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At least 991 records · Page 55Linked to original sources

T cell derived IgE binding factors inhibit IgE-specific rosettes on basophilic leukaemic cells and lymphocytes.

T cell derived IgE-binding factors inhibited the formation of IgE-specific rosettes on both rat basophilic leukaemic cells and lymphocytes. This was demonstrated by IgE-binding factors released from either IgE-pulsed T cells or adherent cells as well as from IgE-binding factor producing hybridomas. The different binding factors inhibited the IgE-rosettes to a different degree when tested on basophils and lymphocytes, most likely due to the higher affinity for IgE of the former cell type. The results indicate that rat basophils and mouse T cells may recognize common epitopes on the Fc portion of IgE.

Animals↗

Persistence of early crosslink-dependent signal transduction events in human basophils after desensitization.

Desensitization of human basophils with anti-IgE antibody or antigen induced an increased sensitivity to the phorbol ester TPA, evidenced as 2-5 fold increase in the potency of TPA to induce histamine release. As noted in previous publications the magnitude of the change in sensitivity to TPA was a function of the extent of cell surface IgE crosslinking. Thus, the density of cell surface antigen-specific IgE determined the magnitude of the curve shift and the multivalent antigen, BPO21-HSA was found to produce a greater curve shift than the simpler bivalent hapten, BPO2, in accord with previous studies which demonstrated that BPO2 was a "weak" stimulus compared to BPO21-HSA. Basophils which had been fully desensitized by prior treatment with anti-IgE or antigen in the absence of calcium also displayed the curve shift for at least 24 h after desensitization. However, the change induced by crosslinking which altered the cells' sensitivity to TPA required the continued presence of crosslinks and was therefore not the result of a permanent alteration induced by desensitization. Basophils which were fully desensitized to BPO7-HSA demonstrated the curve shift provided that the antigen-induced crosslinks were maintained: treatment with monovalent hapten, BPO-EACA, rapidly returned the cell response to TPA to control, non-desensitized, levels. Since TPA is thought to act by activation of protein kinase C (PKC) we demonstrated that BPO-HSA induced crosslinking increased PKC activity and that the increased activity persisted unless antigen-induced crosslinks were dissociated by the addition of monovalent hapten.(ABSTRACT TRUNCATED AT 250 WORDS)

Antigen-Antibody Complex↗

Translocations t(15;17) and t(9;14)(q34;q22) in a case of acute promyelocytic leukemia with increased number of basophils.

A patient with a variant form of acute promyelocytic leukemia (M3 variant) associated with an increased number of basophils was found to present a reciprocal translocation, t(9;14)(q34;q22) in addition to t(15;17)(q22;q12). Although several cases of acute nonlymphocytic leukemia with increased bone marrow and peripheral blood basophils have been reported, this is the first case in which both the t(9;14) and basophilia were observed in a patient with M3. Our findings support the hypothesis that 9q34 may be associated with the chromosomal location of genes regulating the production and maturation of basophils.

Adult↗

In vitro inhibition of histamine release from mouse mast cells and human basophils by an anthranilic acid derivative.

Optimal conditions for in vitro histamine release mediated either by Con A (3 micrograms/ml) or by Ionophore A23187 (1 microgram/ml), were established for mouse peritoneal mast cells and human normal basophils. In both systems N-5' exerts potent inhibiting effects on histamine release after short term (1-5 minutes) in vitro preincubation. At concentrations of 1mM for mouse mast cells and 3mM for normal human basophils, N-5' inhibits up to 95% Con A-induced histamine release and more than 50% ionophore-induced histamine release. Such in vitro effects are more potent than DSCG-mediated inhibition, under similar experimental conditions, and are resistant to challenge with exceeding doses of the two releasing agents, particularly in the Con A system. Interestingly, basophils with apparent normal morphology, from a CML patient, were resistant to both challenges.

Animals↗

Nerve growth factor influences IgE-mediated human basophil activation: functional properties and intracellular mechanisms compared with IL-3.

