Malaria caused by Plasmodium parasites remains a major global health problem, with 3.2 billion people estimated to be at risk of infection. Malaria compromises dendritic cell (DC) function and DC impairments contribute to incomplete immunity characterised by recurrent infection. We sought to determine whether a single P. falciparum infection is sufficient to compromise CD16⁺ or CD141⁺ DC function and determine if specific subsets of circulating DC are impacted. In malaria-naive adult volunteers, peripheral DC subsets were examined before and after intra-venous inoculation of P. falciparum-infected red blood cells. Using flow cytometry, the number and maturation (HLA-DR, CD86) was assessed as well as cytokine production (TNF, IL-12 and IL-10) in response to stimulation with toll like receptor (TLR) ligands and P. falciparum-infected red blood cells before and at peak parasitemia (day 7). A primary P. falciparum infection increased CD16⁺ DC maturation in the circulation and induced production of TNF, IL-12, and IL-10. Upon TLR stimulation CD16⁺ DC increased IL-12 production yet preferentially produced IL-10 after stimulation with P. falciparum-infected red blood cells. In contrast, in the same individuals, P. falciparum infection significantly reduced circulating CD141⁺ DC number and HLA-DR expression. We conclude that P. falciparum expands and activates CD16⁺ DC, which respond to infection by producing TNF, IL-12 and IL-10, while CD141⁺ DC reduce in both number and activation. In the same clinical trials, we recently reported dysfunctional CD1c⁺ DC with impaired HLA-DR expression and skewed TNF pro-inflammatory cytokine production. These data now define CD16⁺ DC as unique early DC responders in a first sub-microscopic blood-stage P. falciparum infection. Additional research is warranted to determine whether CD16⁺ DC activation is associated with patent parasitemia and malaria.