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Prednisone inhibits the appearance of inflammatory mediators and the influx of eosinophils and basophils associated with the cutaneous late-phase response to allergen.

To better define the effect of systemic glucocorticoids on the cutaneous early and late phase response (LPR), nine atopic subjects were examined in a double-blind cross-over study using skin chambers fixed over denuded skin blisters. A challenge was carried out by placing allergen in the chamber for 60 min in subjects who received either a 3-day pretreatment with 60 mg/day of prednisone or placebo. Skin chamber cell counts and inflammatory mediators (histamine, PGD2, and leukotriene C4 (LTC4)) were measured at hourly intervals for 12 h. Prednisone pretreatment did not alter the immediate skin erythema or release of histamine but ablated the late secondary erythema and rise in histamine. The median histamine values during h 10, 11, and 12 in the placebo and prednisone pretreatment visits were 3.73 and 0.22 ng/ml, respectively (p less than or equal to 0.02). Prednisone did not alter PGD2 production; however, LTC4 production was suppressed during the LPR. The cumulative median LTC4 values during h 7, 8, and 9 were 5.6-fold (p less than or equal to 0.05) more after placebo than after prednisone pretreatment. Prednisone altered cellular traffic more dramatically than it did inflammatory mediators. The influx of eosinophils, which peaked during the 9th and 10th h in placebo-treated patients, was completely blocked by prednisone (p less than or equal to 0.02) for every h from 6 through 12. The influx of basophils, which started during the 9th h and peaked during the 12th h in placebo-treated patients, was suppressed at all time points (p less than or equal to 0.02) in prednisone-treated patients. There was no significant alteration in neutrophil transit into the skin chambers induced by prednisone. We suggest that the selective blockade of eosinophil and basophil influx by prednisone and the associated decrease in inflammatory mediators may contribute to the blockade of the clinical expression of the cutaneous LPR.

Acute-Phase Reaction↗

[The use of the direct basophil degranulation test for the diagnosis of allergy].

Basophil degranulation test (BDT) was used in examinations of patients suffering from allergies. A modified technique was employed with toluidine blue staining and basophil scintillation in a counter chamber. The examinees were divided into three groups. Group 1 (n = 34) consisted of patients suffering from atopic bronchial asthma with suspected sensitization to home dust. Group 2 included 118 patients with a history of allergy to penicillin. Group 3 consisted of 51 patients with drug allergies (streptomycin, vitamin B1, procaine, analgin intolerance). In all the cases specific diagnosis was confirmed by skin and provocation test. 71.4 percent of the patients with sensitization to home dust allergen showed positive results in BDT, as did 68.6 percent of Group 3 subjects. Of the cases with penicillin sensitization confirmed by application test 91.7 percent were positive in BDT, of those with positive intradermal tests, 66.7 percent were BDT-positive, this being probably explained by individual sensitivity of the patients. These results recommend BDT, an informative and simple technique, for wide practice.

Asthma↗

[Basophilic adenoma in combination with a prolapse of adenohypophyseal tissue through the capsule].

During the post-mortem examination of a 15-year patient who had died after the operation connected with incision of commissures a distorted form of the hypophysis was noted. Histological, histochemical and morphometric investigations showed that there was basophilic adenoma of adenohypophysis and prolapse of the tissue (beyond the borders of the adenoma) through a fistula against the background of changes characteristic of chronic diseases in childhood. Oxyphilic shift, moreover, was noted not only in the first zone of the adenohypophysis, but in other zones, as well as with a simultaneous increase in number of basophiles in I--III zones. Prolapse of the tissue, the latter being normal as far as the tumours growth was concerned, through the fistula took place at a considerable distance from the adenoma, which justifies the consideration of this process as a result of hyperplasia not connected with adenoma. Edges of the fistula bordering the prolapse, haemorrhages were to be seen.

Adenoma, Basophil↗

Effect of azelastine on activation and release stages of allergic histamine secretion in rabbit basophils.

In this study the effect of azelastine on the activation (antigen-dependent, Ca(2+)-dependent, Stage I) and release (antigen-independent, Ca(2+)-dependent, Stage II) phases of allergen-induced histamine secretion in rabbit mixed leukocytes (basophils) was investigated. Azelastine (5 microM, 10-min) and diltiazem (5 microM: a Ca2+ antagonist, 10 min) inhibited ragweed extract-induced histamine secretion during the Stage II (release) phase. Theophylline (100 microM), a phosphodiesterase inhibitor, added immediately before antigen challenge, inhibited allergic histamine secretion during the Stage I (activation) phase. The data obtained in this study suggest that diltiazem and azelastine act on the Ca(2+)-dependent and antigen-independent phase (Stage II) of allergic histamine secretion in rabbit basophils.

