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PubMed2005

Natural history of Hymenoptera venom allergy in Eastern Spain.

INTRODUCTION: The natural history of stings, the clinical reaction of the patient and in vivo and in vitro tests are necessary parameters to assess before initiating Hymenoptera venom immunotherapy. In the decision to initiate immunotherapy with Hymenoptera venom, it is not usual to evaluate the natural history of the disease, which seems to be self-limiting and therefore of variable clinical significance. OBJECTIVES: Our aim was to determine the natural history of Hymenoptera hypersensitivity over 4 consecutive years in a rural Mediterranean population. METHODS: An epidemiological study of Hymenoptera sting reactions and possible sensitivity was carried out in 145 randomly selected subjects out of a rural Mediterranean population of 600. Seventy-two subjects, including those with a history of anaphylaxis, completed the 4-year study. The nature of their clinical reactions, age, sex, history of atopy, profession, family history of reactions to Hymenoptera insects, time elapsed since the last sting, number of stings and specific IgE and IgG were determined (the latter, to the three most important insects in the area: Apis mellifera, Polistes dominulus, and Vespula germanica). RESULTS: Of the 72 subjects, four subjects had systemic reactions (SR), 23 had large local reaction (LLR) and all the others (117) was minor local reactions. None who had experienced an SR had a repeat SR when re-stung over the 4-year study. Of those with LLR, 12 subjects had the same type of reaction and 11 experienced more mild local reactions when re-stung. In the SR and local reaction groups, IgE to honey bee (Hb) increased significantly during the study period, whereas in those with only LLR, specific IgE to wasp (Polistes) decreased. Specific IgG to Polistes and Vespula (wasps) decreased significantly, whereas there was no change in the specific IgG to Hb in any of the groups. The number of stings per year decreased at the end of the study in all groups, but positive-specific IgG was higher in subjects with the greatest number of stings. On multivariate analysis using logistic regression, the existence of specific IgE to bee venom, male sex, being an agricultural worker, age of over 16 years and more than five stings per year are seen as individual or co-factors for developing SR or LLR. CONCLUSIONS: The prevalence of SR was 2.8% in our Mediterranean population. The evolution of IgE and IgG throughout the 4-year study differed depending on the Hymenoptera studied. Adult male agricultural workers are at greater risk of having SR or LLR, especially when they have specific IgE to Hb and suffer from more than five stings every 4 years.

Adult↗

PubMed2005

Characterization of the human T cell response to antigen 5 from Vespula vulgaris (Ves v 5).

BACKGROUND: The T cell reactivity to the major allergen of bee venom, phospholipase A2, has been thoroughly characterized. In contrast, only little is known about the human cellular response to major allergens from wasp venom. OBJECTIVE: To characterize the human T cell response to antigen 5 from Vespula vulgaris, Ves v 5. METHODS: Recombinant Ves v 5 was used to establish allergen-specific T cell lines (TCL) and T cell clones (TCC) from the peripheral blood of vespid-allergic and non-allergic individuals. Ves v 5-specific TCL were mapped for T cell epitopes using overlapping synthetic peptides representing the complete amino acid sequence of Ves v 5. Ves v 5-specific TCC were analysed for antigen-induced secretion of IL-4, IFN-gamma and IL-10. RESULTS: Seventeen distinct T cell epitopes were recognized by allergic individuals among which Ves v 5(181-192) was identified as a dominant T cell epitope. Partially different epitopes were observed in TCL from non-allergic subjects and the dominant epitope Ves v 5(181-192) was not prevalent in these cultures. Ves v 5-specific TCC isolated from allergic individuals did not show the typical T helper type 2 (Th2)-like cytokine profile in response to specific stimulation, i.e. high amounts of IL-4 and low IFN-gamma. TCC from non-allergic individuals showed a Th1-like cytokine pattern. CONCLUSIONS: Our findings provide evidence that the allergic T cell response to Ves v 5 is not Th2-dominated and that different immunogenic sites on this major wasp venom allergen are recognized by allergic and non-allergic individuals.