NGF and IL-3 play a unique role in supporting human basophil differentiation and mediator secretion. Their importance in allergic disease is underlined further by studies showing elevated levels of these factors in asthmatics. Here, we compared the abilities of IL-3 and NGF to stimulate basophil histamine, IL-4 or IL-13 release, either directly or in conjunction with IgE-dependent stimulation and assessed the intracellular signals responsible. Our results show that the ability of IL-3 and NGF to enhance IgE-dependent histamine release are similar. Both factors also potentiated IgE-dependent IL-13 secretion to a greater degree than the release of histamine or IL-4. At high concentrations (100 ng/ml), IL-3 and NGF alone were capable of releasing cytokines but little histamine. These abilities of IL-3 and NGF to modulate basophil activation were sensitive to blockade by specific inhibitors of PI 3-kinase, p38 MAPK and PLC, but not PKC, suggesting that their effects are mediated considerably by pathways comparable to IgE-dependent signalling.

Basophils↗

Effect of horse gram lectin (Dolichos biflorus agglutinin) on degranulation of mast cells and basophils of atopic subjects: identification as an allergen.

Horse gram (Dolichos biflorus) is widely consumed in the tropical south Asian countries including rural areas of India. Since D. biflorus agglutinin (DBA) is an important dietary lectin in horse gram, we have studied its effect on the degranulation of mast cells and basophils of atopic subjects. Allergy to horse gram lectin has not been reported so far. Skin prick test (SPT) was performed with 100 microg/mL of DBA. DBA-specific IgE was detected by dot-blot, and ELISA. Histamine release (HR) assay was carried out using leukocytes from non-atopic and atopic subjects, and rat peritoneal exudate cells. Among the atopic group, 10 of 48 subjects (21%) were found to be positive for DBA by SPT, and none were positive in the non-atopic group (n=20). Two subjects out of the ten who tested positive for DBA by SPT were found to be sensitized to DBA as revealed by the presence of specific IgE by ELISA and dot-blot. The HR was found to be 2- to 3-fold higher in DBA-allergic subjects than in non-atopic and atopic subjects. Basophil HR by DBA was found to be similar in both non-atopic and atopic subjects. However, DBA induces activation of mast cells in vivo in a sub-population (21%) of atopic subjects. Two subjects have been identified as having food allergy to horse gram based on the presence of DBA-specific IgE with a positive correlation to basophil HR. This is the first report of food allergy to horse gram, and DBA has been identified as an allergen.

Adolescent↗

Loss of syk kinase during IgE-mediated stimulation of human basophils.

BACKGROUND: Ongoing secretion from human basophils is a balance of activation and deactivation events. Recent studies have focused on downregulatory steps that appear to modify the presence of the activated state of various signaling molecules. We now examine downregulation regulated by mechanisms related to proteasome processing. OBJECTIVE: To determine the long-term effects of FcepsilonRI aggregation on expression of syk kinase. METHODS: Peripheral blood basophils were examined for changes in the expression of syk kinase after stimulation with optimal and suboptimal stimulation. RESULTS: Stimulation results in a 20% loss of syk in 1 hour and an 80% loss of syk in longer incubations (>18 hours). Loss of syk in this time frame can occur at levels of stimulation that do not result in observable mediator release. Loss of syk occurs after stimulation with either anti-IgE antibody or antigen. Activation is shown to result in c-Cbl phosphorylation, and its association with syk and immunoblotting reveals the appearance of a ladder of syk species with molecular weights that are consistent with ubiquitylation of syk. Stimulation in the presence of a proteasome inhibitor such as lactacystin A results in the sustained presence of very high-molecular-weight ubiquitylated species, although it does not alter the presence of the syk ladder. CONCLUSIONS: Although the loss of syk is probably too slow to account for downregulation of ongoing secretion of histamine or leukotriene C4 release, it may lead to longer-term alterations in basophil function that explain characteristics of clinical procedures like rapid drug desensitization.

Basophils↗

Potential food allergens in wine: double-blind, placebo-controlled trial and basophil activation analysis.