Animals↗

Effect of an immunostimulatory substance of Klebsiella pneumoniae on inflammatory responses of human granulocytes, basophils and platelets.

RU 41740 (Biostim), a biological response modifier of bacterial origin obtained from the 01.K2 strain of K. pneumoniae, was studied with regard to its effect to modulate the chemiluminescence response, phagocytosis and leukotriene formation from polymorphonuclear leukocytes (PMNs), the histamine release from basophils and the aggregation of human platelets. For histamine release human basophils were analyzed with either the calcium ionophore A23187 or hemolysin producing bacteria. RU 41740 reduced the E. coli K12 pANN5211 induced histamine release whereas the calcium ionophore A23187 induced histamine release was not affected. The studies with regard to adherence and phagocytosis showed a significant increase in phagocytosis for S. aureus in the presence of RU 41740. With regard to the chemiluminescence response of human granulocytes, donor cells were divided into cells with a high and a low response. The chemiluminescence response of high responder cells towards a potent stimulus, e.g. the calcium ionophore A23187, was decreased by RU 41740 whereas weak stimuli (e.g. E. coli K12) were not affected. In contrast to these results the chemiluminescence response of low responder cells to a weak stimulus e.g. E. coli K12 was increased by RU 41740 and the potent stimuli were not affected. RU 41740 also modulates the eicosanoid synthesis of human polymorphonuclear granulocytes. For the calcium ionophore A23187 induced leukotriene production an increase of LTB4 and a decrease of LTC4 by RU 41740 was observed. Furthermore, preincubation of platelets with various concentrations of RU 41740 for 5 min shows an inhibition of platelet aggregation, when an additional stimulation with the calcium ionophore A23187 was carried out.

Adjuvants, Immunologic↗

[Clinical significance of human basophil releasability in bronchial asthma].

Human basophil releasability (HBR) was determined in 50 patients with asthma by modified human basophil degranulation test (HBDT). The result of HBDT was expressed as degranulation index (DI). 1. Anti-IgE-induced HBR: the mean value of DI in extrinsic asthma, intrinsic asthma and control subjects was 50.1 +/- 2.26(%), 32.2 +/- 2.3(%) and 18 +/- 2.74(%) (x +/- sem) respectively. If a DI of 30% or greater was considered as positive, the positive rate in the above three groups would be 96.9%, 60.0% and 20.0% respectively. There was significant difference between any two of the three groups (P less than 0.01). 2. Allergen-induced HBR: HBDT was performed in the asthmatics with sixteen allergens and in the controls with mite allergen. The positive rates were 68% in the patients and 0% in the controls. The coincident rate of the results between HBDT and skin test reached 82.9%. The results showed that anti-IgE-induced HBR could be regarded as an objective parameter for differentiating extrinsic and intrinsic asthma and allergen-induced HBR could be considered as an useful means for in vitro allergen diagnosis of asthma.

Adolescent↗

Characterization of increased K+ permeability associated with the stimulation of receptors for immunoglobulin E on rat basophilic leukemia cells.

Aggregation of immunoglobulin E-receptor complexes on the surface of rat basophilic leukemia cells stimulates an increase in plasma membrane K+ permeability that is monitored as an increase in the rate of efflux of preloaded 86Rb+. A major component of this stimulated 86Rb+ efflux appears to be due to a Ca(2+)-activated K+ channel because it is inhibited by quinidine in parallel with the inhibition of degranulation and membrane potential repolarization, it is blocked by 0.1 mM La3+, and it is dependent on external Ca2+. Depolarization of the plasma membrane by carbonyl cyanide 3-chlorophenylhydrazone inhibits stimulated Ca2+ influx and prevents antigen-induced 86Rb+ efflux, and increased external Ca2+ partially restores 86Rb+ efflux under these conditions. In addition, potentiation of antigen-stimulated Ca2+ influx by pretreatment with cholera toxin increases the initial rate of stimulated 86Rb+ efflux. Another component of antigen-stimulated K+ efflux appears to be mediated by a guanine nucleotide-binding protein because pretreatment of rat basophilic leukemia cells with pertussis toxin decreases the initial rate of antigen-stimulated 86Rb+ efflux to 40% of that for the untreated cells. Stimulated 86Rb+ efflux is also observed when ionomycin is used to increase cytoplasmic Ca2+ and to trigger membrane depolarization. The efflux stimulated by ionomycin is inhibited by quinidine but not by pertussis toxin pretreatment; thus, it appears to occur through the Ca(2+)-activated K+ efflux pathway. It is proposed that these K+ efflux pathways serve to sustain the Ca2+ influx that is necessary for receptor-mediated triggering of cellular degranulation.