Allergens↗

PubMed1995

Parthenogenesis-inducing microorganisms in Aphytis (Hymenoptera: Aphelinidae).

Production of males in uniparental lines of two species in the parasitic wasp genus Aphytis was induced by rifampicin, and male sexual functioning was determined. Wolbachia-specific 16S rDNA primers were used in a PCR in order to: (1) assess correlation between thelytokous reproduction and the presence of Wolbachia; (2) detect the loss of Wolbachia DNA in uniparental A. lingnanensis following antibiotic treatments, with or without the presence of a host; and (3) clone and sequence part of the Wolbachia 16S rDNA from the uniparental Aphytis species for phylogenetic studies. Males produced viable sperm that was transferred to the female spermatheca following mating. However, sperm failure to effect egg fertilization resulted in all-male progeny. Wolbachia were found in the two uniparental (A. lingnanensis and A. diaspidis) but not in the two biparental (A. lingnanensis and A. melinus) Aphytis lines tested. They can be detected in wasps up to 7 days following antibiotic treatments, regardless of the presence of host. The 16S rDNA for the symbionts in the two Aphytis species is virtually identical, and is most closely related to the Wolbachia found in Muscidifurax uniraptor (Pteromalidae).

Animals↗

PubMed1996

Comparison of preservation techniques for DNA extraction from hymenopterous insects.

Two species of parasitic wasp, Venturia canescens and Leptomastix dactylopii, were killed and preserved by various methods used for Hymenoptera and in mass-collecting devices. Total genomic DNA was subsequently extracted and a 524 bp fragment of the mitochondrial 16S ribosomal RNA gene amplified by PCR. Results for these techniques were compared with that for fresh material and museum specimens. Material from -80 degrees C, 100% ethanol, air-drying in a desiccator, and critical-point dried from alcohol all yielded good results after short and long-term storage, as did specimens from ethylene glycol but not formalin (the latter two being commonly used in pitfall and flight intercept traps). Specimens killed in ethyl acetate vapour and air-dried yielded very degraded DNA which did not successfully PCR. The use of this killing agent is a likely reason for previous reports of inconsistent results obtained from museum specimens, and the now widespread use of critical-point drying of wasps and other insects from alcohol is advocated as a potential source of DNA from rare taxa.

Acetates↗

PubMed2005

Genetic variability in the three genomes of Nasonia: nuclear, mitochondrial and Wolbachia.

Nasonia consists of three closely related species of parasitoid wasps that are all infected with the endosymbiotic bacteria Wolbachia, a reproductive parasite common in arthropods. This situation presents the opportunity to compare patterns of variation in three associated genomes, Wolbachia and the nuclear and mitochondrial genomes of its host. Furthermore, although Nasonia wasps are emerging as a model for evolutionary and genetic studies, little is known about their genetic variability. Using amplified fragment length polymorphisms (AFLPs), all three species present a relatively high level of nuclear polymorphism and have different patterns of variation, with one of the species, Nasonia giraulti, being divided into two divergent subgroups. In each species, the mitochondrial pattern of variation is different from the nuclear pattern, possibly due to genetic hitchhiking of the mitochondria during (cytoplasmically inherited) Wolbachia sweeps. Mitochondria in Nasonia show a synonymous substitution rate approximately 10-15-fold higher than nuclear genes, probably reflecting an elevated mitochondrial mutation rate that is among the highest found in insects. Finally, all three species are doubly infected with their own strains of Wolbachia, one each from the two major supergroups (A and B). Sequence analysis reveals that each of the three Nasonia species acquired their A and B bacteria independently by horizontal transfer events from other insects with the exception of B type Wolbachia in N. longicornis and N. giraulti, which were acquired prior to speciation and then codiverged with the host. This represents one of the few clear-cut examples of codivergence of Wolbachia during host speciation.

Animals↗

PubMed2004

Genetic diversity and Wolbachia infection of the Drosophila parasitoid Leptopilina clavipes in western Europe.

