Establishment of Neospora caninum antigen-specific T cell lines of primarily CD4 T cells.

Neosporosis is an important cause of pregnancy loss in cattle worldwide. Protective immunity against Neospora caninum infection may include both cell-mediated (CMI) and humoral immune responses. This study was to establish short-term antigen-specific T cell lines composed of primarily CD4(+)T cells from peripheral blood lymphocytes (PBL) of infected cows, which may be used to identify immunodominant antigens for the development of N. caninum vaccines. Crude N. caninum tachyzoite antigen was prepared from in vitro derived N. caninum tachyzoites. Multiple T cell lines were established and maintained for 11 weeks by weekly re-stimulation with N. caninum antigen and antigen-presenting cells. All cell lines responded highly to antigen between weeks 3 and 11. Phenotypically, these cells were composed primarily of CD4(+)T cells between weeks 2-8, with a gradual expansion of gamma/delta(+)T cells thereafter. The results indicate that N. caninum-specific T cell lines can be established and maintained without exogenous T cell growth factors and may be used to identify N. caninum antigens. This research will enhance our understanding of bovine CMI to neosporosis and may facilitate development of a proven neosporosis vaccine.