the basophils of the pituitaries can be divided, in their cytological properties, into the II- (classical thyrotrophs), II/III-,III, (classical LH-cell), and III/IV-type cell. The typical IV-type cells (classical FSH-cell), however, are scarcely found in the young rats. Following 1-hr infusion of TRH, the II-type cells decrease in number with the advancement of granular release, but morphological changes are not yet concrete on the other types of basophils. The II-type cells are quickly invisible following a 3-hr infusion, while the III- and III/IV-type cells remain without any significant changes. The III- and III/IV-type cells are progressively degranulated after a 24-hr infusion. The diameter of secretory granules is reduced to 100--150 nm. The smallest ones below 50 nm in diameter, are disintegrated to disperse into the ground matrix. After degranutlaion, the III/IV-type cells appear to revert to the polygonal or stellate cells with the identical fine structure with the II-type cells. There is evidence that the thyroidectomy cells may develop from the III/IV-type cells only after a 48-hr infusion. After 72 hr, most basophils are provided with the uniform structure of "reversionary II-type cells". In reference to the high serum TSH concentration and no significant change of pituitary TSH concentration under the same experimental condition (Soji, 1978), the present author conclusively postulates that the degranulation of the III/IV-type cells may mainly reflect the conspicuous elevation of serum TSH concentration. The above morphological results are contradictory a plausible view that TRH acts only upon the thyrotrophs to release TSH. The fact that all the basophils ultimately take an appearance of "reversionary II-type cells" in the gland by the prolonged infusion of TRH may not only suggest the share of responsiveness of all the basophils to TRH, but also support the hypothesis of secretory cycle of the basophils.

Animals↗
PubMed2005

Replication study concerning the effects of homeopathic dilutions of histamine on human basophil degranulation in vitro.

BACKGROUND: Various investigators have observed significant effects of highly diluted histamine on human basophil degranulation in vitro, compared to corresponding water controls. However, active and inactive dilution levels differed in most studies. OBJECTIVE: We aimed to reproduce former studies with flow-cytometry using rigorously controlled experimental conditions to minimise confounding factors. METHODS: In seven independent experiments, basophils of the same human donor were incubated with diluted histamine (up to 10(-34)M) or water controls and activated with anti-IgE antibodies. Basophil activation was determined by using bi-colour flow-cytometry. Experiments were blinded and performed with a randomised arrangement of the solutions on microtiter-plates. RESULTS: Histamine at the dilutions 10(-2)M and 10(-22)M was associated with a significant inhibition of basophil degranulation (p=0.018, Wilcoxon signed rank test) of 23.1% and 5.7%, respectively, if compared to "diluted" water treated in an identical manner. However, if all controls were pooled, only histamine 10(-2)M had a significant effect. Significant effects were seen for row numbers of the microtiter plates. CONCLUSION: We were not able to confirm the previously reported large effects of homeopathic histamine dilutions on basophil function of the examined donor. Seemingly, minor variables of the experimental set up can lead to significant differences of the results if not properly controlled.

Basophil Degranulation Test↗
PubMed2006

Initial Fc epsilon RI-mediated signal strength plays a key role in regulating basophil signaling and deactivation.

BACKGROUND: Mediator releases after high-affinity IgE receptor (Fc(epsilon)RI) cross-linking in basophils and mast cells crucially govern the symptoms of allergic disease and amplify underlying T(H)2-type responses. Interestingly, the dose-response curve for Fc(epsilon)RI activation is bell-shaped, with supraoptimal stimulation leading to reduced mediator release. OBJECTIVE: We sought to characterize the mechanisms responsible for this control of Fc(epsilon)RI-triggered basophil activation. METHODS: Human basophils were purified by means of Ficoll density centrifugation, elutriation, and negative selection with immunomagnetic beads. Various intracellular signal protein activities were assessed by means of Western blotting, and mediator releases were analyzed either spectrofluorometrically (histamine) or by means of ELISA (IL-4 and IL-13). RESULTS: Supraoptimal anti-IgE concentrations led to lower mediator release than optimal concentrations but simultaneously to considerably faster histamine release kinetics. In parallel, basophil signaling proteins (Syk, p38 mitogen-activated protein kinase, and extracellular signal-regulated kinases 1 and 2) were more rapidly phosphorylated at higher anti-IgE concentrations but more transiently activated in the supraoptimal range. This endogenous regulation most likely involved src homology 2 domain-containing inositol 5' phosphatase (SHIP), which was highly phosphorylated after supraoptimal anti-IgE triggering compared with lower stimulus concentrations. Conversely, N-formyl-methionyl-leucyl-phenylalanine-stimulated basophils failed to phosphorylate SHIP in the supraoptimal concentration range and did not display a bell-shaped dose-response curve. CONCLUSION: The kinetics of IgE-mediated signaling and mediator release in primary human Fc(epsilon)RI(+) cells varies substantially according to the magnitude of stimulation, and SHIP most likely plays an important role in terminating these events. CLINICAL IMPLICATIONS: The speed of allergic symptom generation depends on the degree of IgE receptor triggering, which is downregulated by SHIP, a potential target for allergy therapy.

