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[Value of the basophil degranulation test (BDT) in the follow-up of immunotherapy with hymenoptera venoms].

The basophil degranulation test (BDT) with the plasma of 24 patients undergoing venom immunotherapy was usually negative after only 6 months of treatment. Cellular sensitivity, however, could still be proved by BDT on purified cells, i.e. after removal of the plasma which contained blocking IgG antibodies. In 30% of the cases, we found primary desensitization of the basophils, which means that the IgG antibodies did not have any influence on the test results.

Basophils↗

[Basophil degranulation test (BDT) as a parameter of hyposensitization with Hymenoptera venoms].

In a population of 24 insect sting allergy patients undergoing venom immunotherapy the basophil degranulation test (BDT) in the patient sera ("unwashed" BDT) and with washed leukocytes ("washed" BDT) after incubation with bee and wasp (yellow jacket) venom was performed before and during treatment. Venom specific IgE and IgG antibodies, detected by means of RAST, were also monitored. The "unwashed" BDT usually became negative within 6-9 months of beginning immunotherapy, whereas the IgE-RAST was still clearly positive. This was attributed to the blocking influence of the venom specific IgG antibodies induced by the venom therapy. In fact, at this time the BDT with "washed" blood leukocytes, i.e. after elimination of the serum antibodies, was generally still positive. Only during further immunotherapy did cellular sensitization in the "washed" BDT gradually disappear, whereas the IgE-RAST usually turned out weakly positive. A third of the patients showed simultaneously negative results of "unwashed" and "washed" BDT, independently of venom specific IgE and IgG levels. These findings suggest a specific reactivity change of the blood basophils (cellular desensitization) induced by the immunotherapy. The BDT can be used as an immunological parameter for IgG-monitoring of the course of venom immunotherapy and--in addition to skin tests and IgE-RAST--as a further criterion for deciding to stop venom therapy if it turns negative with the washed cells.

Adolescent↗

Conjunctival basophil hypersensitivity lesions in guinea pigs. Analysis of upper tarsal epithelium.

The upper tarsal conjunctival epithelium was analyzed for inflammatory cell profile and accompanying morphological changes in a guinea pig system with histopathology resembling two human ocular diseases: vernal conjunctivitis and contact lens-associated giant papillary conjunctivitis (GPC). Female Hartley strain guinea pigs were immunized intradermally on day 0 with 200 micrograms keyhole limpet hemocyanin (KLH) and challenged on day 6 with varying doses of KLH by injection beneath the conjunctival epithelium of one lid and phosphate-buffered saline in the contralateral lid. Tissues containing the reaction site were examined by light microscopy. The 50 micrograms dose of KLH elicited the maximal accumulation of basophils and eosinophils. These values were significantly higher than in the PBS-injected control. Injection of KLH, PBS, or insertion of a sterile needle into unsensitized animals, and uninjected tissue served as additional controls. Neutrophils were significantly higher in the epithelium of the traumatized tissue (repeated needle insertions) than in the uninjected control. Basophils and mast cells were rarely found in the epithelium of unsensitized animals. Epithelial thickening, quantified by a Zeiss Videoplan2 Image Analysis system (Zeiss, West Germany), was greatest in the traumatized tissue, followed by the KLH-challenged tissue of sensitized animals. These values were significantly greater than that of the PBS-injected lid or of naive animals, uninjected or KLH-injected. These results indicate that epithelial changes can be induced by both antigen and trauma. Such epithelial changes may have a role in both vernal conjunctivitis and giant papillary conjunctivitis.

Animals↗

Human basophil degranulation test in drug allergy.

