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<p>Extracellular vesicles (EVs) from commensal microbiota are emerging mediators of host–microbe communication, yet their immunological roles remain incompletely defined. In this study, we investigated the immunostimulatory potential of EVs derived from <i>Limosilactobacillus reuteri</i> EIR/Spx-2 (LrEVs), a strain isolated from the gut microbiota of the naturally cancer-resistant blind mole rat. LrEVs were isolated by ultracentrifugation and displayed spherical morphology with diameters of 30 to 100&#xa0;nm, confirmed by electron microscopy, atomic force microscopy, and tunable resistive pulse sensing. In RAW 264.7 macrophage cells, a non-cytotoxic concentration of LrEVs (25&#xa0;µg/mL) significantly upregulated the expression of pro-inflammatory mediators (<i>TNF-α</i>,<i> IL-6</i>,<i> IL-1β</i>,<i> iNOS</i>,<i> COX-2</i>), and pattern recognition receptors (<i>TLR2</i>,<i> NOD2</i>). Consistently, ELISA results demonstrated marked increases in TNF-α and IL-6 secretion comparable to lipopolysaccharide stimulation. Flow cytometry analysis further revealed upregulation of CD80 and MHC II, indicating macrophage activation. Proteomic profiling revealed vesicle-associated enzymes involved in the biosynthesis of lipoteichoic acid, peptidoglycan, and muramyl dipeptide, supporting activation of TLR2- and NOD2-mediated immune pathways. Collectively, LrEVs exhibit potent immunostimulatory activity as bioactive mediators of host–microbe interactions. However, given the <i>in vitro</i> nature of the study, further investigation in relevant preclinical models is required to assess their translational relevance.</p>

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Immunostimulatory potential of extracellular vesicles from Limosilactobacillus reuteri EIR/Spx-2, a strain from the gut microbiota of a naturally cancer-resistant host

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月2日

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  • Hazal Kibar Demirhan,
  • Meltem Asicioglu,
  • Tamer Kahraman,
  • Fadime Kiran

摘要

Extracellular vesicles (EVs) from commensal microbiota are emerging mediators of host–microbe communication, yet their immunological roles remain incompletely defined. In this study, we investigated the immunostimulatory potential of EVs derived from Limosilactobacillus reuteri EIR/Spx-2 (LrEVs), a strain isolated from the gut microbiota of the naturally cancer-resistant blind mole rat. LrEVs were isolated by ultracentrifugation and displayed spherical morphology with diameters of 30 to 100 nm, confirmed by electron microscopy, atomic force microscopy, and tunable resistive pulse sensing. In RAW 264.7 macrophage cells, a non-cytotoxic concentration of LrEVs (25 µg/mL) significantly upregulated the expression of pro-inflammatory mediators (TNF-α, IL-6, IL-1β, iNOS, COX-2), and pattern recognition receptors (TLR2, NOD2). Consistently, ELISA results demonstrated marked increases in TNF-α and IL-6 secretion comparable to lipopolysaccharide stimulation. Flow cytometry analysis further revealed upregulation of CD80 and MHC II, indicating macrophage activation. Proteomic profiling revealed vesicle-associated enzymes involved in the biosynthesis of lipoteichoic acid, peptidoglycan, and muramyl dipeptide, supporting activation of TLR2- and NOD2-mediated immune pathways. Collectively, LrEVs exhibit potent immunostimulatory activity as bioactive mediators of host–microbe interactions. However, given the in vitro nature of the study, further investigation in relevant preclinical models is required to assess their translational relevance.