Evaluation of severe reactions to sweat bee stings

Thirteen patients with severe reactions to sweat bee stings were evaluated. Eleven patients had had systemic reactions and two, large local reactions. Skin testing and RAST with venom of other Hymenoptera and whole body extract of sweat bees disclosed negative reactions to both skin tests and RAST of all other insects tested in eight, while three showed sensitivity to honey bee. Only one patient had a positive RAST reaction to whole body extract of sweat bee. Whole body extract of sweat bee causes some irritant reaction when skin testing controls and is not a reliable diagnostic agent even though patients were more reactive than controls. Hypersensitivity reactions to sweat bee stings are distinct and not associated with reactions to other stinging insects. Currently, no reliable antigen is commercially available for diagnosing or treating this condition. Sweat bee venom or other venom antigens might be beneficial.     

Allergenic potency of bee antigens measured by RAST inhibition

The potency of various bee antigens including bee venom, several whole bee body extracts and fractions of bee venom was studied using the RAST inhibition method. As compared to whole bee body extract, bee venom was a much more potent inhibitor of both bee venom and whole body RAST, suggesting that venom has a greater capacity to bind specific bee IgE antibodies. Whole body extracts also varied substantially in their inhibiting activity. Phospholipase A and hyaluronidase were the most potent of the bee venom fractions suggesting their potential use as an assay for standardization of insect extracts.     

Enzyme-linked immunosorbent assay of allergen-specific IgG antibodies in bee sting allergic patients hyposensitized with pure bee venom

Summary An ELISA is presented for detection of IgG antibodies to bee venom. By this method, sera of 11 bee sting allergic patients, who were treated with rapid hyposensitization with pure bee venom, were tested. The highest antibody titers were observed after 30 days of treatment, a maximum rise of 7.4±1.5 log 2-titer steps. Pure bee venom is shown to be more potent immunologically than whole body bee extract. Prediction of the clinical success, measured by tolerance to a bee sting challenge, is not yet possible using venom specific IgG determinations.This work was supported by the Deutsche Forschungsgemeinschaft, grant UR 12/2     

Ultra rush bee venom immunotherapy does not reduce cutaneous weal responses to bee venom and codeine phosphate

Background rapid administration of bee venom in cumulative doses exceeding the quantity contained in one bee sting is well tolerated by most of the patients during 3.5 h of ultra-rush bee venom immunotherapy (VIT). The mechanism of this tolerance is unknown. Objective The aim of the study was to verify the hypothesis that either slow mediator depletion of mast cells or blockade of their surface receptor mechanisms by increasing doses of allergen might be the major mechanisms of tolerance induced by ultra-rush VIT. Methods Nine bee venom allergic patients with a history of severe systemic reactions after a bee sting, positive skin tests and bee venom specific serum IgE antibodies were treated as follows: on the first day a cumulative dose of 111 μg was administered over 3.5 h under intensive care conditions. Further injections were given on day 7, day 21 and thereafter at 4 week intervals, Intradermal tests with codeine phosphate (nonspecific mast cell degranulation) and bee venom were performed before the initiation of VIT and 30 min after the last injection on the same day as well as before the subsequent bee venom injections. Results No significant changes of skin reactivity to both codeine phosphate and bee venom were observed on day I (before initiation of VIT and after the last injection on the same day). Conclusions Ultra-rush VIT does not induce mediator depletion or surface receptor blockade in skin mast cells.     