Animals↗

[Mast cells and basophilic granulocytes: their significance in allergic diseases].

Mast cells, and to a lesser extent basophilic granulocytes, have long determined models in the field of pathogenetic mechanisms in allergic disease. Unfortunately this has not resulted in adequate therapy for these diseases, and other cell types such as eosinophils and lymphocytes are now thought to influence allergic mechanisms too. However, knowledge of mast cells has increased to such an extent over the last decade that there may be a "revival" of their importance for allergic disease. In many organs in the body heterogeneity of mast cell morphology has been demonstrated. These different types of mast cells may also release different types of mediators. Moreover, mast cells have recently been shown to be capable of releasing cytokines, which may have a very important bearing on the pathogenesis of allergic diseases. Because mast cells and basophils are able to release a wide variety of mediators which may be responsible for many different physiological responses, they remain important cell types for allergy.

Basophils↗

Interdigital skin test for evaluation of delayed hypersensitivity and cutaneous basophil hypersensitivity in young chickens.

A skin test to assess T-cell mediated delayed hypersensitivity (DH) and cutaneous basophil hypersensitivity (CBH) was evaluated in the interdigital skin of young chickens. Three-day-old chickens were sensitized with Mycobacterium tuberculosis, and the DH reaction was elicited in the interdigital skin in 10-, 17-, 24-, and 31-day-old chickens by intradermal injection of tuberculin. Cutaneous basophil hypersensitivity was elicited in the interdigital skin of 10- and 14-day-old chickens by a single intradermal injection of phytohemagglutinin-P (200 micrograms). The effect of immunosuppression on the results of interdigital skin test for DH and for CBH was evaluated in chickens that were treated with dexamethasone daily for 4 days before testing. The DH reaction, as indicated by a significant (P less than 0.01) increase in the mean interdigital skin thickness, was detectable in 10-day-old chickens and was consistently evident in 17-, 24-, and 31-day-old chickens. The DH response in the interdigital skin of 24-day-old chickens was comparable with that elicited in the standard wattle test. The CBH reaction, as indicted by a significant increase (P less than 0.005) in skin thickness, was evident in the interdigital skin of 10- and 14-day-old chickens. Treatment with dexamethasone significantly decreased (P less than 0.01) the DH and CBH reactions. Results of the study indicated that the interdigital skin test may be used to evaluate normal and suppressed cell-mediated DH and CBH reactions in chickens as young as 10 and 14 days old.

Animals↗

Gene expression and production of tumour necrosis factor by a rat basophilic leukaemia cell line (RBL-2H3) with IgE receptor triggering.

This study evaluated the gene expression of tumour necrosis factor (TNF) and the molecular weight of the cytotoxic factor in a subline of a rat basophilic leukaemia cell line, RBL-2H3. After IgE receptor triggering with a specific antigen that was associated with histamine release, cytotoxic activity in the cell lysates and supernatants increased for 2 hr during the culture of RBL-2H3 cells. Furthermore, calcium ionophore A23187 could induce release of histamine and cytotoxic activity from RBL-2H3 cells. However, compound 48/80, lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA) were unable to induce the release of either histamine or cytotoxic activity from the cells. These data suggested that, at least in part, there was a common pathway in histamine release and production of cytotoxic activity. A protein synthesis inhibitor, cycloheximide, did not affect histamine release, but inhibited the induction of cytotoxic activity. This cytotoxic activity from RBL-2H3 cells was completely neutralized by anti-mouse TNF rabbit serum. With Northern blot analysis, mouse TNF cDNA probe could hybridize with RNA isolated from RBL-2H3 cells. TNF mRNA was induced as early as 1 hr after stimulation with specific antigen and decreased by 4 hr. Moreover, the molecular weight (MW) of the released cytotoxic factor from RBL-2H3 cells triggered with IgE receptors was approximately 17,000 by SDS-PAGE, which was compatible to that of TNF. Thus, it is concluded that the gene expression and production of TNF occurred in RBL-2H3 cells after IgE receptor triggering in association with histamine release, suggesting that TNF produced by basophils and mast cells may play an important role in allergic reaction through its wide range of biological activity.

Animals↗

Antigen-induced cross-linking of the IgE receptor leads to an association with the detergent-insoluble membrane skeleton of rat basophilic leukemia (RBL-2H3) cells.

