A 24 kDa Excretory-Secretory Protein of Anisakis simplex Larvae Could Elicit Allergic Airway Inflammation in Mice

We have reported that a 24 kDa protein (22U homologous; As22U) of Anisakis simplex larvae could elicit several Th2-related chemokine gene expressions in the intestinal epithelial cell line which means that As22U may play a role as an allergen. In order to determine the contribution of As22U to allergic reactions, we treated mice with 6 times intra-nasal application of recombinant As22U (rAs22U). In the group challenged with rAs22U and ovalbumin (OVA), the number of eosinophils in the bronchial alveolar lavage fluid (BALF) was significantly increased, as compared to the group receiving only OVA. In addition, mice treated with rAs22U and OVA showed significantly increased airway hyperresponsiveness. Thus, severe inflammation around the airway and immune cell recruitment was observed in mice treated with rAs22U plus OVA. The levels of IL-4, IL-5, and IL-13 cytokines in the BALF increased significantly after treatment with rAs22U and OVA. Similarly, the levels of anti-OVA specific IgE and IgG1 increased in mice treated with rAs22U and OVA, compared to those treated only with OVA. The Gro-alpha (CXCL1) gene expression in mouse lung epithelial cells increased instantly after treatment with rAs22U, and allergy-specific chemokines eotaxin (CCL11) and thymus-and-activation-regulated-chemokine (CCL17) gene expressions significantly increased at 6 hr after treatment. In conclusion, rAs22U may induce airway allergic inflammation, as the result of enhanced Th2 and Th17 responses.

Seroprevalence of Antibodies against Anisakis simplex Larvae among Health-Examined Residents in Three Hospitals of Southern Parts of Korea

The present study was performed to estimate the seroprevalence of larval Anisakis simplex infection among the residents health-examined in 3 hospitals in southern parts of Korea. A total of 498 serum samples (1 serum per person) were collected in 3 hospitals in Busan Metropolitan city, Masan city, and Geoje city in Gyeongsangnam-do (Province) and were examined by IgE-ELISA and IgE-western blotting with larval A. simplex crude extract and excretory-secretory products (ESP). The prevalence of antibody positivity was 5.0% and 6.6% with ELISA against crude extracts and ESP, respectively. It was also revealed that infection occurred throughout all age groups and higher in females than in males. A specific protein band of 130 kDa was detected from 10 patients with western blot analysis against crude extract and ESP among those who showed positive results by ELISA. Our study showed for the first time the seroprevalence of anisakiasis in Korea. The allergen of 130 kDa can be a candidate for serologic diagnosis of anisakiasis.

The Clinical Characteristics of Anisakis Allergy in Korea

Anisakidae larvae can cause anisakiasis when ingested by humans. Although several groups have reported a gastrointestinal Anisakis allergy among people in Spain and Japan, our report is the first to summarize the clinical features of 10 Anisakis allergy cases in Korea. We enrolled 10 Korean patients (6 men and 4 women) who complained of aggravated allergic symptoms after ingesting raw fish or seafood. Sensitization to Anisakis was confirmed by detecting serum specific IgE to Anisakis simplex. The most common manifestation of anisakiasis was urticaria (100%), followed by abdominal pain (30%) and anaphylaxis (30%). All patients presenting with these symptoms also exhibited high serum specific IgE (0.45 to 100 kU/L) to A. simplex. Nine patients (90%) exhibited atopy and increased total serum IgE levels. The fish species suspected of carrying the Anisakis parasite were flatfish (40%), congers (40%), squid (30%), whelk (10%), and tuna (10%). Anisakis simplex should be considered as a possible causative food allergen in adult patients presenting with urticaria, angioedema, and anaphylaxis following the consumption of raw fish or seafood.