Allergens in bee venom: I. Separation and identification of the major allergens

Honeybee venom was separated into seven fractions by gel filtration on Sephadex G-75. Allergenic activities of these fractions were assessed by the paper disc radioallergosorbent test (RAST) with a panel of sera from 24 individuals who had systemic reactions to bee stings, 7 who had large local reactions, and 10 control subjects who had reactions of 5 cm or less following bee stings. Three fractions were identified by enzyme or direct hemolytic activity. Twenty-nine of 31 sera from patients having either systemic or large local reactions to bee stings were positive when radioallergosorbent tested with whole bee venom; 22 were positive to phospholipase A, and 28 were positive to both fractions 1 and 2. Thirteen sera combined most strongly with fraction 1, 12 sera most strongly with fraction 2, hyaluronidase, and three sera about equally with fractions 1, 2, and 3. Reactions with other fractions were much weaker. Fractions 1 and 2 were potent inhibitors of RAST with whole venom in the sera reacting most strongly with fractions 1 or 2, respectively. Fraction 3, phospholipase A, and commercial bee venom phospholipase A were significantly less potent inhibitors with the sera tested. In the cases in which IgE antibody bindings to fractions 1, 2, and 3 were of similar magnitude, inhibitions of RAST using the variovs fractions both on the discs and as inhibitors demonstrated substantial cross-reactivity between the fractions. These results strongly indicate that by using RAST with human sera from bee sting-sensitive individuals, fraction 1, the high molecular materials, and fraction 2, hyaluronidase, are the major allergens in honeybee venom. Phospholipase A appears to be of secondary importance.     

Increased specificity of diagnostic tests with recombinant major bee venom allergen phospholipase A2

Background In diagnosis of type I allergy recombinant allergens have potential advantages over conventional allergenic extracts, both regarding specificity and reproducibility. Objectives We therefore decided to study honey bee venom (BV) and its major allergen phospholipase A2 (PLA) in native and recombinant form for diagnosis of bee sting allergy. Method We investigated 85 patients with a history of a recent systemic allergic bee sting reaction and positive intracutaneous skin test to BV, and 21 controls with no history of allergic bee sting reaction and negative skin test to BV. Intracutaneous skin tests and determination of specific IgE by ImmunoCAP^R to BV, native PLA (nPLA) and recomhinant PLA (rPLA) were done in all patients and controls. Results In skin testing 84 (99%) of the 85 patients reacted to nPLA and 81 (95%) to rPLA, while none of the 21 controls was positive with nPLA or rPLA. Specific serum IgE to BV could be detected in 82 of the patients (96%), to nPLA in 73 (86%) and to rPLA in 66 (78%). Four (19%) of the controls had a positive CAP to BV, one (4.8%) to nPLA and none to rPLA. Analysis of discordant results in CAP showed, that most patients with specific IgE to BV, but not to nPLA and rPLA, had positive skin tests to both PLA preparations and low levels of BV specific IgE. Patients with specific IgE to nPLA but not to rPLA were usually sensitized to minor allergens of BV which contaminated the commercial nPLA. Conclusions PLA is the major allergen in BV. While diagnostic tests with BV are more sensitive, the specificity of tests with PLA, especially rPLA is clearly increased as compared with BV.     

Hymenoptera venom allergy: analysis of double positivity to honey bee and Vespula venom by estimation of IgE antibodies to species-specific major allergens Api m1 and Ves v5

Background:  In patients with hymenoptera venom allergy diagnostic tests are often positive with honey bee and Vespula venom causing problems in selection of venoms for immunotherapy.
Methods:  100 patients each with allergic reactions to Vespula or honey bee stings and positive i.e. skin tests to the respective venom, were analysed for serum IgE to bee venom, Vespula venom and crossreacting carbohydrate determinants (CCDs) by UNICAP (CAP) and ADVIA Centaur (ADVIA). IgE-antibodies to species specific recombinant major allergens (SSMA) Api m1 for bee venom and Ves v5 for Vespula venom, were determined by ADVIA. 30 history and skin test negative patients served as controls.
Results:  By CAP sensitivity was 1.0 for bee and 0.91 for Vespula venom, by ADVIA 0.99 for bee and 0.91 for Vespula venom. None of the controls were positive with either test. Double positivity was observed in 59% of allergic patients by CAP, in 32% by ADVIA. slgE to Api m1 was detected in 97% of bee and 17% of Vespula venom allergic patients, slgE to Ves v5 in 87% of Vespula and 17% of bee venom allergic patients. slgE to CCDs were present in 37% of all allergic patients and in 56% of those with double positivity and were more frequent in bee than in Vespula venom allergic patients.
Conclusions:  Double positivity of IgE to bee and Vespula venom is often caused by crossreactions, especially to CCDs. IgE to both Api m1 and Ves v5 indicates true double sensitization and immunotherapy with both venoms.     