Cross-linking of the IgE receptor on the surface of rat basophilic leukemia cells by multivalent Ag (DNP-BSA) causes a rapid conversion to a detergent-insoluble form. There is a concurrent increase in the amount of filamentous actin associated with the plasma membrane. Both the degree of receptor detergent insolubility and the rise in F-actin content are rapid with a half-maximal response of less than 1 min and can be rapidly reversed by the addition of monovalent Ag (DNP-lysine). These two early steps in the triggering of rat basophilic leukemia cells can be dissociated from each other by pretreatment of the cells with either cytochalasin or sodium azide. These reagents block the increase in F-actin but have no effect on receptor detergent insolubility. This indicates that microfilaments are not responsible for detergent insolubility of the receptor and that it may be the membrane skeleton that is interacting with the complex. This was further confirmed by the finding that cross-linking of the IgE receptors on the surface of purified plasma membranes also leads to detergent insolubility of the receptor. Therefore, all of the components necessary for detergent insolubility of the receptor are present in the plasma membrane, and cytoplasmic components are not needed. These results suggest that detergent insolubility and immobility of the cross-linked receptors are caused by multivalent interaction with the membrane skeleton. Actin filaments may then interact with these receptor-membrane skeletal complexes in order to produce large scale clustering and capping. The membrane skeleton may therefore be acting as an intermediate structure between the cell-surface receptors and microfilaments.

Actin Cytoskeleton↗

[Immunocytochemical similarities between basophilic and chromophobe pituitary adenomas. Light and electron microscopic study of 13 cases (author's transl)].

ACTH, beta MSH, beta LPH, beta endorpSin, leucine and methionine enkephalin activities were tested in four Cushing pituitary basophilic adenomas and nine "chromophobe" adenomas with immunoperoxidase methods. The antisera gave various positive reactions in both of these groups. ACTH, alpha MSH and enkephalins were predominant. The reason why these hormones are clinically silent in "chromophobe" or, even, basophilic adenomas is still to be elucidated.

Adenoma, Basophil↗

[An unusual case of a basophilic variant of chronic myelogenous leukemia].

The described basophilic variant of CML in blastic crisis was characterized by a distinct hyperhistaminemia, a moderate hypocoagulation and an insufficiently effective cytostatic therapy. Cell kinetic was characterized by a marked diminution of the whole marketing index (with H3-thymidine) of blasts and immature basophils with simultaneous prolongation of the generation time in comparison to myeloblasts at the chronic stage of the CML. Aneuploid cell clones prevailed in the bone-marrow.

Adult↗

A chymotrypsin-type serine protease in rat basophilic leukemia cells: evidence for its immunologic identity with atypical mast cell protease.

Activity of a chymotrypsin-type serine protease was found in a subline of rat basophilic leukemia (RBL-2H3) cells. The protease was immunologically cross-reactive with anti-atypical mast cell protease immunoglobulin (Ig) G, and its activity was inhibited in a dose-dependent manner by the antibody. The apparent m.w. of the protease that reacted with the antibody was 25,000, which was identical with that of atypical mast cell protease in rat mucosal mast cells. These results show that the chymotrypsin type serine protease in RBL-2H3 cells is immunologically identical with atypical mast cell protease, which was first purified from rat small intestine. Immunohistochemical studies showed that the protease was located not only in intracytoplasmic granules but also in organelles synthesizing protein, such as cisternae of the rough endoplasmic reticulum, perinuclear spaces, and the Golgi apparatus. However, no immunoreactivity was demonstrated in rat basophils. The activity of the protease increased in the exponential phase of growth of RBL-2H3 cells in which some activity was also detected in the medium, and it decreased in the late stationary phase.

Animals↗

Biochemical and phenotypic characterization of human basophilic cells derived from dispersed fetal liver with murine T cell factors.