We have evaluated the usefulness of HBDT as an in vitro method for the diagnosis of drug allergy. Two hundred and thirty six patients with suspected drug sensitization to penicillin, streptomycin, sulfamides, pyrazolones and A.S.A. were analyzed. Seventy-nine of them were allergic; in 43 cases it was confirmed by in vivo methods. Other patients were diagnosed by clinical history only if they had more than two reactions to the same drug. In order to be included in this group patients with reactions to pyrazolones and A.S.A. had to have tolerated other NSAI, therefore these patients were allergic to one compound only. All patients were considered non-allergic were determined by a negative provocation test. In the group of allergic patients we obtained 63 (79%) positive degranulations and 16 (21%) negative. One hundred and thirty two (84%) negative degranulations and 25 (16%) positive were obtained in the group of non-allergic patients. Once having analyzed 10 statistical parameters with each drug, the HBOT appears to be a useful method for these drugs except for streptomycin. In 16 (80%) out of 20 aspirin sensitive asthmatic patients we found that their basophils were degranulated. In 7 patients with urticaria and/or angioedema by A.S.A. and other NSAI the degranulation was negative, confirming the absence of the involvement of basophils in this reactions.

Anti-Inflammatory Agents↗

In vitro IgE production by peripheral blood lymphocytes and rectal mucosal biopsies and antigen-induced basophil degranulation in patients with inflammatory bowel disease.

IgE production by peripheral blood lymphocytes (PBL) and rectal mucosal biopsies in vitro has been examined in patients with ulcerative colitis (UC) and Crohn's disease (CD). The degranulation of peripheral blood basophils to food antigens has also been investigated in these patients. Mitogen-induced IgE production was reduced in patients with UC, but enhanced in PBL cultures from patients with CD. Increased numbers of basophils degranulated in the presence of cows' milk in UC patients, but normal responses occurred in patients with CD. This evidence supports the suggested role of IgE-mediated mechanisms involving mast cell/basophil interactions in the pathogenesis of inflammatory bowel disease.

Antigens↗

[Basophil histamine and peripheral blood serotonin in metal and toxic gas poisoning].

Thirty-six persons in non-occupational exposure to air polluted by sulphur and nitrogen oxides and metallic lead and cadmium dust have been examined. The levels of basophil histamine and whole blood serotonin have been determined. A decrease in the basophil histamine level observed in our studies may be indicative of allergic effects of industrial pollution in that region. Simultaneously, we observed changes in the ratios of biogenic amines responsible for immunological response, i.e. changes in the histamine-serotonin ratio.

Adolescent↗

The effect of short-term immunotherapy with Pollinex on anti-IgE and antigen-induced histamine release from sensitive human basophils.

Anti-IgE and antigen-induced histamine release from basophils isolated from 30 atopic patients sensitive to grass pollen allergens was evaluated. The studies were made before and after short-term immunotherapy with glutaraldehyde-pollen-tyrosine adsorbate (Pollinex). It was shown, that after immunotherapy a significant drop in anti-IgE and specific antigen-induced histamine release from isolated basophils occurs. The investigations confirm that the diminution of the target cell sensitivity and releasability may be one of the effects of specific immunotherapy.

Aldehydes↗

Inflammatory mediators and modulators released in organ culture from rabbit skin lesions produced in vivo by sulfur mustard. I. Quantitative histopathology; PMN, basophil, and mononuclear cell survival; and unbound (serum) protein content.

When applied topically to the skin of rabbits in vivo, sulfur mustard (SM), the vesicant used in World War I, produced a slowly developing inflammatory response, which peaked in size at 1 and 2 days, ulcerated within 3 days, and reepithelialized by 10 days. Histologically, basophils and polymorphonuclear leukocytes (PMNs) were common in both early and late lesions, and the crust over the ulcers was composed of dead epidermal cells, fibrin, and large numbers of PMNs. Healing occurred under the crust by migration of epidermal cells from the margins of the lesions and from the hair follicles. In organ culture, the lesion explants survived well, and reepithelialization even took place. Their excellent survival enabled us to compare the life spans of the infiltrating leukocytes within an inflammatory site. PMNs within the explants began disappearing during the first day of culture, and almost all had disappeared by 3 days. In contrast, over half of the basophils and the mononuclear cells within the explants were still present after 3 days of culture. The 1-, 2-, 3-, 6-, and 10-day (1.0-sq cm) SM lesion biopsies showed a 30-45% increase in weight (when compared with normal skin), presumably due to the extravasation of serum proteins and the fluids retained by them. When the biopsies were organ-cultured for 3 days, the 1-, 2-, and 3-day lesions lost weight, and the 6- and 10-day lesions (and normal skin) gained weight. These weight differences were not due to the amount of unbound protein extractable into the culture fluids, because both the early lesions and the late lesions contained about the same amount of unbound protein. The most likely explanation for these weight differences is that the newly formed ground substances of late lesions absorbed culture fluid, because the ground substance had changed from the sol state of acute inflammation (in which it was extractable) back to its normal gel state (in which it was not extractable). The unbound protein extractable into the culture fluids was mostly of serum origin. This protein averaged 1.9 mg for 1.0 sq cm normal skin explants (with a mean weight of 215 mg), and 6.4 mg for 1-day SM lesions (with a mean weight of 313 mg). Because rabbit serum contains about 60 mg protein/ml, these figures indicate that normal skin contained about 15% (unbound) serum by weight.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