Basophil Degranulation Test↗
PubMed2001

Expression and modulation of FcepsilonRIalpha and FcepsilonRIbeta in human blood basophils.

BACKGROUND: The IgE receptor (FcepsilonRI) may exist as a tetramer (alphabetagamma2) or a trimer (alphagamma2) because FcepsilonRIbeta is dispensable for membrane expression of FcepsilonRIalpha. FcepsilonRIbeta amplifies signaling of FcepsilonRI so that regulation of FcepsilonRIalpha:beta stoichiometry would affect cellular responsiveness. OBJECTIVE: We examined basophils from a variety of donors for differences in their expression of FcepsilonRIalpha and FcepsilonRIbeta protein. METHODS: Enriched blood basophils were assessed at baseline and after IL-3 culture for FcepsilonRIalpha and FcepsilonRIbeta protein by Western blotting, surface FcepsilonRIalpha by flow cytometry, and FcepsilonRIbeta mRNA by real-time PCR. Basophil functional response was measured by allergen-triggered histamine release. RESULTS: For the FcepsilonRIalpha subunit, 2 protein bands with molecular weights of 50 kd and 60 kd were identified by Western blots. The 60-kd band correlated to surface-expressed FcepsilonRIalpha detected by flow cytometry (Spearman R = 0.78, P <.01). Surface FcepsilonRIalpha also correlated with FcepsilonRIbeta protein (Spearman R = 0.92, P <.01). FcepsilonRIbeta protein levels increased disproportionately with higher surface FcepsilonRIalpha expression. The ratio of FcepsilonRIbeta to FcepsilonRIalpha varied 10-fold among donors and correlated with surface FcepsilonRIalpha. Basophil 50-kd alpha protein levels were similar despite a 10-fold range in surface FcepsilonRIalpha expression, implying stores of this protein such as those found in eosinophils. Unlike eosinophils, the basophil 50-kd protein was lost with culture and was absent from supernatants. Levels of beta protein and mRNA were enhanced by IL-3 culture, whereas FcepsilonRIalpha expression (by flow cytometry and 60 kd) was not. CONCLUSION: These findings demonstrate variable stoichiometry of FcepsilonRIalpha:beta in whole cells and that this stoichiometry can be altered by IL-3 culture. With the assumption that all detected beta protein is surface expressed, these findings suggest a variable stoichiometry for FcepsilonRIalpha:beta that is also related to FcepsilonRIalpha surface expression.

Adult↗
PubMed2006

Anaphylaxis to Gelofusine confirmed by in vitro basophil activation test: a case series.

The plasma expander Gelofusine (succinylated gelatin) is a recognised cause of peri-operative anaphylaxis. Current diagnosis of Gelofusine sensitivity is by skin testing, a procedure that itself carries a risk of allergic reaction. We evaluated the reliability of the in vitro basophil activation test as a diagnostic assay for Gelofusine sensitivity in subjects with a clinical history highly suggestive of Gelofusine allergy. Six patients with peri-operative anaphylaxis clinically attributed to Gelofusine were skin tested to confirm sensitivity. Control subjects included three healthy subjects and five subjects allergic to a neuromuscular blocking drug, all negative on Gelofusine skin testing. Whole blood basophil activation to Gelofusine was analysed by flow cytometry for CD63 surface expression. All of the Gelofusine sensitive patients and one of the control allergic subjects showed positive basophil activation to Gelofusine. In this series of subjects, the basophil activation test for Gelofusine allergy had a sensitivity of 100% and a specificity of 87.5%. Our findings suggest that basophil activation testing is a safe and reliable in vitro assay for prediction or confirmation of Gelofusine sensitivity in patients with high clinical suspicion of Gelofusine-induced anaphylaxis.

