Anisakis simplex sl and its interaction with human caco-2 cells

Background: Anisakis is a marine parasite nematode characterized by a complex life cycle. Humans can accidentally develop a zoonosis called anisakiasis, eating raw fish with parasitic infective larvae. The larval mechanical action in the gastrointestinal tract and the release of excreted/secreted products and extracellular vescicles (EVs) can progressively determine hemorrhagic, erosive and ulcerous lesions, granuloma and chronic inflammation. Anisakiasis can be accompanied by mild or severe allergic symptoms ranging from urticaria, angioedema to anaphylaxis. Several studies characterized the parasite allergens and the associated Th2-mediated immune response, resulting in the production of the IgE-mediated allergic reaction. However, the role of host's immune response during the infection and the potential implications are still unknown. Objectives: The aim of this project is to develop a suitable in-vitro model to study Anisakis-human intestinal cells interactions, focusing on the host cell inflammatory response and how such molecules are involved in the disease progression. Intestinal Caco-2 cell monolayers will be challenged with the live larvae (L), the crude extract (CE) and the exosome-enriched fraction (Evs). After 24 h of incubation, cell supernatants will be tested for IL-8 and IL-6 production and cells will be tested for NFKB and MAPK signaling. Expected results: Previous studies on human dendritic cells, fibroblasts and cancer colonic cells have shown signs of inflammatory response, characterized by an induction of oxidative stress and an inhibition of apoptosis with an alteration in the expression of important cell cycle check points, as P53. Similarly, we expect that the alive parasite or its products can fine tune the expression of host inflammatory products, in association with a potential alteration of intracellular signaling pathways as the ones described above. Future perspective: This study can highlight the pathogenetic mechanisms that charachterize the accidental interactions between Anisakis and humans, opening new ways for the identification of new diagnostic biomarkers and to deepen eventual potential implications and consequences of the infection in the host.