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.     

Serological investigations in hymenoptera sting allergy: IgE and haemagglutinating antibodies against bee venom in patients with bee sting allergy, bee keepers and non-allergic blood donors

Specific IgE-antibodies and haemagglutinins to bee venom were determined in fifty-five bee sting allergic patients, fifty-seven bee keepers and fifty-two blood donors without evident allergy to hymenoptera stings. IgE-antibodies were detected by RAST in 70% of allergic patients, 40% of bee keepers and 12% of blood donors. Most bee keepers with detectable IgE-antibodies to bee venom recorded severe local or even general reactions to bee stings. Most blood donors with detectable specific IgE had been stung by hymenoptera in the past. High titres of haemagglutinating antibodies against phospholipase A were found in most bee keepers, occasionally in bee sting allergic patients but only rarely in blood donors. The determination of specific IgE-antibodies to insect venoms by RAST seems to be a valuable method for the diagnosis of hymenoptera sting allergy.     

Comparison of the allergenic properties of bee venom and whole bee body extract

The allergenic properties of bee venom and whole bee body extract were compared by in vivo and in vitro tests. The majority of patients with known bee sting sensitivity had positive intracutaneous skin test reactions with bee venom and had bee venom specific IgE in their sera. Of seventeen patients with positive bee venom skin tests, nine had positive tests with whole bee body extract. Of thirty sera containing elevated levels of bee venom specific IgE obtained from untreated patients, fourteen sera contained whole body specific IgE but in much lower titres. In RAST inhibition experiments using both bee venom and whole bee body extract as coupling antigens, bee venom was a more potent inhibiting antigen than whole body extract. From these experiments we conclude that bee venom is a more potent allergen than whole bee body extract.     

Clinical application of measurements of serum levels of bee venom-specific IgE and IgG

Bee venom-specific IgE and IgG antibodies were measured in the serum of beekeepers, bee-allergic persons, and normal persons infrequently stung without adverse effects. Beekeepers, who are stung frequently and relatively “immune” to bee stings, are characterized by high serum levels of IgG- and low serum levels of IgE-specific antibodies. Bee-allergic individuals have high titers of bee venom-specific IgE and generally low titers of bee venom-specific IgG. Following a bee sting, allergic individuals develop a rising titer of IgE antibodies, accompanied occasionally by a rise in IgG antibodies. Therapy with whole bee body extracts fail to effect IgE or IgG antibody titers. During the course of venom immunotherapy IgG-specific antibodies are stimulated and IgE-specific antibodies continue to decline. These observations suggest that: (1) bee sting allergy is a function of bee venom-specific IgE; (2) bee sting immunity is a function of bee venom-specific IgG; and (3) bee venom is an appropriate therapeutic antigen.     

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.     

An autopsy approach to bee sting-related deaths

Although severe reactions to the sting of the common honey bee (Apis mellifera) are a common problem in Australia, reported deaths are uncommon, with the estimated mortality varying from one to four persons each year. The following study presents the postmortem findings in three cases of bee sting fatality, including one in which no observable sting was found. An autopsy approach to such cases is detailed. Overreporting of bee sting-related deaths may occur due to the inclusion of deaths unrelated to a reaction to bee venom, while under-reporting may be due to unexplained deaths where a history of a bee sting is not available or apparent at autopsy. A classification of bee sting-related deaths is proposed, which would allow more accurate reporting of bee sting-related fatalies. A serum tryptase and specific IgE to bee venom on serum obtained at autopsy can assist in confirming anaphylactic reaction to bee venom as the cause of death, particularly in the absence of observable stings. Although there are limitations to the usefulness of serum tryptase tests in the postmortem situation, it may still be useful to confirm suspected anaphylaxis in autopsy cases with an undetermined cause of death.     

