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Receptor for advanced glycation end products (RAGE) activation is known to play an important role in the development of diabetes complications by amplifying the inflammatory process. Herein, we examined the effect of polyclonal antibodies against human RAGE (Ab anti-hRAGE) on “primed” peripheral blood mononuclear cells (PBMNC by hyperglycemia in type 2 diabetes mellitus (T2DM) patients in comparison to healthy control. ROS generation and IL1β, IL-6 and TNF-α secretion were studied. PBMNCs were purified utilizing Ficoll-hypaque gradient. ROS was quantified by luminol-dependent chemiluminescence. Antibodies antihuman RAGE (AGE-like) was purched from Sigma Co. The cytokines were from PBMNC culture in presence or in the absence of anti-hRAGE. IL-1β, IL-6 and TNF-α were quantified in the supernatant of Ab anti-hRAGE-stimulated PBMNCs trough ELISA. Ab anti-hRAGE (AGE-like) significantly inhibited ROS production in unstimulated or stimulated PBMNCs from T2DM and healthy controls in a similar way. The percentage of inhibition was greater in primed PBMNC by hyperglycemia in diabetes (T2DM patients). In contrast, anti-hRAGE increased secretion of IL-1β, IL-6 and TNF-α (p < 0.05) in the supernatant of Ab anti-hRAGE-stimulated PBMNCs from T2DM patients compared with healthy controls (p < 0.05). The effect of antibody anti-hRAGE (AGE-like) in primed cells by hyperglycemia in diabetes may activates different signaling network when interact with RAGE on cells surface. Dual results induced by anti-hRAGE (AGE-like) associated with oxidizing response and pro-inflammatory cytokine secretion suggest activation of several signaling network in AGE-RAGE interaction. It may have consequences on innate immunity.

IL-1β, IL-6 and TNF-Alpha Secretion by PBMNC Stimulated with Polyclonal Antibodies Anti-Hrage (AGE-Like) is Enhanced by Hyperglycemia in Diabetes