Metachromatically granulated cells were generated from human fetal liver stem cells cultured in heterologous mouse conditioned medium rich in interleukin 3. After 2 to 3 wk of culture with biweekly changes of medium and selection of nonadherent cells, all cells present in five cultures had cytoplasmic granules, and 60 to 95% of the cells stained metachromatically with toluidine blue or with alcian blue but not with the safranin counterstain. Ultrastructurally, many granules contained fibrillar material or electron-dense cores with fibrils and vesicular fragments. In addition, the granules of many cells were filled with electron-dense material, which in some cases had a fine structure consisting of concentric whorls or a reticular pattern. Analysis of high-affinity IgE receptors on the cultured cells by flow cytometry demonstrated a unimodal fluorescence pattern, suggesting that most cells were in the basophil or mast cell lineage. The cultured cells lacked the lymphoid cell surface determinants B1, B4, T3, and T11, the myeloid determinants Mo2 and MY9, the natural killer cell determinant 901, and Ia histocompatibility antigens, but expressed the myeloid determinant MY7. The cells contained 52 ng/10(6) cells of histamine and incorporated [35S]sulfate at an average rate of 31,300 cpm/10(6) cells/4 hr into 175,000 m.w. chondroitin sulfate A proteoglycans. Upon activation with 1 microM calcium ionophore A23187, the cultured cells released 53% of their cell-associated histamine and metabolized arachidonic acid to 15.0 ng/10(6) cells of immunoreactive leukotriene C4 equivalents, 0.5 ng/10(6) cells of leukotriene B4, and 3.1 ng/10(6) cells of prostaglandin D2 (means, n = 3). Thus, stem cells present in human fetal liver give rise, as do stem cells in mouse fetal liver, to metachromatically granulated cells when cultured in the presence of mouse interleukin 3. In both species, the cultured cells bear IgE receptors, lack characteristic lymphoid and most myeloid cell surface determinants, and contain histamine and chondroitin sulfate proteoglycans. The human fetal liver-derived cells are similar in morphology and T cell factor dependence to basophil-like cells derived from umbilical cord blood, but are novel in their capacity to generate leukotrienes and prostaglandin D2.

Animals↗

Basophil histamine release induced by a substance from stimulated human platelets.

Platelet activation may occur during immunoglobulin E antibody (IgE)-mediated reactions. In these studies, we confirm that platelet-derived supernatants (PDS) induce histamine release from human mixed leukocytes containing basophils, one of the initial target cells in IgE-mediated reactions. In extending this observation, we have shown that this PDS-induced histamine release is both temperature- and calcium-dependent. Kinetic studies of release induced by PDS indicate that release is more rapid than that associated with IgE-dependent mechanisms. This platelet-derived, histamine-releasing activity is produced by platelet stimulation with collagen (5 micrograms/ml) and acetylglyceryl ether phosphorylcholine (10(-7)), as well as thrombin (1 U/ml). Initial characterization has shown that it is stable to acid and to freeze-thawing but not to boiling for 10 min. In addition, although this histamine-releasing activity is nondialyzable (i.e., greater than 3500 m.w.), it cannot be attributed to platelet factor 4. Thus, platelets, once activated, can produce a soluble substance or substances which can initiate basophil-mediated reactions, further suggesting that platelet activation can enhance allergic and inflammatory reactions.

Antigens↗

[Clinical performance of the basophil degranulation test. Correlation with other diagnostic technics in allergy (II)].

Although the clinical history is the fundamental method for the diagnosis of reaginic diseases, the use of complementary techniques that gears on the detection of the humoral and cellular effector cells intervening in such affectation is necessary. Thus, we can detect the existence of specific IgE. It can be joined to the basophil membrane by means of techniques "in vivo" as in cutaneous tests, or by means of techniques "in vitro" through cellular techniques in which we apply a biochemical pattern (release of histamine) or a morphological one (HBDT). It can also be free in serum by means of the P-K test or "in vitro" by means of techniques of double antibody. We can also determine, by this last technique, the quantity of the total IgE in serum. In the present work, we review the human basophil degranulation test (HBDT) and its correlation with the prick test, the peripheral blood eosinophilia, the total serum IgE quantification by enzymatic technique and the specific IgE detection by isotope method. In our technique the test was considered positive when the degranulation index with known allergen concentration was greater than 30% for pollens and greater than 25% for D. pteronyssinus with a p less than 0.001. As reported in a previous work, we explain that the normal limits are slightly inferior to those used by some other authors, because in our series, the degranulation average of healthy controls is inferior to that reported by those authors which makes us consider the convenience that the normal limits of degranulation index should be established in a particular way by each technique and allergen.(ABSTRACT TRUNCATED AT 250 WORDS)

Basophils↗

[Atypical urticaria disclosing acute basophilic leukemia. Apropos of a case associated with hyperhistaminemia].

Acute basophilic leukaemia (ABL) is a rare disease characterized by high fever, anaemia and haemorrhagic diathesis. Its prognosis is somber and its response to therapy mediocre. Death is mainly due to cerebral or digestive hemorrhage and coronary disease. In a very small number of cases ABL, like all diseases accompanied by an increase in basophils, may be associated with hyperhistaminemia responsible for cutaneous and gastric symptoms.

Acute Disease↗