IgE mediated triggering of rat basophil leukemia cells: lack of evidence for serine esterase activation.

Inhibition of mediator release from mast cells and basophils by diisopropylfluorophosphate (DFP) and other organophosphorus compounds known to inhibit serine esterases has in the past led to the hypothesis that immunologic triggering of these cells involves an activatable serine esterase. In this study we have shown that two nonphosphorylating or poorly phosphorylating structural analogs of two potent phosphorylators inhibit release of incorporated serotonin from cultured rat basophil leukemia cells. We conclude that, by itself, inhibition of immunologic mast cell triggering by phosphorylating organophosphorus compounds can no longer be considered evidence for involvement of an activatable serine esterase in mast cell triggering.

Animals↗

Mechanism of action of concanavalin A on human basophils.

Concanavalin A (Con A) activates human basophils to release histamine. This reaction has great similarities to that of antigen-or anti-IgE-induced release. It is characterized by a two stage reaction: the initial stage is the binding of Con A to the cell and the second stage is a secretory process of the cell. Cells desensitized by incubation with excess amounts of Con A or by prolonged incubation in the absence of cations cannot be activated to release with antigen or anti-IgE. When the native (tetravalent) Con A molecule is modified by succinic anhydride to become divalent it is much less active in releasing histamine. There is wide variation in the response of cells from different individuals to Con A. However, a close correlation exists between the histamine release induced by Con A and anti-IgE. Therfore, in genral, cells from allergic donors release more histamine with Con A than those from non-allergic individuals. Evidence is presented to demonstrate that Con A acts by binding to carbohydrate groups on the IgE molecule located on the basophil membrane. On a molar basis, IgE is 4000-fold more active than alpha-methyl-D-mannoside in inhibiting the Con A-induced histamine release. Succinyl Con A blocks the ability of Con A and anti-IgE equally well and to a lesser extent the ability of antigen to activate cells to release histamine.

Allergens↗

Blood histamine levels and eosinophil, basophil counts in urticaria.

The blood histamine levels, basophil and eosinophil counts and the percentage of vacuolated eosinophils were observed in 30 controls and 15 patients of urticaria. There was a definite rise in eosinophil and basophil count during the acute stage of the disease which decreased in the quiescent or the symptom-free stage. The behavior of blood histamine level was similar. The mechanisms involved are discussed.

Adult↗

Basophil kallikrein of anaphylaxis (BK-A)--purification and characterization.

These studies describe the IgE-mediated release of a basophil kallikrein of anaphylaxis (BK-A) that has arginine esterase activity and is inhibited by plasma, DFP, and Trasylol. The interaction of BK-A active fractions from ion exchange (DEAE-Sephacel) and gel filtration (Sepharose 6B) chromatography, with human plasma kininogen generates immunoreactive kinin. The BK-A and kinin-generating activities co-chromatograph on DEAE-Sephacel and Sepharose 6B columns, and the quantity of kinin generated is, in general, proportional to the BK-A activity of the column fractions, suggesting that these two activities are subserved by the same protease. These data suggest that kallikrein-like activity can be generated from human basophils as a direct result of a primary IgE-mediated immune reaction, thus providing a potential link between reactions of immediate hypersensitivity and the plasma and/or tissue kinin-generating systems.

