title: Effects of Toxocara larvae on brain cell survival by in vitro model assessment.
abstract: Neuroinvasive larvae of the common dog and cat roundworms, Toxocara canis and Toxocara cati, may cause severe neurological and neuropsychological disturbances in humans. Despite their pathogenic potential and high prevalence worldwide, little is known about their cell-specific influences and cerebral host-pathogen interactions in neurotoxocarosis. To address this discrepancy, a co-culture system of viable larvae with murine neuronal (CAD), oligodendrocytal (BO-1) and microglial (BV-2) cell lines has been established. Additionally, murine adult brain slices have been co-cultured with Toxocara larvae to consider complex organotypic cell-cell interplay. Cytotoxicity of larval presence was measured enzymatically and microscopically. Microscopic evaluation using trypan blue exclusion assay revealed to be less reliable and sensitive than the lactate dehydrogenase activity assay. Ultimately, even low numbers of both T. canis and T. cati larvae have impaired survival of differentiated CAD cells, which morphologically resemble primary neurons. In contrast, viability of oligodendrocytal and microglial cells as well as brain slices was not impaired by larval presence. Therefore, immune-mediated mechanisms or trauma by migrating larvae presumably induce the in vivo pathology rather than acute cytotoxic effects. Conclusively, the helminthic larvae co-culture system presented here is a valuable in vitro tool to study cell-specific effects of parasitic larvae and their products.
authors.affiliation: Institute for Parasitology,University of Veterinary Medicine Hannover,Buenteweg 17,30559 Hannover,Germany.
authors.forename: Lea
authors.initials: L
authors.lastname: Heuer
authors.affiliation: Institute for Parasitology,University of Veterinary Medicine Hannover,Buenteweg 17,30559 Hannover,Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Haendel
authors.affiliation: Department of Pathology,University of Veterinary Medicine Hannover,Buenteweg 17,30559 Hannover,Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Beineke
authors.affiliation: Institute for Parasitology,University of Veterinary Medicine Hannover,Buenteweg 17,30559 Hannover,Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Strube
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000469:parasitology; D002460:Cell Line; D002470:Cell Survival; D018920:Coculture Techniques; D054884:Host-Pathogen Interactions / Q000502:physiology*; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007814:Larva; D051379:Mice; D014119:Toxocara / Q000502:physiology*; D014120:Toxocariasis / Q000469:parasitology / Q000473:pathology*; D014343:Trypan Blue / Q000378:metabolism
chemical_list: D007770:L-Lactate Dehydrogenase; D014343:Trypan Blue
keywords: BO-1 cells; BV-2 cells; CAD cells; Toxocara canis; Toxocara cati; brain slices; neurotoxocarosis; parasitic zoonosis