Clinical and immunological studies of beekeepers

Thirty-four beekeepers were interviewed and their blood assayed for the presence of antibodies reacting with bee venom, bee venom phospholipase A (PLA), and whole bee body extract. Following a bee sting, most beekeepers experienced only minimal local tissue reaction. Their serum contained high levels of total antibodies (primarily IgG) reacting to bee venom and phospholipase A. These antibody titres correlated with the frequency of bee stings. Bee venom and PLA specific IgE antibodies were present in serum of some beekeepers. Beekeepers who had experienced allergic reactions were characterized by low total antibody and high venom specific IgE antibody titres. Bee body IgE antibodies were found in varying degree and did not correlate with levels of venom IgE antibodies. There was no difference in the titres of bee body IgE in the sera of beekeepers with and without systemic reactions. The data suggest that allergic reactions are mediated by venom specific IgE and immunity is at least in part a function of other antibodies, probably primarily IgG.     

Antibodies to purified bee venom proteins and peptides. I. Development of a highly specific RAST for bee venom antigens and its application to bee sting allergy

IgE antibodies to purified proteins and peptides from honeybee venom have been measured by the RAST. Trace amounts (less than 0.1%) of the major venom protein phospholipase A2 (PLA2) grossly distorted the measurement of IgE antibody to the other venom proteins, acid phosphatase (Acid P) and hyaluronidase (HYAL), and overemphasized their importance. Reduction of antigen coupled to the cellulose paper discs, which were used in the assay, diluted out the contaminating PLA2 without apparent loss in sensitivity. The reduction of disc-bound antigen increased the competition between IgE and IgG antibodies but did not affect measurement of IgE antibodies in sera taken from 35 untreated patients who had a history of general allergic reactions to bee stings. In 54% of sera from bee venom--allergic patients, the greatest IgE antibody response was to PLA2. In all, IgE antibodies to PLA2 were present in 91% of these sera. IgE antibodies to Acid P, HYAL, or melittin were present in 60%, 51%, and 31% of sera, respectively, and accounted for the highest level of binding in 17%, 17%, and 6% of these. Only 6% of sera were positive for whole venom but negative for the isolated antigens. A low level of IgE antibody was found to peptide 401 in 6% of sera. No IgE antibodies were found to apamin. While confirming the central role played by PLA2 in bee sting allergy, these results show that other venom components are also important in some patients.     

Studies of the antigenicity and allergenicity of phospholipase A2 of bee venom.

The antigenic and allergenic properties of phospholipase A2 (PLA2) and whole bee venom were compared by measuring the IgG and IgE antibody responses in animals and man. Precipitating antibodies raised in rabbits and reaginic and other antibodies raised in mice reacted about equally with both bee venom and PLA. The majority of human sera containing bee venom-specific IgE also contained PLA-specific IgE, although in somewhat lower titers. Similarly, most human sera with significant amounts of total antibodies reacting with bee venom also had antibodies reacting with PLA. Histamine and SRS-a release from leukocytes of sensitive patients followed challenge with whole bee venom and PLA in the majority of instances. However, mediator release from several patients' cells was obtained with bee venom only. These studies suggest that although PLA is a major allergen and antigen in bee venom, significant exceptions in patients' reactivity may limit its potential diagnostic and therapeutic usefulness.     

Allergy to honey: relation to pollen and honey bee allergy

To identify the allergenic components of honey we studied 22 patients with a history of systemic allergic symptoms following honey ingestion. The group of honey-allergic patients was compared with three control groups: 10 subjects sensitized to artemisia, 10 with honey bee venom allergy and 10 without a history of atopy or bee sting reactions. The allergological tests included skin tests and RAST with three different kinds of Swiss honey (dandelion, forest and rape), pollen of compositae species, celery tuber, extract of bee pharyngeal glands, honey bee venom and bee whole body extract. The results show that 3/4 of honey-allergics are sensitive to dandelion honey and 13 of 22 also to compositae pollen. Nine of the honey allergic patients were sensitized to honey bee venom, 3 also to bee pharyngeal glands and to bee whole body extract. Analysis of diagnostic tests and RAST inhibition studies suggest that besides compositae pollen other allergens, most likely of bee origin are important. In honey allergics primary sensitization may be due either to the honey itself, to airborne compositae pollen or even to cross-reacting bee venom components.     