Aged↗
PubMed1995

Histamine distribution in human basophil secretory granules undergoing FMLP-stimulated secretion and recovery.

We examined subcellular histamine localizations in purified human basophils that were stimulated to degranulate with FMLP using an ultrastructural enzyme-affinity technique. Basophils were collected at early (0, 20 seconds, 1 minute) and late (10 minutes to 6 hours) time points poststimulation and were prepared for routine ultrastructural and diamine oxidase-gold (DAO-gold) cytochemical analysis. Histamine was present in unaltered cytoplasmic secretory granules (30.77 gold particles per square micrometer; P < .001 compared with background); specificity controls (histamine absorption, diamine oxidase digestion) abrogated granule labeling for histamine. Altered granules in stimulated cells were not significantly labeled for histamine, as compared with background (P = not significant); unaltered granules in the same cells contained more histamine than altered granules (P < .05). During recovery times, spanning 10 minutes to 6 hours, granules again appeared to be electron-dense and contained histamine (33.49/microns2; P = not significant as compared with unaltered granules in 1-minute FMLP-stimulated cells, and P < .05 as compared with altered granules in 1-minute FMLP-stimulated samples). Other structures devoid of histamine in actively secreting cells included extruded granules and intragranular and extruded Charcot-Leyden crystals. Recovering basophils displayed morphologic evidence of material and membrane conservation, granule content condensation, and biosynthesis. Subcellular histamine-rich sites in actively recovering basophils included condensing granules and collections of cytoplasmic vesicles in three locations: beneath the plasma membrane, adjacent to granules, and in the Golgi region. These studies show that unaltered granules of actively releasing human basophils, as well as similar granules that are reconstituted after FMLP-stimulated degranulation, contain histamine, but that altered granules in stimulated cells undergoing degranulation are devoid of histamine. Reconstitution of histamine-rich granules is associated with DAO-gold-positive cytoplasmic vesicles, suggesting transport of histamine derived from either new synthesis, re-uptake of released histamine, or both, to reconstituted granules.

Amine Oxidase (Copper-Containing)↗
PubMed1998

Metal ion-induced toxic histamine release from human basophils and mast cells.

Recent data suggest that distinct metal ions can be released from dental alloys or other biomaterials, and may cause toxic effects on various cells. In this study, the effects of 14 metal ions on histamine release from human blood basophils (n = 4), isolated tissue mast cells (lung n = 8, uterus n = 2, skin n = 1, gingiva n = 1), the basophil cell line KU-812, and the mast cell line HMC-1 were analyzed. Of the 14 metal ions, Ag+ (0.33 mM) and Hg2+ (0.33 mM) were found to induce release of histamine in blood basophils, KU-812, mast cells, and HMC-1. The effects of Ag+ and Hg2+ were dose dependent and were observed within 60 min of incubation. In primary mast cells and basophils, AU3+ (0.33 mM) also induced histamine release, whereas no effects of Au3+ on HMC-1 or KU-812 cells were seen. The other metal ions showed no effects on primary or immortal cells within 60 min. However, Pt4+ (0.33 mM) induced histamine liberation in HMC-1 and lung mast cells after 12 h. The Ag+- and Hg2+-induced rapid release of histamine from HMC-1 was associated with ultrastructural signs of necrosis, but not apoptosis. In contrast, prolonged exposure to Pt4+ (0.33 mM, 14 h) induced apoptotic cell death in HMC-1 cells, as assessed by electron microscopy and DNA analysis. Together, certain metal ions induce distinct cytopathogenic effects in mast cells and basophils. Whereas Ag+, Hg2+, and Au3+ cause direct toxicity, Pt4 causes cell death through induction of apoptosis. Whether such effects contribute to local adverse reactions to metal-containing biomaterials in vivo remains to be determined.

Basophils↗