EFFECTS OF FULLERENOL C60(OH)22-24 ON LUMINOL- AND LUCIGENIN-DEPENDENT CHEMILUMINESCENCE OF HUMAN LEUKOCYTES

Fullerenol C₆₀(OH)_22-24 is a promising fullerene derivative characterized by high water solubility, biocompatibility, and antioxidant properties. Despite extensive studies of its reactive oxygen species-scavenging effects in vitro and in vivo, its impact on the oxidative response of immune phagocytes remains unclear. This study aimed to assess reactive oxygen species production by human peripheral blood leukocytes exposed to fullerenol C₆₀(OH)_22-24 using luminol- and lucigenin-dependent chemiluminescence. Fullerenol C₆₀(OH)_22-24 (MST-WS60-Bio, 99.99% purity) formed monodisperse aggregates (~130 nm) with a zeta potential of -26 ± 6.31 mV (mean ± SD). Leukocytes (10⁶/mL), isolated from healthy female donors (n=4, aged 18–25 years) by sedimentation, were stimulated with opsonized zymosan (1.5 μg/mL). Fullerenol C₆₀(OH)_22-24 did not affect spontaneous luminol-dependent chemiluminescence at 30 and 60 min but nanoparticle’s high concentrations (50–200 μg/mL) reduced it after 90 min. In opsonized zymosan-stimulated luminol assays, inhibition occurred from 30 min across 5–200 μg/mL and persisted to 90 min (25–200 μg/mL). Spontaneous lucigenin-dependent chemiluminescence remained unchanged, but opsonized zymosan-stimulated signals were suppressed from 12.5 to 200 μg/mL at 30–60 min, extending to 90 min at 50–200 μg/mL. In cell-free H₂O₂ systems (0.039–0.625%) fullerenol did not quench either signal but enhanced luminol-dependent chemiluminescence (5–200 μg/mL), indicating peroxidase-like activity. Fullerenol C₆₀(OH)_22-24 (5–200 μg/mL) significantly suppresses luminol- and lucigenin-dependent chemiluminescence in stimulated human leukocytes, consistent with its antioxidant properties and suggesting superoxide dismutase- and/or catalase-like activity via hydroxyl hydrogen bonding and π-electron interactions.