Observations on the aetiology and natural history of stinging insect sensitivity: application of measurements of venom-specific IgE

Measurements of serum venom specific IgE were used to gain further insight into the aetiology and natural history of stinging insect allergy. Several aspects were studied. (a) Following sting reactions, sequential sera were examined in the absence of venom immunotherapy. There was a marked individual patient variation in the persistence of venom specific IgE. In nine patients, antibody levels dropped to insignificant levels, suggesting the possibility that sensitivity can be lost spontaneously. (b) Details of four patients with anamnestic IgE antibody responses after sting reactions are presented. These responses occurred even when prior antibody titres were negligible, suggesting that patients mentioned above (No. 1) will require careful monitoring. The anamnestic responses are insect specific. (c) Eighteen patients were studied who had typical insect sting anaphylaxis and subsequent insignificant serum levels of venom specific IgE. One third also had negative direct skin tests with venom. These observations suggest that the possibility for a non-IgE pathogenesis for some insect sting reactions may have to be considered. (d) One patient was identified who became sensitized to non-venom protein in whole body insect extracts as the result of whole body extract therapy.     

Anaphylaxis caused by the new ant, Pachycondyla chinensis: Demonstration of specific IgE and IgE-binding components

Background: There have been no reports dealing with the pathogenic mechanism and IgE-binding components in patients with anaphylaxis caused by a sting from Pachycondyla chinensis. Objectives: This study was conducted to observe the clinical features of patients with P chinensis–induced anaphylaxis. The roles of specific (s) IgE and sIgG4 antibodies were evaluated, and IgE-binding components were identified. Methods: Seven patients with P chinensis–induced anaphylaxis and 15 unexposed control subjects were enrolled. P chinensis ants were collected at the patients’ homes, and venom was prepared as P chinensis extract. Five patients complained of bee venom–induced anaphylaxis and had positive sIgE levels to yellow jacket venom, wasp venom, or both as well. Serum sIgE and sIgG4 were detected by means of ELISA. To identify IgE-binding components within P chinensis extracts, 12% SDS-PAGE with immunoblot analysis was applied. Results: All patients had positive skin prick test responses to P chinensis antigen and positive sIgE levels. Five (71%) patients had positive sIgG4 levels. Eight IgE-binding components (58, 46, 3l, 29, 27, 25, 22, and 12 kd) were noted, and the component at 12 kd was the most frequently found allergen (85%). IgE ELISA inhibition tests were performed on 2 groups of sera: one from patients with anaphylaxis induced by both P chinensis and bee venom (group A) and the other from patients with anaphylaxis induced by P chinensis venom alone without bee venom allergy (group B). ELISA inhibition tests with serum from group A showed significant inhibitions with addition of P chinensis extract, partial inhibitions with yellow jacket antigen, and minimal inhibitions with wasp or imported fire ant antigens. However, ELISA inhibition tests with serum from group B showed significant inhibitions with P chinensis antigen but no inhibition with wasp, yellow jacket, or imported fire ant antigens. Conclusions: IgE-mediated reactions contributed to the development of P chinensis–induced anaphylaxis. Eight IgE-binding components and one major allergen (12 kd) were identified. Further studies will be needed to clarify the role of sIgG4 and to identify allergenic relationships with major bee and wasp allergens. (J Allergy Clin Immunol 2001;107:1095-9.)     

Three cases of centipede allergy--analysis of cross reactivity with bee allergy]

BACKGROUND: We experienced three cases of patients who suffered from systemic urticaria and systemic symptoms such as general fatigue and dyspnea which occurred just after the sting of centipede. The result of prick test by centipede venom revealed positive reaction in all three patients, so we diagnosed the symptoms of them as immediate type allergic reaction against centipede venom. And more there have been some reports documented the relationship between centipede allergy and bee allergy, so we added the examination about bee allergy on these three patients. METHODS: The measurement of bee venom specific IgE was practiced on all three patients and intradermal skin test about bee venom was done only on one patient. Moreover we also practiced prick test of centipede venom against another three patients who were diagnosed as having bee allergy. RESULTS: All patients of centipede allergy showed positive reaction for the specific IgE antibody of Wasp and/or Honeybee and one patient who was practiced skin test also reacted positively against Wasp and Yellow Jacket venom. But none of three patients of bee allergy showed positive reaction against centipede venom. CONCLUSION: According to these results we suspected that there lies some relationship between centipede allergy and bee allergy. But it is supposed that only little involvement of centipede allergy was concerned among the patients of bee allergy. Until now there have been only a few reports as to the centipede allergy. But actually we guess it may be the phenomenon that could occur frequently.     

