Sexual dimorphism is shown to affect the immune response, with females presenting an increased immune reactivity in comparison to males. The difference in immune response is impacted by an intricate interplay of sex chromosomes and sex hormones. Plasmacytoid dendritic cells (pDCs), recognised for their vital role in antiviral immunity, are influenced by these factors. However, the exact mechanism beneath this influence is still unknown. This study explores the sexual dimorphic antiviral responses of pDCs, specifically within the role of Toll-Like Receptor 7 and type 1 interferon signalling. Peripheral blood mononuclear cells (PBMCs) and pDCs were isolated from the blood of healthy human donors obtained from Sahlgrenska University Hospital and stimulated with gardiquimod (a Toll-like receptor 7 agonist) and lipopolysaccharide (a Toll-like receptor 4 agonist). Quantitative polymerase chain reaction (qPCR) and fluorescence activated cell sorting (FACS) were used to analyse the gene expression and protein levels. The findings in this study illustrate sex-specific differences in the immune-related genes. Gene expression analysis displayed higher IFNα2 levels in male pDCs than females. Conversely, female pDCs exhibited increased IFNα2 protein levels than males. IRF7 and TLR7 expression levels were increased for female pDCs but higher levels of TNF expression were displayed for male pDCs. These findings allow us to understand the complex interaction between sex hormones, sex chromosomes and the immune response, specifically in the antiviral functions of pDCs. This also provides insight into the possible sex-specific differences in disease susceptibilities and therapeutic approaches.