Neutrophils play a key role in the defense against microbial infections. One of their primary weapons to destroy microbes is the production of reactive oxygen species (ROS). This paper shows how 2', 7'-dichlorodihydrofluorescein-diacetate (DCFH-DA) was used to measure the effects of polysaccharides on the production of ROS by polymorphonuclear leukocytes (PMNs). The DCFH-DA method has been designed to provide a highly sensitive, quantifiable, real-time assessment of ROS production by PMNs.
Use of carbohydrates as elicitors is a novel technique for enhancement of the production of industrially important microbial products. The relation between the levels of ROS (reactive oxygen species) and overproduction of antibiotics in microbial cultures has already been established. In the present study, we aimed to exploit the ROS response to develop a fast technique to assess the potential of oligosaccharides [oligoguluronate, oligomannuronate and MO (mannan oligosaccharides)] and polysaccharides [alginate and LBG (locust-bean gum)] as elicitors for overproduction of secondary metabolites in Streptomyces rimosus and Penicillium chrysogenum. We have also investigated changes in the production of ROS in neutrophils as a result of the action of the same elicitors. LBG-derived oligosaccharides (MO) were most potent inhibitors of ROS in all systems investigated. This correlates with overproduction of secondary metabolites in microbes and enhancement of a number of mammalian systems. We believe that the effects of oligosaccharides and polysaccharides on ROS production by mammalian and microbial cells can be correlated predicatively with overproduction. The underlying methodology offers a fast screening of elicitors that can be applied across the different systems.
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