Systemic and mucosal IgE antibody responses of horses to infection with Anoplocephala perfoliata.

Abstract: Infection of horses with Anoplocephala perfoliata induces a severe inflammatory reaction of the caecal mucosa around the site of parasite attachment adjacent to the ileocecal valve. Lesions show epithelial erosion or ulceration of the mucosa with infiltration by eosinophils, lymphocytes and mast cells leading to oedema, gross thickening and fibrosis of the caecal wall. Despite this evidence of an inflammatory reaction to A. perfoliata within the mucosa of the caecum there is little information about the nature of the local immune response to A. perfoliata. An ELISA which assays serum IgG(T) antibodies to A. perfoliata excretory/secretory antigens has been developed as a diagnostic test. However, the specificity of the ELISA remains sub-optimal and the role of other isotypes in the immune response to A. perfoliata has not been reported. This study measured IgA, IgE and IgG(T) antibody responses to A. perfoliata excretory/secretory antigens in sera of 75 horses presented for slaughter. The prevalence of A. perfoliata infection, as confirmed by the presence of parasites in the terminal ileum, caecum or proximal colon, was 55%. A. perfoliata-specific IgG(T) and IgE antibodies were significantly elevated in infected horses compared to controls; IgA antibodies were also detected but did not differ between infected and control horses. Diagnosis by serum IgG(T) ELISA had a sensitivity of 78% and a specificity of 80%, by comparison the serum IgE ELISA had a sensitivity of just 44% with a specificity of 82% and therefore did not provide an improved diagnostic test. Western blots with sera from infected horses demonstrated IgE-binding to at least 10 separate components of excretory/secretory (E/S) antigens. A similar pattern was also found with IgG(T). Around 30% of horses had high levels of serum IgE which bound fucose-containing carbohydrate antigens on the parasite surface but this was unrelated to the presence of A. perfoliata infection. Immunoperoxidase staining detected numerous IgE-positive cells within lymphoid follicles in the caecal mucosa close to the site of A. perfoliata attachment and quantitative RT-PCR detected high levels of IgE transcription in the caecal mucosa of all horses. Mucosal synthesis of antibodies was confirmed by the demonstration of A. perfoliata-specific IgG(T) and IgE in the supernatant of lamina propria explant cultures that discriminated clearly between infected and uninfected horses. We conclude that there is an active immune response to A. perfoliata within the caecal mucosa involving local production of both IgG(T) and IgE antibody isotypes; but it remains unclear whether this immune response can reduce or eliminate parasite burden.