Polymerized soluble venom--human serum albumin

Extensive previous studies have demonstrated that attempts to produce polymers of Hymenoptera venoms for human immunotherapy resulted in insoluble precipitates that could be injected with safety but with very limited immunogenicity in allergic patients. We now report soluble polymers prepared by conjugating bee venom with human serum albumin with glutaraldehyde. The bee venom-albumin polymer (BVAP) preparation was fractionated on Sephacryl S-300 to have a molecular weight range higher than catalase. 125I-labeled bee venom phospholipase A was almost completely incorporated into BVAP. Rabbit antibody responses to bee venom and bee venom phospholipase A were induced by BVAP. Human antisera against bee venom were absorbed by BVAP. No new antigenic determinants on BVAP were present as evidenced by absorption of antisera against BVAP by bee venom and albumin. BVAP has potential immunotherapeutic value in patients with anaphylactic sensitivity to bee venom.     

Clinical and immunological surveys in bee keepers

Two hundred and fifty bee keepers in the South of France, working seasonally, were clinically investigated by means of a questionnaire. Forty-three percent had presented anaphylactic symptoms and 7.0%, toxic reactions when stung by bees. The personal atopic history was found to be significantly (P < 0.01) elevated in bee keepers who experienced anaphylaxis. Total serum IgE and bee venom-specific IgE were titrated in 100 subjects. Total serum IgE was significantly elevated in allergic bee keepers (P= 0.02). Although bee venom-specific IgE were significantly (P < 0.01) higher in allergic bee keepers this parameter cannot discriminate between allergic and non-allergic bee keepers owing to a considerable overlap. Bee venom-specific IgG was assayed in seventy subjects. Their level was significantly (P < 0.001) higher in allergic and non-allergic bee keepers as compared with non-allergic blood donors and non-bee-keeping allergic patients. In both bee keeper groups there was no difference in bee venom-specific IgG titres.     

Distribution of RNA-containing bee viruses in honey bee (<Emphasis Type="Italic">Apis mellifera</Emphasis>) in several regions of Russia

In several regions of Russia, broad distribution of RNA-containing bee viruses is found at apiaries of honey bee Apis mellifera using RT-PCR. Detected RNA-containing bee viruses are transferred simultaneously with invasion of mite Varroa destructor and lead to mass bee mortality that results in economic losses in bee breeding. In samples of Varroa destructor, bee viruses DWV and ABPV are found. A high degree of RNA-containing virus (BQCV, DWV SBV, ABPV, CBPV, and KBV) infection is revealed: on average, at least 50% for bee families with mite infection. In the bee families studied in this work, mixed infection with two to six viruses simultaneously is detected. The amplified fragments of viruses BQCV, DWV, and SBV obtained using RT-PCR are sequenced and registered in GenBank.     

Comparison of Skin Tests and RAST for the Diagnosis of Bee Sting Allergy

Skin tests and estimation of specific IgE-antibodies by the RAST are evaluated as diagnostic procedures in bee sting hypersensitivity with the followiny extracts. (2) With bee venom results of skin tests and RAST correspond in 82%, with bee wholebody extracts in 68%. (3) Both with skin tests and the RAST with bee venom a good discrimination between patients with bee sting hypersensitivity and non-allergic controls is observed. Skin tests with bee venom are somewhat more sensitive than bee venom-RAST. (4) With the careful skin test procedure chosen, no adverse reactions were observed.     

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.     

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     

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.     

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.     

Bee pollen sensitivity in airborne pollen allergic individuals.

BACKGROUND: Physicians who practice alternative medicine often prescribe bee pollen as a food supplement and a treatment for various ailments. OBJECTIVES: To determine the qualitative and quantitative composition of bee pollen and to investigate the cutaneous reactivity of atopic patients to bee pollen extracts. METHODS: The absolute number of pollen grains per gram of bee pollen was calculated, and morphologic identification of the botanical family was performed. Five extracts of bee pollen were prepared for skin prick testing, according to standard methods. Two hundred two volunteers participated in the study; 145 were atopic patients with respiratory allergy. The remaining 57 were healthy volunteers or nonatopic patients and served as a control group. All participants underwent skin prick testing with a standard battery of 6 aeroallergens (olive, grasses mix, Parietaria, mugwort, Dermatophagoides pteronyssinus, and Dermatophagoides farinae) and with all homemade bee pollen extracts. RESULTS: All samples of bee pollen contained Oleaceae pollen in high concentrations. Small amounts of anemophilous pollen (Compositeae, Chenopodiaceae) were detected in various samples. A strong positive correlation was observed between cutaneous reactivity to bee pollen extracts and olive, grasses, and mugwort. CONCLUSIONS: Bee pollen contains a large amount of pollen, which belongs to various allergenic families of plants. Bee pollen retains its allergenic potential as demonstrated by strong cutaneous responses to bee pollen extracts observed in atopic patients in contrast to nonatopic subjects. Regarding pollen allergic individuals, further studies are needed to evaluate the safety of ingesting large amounts of bee pollen.     