Reactividad alérgica a Anisakis simplex y su asociación con asma bronquial en niños escolares del estado Nueva Esparta, Venezuela / Anisakis simplex allergic reactivity and its association with bronchial asthma in scholar children from Nueva Esparta state, Venezuela

La reactividad alérgica frente a Anisakis simplex se ha convertido en un problema de salud pública en muchos países del mundo. En Venezuela, se ha reportado la presencia de mamíferos marinos infectados y se ha evidenciado la presencia del parásito en pescados de consumo masivo procedentes de las zonas costeras de oriente y occidente del país. No obstante, la importancia de este helminto en el desarrollo y severidad de patologías alérgicas no ha sido evaluada en el país. El objetivo de este estudio fue evaluar la reactividad alérgica al An. simplex en un grupo de 144 niños del estado de Nueva Esparta, así como su posible relación con la severidad de asma bronquial y rinitis alérgica. Todos los niños fueron evaluados utilizando un cuestionario socioeconómico validado y modificado de Graffar y un cuestionario validado según los criterios de ARIA (International Consensus Report) para rinitis alérgica, GINA (WHO/NIH Global Initiative for Asthma) para asma bronquial y Hannifin para dermatitis atópica, respectivamente. Se realizaron encuestas para evaluar las principales fuentes de alergenos a los cuales se expone la población de estudio. Se realizó la evaluación física de los niños y se practicaron pruebas cutáneas, así como pruebas de funcionalismo pulmonar. Encontramos un alto porcentaje (45%) de niños con pruebas de piel positiva al An. simplex. Se observó una mayor prevalencia y severidad, de asma y rinitis en aquellos niños con pruebas de piel positivas a An. simplex. Dado que ha sido previamente reportada la reactividad cruzada entre Ascaris lumbricoides y An. simplex, es importante considerar que la alta reactividad encontrada frente al An. simplex pueda deberse a la co-infección por Ascaris. De igual forma, la sensibilización frente a An. simplex, ya sea especifica o por reactividad cruzada existe, lo que sugiere que las autoridades locales deben considerar ciertas medidas preventivas, para evitar el consumo de pescados crudos o posiblemente parasitados con este helminto, que podrían exacerbar la patología respiratoria.

Allergic reactivity to Anisakis simplex has become in a health problem in many parts of the World. In Venezuelan coasts, it has been found infecting marine mammals and also different parasites fishes commonly consumed by human. However, the association of this helminth with the development and severity of allergic reactivity has not been previously evaluated in Venezuela. The aim of this study was to evaluate the allergic reactivity to An. simplex in a group 144 children from Nueva Esparta, state, and its possible relation with bronquial asthma and rhinitis allergic severity. All children were evaluated using a validated socioeconomic questionnaire modified by Graffar, and a validate questionnaire according to ARIA (International Consensus Report) for allergic rhinitis, GINA (WHO/NIH Global Initiative for Asthma) for bronquial asthma and Hannifin for atopic dermatitis. Surveys were performed in order to evaluate the main sources of allergens to which the study population exposes itself. Children physical evaluation and skin prick test were performed, as well as pulmonary function test. We found a high percentage (45%) of children with positive skin prick test to An. simplex. Moreover, a greatest prevalence and severity of asthma and rhinitis, was observed in those children with positive skin prick test to An. simplex. Since it has been reported the presence of the crossed reactivity between Ascaris lumbricoides and An. simplex, it’s important to consider that the high reactivity found to An. simplex might be due to the Co-infection with A. lumbricoides. Nevertheless, the sensibilization to An. simplex, due by specific or by crossed reactivity exists, and it should be considered by local health authorities in order to prevent the exacerbation of the respiratory pathologies mediated by the consumption of fresh fishes potentially infected.

