Isotype‐Specific Immune Responses in Murine Experimental Anisakiasis

Abstract

A murine experimental model of anisakiasis has been developed in BALB/c and C57BL/10 mice orally inoculated with an A n i d i s sihpfex living third stage larva (L3) in order to investigate isotype-specific immune responses against excretory-secretory (ES) products and crude extracts (CE) from L3. Specific antibody production showed similar patterns against both ES and CE antigens with higher levels against the latter. The dynamics of the production showed the earliest responses in BALB/c, in which antibodies were principally of the immunoglobulin (I@M isotype. Responses to the IgGl subclass were mainly produced in the C57BL/10 strain. Levels of IgG2a were practically undetectable. With sera from C57BL/10 mice high levels of the IgG2b isotype were detected. A slight IgG3 response was only detected against the CE antigen in the C57BL/10 strain by the end of the experiment and IgA responses were very low. Humoral responses against A. simpkx antigens are dlfferent depending on individual characteristics and thymus-independent epitopes might be represented into A. sihpfex antigens and their stimuli could be different regarding the murine strain used.

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

“…Parasites -A. simplex L3 were picked up manually from the viscera, flesh, and body cavities of naturally infected blue whiting (Micromesistius poutassou) and exhaustively washed in water (Perteguer & Cuéllar 1998). A. suum were obtained from swine natural infections (Guillén et al 1986).…”

“…The supernatant was dyalised overnight at 4ºC in PBS. The protein content of the extract was estimated and the extract was frozen at 20ºC until used (Perteguer & Cuéllar 1998). The antigen from A. suum adults (A. suum CE antigen or AS antigen) was obtained using a modification of the Welch et al (1983) method by homogenisation and extraction in PBS at 4ºC overnight (instead of ultrasonic burst).…”

Evaluation by ELISA of Anisakis simplex Larval Antigen Purified by Affinity Chromatography

“…Parasites -A. simplex L3 were picked up manually from the viscera, flesh and body cavities of naturally infected blue whiting (Micromesistius poutassou) and exhaustively washed in water (Perteguer & Cuéllar 1998). A. suum and T. canis were obtained from natural infections of swine or dogs, respectively.…”

“…The supernatant was dialysed overnight at 4 o C in PBS. Protein content of the extract was estimated and the extract was frozen at -20 o C until used (Perteguer & Cuéllar 1998). CE antigen from A. suum adults (A. suum CE antigen) was obtained using a modification of the Welch et al (1983) method by homogenization and extraction in PBS at 4 o C overnight (instead of ultrasonic burst).…”

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

“…Since the specific antibodies showed by the murine BALB/c strain (susceptible strain) indicated a Th2 response after the Anisakis infection (Perteguer & Cuéllar 1998), we proposed the investigation of the interleukin (IL-4) levels (marker of Th2 responses) in mice immunized with A. simplex crude extract (CE) in a single dose or in multiple doses, as well as, in mice orally infected with a living third stage larva (L3).…”