Wolbachia are maternally transmitted bacteria that alter their arthropod hosts' reproduction in various ways, including parthenogenesis induction (PI). Wolbachia-induced parthenogenesis can have drastic effects on the genetic structure of its host because it potentially reduces populations to clones without genetic exchange. However, Wolbachia-induced parthenogenesis does not inevitably result in a reduction of genetic variation of infected populations vs. uninfected populations, because the parthenogenetic populations are initially derived from uninfected populations and can thus show similar genetic variation. Here we investigate these issues in infected and uninfected populations of the Drosophila parasitoid Leptopilina clavipes in western Europe. Wasps from 19 sites in the Netherlands, France and northern Spain were screened for Wolbachia and analysed using amplified fragment length polymorphism (AFLP) markers. All the populations from the Netherlands and mid-France were infected with the same two strains of Wolbachia, whereas populations from the Pyrenees were not infected. The infected and uninfected populations show identical levels of genetic variation, but have clearly diverged genetically, indicating the presence of a barrier that prevents gene flow. Within the infected wasps two distinct genotypes were found at multiple localities, indicating the coexistence of multiple clones. The conditions promoting clonal coexistence in L. clavipes are discussed.

Animals↗

PubMed1992

Comparison between two different assays for measurements of allergen-specific IgE and skin prick test in the diagnosis of insect venom allergy.

Forty-one patients stung by wasp and 29 by bee were evaluated by comparing the correlation between skin prick test (SPT), Phadebas radioallergosorbent test (P-RAST) and Magic Lite SQ Specific IgE assay (LIA). All patients had had a systemic reaction to insect sting. In the patients stung by bee we found agreement between SPT and P-RAST, and SPT and LIA. Similarly, there were no significant differences in sensitivity between P-RAST and LIA (p > 0.05). In patients stung by wasp, SPT was found to be more sensitive than P-RAST. There were no differences between SPT and LIA or between P-RAST and LIA.

Allergens↗

PubMed1997

Insect-venom allergy in Greek adults.

Relatively few studies have investigated the prevalence of insect-sting allergy and the results of diagnostic procedures in unselected populations. The prevalence of insect-sting reactions and of venom sensitization in Greece is unknown. We report the results from a stratified random sample of 480 subjects (404 men, 76 women), aged 20-60 years. They all belonged to the ground personnel of the Hellenic Air Force. A detailed history particularly focused on the reactions to Hymenoptera stings was taken in all subjects. Intradermal skin tests (concentration: 1 microgram/ml) with three venoms (honeybee, paper wasp, common wasp) were performed. The prevalence of venom sensitization (one or more positive skin tests) was 32.7%. Sensitization appears to be more common (2.69 times) in those living in rural areas than in those living in the capital (Athens). The prevalence of systemic reactions was 3.1% (86.7% of them had positive skin tests). Large local reactions were reported by 4.6% of the subjects (77.3% of them had positive skin tests). Asymptomatic sensitization (positive skin tests to venoms) was observed in 28.7% of subjects with no history of an allergic sting reaction. We concluded that the prevalence of Hymenoptera allergy and venom sensitization in Greece is rather high compared to that of other countries.

Adult↗

PubMed2005

High sensitivity of basophils predicts side-effects in venom immunotherapy.

BACKGROUND: Systemic side-effects of venom immunotherapy (VIT) represent a considerable problem in the treatment of patients allergic to Hymenoptera venom. We examined the hypothesis whether basophil responsiveness might be connected with the adverse reactions to VIT. METHODS: Basophil surface expression of activation marker CD63 induced by different concentrations of honeybee and wasp venom (0.1 and 1 mug/ml) was measured by flow cytometry in 34 patients with history of systemic anaphylactic reactions to Hymenoptera sting just before rush honeybee or wasp VIT. RESULTS: Eleven of 34 patients had systemic anaphylactic reaction (Mueller grades I-III) and one patient a large local reaction to VIT. In those 12 patients, median percentage of activated basophils after stimulation with VIT-specific venom in concentration of 0.1 microg/ml was 99% (range: 17-195) of value reached with stimulation with 1 microg/ml. Side-effects occurred in all patients with 0.1/1 ratios over 92% (eight of 12). In contrast, in 22 patients with no side-effects, the median 0.1/1 ratio was 25% (range: 2-92). These concentration-dependent activation ratios were significantly different between the groups with and without side reactions (P < 0.0001). We also show significant positive correlation of the occurrence/clinical grade of the side-effects with individual ratios of CD63 basophil response (r = 0.73, P < 0.0001). CONCLUSION: The results suggest that increased basophil sensitivity to allergen-specific in vitro stimulation is significantly associated with major side-effects of VIT.