Evaluation of bee sting allergy by skin tests and serum antibody assays.

We studied 55 subjects who had had anaphylactic reactions to bee stings within the previous 3 years. 38 out of 54 tested had IgE antibody to honey bee venom (HBV) as measured by radioallergosorbent test (RAST). On skin testing, 30 out of 34 had a positive test to HBV. Of these, 26 had a positive RAST. A positive skin test to HBV at high dilution or else a high anti-HBV RAST score appeared to identify those who, in a 6-month follow-up period, were at risk of developing further anaphylaxis following bee stings or immunotherapy. Of the two tests, RAST appeared to be the less sensitive. Measurements of IgG antibody to phospholipase A were seldom available for the period immediately preceding an anaphylactic episode and proved to be a poor means of predicting the liability to bee sting anaphylaxis in subsequent months.     

IgE antibodies to bee venom, phospholipase A, melittin and wasp venom

Specific IgE antibodies against bee venom, phospholipase A, melittin and wasp venom have been examined in fifty patients with an unusually severe reaction after bee or wasp sting. Two thirds of the bee venom-sensitive patients also have detectable IgE antibodies to wasp venom. More than 50% of the wasp venom-sensitive patients are also allergic to bee venom. Phospholipase A and melittin IgE antibodies were found, respectively, in two thirds and one third of the bee venom-sensitive cases. Specific IgE antibody determinations by the Radioallergosorbent test play an essential role in the diagnostic work. After a reaction to hymenoptera stings both bee and wasp venom tests are necessary due to the high incidence of a false or incomplete identification of the stinging insect. Melittin, known for its potent pharmacological activity and possibly responsible for most of the side effects in bee venom immunotherapy, can probably not be excluded from therapeutic venom preparations since IgE antibodies to the melittin preparation were detected in one third of the cases.     

Prevalence of precipitating antibodies to different extracts of Aspergillus fumigatus in a North American asthmatic population

This study was carried out to find the prevalence of precipitin reactions in the sera of 200 North American asthmatic subjects. Precipitins were detected by the double diffusion technique using different extracts of Aspergillus fumigatus, including a reference ‘home produced’ extract and five commercial extracts from three different suppliers. In addition, antigenicity of these extracts was assessed by crossed immunoelectrophoresis (XIE). Of the sera, 13.5% reacted to the reference extract and from 2.5 to 12% to the different commercial extracts; 22.5% of the sera reacted to at least one extract. No one serum reacted to all the extracts. Two of fifty-one (4%) non-atopic patients with a negative immediate prick test to A. fumigatus, six of eighty-seven (6.9%) atopic patients with a negative immediate reaction to A. fumigatus, and thirty-seven of sixty two (59%) atopic patients with a positive immediate reaction to A. fumigatus had precipitins to at least one of the extracts used, the skin tests being performed using the A. fumigatus reference extract. The prevalence of precipitin reactions bore a strong correlation with the antigenicity of the extracts by XIE. The same reference extract was also used for specific IgE measurements (Brompton extract, Malo & Paquin, 1979). It was found that patients with precipitins had significantly (P < 0.001) higher specific and total IgE values than patients without precipitins. In the group of patients with positive skin test, those with precipitins had significantly (P<0.05) higher specific IgE values than those without. The authors conclude that different extracts of A, fumigatus should be used to assess the presence of precipitins. The antigenicity of these extracts should also be assayed.     

