To date, this has only been achieved with attenuated N. caninum isolates used as live vaccines (10,11). However, application of a live vaccine poses a series of logistic and economical problems, which render inactivated and/or subunit vaccines much more attractive, provided a reasonable degree of protection against infection and disease can be achieved. Several research groups have reported promising results using recombinant antigens for vaccination studies, but others have reported failures or even anti-protective effects (3,9). This shows that the antigen repertoire of N. caninum contains both protective and immunomodulatory or

even immunosuppressive molecules, and these need to be defined and investigated. In addition, the route of antigen delivery and Raf inhibitor the type of adjuvant employed also need further investigation, SCH772984 considering that they can also alter the efficacy of a given vaccine candidate (41,43,44). Infection studies in cattle do not represent a cost-effective system to work with, and only

a limited number of research groups have taken up the enormous task to work with cattle directly (9,12). Accordingly, murine models have been extensively used for proof-of-concept studies on how an immune response against a vaccine could limit parasite dissemination and pathology. The currently used experimental murine models include (i) cerebral infection models with challenge infections of nonpregnant mice leading to cerebral disease and death, (ii) foetal infection models where mice are challenged during pregnancy and (iii) transplacental transmission of N. caninum tachyzoites leading to stillbirth, abortion or birth of infected offspring (9,49). In

the present study, we employed the acute disease model of cerebral infection in nonpregnant animals. For the vaccine, we employed an innovative approach by analysing the relative efficacy of recNcPDI vaccine antigen associated with nanogel vaccine delivery formulations. RecNcPDI has been previously shown to be ineffective when applied i.p. emulsified in SAPs, but highly effective and mediating protection against cerebral infection and disease when applied i.n. in the presence of cholera toxin (19). The purpose of the current work was to use chitosan-based nanogels, combined with different adjuvants (saponin and CT), as carriers for the E. coli Progesterone expressed recNcPDI antigen. Thereby, the aims were to investigate whether this nanogel association would influence the immunogenic and efficacy characteristics of the vaccine antigen upon i.p. and i.n. vaccination. SDS–PAGE and immunoblotting showed that recNcPDI was efficiently associated with both types of nanogels employed – alginate-coated chitosan nanogels and mannosylated, alginate-coated nanogels. The vaccine antigen was well associated with the nanogels, in terms of no nanogel-free material being detected. It also retained its antigenic reactivity with a polyclonal anti-recNcPDI antiserum.