Adolescent↗

PubMed2006

Comparison of basophil activation tests using CD63 or CD203c expression in patients with insect venom allergy.

BACKGROUND: Flow cytometric basophil activation tests have been developed as cellular tests for in vitro diagnosis of IgE-mediated reactions. Different activation markers (CD63 or CD203c) with distinct ways of regulation have been used after stimulation with various allergens. OBJECTIVE: It was the aim of the present study to compare basophil activation tests by measuring both CD63 and CD203c upregulation in patients with insect venom allergy. MATERIALS AND METHODS: 43 patients with a history of insect venom anaphylaxis were examined. A careful allergy history was taken, and skin tests and determination of specific IgE-antibodies were performed. Basophil activation tests (BAT) using CD63 or CD203c expression were done after stimulation with different concentrations of bee and wasp venom extracts. 25 healthy subjects with negative history of insect venom allergy were studied as controls. RESULTS: The CD203c protocol showed a slightly higher sensitivity than the CD63 protocol (97% vs. 89%) with regard to patients' history. The magnitude of basophil response was higher with CD203c in comparison to CD63 for both insect venoms. Specificity was 100% for the CD63 protocol and 89% for the CD203c protocol with regard to controls with negative history and negative RAST. CONCLUSION: These results support the reliability of basophil activation tests using either CD63 or CD203c as cellular tests in the in vitro diagnosis of patients with bee or wasp venom allergy with a slightly higher sensitivity for the CD203c protocol.

Allergens↗

PubMed2005

Assembly and breakdown of Cajal bodies in accessory nuclei of Hymenoptera.

In some species of insects, oocytes have vesicular organelles, termed accessory nuclei (ANs). The ANs form by budding off from the nuclear envelope of the oocyte and are filled with translucent matrix containing dense inclusions. One type of these inclusions contains coilin and small nuclear ribonucleoproteins (snRNPs) and is homologous to Cajal bodies. We describe the early events in the morphogenesis of Cajal bodies in the ANs (ANCBs) of the common wasp, Vespula germanica, and show that they contain survival of motor neurons (SMN) protein. We present evidence that in the wasp, ANCBs form by the gradual accumulation of aggregates composed of SMN and small nuclear RNAs. We also show that ANCBs break down and disperse within the ANs as the ANs, which initially surround the oocyte nucleus, localize to the oocyte cortex. The components of dispersed ANCBs are retained within ANs until the end of oogenesis, which suggests that their function may be required at the onset of embryonic development. Because the morphology and behavior of ANs and their Cajal body-like inclusions are conserved in two other hymenopteran species, these features might be characteristic of all hymenopterans.

Animals↗

PubMed2004

Genome sequence of a polydnavirus: insights into symbiotic virus evolution.

Little is known of the fate of viruses involved in long-term obligatory associations with eukaryotes. For example, many species of parasitoid wasps have symbiotic viruses to manipulate host defenses and to allow development of parasitoid larvae. The complete nucleotide sequence of the DNA enclosed in the virus particles injected by a parasitoid wasp revealed a complex organization, resembling a eukaryote genomic region more than a viral genome. Although endocellular symbiont genomes have undergone a dramatic loss of genes, the evolution of symbiotic viruses appears to be characterized by extensive duplication of virulence genes coding for truncated versions of cellular proteins.

Amino Acid Motifs↗

PubMed2003

Activities of temporin family peptides against the chytrid fungus (Batrachochytrium dendrobatidis) associated with global amphibian declines.

