In summary, we demonstrated that co-immunizations with Ad5-pIX-ASP-M and Ad5-gp83 would be useful in the development of vaccines against Chagas disease, due to the ability of the vector to trigger robust ASP-M-activated CD8+ T lymphocytes that reduce T. cruzi parasitism by secreting IFNγ and TNFα induced by Ad5-pIX-ASP-M and eliciting neutralizing antibodies via Ad5-gp83 to reduce parasitemia. Here, TNF is linked to Chagas disease.