Detection of <i>Anisakis simplex</i>‐induced basophil activation by flow cytometry

González‐Muñoz, Miguel; Luque, R.; Nauwelaers, Frans; Moneo, Ignacio

doi:10.1002/cyto.b.20070

Cited by 26 publications

(24 citation statements)

References 37 publications

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“…Recently, detection of allergen-induced basophil activation (BAT) by flow cytometry has been proposed as a useful diagnostic technique for allergy. This test uses whole-blood samples and live basophils, detecting activation-associated membrane markers (CD63) of antigen binding to the IgE high-affinity receptor (FcεRI-bound IgE) (105). A significant activation of basophils was found for patients allergic to A. simplex versus controls (M. T. Audicana and N. Longo, unpublished data).…”

Section: A Simplex Allergy In Humans (I) General Aspects the Potenmentioning

confidence: 99%

Anisakis simplex: from Obscure Infectious Worm to Inducer of Immune Hypersensitivity

2008

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SUMMARY Infection of humans with the nematode worm parasite Anisakis simplex was first described in the 1960s in association with the consumption of raw or undercooked fish. During the 1990s it was realized that even the ingestion of dead worms in food fish can cause severe hypersensitivity reactions, that these may be more prevalent than infection itself, and that this outcome could be associated with food preparations previously considered safe. Not only may allergic symptoms arise from infection by the parasites (“gastroallergic anisakiasis”), but true anaphylactic reactions can also occur following exposure to allergens from dead worms by food-borne, airborne, or skin contact routes. This review discusses A. simplex pathogenesis in humans, covering immune hypersensitivity reactions both in the context of a living infection and in terms of exposure to its allergens by other routes. Over the last 20 years, several studies have concentrated on A. simplex antigen characterization and innate as well as adaptive immune response to this parasite. Molecular characterization of Anisakis allergens and isolation of their encoding cDNAs is now an active field of research that should provide improved diagnostic tools in addition to tools with which to enhance our understanding of pathogenesis and controversial aspects of A. simplex allergy. We also discuss the potential relevance of parasite products such as allergens, proteinases, and proteinase inhibitors and the activation of basophils, eosinophils, and mast cells in the induction of A. simplex-related immune hypersensitivity states induced by exposure to the parasite, dead or alive.

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Section: A Simplex Allergy In Humans (I) General Aspects the Potenmentioning

confidence: 99%

Anisakis simplex: from Obscure Infectious Worm to Inducer of Immune Hypersensitivity

2008

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“…Patients blood (100 mL) was incubated as described (Gonzalez-Munoz et al, 2005), with 1 mg=mL of A. simplex crude and mature hake (fresh, boiled, or frozen), red mullet and juvenile hake (weight 150 g) extracts, and with hake stock. To analyze basophil activation, cells were stained with 20 mL CD63(IgG1)-FITC=CD123(IgG1)-PE=HLA-DR(IgG2a)-PerCP reagent mixture (BD Biosciences, Erembodegem, Belgium) for 20 minutes.…”

Section: Basophil Activation Testmentioning

confidence: 99%

Quantification ofAnisakis simplexAllergens in Fresh, Long-Term Frozen, and Cooked Fish Muscle

Rodríguez-Mahillo

González‐Muñoz²,

Heras

et al. 2010

Foodborne Pathogens and Disease

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Fish-borne parasitic zoonoses such as Anisakiasis were once limited to people living in countries where raw or undercooked fish is traditionally consumed. Nowadays, several factors, such as the growing international markets, the improved transportation systems, the population movements, and the expansion of ethnic ways of cooking in developed countries, have increased the population exposed to these parasites. Improved diagnosis technology and a better knowledge of the symptoms by clinicians have increased the Anisakiasis cases worldwide. Dietary recommendations to Anisakis-sensitized patients include the consumption of frozen or well-cooked fish, but these probably do not defend sensitized patients from allergen exposure. The aim of our work was to develop a sensitive and specific method to detect and quantify Anisakis simplex allergens in fish muscle and its derivatives. Protein extraction was made in saline buffer followed by preparation under acid conditions. A. simplex antigens were detected by IgG immunoblot and quantified by dot blot. The allergenic properties of the extracts were assessed by IgE immunoblotting and basophil activation test. We were able to detect less than 1 ppm of A. simplex antigens, among them the allergen Ani s 4, in fish muscle with no cross-reactions and with a recovery rate of 82.5%. A. simplex antigens were detected in hakes and anchovies but not in sardines, red mullets, or shellfish. We detected A. simplex allergens in cooked hakes and also in hake stock. We proved that A. simplex allergens are preserved in long-term frozen storage (-20 degrees C +/- 2 degrees C for 11 months) of parasitized hakes. Basophil activation tests have proven the capability of the A. simplex-positive fish extracts to induce allergic symptoms.

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“…However, few studies applied flow cytometry to diagnose Anisakis allergy, but using samples other than lymphocytes. Gónzalez-Muñoz et al 2005 [33] detected allergeninduced basophil activation (BAT) by flow cytometry, using wholeblood samples and live basophils, detecting activation-associated membrane markers (CD63) of antigen binding to the IgE high-affinity receptor in response to Anisakis simplex crude extract. A significant activation of basophils (98 %) was found for patients allergic to Anisakis simplex versus controls (3 %).…”

Section: Discussionmentioning

confidence: 99%

A Study of Allergic Sensitization to Anisakis Species in Experimental Mice

Diab¹,

Temsahy²,

Elkerdany³

et al. 2011

J Bacteriol Parasitol

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Background: Hypersensitivity to Anisakis species is a worldwide medical problem. The aim of this study was to detect the immunological response in experimental mice, through measurement of Immunoglobulin E (IgE) antibodies in their lymphocytes by flow cytometry, following the ingestion of Anisakis crude antigen.Method: Sixty Swiss albino mice were divided equally into control and experimental groups and each of them were further subdivided into five subgroups, six mice each. The percentage of IgE antibodies was measured in the lymphocytes of their splenic suspensions at zero, 1 st , 3 rd , 5 th and 7 th weeks post inoculation using Fluorescein isothiocyanate (FITC) anti -mouse IgE and were analyzed on a FACS Calibur flow cytometer Becton Dickinson equipped with an argon-ion laser apparatus operating at 488 nm. Results:The percentage of IgE antibodies was enhanced in lymphocytes of animals exposed to Anisakis antigen from the first week, peaking three weeks following initial exposure, starting a decline by week five and decreased more by the seventh week, as compared to the control group. Conclusion:The results obtained from this study proved the efficacy of flow cytometry in detecting the sensitization against Anisakis species through the measurement of IgE antibodies.

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Detection of Anisakis simplex‐induced basophil activation by flow cytometry

Cited by 26 publications

References 37 publications

Anisakis simplex: from Obscure Infectious Worm to Inducer of Immune Hypersensitivity

Anisakis simplex: from Obscure Infectious Worm to Inducer of Immune Hypersensitivity

Quantification ofAnisakis simplexAllergens in Fresh, Long-Term Frozen, and Cooked Fish Muscle

A Study of Allergic Sensitization to Anisakis Species in Experimental Mice

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