title: Transgenic Neospora caninum strains constitutively expressing the bradyzoite NcSAG4 protein proved to be safe and conferred significant levels of protection against vertical transmission when used as live vaccines in mice.
abstract: At present, there is no effective treatment or vaccine to prevent vertical transmission or abortion associated with Neospora caninum infection in cattle. Different vaccine formulations have been assayed, and live vaccines have shown the most promising results in terms of protection. Previously, transgenic N. caninum tachyzoites expressing the bradyzoite stage-specific NcSAG4 antigen in a constitutive manner (Nc-1 SAG4(c)) were obtained and showed a reduced persistence of parasite in inoculated mice. Thus, the present study evaluates the Nc-1 SAG4(c)1.1 and Nc-1 SAG4(c)2.1 transgenic strains and the Nc-1 wild-type (WT) strain to determine their protective efficacy against vertical transmission and cerebral neosporosis in mice. Consequently, dams were immunized twice with 5 × 10(5) tachyzoites of each strain and challenged with 2 × 10(6) tachyzoites of a heterologous and virulent isolate at 7-10 days of gestation. The Nc-1 SAG4(c)1.1 strain offered less protection than the other transgenic strain (Nc-1 SAG4(c)2.1) or their ancestor (Nc-1 WT). Indeed, 40%, 7% and 5.6% of the postnatal deaths corresponded to pups from dams vaccinated with Nc-1 SAG4(c)1.1, Nc-1 SAG4(c)2.1 and Nc-1 (WT) strains, respectively. In comparison, the non-immunized challenge group had a 100% mortality rate. In addition, mice were protected against congenital transmission; vertical transmission rates were 45%, 11.1% and 10.8% in the Nc-1 SAG4(c)1.1, Nc-1 SAG4(c)2.1 and Nc-1 WT immunized groups, respectively, vs. 94.9% in the non-vaccinated infected group. However, this protection against the postnatal mortality and the vertical transmission was not associated with a consistent Th1 or Th2-type immune response. Nonetheless, the Nc-1 SAG4(c)2.1 strain appears to be the best candidate for use as a live vaccine, as evidenced by results demonstrating its high levels of protection against vertical transmission and its lower persistence in mice, making this transgenic strain safer than Nc-1 WT.
authors.affiliation: SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain. vmarugan@vet.ucm.es
authors.forename: V
authors.initials: V
authors.lastname: Marugán-Hernández
authors.forename: L M
authors.initials: LM
authors.lastname: Ortega-Mora
authors.forename: A
authors.initials: A
authors.lastname: Aguado-Martínez
authors.forename: E
authors.initials: E
authors.lastname: Jiménez-Ruíz
authors.forename: G
authors.initials: G
authors.lastname: Alvarez-García
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000276:immunology*; D003048:Coccidiosis / Q000517:prevention & control* / Q000635:transmission; D005260:Female; D015870:Gene Expression; D018445:Infectious Disease Transmission, Vertical / Q000517:prevention & control*; D051379:Mice; D018688:Neospora / Q000235:genetics / Q000276:immunology*; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000276:immunology; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000276:immunology; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000276:immunology*; D016052:Protozoan Vaccines / Q000008:administration & dosage / Q000276:immunology*; D016019:Survival Analysis; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D014613:Vaccines, Attenuated