title: Characterization of an IgG monoclonal antibody targeted to both tissue cyst and sporocyst walls of Toxoplasma gondii.
abstract: Toxoplasma gondii infects animals habiting terrestrial and aquatic environments. Its oocysts and tissue cysts are important for the horizontal transmission of this parasite. The oocyst and tissue cyst walls are crucial for the ability of the parasite to persist in the environment or in animal tissues, respectively. However, the composition of these walls is not well understood. We report the generation of monoclonal antibodies directed against wall components using mice immunized with oocyst antigens of T. gondii. One monoclonal antibody (mAb) G1/19 reacted solely with T. gondii sporozoites. The respective antigen had a relative molecular weight (Mr) of 30 kDa. MAb G1/19 failed to react with sporozoites of any other coccidian parasite species tested (Hammondia hammondi, Hammondia heydorni, Cystoisospora felis, Eimeria bovis, Sarcocystis sp.). Another mAb, designated K8/15-15, recognized antigens in sporocyst walls of the parasite and in the walls of in vivo or in vitro produced tissue cysts, as demonstrated by immunofluorescence and immunoblot assays. Antigens of 80 to a high molecular weight protein of about 350 kDa Mr were recognized by this antibody using antigen extracts from sporocysts, and from in vitro or in vivo generated tissue cysts of the parasite. Tissue cyst and sporocyst walls of H. hammondi and H. heydorni, and tissue cysts of Neospora caninum were also recognized by mAb K8/15-15. Sporocyst walls of C. felis also reacted to this mAb. The cyst walls of Sarcocystis sp. and Besnoitia besnoiti were not recognized by mAb K8/15-15. Reactivity by a single mAb against T. gondii antigens in tissue cysts and sporocysts had not been reported previously. MAb K8/15-15 may be a practical tool for the identification of both cysts and sporocysts of the parasite, and may also be potentially employed in proteomic studies on the identification of new components of the cyst and sporocyst walls of T. gondii.
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493, Greifswald-Insel Riems, Germany; Universidade Federal da Bahia, Escola de Medicina Veterinária e Zootecnia, Departamento de Anatomia, Patologia e Clínicas, Av. Adhemar de Barros 500, 40170-110, Salvador, Bahia, Brazil. Electronic address: pita@ufba.br.
authors.forename: Luís F P
authors.initials: LF
authors.lastname: Gondim
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493, Greifswald-Insel Riems, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Wolf
authors.affiliation: IDEXX Laboratories, D-71636, Ludwigsburg, Germany.
authors.forename: Majda G
authors.initials: MG
authors.lastname: Vrhovec
authors.affiliation: IDEXX Laboratories, D-71636, Ludwigsburg, Germany.
authors.forename: Nikola
authors.initials: N
authors.lastname: Pantchev
authors.affiliation: Institute of Parasitology, Justus Liebig University Giessen, Giessen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Bauer
authors.affiliation: Institute of Veterinary Pathology, Ludwig Maximilians University Munich, Munich, Germany; Institute for Infectious Diseases and Zoonoses, Ludwig Maximilians University Munich, Munich, Germany.
authors.forename: Martin C
authors.initials: MC
authors.lastname: Langenmayer
authors.affiliation: Department of Microbiology, University of Gottingen, Kreuzbergring 57, D-37075 Göttingen, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Bohne
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Department of Experimental Animal Facilities and Biorisk Management, 17493, Greifswald-Insel Riems, Germany.
authors.forename: Jens P
authors.initials: JP
authors.lastname: Teifke
authors.affiliation: Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, US Department of Agriculture, Beltsville, MD 20705, USA.
authors.forename: Jitender P
authors.initials: JP
authors.lastname: Dubey
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493, Greifswald-Insel Riems, Germany.
authors.forename: Franz J
authors.initials: FJ
authors.lastname: Conraths
authors.affiliation: Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, Südufer 10, 17493, Greifswald-Insel Riems, Germany.
authors.forename: Gereon
authors.initials: G
authors.lastname: Schares
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000953:Antigens, Protozoan / Q000008:administration & dosage / Q000276:immunology*; D002415:Cats; D002417:Cattle; D003044:Coccidia / Q000145:classification / Q000276:immunology; D004285:Dogs; D005455:Fluorescent Antibody Technique; D006825:Hybridomas; D007117:Immunization, Secondary; D007074:Immunoglobulin G / Q000276:immunology*; D007275:Injections, Intravenous; D007279:Injections, Subcutaneous; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D033661:Oocysts / Q000276:immunology; D012756:Sheep; D014122:Toxoplasma / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D007074:Immunoglobulin G
keywords: Immunoblot; Monoclonal antibody; Oocyst; Sporocyst; Tissue cyst; Toxoplasma