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Research Article: Effect of bacteriophages on cytokine production by human peripheral blood mononuclear cells and their subpopulations

Date Published: 2026-10-13

Abstract:
Fulfilling a vision that extends beyond their role as antibiotic alternatives, bacteriophages are now recognized as active immunomodulators, a concept aligning with early hypotheses of their biological activity. In addition to their bactericidal action, these properties require detailed investigation. This study aims to characterize the immunomodulatory effects of bacteriophages on pro- and anti-inflammatory cytokine production by human immune cells isolated from healthy blood donors. We assessed the secretion of a comprehensive panel of 11 cytokines (including IFN-?, IL-6, TNF-?, and IL-10) by peripheral blood mononuclear cells and their isolated subpopulations (B and T lymphocytes, helper and cytotoxic T lymphocytes, natural killer cells, and monocytes). The cells were incubated for 24 hours with purified Escherichia coli phage T4 and therapeutic staphylococcal phage A5/80. Reference cells were incubated with lipopolysaccharide from E. coli B at a concentration corresponding to its activity in the applied phage preparations. For comparison, we also assessed cytokine responses to human adenovirus 5, used as a benchmark for a virus capable of infecting human cells. Cell-specific analysis revealed a distinct hierarchy in immune responsiveness: our data suggest that monocytes may act as the primary sensors driving cytokine signatures, while we did not confirm a discernible direct response in B, natural killer, and cytotoxic T cells. Importantly, the magnitude of these responses was markedly lower than the robust inflammatory cytokine secretion triggered by human adenovirus 5, providing context for the phage-induced effects relative to a viral infection of human cells and underscoring the limited inflammatory footprint of therapeutic phages. These results improve our understanding of phage–human immune system interactions, which is important for appreciating their broader impact on human homeostasis and for assessing the safety of phage therapy.

Introduction:
Fulfilling a vision that extends beyond their role as antibiotic alternatives, bacteriophages are now recognized as active immunomodulators, a concept aligning with early hypotheses of their biological activity. In addition to their bactericidal action, these properties require detailed investigation. This study aims to characterize the immunomodulatory effects of bacteriophages on pro- and anti-inflammatory cytokine production by human immune cells isolated from healthy blood donors.

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