Anaphylaxis↗

[Basophil "degranulation" as an index of the body stress state].

A regular "degranulation" development of basophilic leucocytes in the blood has been established not only in cases of the joint and skin allergic reactions but in acute and chronic experimental hemorrhage, skin burns and acute radiation disease. Basophilic "degranulation" denotes the stress state of the organism as a result of stress situations.

Acute Disease↗

The basophil response in blood and bone marrow of tick-infested guinea pigs.

The concentrations of basophils in blood and bone marrow were monitored in guinea pigs subjected to primary and secondary infestations with larvae of Dermacentor andersoni. In these guinea pigs a significant increase of basophils was evident in peripheral blood. The response following the secondary infestation was more rapid and more intense than that following the primary infestation. Similar changes were also shown to occur in bone marrow. The possible significance of these responses in relation to acquired tick resistance in guinea pigs is discussed.

Animals↗

Complement-induced histamine release from human basophils. II. Mechanism of the histamine release reaction.

The activation of human serum complement by incubation with zymosan generates C5a which releases histamine from autologous basophils. The characteristics of the C5a-induced histamine release were investigated. It is similar to IgE-mediated reactions in requiring Ca++ and in being inhibited by EDTA. However, it has marked differences from IgE-mediated reactions. C5a, at all concentrations, released histamine completely in less than 2 min. The C5a reaction has a narrow pH optimum that antigen-induced release and occurs well at 17 degrees to 37 degreesC but not at 0 degreesC. The optimal reaction temperature is 25 degrees to 30 degrees C. Unlike the antigen-induced release, no two-stage activation with C5a for the release of histamine could be demonstrated. There was additive release between C5a- IgE-mediated reactions. Leukocytes could be desensitized to the C5a-mediated reaction by 1) incubating the cells at 37 degrees C for 45 min, 2) pretreating the leukocytes with activated serum in the presence of EDTA, and 3) adding the activated serum to the leukocytes at 0 degrees C before transferring to the optimal reaction temperatures. Cells desensitized to the complement-induced release have normal reactions to IgE-mediated histamine release. In parallel experiments, cells from allergic donors desensitized for IgE-mediated reactions by incubation with antigen under sub-optimal conditions release histamine normally upon the addition of C5a. The results indicate that histamine release by C5a involves a mechanism of basophil activation that is different from the pathway involved in the IgE-induced reaction.

Antigens↗

The inhibition of cutaneous basophil hypersensitivity reactions by a heterologous anti-guinea pig T cell serum.

Systemic treatment with a heterologous anti-T cell serum of guinea pigs immunized with EA in IFA markedly suppressed CBH reactivity to specific antigen and T cell mitogens, as judged by gross reactivity, histology, and skin histamine. The antiserum produced a marked drop in circulating lymphocytes, mainly at the expense of T cells, as indicated by the ability of surviving lymphocytes to rosette with rabbit RBC. It was postulated that the suppression of CBH reactivity is due to the depletion of T cells, which would have released a factor chemotactic for basophils. The data therefore provide further evidence that cutaneous reactions rich in basophils are primarily dependent on a population of T cells.

Animals↗

Histamine release from cultured human basophils: lack of histamine resynthesis after antigenic release.

Human peripheral basophils can be maintained in cultured for up to 72 hr. These cells retain their functional integrity as judged by total histamine content and by their ability to release histamine by an IgE-mediated reaction in response to a specific antigen challenge. Cells cultured after suboptimal antigenic challenge could be activated to release histamine upon the addition of antigen. In contrast, cells culture after supra-optimal antigenic challenge did not fully recover their ability to release histamine even after 24 hr. With a variety of culture conditions, it was impossible to demonstrate any net synthesis of histamine in cells. Cells cultured after depletion of their stores by reaction with antigen did not show any net histamine formation. The experiments suggest that the basophil in peripheral blood is functionally an end-stage cell and can participate in the histamine release reaction only once.

Antigens↗