Background: Alpha-type 1 polarized dendritic cells (αDC1s) are specialized dendritic cells characterized by their enhanced ability to induce Th1-polarized immune responses and activate antigen-specific cytotoxic T lymphocytes, making them promising candidates for therapies in different infectious contexts. Despite their therapeutic potential, the temporal dynamics of antiviral gene expression in αDC1s remain incompletely understood.

Methods: Monocyte-derived αDC1s from three healthy donors were generated and matured using a specific cytokine cocktail containing IL-1β, TNF-α, IFN-α, IFN-γ, and poly(I:C). The cells were evaluated by qPCR to investigate the kinetic expression profile of interferon-stimulated and inflammatory-associated genes at 4 h, 6 h, 8 h, 24 h, and 48 h after stimulation. Relative mRNA expression of MxA, ISG15, and CCL5 was analyzed using the 2-ΔΔCt method.

Results: Distinct temporal and donor-dependent transcriptional patterns were observed among the evaluated genes. Expression of MxA and ISG15 was observed predominantly during the early time points, specifically between 4 h and 8 h after stimulation, suggesting the activation of antiviral interferon pathways. Additionally, CCL5 expression was detected across the evaluated periods, with fluctuations observed between donors and time points. Overall, αDC1s displayed a dynamic transcriptional response characterized by the transient expression of interferon-responsive genes and interindividual variability.

Conclusions: Our findings demonstrate that αDC1s exhibit a temporally regulated antiviral transcriptional response following stimulation. The observed donor-to-donor variability highlights the importance of considering individual immune responsiveness in the development of dendritic cell-based therapeutic strategies. These results contribute to the understanding of αDC1 functional biology and may support the optimization of future immunotherapeutic approaches.

Financial Support: Capes 88887.196649/2025-00 and Fapesp 2025/13425-7.