ABSTRACT In seeking substitutions for the current Chagas disease treatment, which has several relevant side effects, new therapeutic candidates have been extensively investigated. In this context, a balanced interaction between mediators of the host immune response seems to be a key element for therapeutic success, as a proinflammatory microenvironment modulated by interleukin-10 (IL-10) is shown to be relevant to potentiate anti- Trypanosoma cruzi drug activity. This study aimed to identify the potential immunomodulatory activities of the anti- T. cruzi K777, pyronaridine (PYR), and furazolidone (FUR) compounds in peripheral blood mononuclear cells (PBMC) from noninfected (NI) subjects and chronic Chagas disease (CD) patients. Our results showed low cytotoxicity to PBMC populations, with 50% cytotoxic concentrations (CC ₅₀ ) of 71.0 μM (K777), 9.0 μM (PYR), and greater than 20 μM (FUR). In addition, K777 showed no impact on the exposure index (EI) of phytohemagglutinin-stimulated leukocytes (PHA), while PYR and FUR treatments induced increased EI of monocytes and T lymphocytes at late stages of apoptosis in NI subjects. Moreover, K777 induced a more prominent proinflammatory response (tumor necrosis factor alpha-positive [TNF-α ⁺ ] CD8 ⁺ /CD4 ⁺ , gamma interferon-positive [IFN-γ ⁺ ] CD4 ⁺ /CD8 ⁺ modulated by interleukin-10-positive [IL-10 ⁺ ] CD4 ⁺ T/CD8 ⁺ T) than did PYR (TNF-α ⁺ CD8 ⁺ , IL-10 ⁺ CD8 ⁺ ) and FUR (TNF-α ⁺ CD8 ⁺ , IL-10 ⁺ CD8 ⁺ ). Signature analysis of intracytoplasmic cytokines corroborated the proinflammatory/modulated (K777) and proinflammatory (PYR and FUR) profiles previously found. In conclusion, the lead compound K777 may induce beneficial changes in the immunological profile of patients presenting the chronic phase of Chagas disease and may contribute to a more effective therapy against the disease.