RESUMEN
Numerous factors contribute to perinatal risk, many of which remain undefined. This study sought to determine the frequency of fish intake in postpartum women, and to establish a relationship between the rates of immunoreactivity for antigens from Anisakis spp. and high-risk pregnancy. In this prospective noninterventional study, a structured questionnaire was administered and serum was collected from postpartum women at two perinatal centers (a high-risk birth unit [HRBU] and a low-risk birth unit [LRBU]) in the Niteroi municipality of Brazil. Anisakis species-specific IgG and IgE were measured by ELISA. The chisquared test was performed, and odds ratios (ORs) with their 95% confidence intervals were estimated. The t-test or Mann-Whitney test was applied to continuous, normally distributed variables. In total, 309 women (170 from HRBU, 139 from LRBU) between 24.8 and 26.7 years old with a median of 6 to 8 prenatal visits were enrolled. Women in the two units exhibited differences in some variables, including prenatal care (p = 0.01), maternal and fetal risk (p = 0.00; OR = 6.17), and gestational age (p = 0.00), but no differences in fish consumption (p = 0.29), frequency of fish intake (p = 0.40), allergic symptoms (p = 0.51), or frequency of anti-Anisakis reactivity (p = 0.22). Logistic regression analysis revealed that only age was independently associated with postpartum anti-Anisakis reactivity. This study confirmed a low prevalence of fish intake and suggested that Anisakis spp. had no impact on high-risk pregnancies among this postpartum study population.
Asunto(s)
Anisakiasis/diagnóstico , Anisakis/aislamiento & purificación , Antígenos Helmínticos/sangre , Complicaciones Parasitarias del Embarazo/diagnóstico , Embarazo de Alto Riesgo , Animales , Anisakiasis/epidemiología , Anisakiasis/parasitología , Anisakis/inmunología , Brasil/epidemiología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Embarazo , Complicaciones Parasitarias del Embarazo/parasitología , Prevalencia , Estudios Prospectivos , Encuestas y CuestionariosRESUMEN
BACKGROUND: Nematode secreted haemoglobins have unusually high affinity for oxygen and possess nitric oxide deoxygenase, and catalase activity thought to be important in protection against host immune responses to infection. In this study, we generated a monoclonal antibody (48Eg) against haemoglobin of the nematode Anisakis pegreffii, and aimed to characterize cross-reactivity of 4E8g against haemoglobins of different nematodes and its potential to mediate protective immunity against a murine hookworm infection. METHODOLOGY/PRINCIPAL FINDINGS: Immunoprecipitation was used to isolate the 4E8g-binding antigen in Anisakis and Ascaris extracts, which were identified as haemoglobins by peptide mass fingerprinting and MS/MS. Immunological cross-reactivity was also demonstrated with haemoglobin of the rodent hookworm N. brasiliensis. Immunogenicity of nematode haemoglobin in mice and humans was tested by immunoblotting. Anisakis haemoglobin was recognized by IgG and IgE antibodies of Anisakis-infected mice, while Ascaris haemoglobin was recognized by IgG but not IgE antibodies in mouse and human sera. Sequencing of Anisakis haemoglobin revealed high similarity to haemoglobin of a related marine nematode, Psuedoterranova decipiens, which lacks the four -HKEE repeats of Ascaris haemoglobin important in octamer assembly. The localization of haemoglobin in the different parasites was examined by immunohistochemistry and associated with the excretory-secretary ducts in Anisakis, Ascaris and N. brasiliensis. Anisakis haemoglobin was strongly expressed in the L3 stage, unlike Ascaris haemoglobin, which is reportedly mainly expressed in adult worms. Passive immunization of mice with 4E8g prior to infection with N. brasiliensis enhanced protective Th2 immunity and led to a significant decrease in worm burdens. CONCLUSION: The monoclonal antibody 4E8g targets haemoglobin in broadly equivalent anatomical locations in parasitic nematodes and enhances host immunity to a hookworm infection.
Asunto(s)
Anisakis/inmunología , Anticuerpos Antihelmínticos/inmunología , Anticuerpos Monoclonales/inmunología , Reacciones Cruzadas , Hemoglobinas/inmunología , Nippostrongylus/inmunología , Infecciones por Strongylida/inmunología , Animales , Anisakis/genética , Anticuerpos Antihelmínticos/administración & dosificación , Anticuerpos Antihelmínticos/aislamiento & purificación , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales/aislamiento & purificación , Antígenos Helmínticos/química , Antígenos Helmínticos/genética , Antígenos Helmínticos/inmunología , Ascaris/inmunología , Modelos Animales de Enfermedad , Hemoglobinas/química , Hemoglobinas/genética , Humanos , Inmunización Pasiva , Immunoblotting , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Inmunohistoquímica , Inmunoprecipitación , Ratones , Ratones Endogámicos BALB C , Nematodos , Análisis de Secuencia de ADN , Espectrometría de Masas en Tándem , Resultado del TratamientoRESUMEN
Although the incidence of anisakiasis is rising worldwide, its frequency is still unknown in Brazil. The aim of this study was to verify immunoreactivity to Anisakis simplex antigens in healthy adults and determine its possible relationship with frequency of fish consumption and allergy symptoms. A prospective cross-sectional study was carried out with 67 volunteers recruited from a military facility in Niterói, Brazil. The subjects completed a structured questionnaire and serum titers of specific anti-Anisakis IgE and IgG antibodies were measured. The association between frequency of fish intake and IgE reactivity was evaluated by Fisher's exact test. Almost all subjects (97.0%, 65/67) that consumed seafood; 64.6% (42/65) ate fish at least once weekly. Of all seafood consumers, 56.9% (37/65) reported allergy symptoms, being gut allergies most often cited (35.5%). IgE seroreactivity to Anisakis simplex was found in 20.9% of subjects (14/67), with 13.4% (9/67) reacting exclusively to somatic antigen, 3.0% (2/67) exclusively to excretory/secretory antigens and 4.5% (3/67) to both antigens. There was a significant association between frequency of fish consumption and positive serology (p = 0.019). An immunoblot assay for Anisakis antigens showed different positive bands for IgG. The direct relationship between ELISA reactivity and frequency of fish intake and absence of association with allergy symptoms suggests previous contact with Anisakis simplex antigens.