OBJECTIVE: Recent Australian and international legislation requires labeling of wines made by using the potentially allergenic food proteins casein, milk, egg white, or isinglass (fish-derived) where "there is a detectable residual processing aid." We investigated whether wines fined using these proteins or non-grape-derived tannins (tree-nut derived) can provoke significant clinical allergic reactions (anaphylaxis) in patients with confirmed immunoglobulin E-mediated relevant food allergy. METHODS: A double-blind, placebo-controlled trial was performed to determine whether allergic reactions followed consumption of Australian commercial wines fined using one or more of the legislation-targeted food proteins. In addition, allergenicity of a larger panel of these wines was evaluated by blood basophil activation. RESULTS: No anaphylaxis was induced by wine consumption. Three mild clinical reactions to protein-fined wine and two mild reactions to unfined wine occurred, but there was no statistically significant difference in reaction parameters between subject groups or between processing aids. No pattern of basophil activation correlated with wine type, processing aid, or subject group. CONCLUSION: Wines fined with egg white, isinglass, or non-grape-derived tannins present an extremely low risk of anaphylaxis to fish-, egg-, or peanut-allergic consumers. Although consumption of milk protein-fined wine did not induce anaphylaxis, there were insufficient subjects to determine statistically whether wines fined with milk proteins present a risk to the very rare milk-allergic consumers. In summary, the observed lack of anaphylaxis and basophil activation induced by wines made using the legislation-targeted food proteins according to good manufacturing practice suggests negligible residual food allergens in these wines.

Adult↗

Peroxisome proliferator-activated receptor ligands negatively regulate the expression of the high-affinity IgE receptor Fc epsilon RI in human basophilic KU812 cells.

The high-affinity IgE receptor Fc epsilon RI is expressed on the cell surface of mast cells and basophils, and plays a central role in IgE-mediated inflammatory reactions. Recently, peroxisome proliferator-activated receptors (PPARs) have been implicated in the anti-inflammatory response. To investigate a possible role for PPAR in human basophils, the effect of PPAR ligands on Fc epsilon RI expression in human basophilic KU812 cells was studied. The PPARalpha ligand, leukotriene B(4), did not affect the cell surface expression of Fc epsilon RI. However, prostaglandin (PG) A(1) and 15-deoxy-Delta(12,14) PGJ(2) (15d-PGJ(2)), which are PPARbeta and gamma ligands, respectively, were both able to decrease Fc epsilon RI expression. Treatment with PGA(1) or 15d-PGJ(2) separately also reduced histamine release from KU812 cells in response to cross-linkage of Fc epsilon RI. In addition, RT-PCR analysis showed that KU812 cells expressed the mRNA for PPARalpha, beta, and gamma, indicating that PPARbeta or gamma may negatively regulate the cell activation via Fc epsilon RI. Cells treated with 15d-PGJ(2) expressed lower levels of Fc epsilon RI alpha and gamma mRNA, and PGA(1) treatment decreased the level of Fc epsilon RI gamma mRNA. These results suggest that the suppression of Fc epsilon RI expression by PPARs may be due to the down-regulation of Fc epsilon RI alpha or gamma mRNA.

Basophils↗

Specific inhibition of phorbol ester and DiC8-induced histamine release from human basophils by the protein kinase C inhibitors, H-7 and H-9.

The release of histamine and other inflammatory mediators from human basophils is triggered by numerous stimuli, including chemical, physical and receptor-mediated activators. Several mechanisms of cell activation including protein kinase C activation have been proposed to operate in these cells. We used phorbol ester and DiC8 to induce histamine release from human basophils and the protein kinase C inhibitors H-7 and H-9 to inhibit this release. Both DiC8 and TPA induced histamine release were inhibited by H-7 (ID 50 = 37 mcM) and H-9 (IC 50 = 20 mcM). However, anti-IgE, fmlp and A23187-induced histamine release were unaffected. In contrast, the calmodulin antagonists W-7 and perphenazine effectively inhibited histamine release by all five stimuli. Therefore, different biochemical pathways appear to be critical for basophil activation depending on the nature of the stimulus used.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

Role of nitric oxide in histamine release from human basophils and rat peritoneal mast cells.