Enzyme-linked immunosorbent assay and Western blot antibody determination in sera from patients diagnosed with different helminthic infections with Anisakis simplex antigen purified by affinity chromatography

An evaluation of the sensitivity and the specificity of the Anisakis simplex antigens purified by affinity chromatography was performed using sera from patients diagnosed with Anisakis sensitisation and sera from patients previously diagnosed with different helminthic infections. Only the sera of the patients diagnosed with Schistosoma mansoni or Onchocerca volvulus parasitic infections were negative against the A. simplex antigen and its purified fractions (PAK antigen: A. simplex antigen purified using columns prepared with anti-A. simplex rabbit IgG and PAS antigen: PAK antigen purified using columns prepared with anti-Ascaris suum rabbit IgG). However all the sera were positive against the A. suum antigen. In all the sera from the patients diagnosed with Anisakis sensitisation, the antibody levels detected using the purified antigens (PAK and PAS antigens) were lower than the observed using the A. simplex crude extract with the highest diminution in the case of the IgG. When these same sera were tested against the A. simplex crude extract by Western blot, several bands of high molecular masses were observed as well as, intense bands at 60 and/or 40 kDa. A concentration of these last proteins was observed in the PAK and the PAS antigens. When the sensitivity and the specificity determinations were performed, only seven of the 38 patients diagnosed of Anisakis sensitisation were positive, as well as, the sera from the patients diagnosed with parasitisms by Echinococcus granulosus or Fasciola hepatica.

Interleukin-4 production in BALB/c mice immunized with Anisakis simplex

We investigated the interleukin (IL-4) levels in BALB/c mice immunized with Anisakis extract in single or multiple doses and in mice orally infected with a larva. From animals immunized maximum responses were obtained with the multiple doses with an only IL-4 peak. Conversely, in the mice inoculated with a larva per os, the IL-4 levels showed two peaks of different rates

Alergia a Anisakis simplex: estudios clínicos e inmunológicos y reactividad cruzada con otros parásitos / Allergy by Anisakis simplex: clinical and immunological study and cross-reactivity with other parasites

El Anisakis simplex es un parásito perteneciente a la familia Anisakidae, de la clase nematodos, que vive en el tubo digestivo de grandes mamíferos marinos y parasita accidentalmente al hombre al ingerir pescado crudo o poco cocinado. Recientemente se ha implicado como posible factor etiológico de reacciones alérgicas inducidas tras la ingesta de pescado parasitado. En el presente estudio investigamos la posible sensibilización a antígenos de A. simplex en 11 pacientes que presentaron reacciones alérgicas tras la ingesta de pescado. También se estudió la existencia de reactividad cruzada con otros parásitos. En todos los pacientes se demostró, mediante pruebas inmunológicas "in vivo" e "in vitro", la existencia de un mecanismo de hipersensibilidad inmediata a antígenos de A. simplex. Las pruebas cutáneas con el extracto comercial, así como la determinación de IgE específica por técnica CAP y la liberación de histamina frente a este parásito fueron positivos, confirmando la sensibilización mediada por anticuerpos IgE a antígenos de Anisakis. Al estudiar la posible reactividad cruzada con otros parásitos (Ascaris lumbricoides, Echinococcus granulosus) por medio del inhibición del CAP, ésta resultó muy escasa para Ascaris lumbricoides, por lo que en estos pacientes dicha reactividad cruzada parece poco relevante. En cambio la inhibición del CAP a Eschinococcus granulosus con A. simplex llegó hasta el 30-60 por ciento, sugiriendo la existencia de cierto grado de reactividad cruzada entre ambos parásitos, aunque estén más alejados taxonómicamente. Debido a que el Anisakis simplex es un parásito ubicuo, ya que infecta a pescados y mariscos en mares de todo el mundo, sugerimos que ante la historia de manifestaciones alérgicas tras la ingesta de esta clase de alimentos, se investigue la sensibilidad a este parásito, además de los antígenos propios de los frutos de mar. Por otra parte, se debería estudiar la posible sensibilización a A. simplex en los pacientes a los cuales se les detecta IgE específica a E. granulosus, descartándose con anterioridad la enfermedad hidatídica.