Temporin A and structurally related peptides produced in amphibian dermal granular glands and in wasp venom were tested for growth inhibition of Batrachochytrium dendrobatidis, a pathogen associated with global amphibian declines. Two natural amphibian temporins, a wasp temporin, and six synthetic analogs effectively inhibited growth. Differences in potency due to amino acid substitution suggest that ability to penetrate membranes and form an alpha-helical structure is important for their effectiveness against this pathogen.

Amino Acid Substitution↗

PubMedDate not supplied

Continuous immunotherapy for hymenoptera venom allergy using six month intervals.

BACKGROUND: Specific immunotherapy for hymenoptera venom allergy (venom immunotherapy [VIT]) is safe and effective. The duration of treatment is still open for discussion because there is no reliable routine test to determine the real risk of serious anaphylactic reactions. This prospective study, which spans more than 25 years, was conducted to ensure unlimited protection through continuous VIT. To reduce workload and cost, the maintenance interval was increased to 6 months without increasing the rate of adverse events. Only patients with continuous follow-up by our service were included in this study. PATIENTS AND METHODS: VIT was conducted in 176 patients (125 allergic to wasp venom, 20 to bee venom, 31 to both) over a mean of 7.14 years (1.16-25.49). Total and specific IgE were determined on a regular basis. RESULTS: A total of 162 re-stings were reported. Of these, 154 produced a strictly local reaction, seven produced non-serious reactions and one produced a systemic reaction similar to that produced by the initial sting before VIT. Total and specific IgE diminished during VIT. Substantial adverse effects were rare and never life-threatening. The effects occurred during observation in the ward and were controlled according to the treatment protocol recommended by a German expert consensus meeting on the treatment of anaphylactoid reactions. VIT was terminated in only one patient due to recurrent adverse effects. CONCLUSION: Continuous VIT at 6-monthly maintenance intervals conferred permanent protection in patients allergic to bee and wasp venoms and was found to be a safe and effective alternative to current standard protocols.

Adolescent↗

PubMed2005

[Anaphylaxis to hymenoptera sting: study of 113 patients].

BACKGROUND AND OBJECTIVE: The purpose of this study was to describe the clinical and epidemiological features of anaphylactic reactions to hymenoptera stings, with a case-history analysis according to severity. PATIENTS AND METHOD: We conducted an observational descriptive study of patients aged between 10 and 80 years who suffered a systemic reaction after hymenoptera sting. All of them showed specific serum IgE to venoms from Apis, Vespula and/or Polistes. A questionnaire including history of atopy, past reactions and characteristics of the reaction, was performed by individual interview. Anaphylactic reactions were classified into two levels of severity according to Müller's classification. An analysis of independence was carried out in order to relate each level with several factors: age, gender, atopy, type of previous reactions, area of sting and time sequence. RESULTS: 113 patients were included (63 male; mean age [standard deviation]: 40.1 [15.9] years). Reactions were accounted for bee venom in 10.6% of patients, and wasp in 89.4%. Specific IgE was positive to Vespula in 91.9% of subjects, Polistes in 71.4%, and Apis in 28.7%. Furthermore, 50.4% were sensitive to both Vespula and Polistes. Personal history of atopy was found in 20.3%. Among the 106 patients who reminded previous stings, local large reactions were referred by 35.9% and systemic reactions by 16.5%. Upper limb was the most frequent area of sting (38.9%). Most common symptoms were: pruritus (77.8%), hives (57.5%), edema (54.8%), erythema (52.2%), dizziness (51.3%) and dyspnea (49.5%). Severe reactions occurred in 65.5% of patients. Age, gender, atopy, type of previous reactions, area of sting and restoration time were not significantly associated with severity. Time elapsed to first symptom was proportionally shorter in severe cases (p < 0.05). CONCLUSIONS: There is a high frequency of hypersensitivity to wasp venom (Vespula) in the studied population. Except for immediacy, severity-associated data could not be established.

Adolescent↗

PubMed2006

Diagnostic and subdiagnostic accumulation of mast cells in the bone marrow of patients with anaphylaxis: Monoclonal mast cell activation syndrome.

