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Previous studies in animal models have revealed an association between interferon-γ (IFN- γ), produced by CD8+ T cells and irradiated sporozoite-induced sterile immunity. To determine whether IFN-γ can serve as a marker of pre-erythrocytic protective immunity in individuals naturally exposed to malaria, we characterized IFN-γ and lymphocyte proliferative responses to previously defined CD8+ cytotoxic T lymphocyte (CTL) epitopes from six pre-erythrocytic stage antigens in 107 children six months to two years old from a community-based birth cohort in western Kenya. We found that IFN- γ positive responders had higher hemoglobin (Hb) levels and significantly reduced prevalence of severe malarial anemia one month after the test compared with IFN- γ non-responders, suggesting that IFN- γ immune responses to these pre-erythrocytic antigens were associated with protection against malarial anemia. Children who responded by lymphocyte proliferation had a significantly longer time to first documented malaria parasitemia after birth; however, there was no correlation between the presence of lymphocyte proliferative response and higher Hb levels. We propose that IFN- γ production could be used as a potential marker of protective immunity against malaria associated anemia in young children living in malaria holoendemic areas.