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.     

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.     

The nociceptive and anti-nociceptive effects of bee venom injection and therapy: A double-edged sword

Bee venom injection as a therapy, like many other complementary and alternative medicine approaches, has been used for thousands of years to attempt to alleviate a range of diseases including arthritis. More recently, additional theraupeutic goals have been added to the list of diseases making this a critical time to evaluate the evidence for the beneficial and adverse effects of bee venom injection. Although reports of pain reduction (analgesic and antinociceptive) and anti-inflammatory effects of bee venom injection are accumulating in the literature, it is common knowledge that bee venom stings are painful and produce inflammation. In addition, a significant number of studies have been performed in the past decade highlighting that injection of bee venom and components of bee venom produce significant signs of pain or nociception, inflammation and many effects at multiple levels of immediate, acute and prolonged pain processes. This report reviews the extensive new data regarding the deleterious effects of bee venom injection in people and animals, our current understanding of the responsible underlying mechanisms and critical venom components, and provides a critical evaluation of reports of the beneficial effects of bee venom injection in people and animals and the proposed underlying mechanisms. Although further studies are required to make firm conclusions, therapeutic bee venom injection may be beneficial for some patients, but may also be harmful. This report highlights key patterns of results, critical shortcomings, and essential areas requiring further study.     

Hymenoptera sting hypersensitivity: IgE, IgG and haemagglutinating antibodies to bee venom constituents in relation to exposure and clinical reaction to bee stings

High levels of IgE antibodies (Ab) to bee venom constituents were mainly found in patients with bee sting hypersensitivity, whereas the sera of bee keepers usually contained high levels of IgG Ab and haemagglutinins, representing blocking antibody activity. In bee keepers there was a positive correlation between the degree of severity of an adverse reaction to bee stings and IgE Ab, a negative one between the severity of reaction and IgG Ab and haemagglutinins, a positive one between the number of stings per season and IgG Ab and haemagglutinins, and a negative one between the number of stings and IgE Ab. This suggests a mutual dependence of the production of IgE Ab and blocking Ab in a regularly exposed population. No such correlation was found in bee sting-allergic patients, indicating that other factors influence the severity of reaction in this only sporadically exposed group. The values of IgG Ab and haemagglutinins correlated well with each other, with IgG Ab giving a better correlation with the severity of reaction and number of stings than the haemagglutinins.     

IgE and T-cell responses to high-molecular weight allergens from bee venom

BACKGROUND: Bee venom contains multiple allergens with a wide distribution of molecular weight. In contrast with conventional bee venom desensitization, peptide or recombinant allergen immunotherapy may have to take into account patients' individual patterns of humoral or cellular response. OBJECTIVE: To study immunoglobulin (Ig)E and T-cell responses to high-molecular weight bee venom allergens >/= 50 kDa. METHODS: Bee venom proteins were separated by size exclusion chromatography and fractions were characterized by one and two-dimensional gel electrophoresis. IgE antibody binding to bee venom fractions was analysed by immunoblotting and T-cell responses by proliferation assay. RESULTS: Among 38 bee venom-hypersensitive patients, IgE recognition pattern of bee venom allergens varied greatly. IgE bound mainly to phospholipase A2 and furthermore to several proteins >/= 50 kDa (50, 54, 69, 84 and 94 kDa). N-terminal sequences of these proteins showed no homology with known proteins. In addition, peripheral mononuclear cells from patients as well as from nonatopic donors strongly proliferated in response to those proteins. CONCLUSIONS: Although present in low amounts, high-molecular weight allergens from bee venom elicit strong IgE and T-cell responses, and may need to be considered as clinically relevant. Therefore, the development of peptide or recombinant protein-based immunotherapy for bee venom allergy may require careful characterization of such allergens.