Asunto(s)
Anisakis/inmunología , Anticuerpos Antihelmínticos/sangre , Adulto , Animales , Brasil/epidemiología , Estudios Transversales , Conducta Alimentaria , Femenino , Voluntarios Sanos , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Estudios Seroepidemiológicos , Suero/química , Encuestas y Cuestionarios , Adulto JovenRESUMEN
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, its 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.
Asunto(s)
Humanos , Masculino , Adolescente , Femenino , Preescolar , Niño , Alergia e Inmunología/estadística & datos numéricos , Asma/inmunología , Asma/microbiología , Asma/parasitología , Anisakis/inmunología , Anisakis/parasitologíaRESUMEN
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.
Asunto(s)
Anisakis/inmunología , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , Helmintiasis/diagnóstico , Animales , Anisakiasis/diagnóstico , Antígenos Helmínticos/aislamiento & purificación , Western Blotting , Cromatografía de Afinidad , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Humanos , Inmunoglobulina G/inmunología , Conejos , Sensibilidad y EspecificidadRESUMEN
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.
Asunto(s)
Animales , Humanos , Conejos , Anisakis/inmunología , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , Helmintiasis/diagnóstico , Anisakiasis/diagnóstico , Antígenos Helmínticos/aislamiento & purificación , Western Blotting , Cromatografía de Afinidad , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/inmunología , Sensibilidad y EspecificidadRESUMEN
In order to improve the specificity and sensitivity of the techniques for the human anisakidosis diagnosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis simplex third-stage larvae (L3) has been developed. New Zealand rabbits were immunized with A. simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG specific antibodies were isolated by means of protein A-Sepharose CL-4B beads columns. IgG anti-A. simplex and -A. suum were coupled to CNBr-activated Sepharose 4B. For the purification of the larval A. simplex antigens, these were loaded into the anti-A. simplex column and bound antigens eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. simplex specific proteins were loaded into the anti-A. suum column. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis were carried out. Further, we studied the different responses by ELISA to the different antigenic preparations of A. simplex used, observing their capability of discriminating among the different antisera raised in rabbits (anti-A. simplex, anti-A. suum, anti-T. canis). The discriminatory capability with the anti-T. canis antisera was good using the larval A. simplex crude extract (CE) antigen. When larval A. simplex CE antigen was loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with A. simplex CE antigen, its capability for discriminate between A. simplex and A. suum was improved, increasing in the case of T. canis. The best results were obtained using larval A. simplex CE antigen loaded into a CNBr-activated Sepharose 4B coupled to IgG from rabbits immunized with adult A. suum CE antigen. When we compared the different serum dilution and antigenic concentration, we selected the working serum dilution of (1/4)00 and 1 microg/ml of antigenic concentration.
Asunto(s)
Anisakiasis/diagnóstico , Anisakis/inmunología , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/inmunología , Antígenos Helmínticos/aislamiento & purificación , Animales , Anticuerpos Antihelmínticos/inmunología , Cromatografía de Afinidad , Perros , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Sueros Inmunes/inmunología , Conejos , Sensibilidad y Especificidad , PorcinosRESUMEN
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.
Asunto(s)
Anisakiasis/metabolismo , Anisakis/metabolismo , Interleucina-4/biosíntesis , Animales , Anisakiasis/inmunología , Anisakis/inmunología , Antígenos Helmínticos/inmunología , Antígenos Helmínticos/metabolismo , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/biosíntesis , Interleucina-4/sangre , Larva/inmunología , Ratones , Ratones Endogámicos BALB CRESUMEN
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
Asunto(s)
Animales , Ratones , Anisakiasis/inmunología , Anisakis/inmunología , Interleucina-4/biosíntesis , Antígenos Helmínticos/inmunología , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/biosíntesis , Interleucina-4/sangre , Larva/inmunología , Ratones Endogámicos BALB CRESUMEN
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. (AU)
Asunto(s)
Humanos , Masculino , Femenino , Ascaridoidea/inmunología , Hipersensibilidad a los Alimentos/etiología , Anisakis/inmunología , Reacciones Cruzadas/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Urticaria/etiología , Angioedema/etiología , Anafilaxia/etiología , Helmintiasis/diagnóstico , Helmintiasis/inmunología , Peces/parasitología , Echinococcus/inmunología , Pruebas Cutáneas/normas , Pruebas Inmunológicas/normas , Anisakiasis/diagnóstico , Anisakis/patogenicidadRESUMEN
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.