The effects of a range of nitric oxide (NO)-related compounds on histamine release from human basophils and rat peritoneal mast cells were studied. Basal and immunologic histamine releases from human basophils were not affected by N(omega)-nitro-L-arginine, N(omega)-nitro-L-arginine methyl ester, aminoguanidine or methylene blue (all inhibitors of NO production), sodium nitroprusside (an NO donor), L-arginine (a substrate for NO synthase) or D-arginine (the inactive enantiomer of L-arginine). In rat peritoneal mast cells, NO donors such as sodium nitroprusside, sodium nitrite and sodium nitrate, and lipopolysaccharide (an inducer of NO synthase) had little effect on basal histamine release, while 3-morpholino-sydnonimine (SIN-1, an NO donor), L-arginine and D-arginine increased this release by up to threefold. None of the inhibitors of NO production had any striking effect on histamine release induced by anti-rat immunoglobulin E (IgE), compound 48/80, sodium fluoride, phospholipase C, 1,2-dioctanoyl-sn-glycerol or ionophore A23187. However, haemoglobin was found to inhibit histamine release by anti-rat IgE or A23187 by ca. 40%. Alone of the NO donors, low concentrations of L-arginine produced a mild inhibition of histamine release induced by anti-IgE, compound 48/80 and A23187, but not other ligands, while sodium nitroprusside dose-dependently inhibited (by a maximum of ca. 30%) histamine release by anti-rat IgE, sodium fluoride or A23187. Stimulation with a variety of secretagogues or treatment with L-arginine, D-arginine, lipopolysaccharide, SIN-1 or sodium nitroprusside had no effect on NO production. Similarly, L-arginine, D-arginine or sodium nitroprusside did not change intracellular cGMP levels. On the basis of these results, it is suggested that NO does not play a significant role in the modulation of histamine release from human basophils or rat peritoneal mast cells. The effects of L-arginine, D-arginine and sodium nitroprusside may involve mechanisms unrelated to NO.

Animals↗

Histamine immunocytochemistry: a new method for detection of basophils in peripheral blood.

We report that basophils in peripheral blood can be stained using histamine immunocytochemistry. The staining is based on the fixation of leucocytes with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (CDI) and the subsequent incubation of these cells with antisera raised against histamine conjugated to different carrier proteins using CDI. The staining appears to be specific for basophils and stained cells can be examined using both fluorescence microscopy and flow cytometry. In addition, histamine immunocytochemistry can be combined with conventional immunocytochemistry by incubating leucocytes with antibodies to cell surface antigens prior to or following fixation of the cells with CDI. Thus, histamine immunocytochemistry may be a valuable tool in future studies of human basophils.

Alcian Blue↗

Purification of morphologically and functionally intact human basophils to near homogeneity.

Certain studies on basophils require highly purified functionally intact cell preparations. Here, a three-step procedure is described that meets these requirements. The procedure consists of a Ficoll density gradient step, counterflow elutriation and negative selection by magnetic cell sorting (MACS). The mean purity of basophils obtained from 30 donors was 97.6+/-3.96% with a viability of 99.6+/-0.83%. The recovery rate was 49.7+/-15.6%. The cells had a normal morphological appearance as assessed by May-Grünwald-stained cytospins and were functionally intact as shown by their unaltered capacity to release histamine and interleukin 4 (IL-4) following immunological activation. This procedure is a clear improvement over currently available techniques and should facilitate future investigations on basophils.

Basophils↗

Detection of homocytotropic antibody in lambs infested with the louse, Bovicola ovis, using a basophil histamine-release assay.

The utility of a basophil histamine-release assay using washed whole blood cells was examined in lambs and was used to determine if homocytotropic antibody with specificity for Bovicola ovis was produced in response to infestation with the louse. Maximal histamine release in the assay in response to Concanavalin A, anti-ovine IgE monoclonal antibody and, in sensitized lambs, to B. ovis antigen ranged from 18 to 48%. Histamine release from blood cells in response to B. ovis antigen was significantly higher in louse-infested lambs than in louse-naive lambs and was significantly correlated with louse and cockle scores. Passive cutaneous anaphylaxis (PCA) tests were negative with sera obtained from the lambs at the same time as blood for the basophil histamine-release assay. Serum histamine levels also were significantly higher in the louse-infested lambs than in louse-naive lambs and were significantly correlated with louse and cockle scores. The present results support a role for B. ovis-specific homocytotropic antibody in the development of cockle and indicate that the basophil histamine-release assay is more sensitive than the PCA test.

Animals↗

A comparison of the inhibitory effect of cromoline and nedocromil Na on histamine release from airway metachromatic cells and from peripheral basophils.