Insect Allergy

One hundred and seventeen persons all stung by yellow jacket (YJ) and/or bee were examined by means of skin prick test with venom of these insects, skin prick test with 10 inhalant allergens and analyses of total IgE. Specific IgE and IgG against honey bee and YJ venom. Eighty-seven persons had had a systemic reaction. Positive correlations ( P < 0.05) were found between results of skin prick tests and specific IgE against venoms and, for YJ, between the severity of symptoms after sting and the size of skin prick test with the venom. That some of the more severe symptoms could have been caused by non-immunological mechanisms could explain why a significant correlation was present only between the results of the prick test and specific IgE and not between these tests and the clinical symptoms. Specific IgE values against YJ and honey bee venom showed convariation, although no correlation could be demonstrated between the clinical symptoms after stings from these insects, or between skin prick test results using the two different extracts. The severity of the sting reactions was not correlated to age, atopic disposition, amount of total IgE, number of stings during life, or positive skin prick test to inhalant allergens. It is concluded that in insect allergy, specific IgE analysis and skin prick tests are supplementary.     

The use of the radioallergosorbent test in the diagnosis of Hymenoptera anaphylaxis

IgE antibody levels to Hymenoptera (honey bee, hornet, wasp or yellow jacket) venom or venom sac were measured in the serum of ninety-six patients with a history of immediate hypersensitivity reactions to these insect stings. Normal levels of IgE antibody were found in fourty-four of these patients tested. The severity of the systemic reaction was similar in those patients with normal and elevated IgE antibody levels, and the interval from the systemic reaction to antibody determination was also similar. In twelve patients with a definite reaction to honey bee venom, the IgE antibody was elevated in all and correlated with a positive venom skin test in nine out of the twelve. It is not possible to determine if the other patients with normal IgE antibody and a systemic reaction represent a false negative RAST value, loss of sensitivity to the Hymenoptera venom, or a lack of RAST sensitivity with some venom or venom sac antigens. Without the ready availability of venom skin tests to all Hymenoptera antigens or other in vitro tests, a definite assessment of insect anaphylaxis remains in doubt for the patient and physician.     

Anaphylaxis caused by the new ant, Pachycondyla chinensis: demonstration of specific IgE and IgE-binding components

BACKGROUND: There have been no reports dealing with the pathogenic mechanism and IgE-binding components in patients with anaphylaxis caused by a sting from Pachycondyla chinensis. OBJECTIVES: This study was conducted to observe the clinical features of patients with P chinensis -induced anaphylaxis. The roles of specific (s) IgE and sIgG4 antibodies were evaluated, and IgE-binding components were identified. METHODS: Seven patients with P chinensis -induced anaphylaxis and 15 unexposed control subjects were enrolled. P chinensis ants were collected at the patients' homes, and venom was prepared as P chinensis extract. Five patients complained of bee venom-induced anaphylaxis and had positive sIgE levels to yellow jacket venom, wasp venom, or both as well. Serum sIgE and sIgG4 were detected by means of ELISA. To identify IgE-binding components within P chinensis extracts, 12% SDS-PAGE with immunoblot analysis was applied. RESULTS: All patients had positive skin prick test responses to P chinensis antigen and positive sIgE levels. Five (71%) patients had positive sIgG4 levels. Eight IgE-binding components (58, 46, 3l, 29, 27, 25, 22, and 12 kd) were noted, and the component at 12 kd was the most frequently found allergen (85%). IgE ELISA inhibition tests were performed on 2 groups of sera: one from patients with anaphylaxis induced by both P chinensis and bee venom (group A) and the other from patients with anaphylaxis induced by P chinensis venom alone without bee venom allergy (group B). ELISA inhibition tests with serum from group A showed significant inhibitions with addition of P chinensis extract, partial inhibitions with yellow jacket antigen, and minimal inhibitions with wasp or imported fire ant antigens. However, ELISA inhibition tests with serum from group B showed significant inhibitions with P chinensis antigen but no inhibition with wasp, yellow jacket, or imported fire ant antigens. CONCLUSIONS: IgE-mediated reactions contributed to the development of P chinensis -induced anaphylaxis. Eight IgE-binding components and one major allergen (12 kd) were identified. Further studies will be needed to clarify the role of sIgG4 and to identify allergenic relationships with major bee and wasp allergens.