Electrochemical Cytokine Profiling Reveals Dual Immunomodulatory Effects of Human Milk Oligosaccharides in THP-1 Macrophages

Abstract Human milk oligosaccharides (HMOs) are bioactive carbohydrates that govern host–microbe interactions and immune regulation. Beyond their antimicrobial and antibiofilm functions, HMOs have been shown to modulate inflammatory pathways by altering cytokine expression, including interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), which are key mediators in infection and inflammation associated complications. In this study, we characterize the effects of HMOs on cytokine secretion in THP-1 macrophages stimulated with lipopolysaccharides using a validated magnetic bead electrochemical sandwich assay (MBESA). The electrochemical assay enabled precise quantification of cytokine responses and revealed that HMO treatment attenuated proinflammatory signaling in LPS stimulated macrophages. These findings provide new insights into the immunomodulatory properties of HMOs and demonstrate the utility of electrochemical biosensing platforms for rapid, sensitive assessment of cellular inflammatory responses. Future studies will expand this approach to microphysiological systems to further elucidate HMO mediated regulation of immune signaling.