Metachromatic cells are increased in the airways of asthmatic patients. We obtained metachromatic cells from asthmatic airways using an induced sputum technique. The histamine release following ConA, anti-IgE and anti-IgE + IL3 from those cells were evaluated before and following the addition of cromoline Na and nedocromil Na. Metachromatic cells had a higher rate of spontaneous histamine release when compared to peripheral basophils. Cromoline Na and nedocromil Na inhibited histamine release from triggered metachromatic cells but not from peripheral basophils. It is concluded that airway metachromatic cells from asthmatics behave differently than peripheral basophils.

Adult↗

Massive infiltration of basophilic cells in inflamed tissue after injection of RANTES.

Regulated upon activation normal T expressed and secreted (RANTES) is a new inducible protein member of the human C-C branch of chemokines. RANTES is a potent monocyte and lymphocyte chemoattractant and is a mediator of inflammatory responses. In these studies we found that RANTES 10 ng/50 microl chemoattracts basophilic cells in a dose-dependent manner 4 h after an intradermal injection in rat skin sites, as revealed by optic microscopy. Moreover, in biopsy specimens from rat skin injection sites histamine release was significantly higher (P < 0.05) than in controls (PBS 50 microl) after 4 h from RANTES treatment. The presence of basophilic cells in rat skin injection sites after RANTES-treatment was also confirmed by electron microscopy studies. In addition, histidine decarboxylase (HDC) mRNA was increased in rat skin sites injected with RANTES compared to sites injected with PBS (controls). Our report describes additional biological activities for RANTES, suggesting that this human chemoattractant protein may play a fundamental role in histamine and HDC generation, along with basophilic cell recruitment.

Animals↗

Increases in cytosolic Ca2+ accompany basophil activation by eosinophil granule major basic protein.

The response of basophil cytosolic free Ca2+ concentrations ([Ca2+]i) to stimulation by eosinophil granule major basic protein (MBP) was assessed using digital video microscopy. MBP stimulated an elevation in basophil [Ca2+]i that, in general, developed slowly and increased monotonically. In some experiments, however, the average rise in [Ca2+]i was biphasic, with an initial transient increase followed by a larger and more sustained elevation. At the single cell level MBP stimulation caused frequent, asynchronous oscillations in [Ca2+]i that ranged up to 300 nM in amplitude. Chelation of extracellular Ca2+ selectively blocked the second, sustained rise in [Ca2+]i as well as MBP-induced histamine release. Within individual experiments, the increase in [Ca2+]i paralleled the level of histamine release. These results demonstrate that, unlike the activation of neutrophils or eosinophils, basophil activation by MBP correlates with an elevation in [Ca2+]i. Further, the MBP induced [Ca2+]i response exhibits several features in common with the IgE-mediated [Ca2+]i response.

Basophils↗

[Human basophil degranulation test in the diagnosis and prevention of peranesthetic anaphylactoid complications].

The human basophil degranulation test (HBDT) studies the allergic response of effector cells. In this work, it was used in the diagnosis of anaphylactoid reactions during anaesthesia. Thirteen patients having had an anaphylactoid shock during general anaesthesia were investigated. An HBDT was performed with every suspected drug used during the anaesthesia. Ten patients had a positive HBDT and in five cases the drug involved was suxamethonium. In one case, the IgE-dependency of the accident was asserted by the ability of the plasma to sensitize in vitro alien basophils. These sensitized basophils degranulated in the presence of Althesin and suxamethonium. Passive sensitization was abolished when using plasma heated for 2 h at 56 degrees C. Two patients had a negative HBDT to 11 drugs tested. To assess the validity of these results, and to rule out unspecific degranulation, thirteen control patients who were to undergo general anaesthesia were investigated. HBDT was performed immediately before anaesthesia in a double-blind method with every drug to be used during the anaesthesia. The twelve patients who had a negative HBDT to every drug had an uneventful anaesthesia. One patient had a positive test to thiopentone and displayed bronchospasm and generalized urticaria during the anaesthesia. In conclusion, the HBDT appears to be a reliable test in the diagnosis of anaphylactoid reactions during anaesthesia.

Anaphylaxis↗