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. 2009 Jun;156(3):471-8.
doi: 10.1111/j.1365-2249.2009.03919.x.

The Anisakis simplex Ani s 7 major allergen as an indicator of true Anisakis infections

A M Anadón  1 , F Romarís, M Escalante, E Rodríguez, T Gárate, C Cuéllar, F M Ubeira
Affiliations

The Anisakis simplex Ani s 7 major allergen as an indicator of true Anisakis infections

A M Anadón et al. Clin Exp Immunol. 2009 Jun.

Abstract

Ani s 7 is currently the most important excretory/secretory (ES) Anisakis simplex allergen, as it is the only one recognized by 100% of infected patients. The allergenicity of this molecule is due mainly to the presence of a novel CX(17-25)CX(9-22)CX(8)CX(6) tandem repeat motif not seen in any previously reported protein. In this study we used this allergen as a model to investigate how ES allergens are recognized during Anisakis infections, and the usefulness of a recombinant fragment of Ani s 7 allergen (t-Ani s 7) as a marker of true Anisakis infections. The possible antigenic relationship between native Ani s 7 (nAni s 7) from Anisakis and Pseudoterranova decipens antigens was also investigated. Our results demonstrate that nAni s 7 is secreted and recognized by the immune system of rats only when the larvae are alive (i.e. during the acute phase of infection), and that this molecule is not present in, or is antigenically different from, Pseudoterranova allergens. The t-Ani s 7 polypeptide is a useful target for differentiating immunoglobulin E antibodies induced by true Anisakis infections from those induced by other antigens that may cross-react with Anisakis allergens, including P. decipiens. The results also support the hypothesis that the Ani s 7 major allergen does not participate in maintaining the antigenic stimulus during chronic infections.

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Figures

Fig. 1
Fig. 1
Kinetics of immunoglobulin (Ig)E, IgA and IgM antibodies induced in rats after intraperitoneal inoculation with live (closed circles) or dead (open circles) L3 Anisakis larvae. Enzyme-linked immunosorbent assays were carried out with recombinant Ani s 7 allergen (t-Ani s 7), native Ani s 7 (nAni s 7) or crude extract (CE) as target antigens. Each serum sample was tested in duplicate. Cut-off values are represented by a horizontal dashed line.
Fig. 2
Fig. 2
Kinetics of immunoglobulin (Ig)G (IgG1, IgG2a, IgG2b and IgG2c) antibodies induced in rats after intraperitoneal inoculation with live (closed circles) or dead (open circles) L3 Anisakis larvae. Enzyme-linked immunosorbent assays were obtained with recombinant Ani s 7 allergen (t-Ani s 7), native Ani s 7 (nAni s 7) or crude extract (CE) as target antigens. Each serum sample was tested in duplicate. Cut-off values are represented by a horizontal dashed line.
Fig. 3
Fig. 3
Kinetics of immunoglobulin (Ig)G1, and IgE antibodies induced in rats after intraperitoneal challenge (secondary response) with live (closed circles) or dead (open circles) L3 Anisakis larvae. All the enzyme-linked immunosorbent assays were carried out with recombinant Ani s 7 allergen (t-Ani s 7) as target antigen. Each serum sample was tested in duplicate. Cut-off values are represented by a horizontal dashed line.
Fig. 4
Fig. 4
Analysis of cross-reactivity between crude extract (CE) antigens of Anisakis simplex and Pseudoterranova decipiens. Mice were injected intramuscularly with CE antigens from P. decipiens (crossed bars) or A. simplex (white bars) and their IgG1 serum antibodies were tested (in duplicate) by enzyme-linked immunosorbent assay against P. decipiens CE antigen (a), recombinant Ani s 7 allergen (t-Ani s 7) (b) and A. simplex CE antigen (c). Values are represented as mean optical density plus standard deviation.
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