BACKGROUND: Patients with mastocytosis may suffer from severe hypotension after wasp or bee stings. In these patients, no specific IgE is detectable, but they usually have skin lesions and an elevated serum tryptase level. METHODS: We report on 6 patients who were referred to our department because of severe hypotension following bee or wasp stings without cutaneous lesions. RESULTS: In 3 patients, the baseline serum tryptase level was elevated (26, 36, and 67 ng/ml, respectively), and investigation of their bone marrow revealed systemic mastocytosis (SM). In the remaining 3 patients, serum tryptase levels were <20 ng/ml, and bone marrow histology and tryptase immunohistochemistry did not reveal diagnostic mast cell infiltrates. However, in 1 patient, three minor SM criteria were demonstrable leading to the diagnosis SM, and in the 2nd patient, two minor SM criteria, including an aberrant mast cell phenotype, were found. In the 3rd patient, no minor SM criteria were detected. CONCLUSIONS: All patients with unexplained hypotension after hymenoptera stings should undergo a thorough investigation for major and minor SM criteria regardless of the tryptase level or presence of skin lesions, in order to diagnose or exclude SM or a related subdiagnostic condition (1 or 2 minor SM criteria) tentatively termed monoclonal mast cell activation syndrome.

Anaphylaxis↗

PubMed2004

Insect oviposition induces volatile emission in herbaceous plants that attracts egg parasitoids.

The egg parasitoid Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae) responded to synomones emitted by leguminous plants induced by feeding and oviposition activity of the bug Nezara viridula (L.) (Heteroptera: Pentatomidae). This was shown by laboratory bioassays using a Y-tube olfactometer. Broad bean leaves (Vicia faba L.) damaged by feeding activity of N. viridula and on which host egg mass had been laid produced synomones that attracted T. basalis. By contrast, undamaged leaves or feeding-damaged leaves without eggs did not attract wasp females. French bean plants (Phaseolus vulgaris L.) also emitted attractive synomones when they were damaged by host feeding and carrying egg masses. Thus, release of feeding- and oviposition-induced synomones does not seem to be plant-specific. Synomone production was shown to be a systemically induced plant physiological response to feeding damage and oviposition. Also, parts of the plant that were left undamaged and did not carry host eggs emitted attractive synomones when other parts of the plant were damaged by feeding and carrying eggs. Furthermore, wasps were not attracted by N. viridula egg masses offered alone or combined with damaged broad bean leaves. Thus, the attractiveness of feeding-damaged leaves carrying eggs is due to induction by feeding and oviposition rather than due to a combined effect of attractive volatiles released from eggs and damaged leaves. The production of synomones was influenced by the age of the host egg mass, because feeding-damaged leaves bearing egg masses attracted the parasitoid until the eggs were approximately 72-96 h old but not once the larvae had hatched from the eggs (approximately 120 h old). These results show that annual plants are able to produce synomones as a consequence of feeding and egg mass oviposition by a sucking insect.

Animals↗

PubMed2006

Novel biologically active peptides from the venom of Polistes rothneyi iwatai.

Four novel peptides, polistes-mastoparan-R1, 2, 3, and polistes-protonectin, were isolated from the venom of a paper wasp, Polistes rothneyi iwatai. MALDI-TOF MS analysis of a small amount of the crude venom gave six molecular-related ion peaks. Among them, m/z 1565 was expected to be a novel peptide. Purification of the crude venom by HPLC gave two known kinins, Thr6-bradykinin and Ala-Arg-Thr6-bradykinin, and four novel peptides named polistes-mastoparan-R1, 2, and 3, and polistes-protonectin. Polistes-mastoparan-R1, 2, and 3 (Pm-R) were tetradecapeptides that possess high sequence homology with that of mastoparan. The sequence of polistes-protonectin was similar to that of protonectin isolated from a Brazilian paper wasp. Histamine-releasing activities of Pm-R1, 2, and 3 were more potent than that of mastoparan. Polistes-protonectin exhibited the most potent hemolytic activity in comparison with the four novel peptides and mastoparan.

Amino Acid Sequence↗