Diagnostic use of recombinant Tha p 2 in the allergy to Thaumetopoea pityocampa.

Thaumetopoea pityocampa causes allergies and skin and ocular lesions. No commercial tools are currently available for the clinical diagnosis of this allergy. We aimed to develop an in vitro method for the diagnosis of this allergy to avoid patients undergoing in vivo tests with insect extracts. Recombinant Tha p 2 was produced and used in an ELISA validated with 15 allergic patients. Subsequently, 42 subjects recruited from a random sampling cross-sectional study were analysed. The ELISA sensitivity and specificity were 93.3% and 100%, respectively, for the allergic patients and 71.4% and 95.3%, respectively, for the epidemiological study. The positive ELISA results correlated with the skin prick test areas with the whole body and the setae extracts. Professional exposure and short latency of symptoms onset were risk factors for a positive result in the ELISA. In conclusion, our ELISA is very useful for T. pityocampa allergy diagnosis and for epidemiologic testing.

Anisakis/Ascaris IgE ratio improves specificity for the diagnosis of Anisakis simplex sensitization in travellers and immigrants.

Anisakis simplex is a fish parasite responsible for human infection and is able to induce IgE-mediated reactions with several clinical manifestations. Laboratory diagnosis of Anisakis allergy is based on the detection of specific IgE using parasite whole antigen. Unfortunately, these diagnostic tools detect cross-reactivities with other nematodes and micro-organisms leading to low specificity of the diagnostic tests. The aim of this retrospective study was to assess the diagnostic value of specific IgE to Anisakis for diagnosis of A. simplex-sensitization in native Spanish residents (IMM, n=766) and subjects coming from tropical and sub-tropical geographic areas (TRO, n=233). Since Ascaris is the human parasite most closely related to Anisakis, specific IgE to Ascaris was also determined to assess Anisakis cross-reaction with other nematodes and the diagnostic value of Anisakis/Ascaris IgE ratio for Anisakis allergy was examined. IMM and TRO groups showed similar specific IgE to Anisakis levels, while TRO had higher levels of specific IgE to Ascaris than IMM group (p=0.001). ROC curve analysis determined that an Anisakis specific IgE threshold of 0.71 kU/L yielded 93% and 82% specificities in IMM and TRO groups, respectively. A cut-off value ≥4.4 for Anisakis/Ascaris IgE ratio increased specificity to 95% for samples having IgE to Ascaris ≥0.35. In conclusion, the ratio of specific IgE to Anisakis and Ascaris improved remarkably the specificity and this parameter easily obtained from the commercially available system could be useful in the diagnosis of hypersensitivity to A. simplex.

Different Th1/Th2 responses to Anisakis simplex are related to distinct clinical manifestations in sensitized patients.

Anisakis simplex is a fish parasite capable of inducing inflammatory and allergic reactions in humans who eat raw or undercooked fish. The aim of this study was to characterize the T helper type 1 (Th1)/Th2 immune response to parasite crude (CE) and thermostable (TsE) extracts in A. simplex-sensitized patients. Cytokines were quantified by a multiplex flow cytometric method in short-term whole blood cultures. Higher concentrations of IL-2, IL-4 and IL-5, measured with the CE and TsE, were found in patients than in controls. Patients showing urticaria-angio-oedema or anaphylaxis (UA/A) had higher total and specific IgE levels than those with gastrointestinal symptoms (GI). The UA/A group showed high levels of IL-5 and IL-4 and low expression of IFN-gamma than the GI group. The GI group had significantly higher IFN-gamma/IL4 ratio than the UA/A group. Four patients with severe GI symptoms reporting a delayed skin test reaction had very low values of specific IgE to A. simplex and higher IFN-gamma/IL4 ratios than that observed in other patients belonging to the GI group. This short-term whole blood test can be useful for immune response characterization in Anisakis infection and showed that heated parasite antigens are still capable of inducing cellular immune response in sensitized patients.