Selective neutrophil activation via a programmable stopped-flow injection approach: Multiple evidences of priming...

Activated polymorphonuclear neutrophils play an important role in host defense system against invading pathogens via releasing reactive oxygen species through respiratory bursts. Nowadays, neutrophil activation process has been found to be modeled as a two-stages continuum from dormant to primed, and to fully activated. Although the differences of salivary and circulatory neutrophils have been investigated in various experimental designs, priming state of salivary polymorphonuclear neutrophils with respiratory burst has not been comprehensively studied. Here, for the first time so far, a programmable flow injection analysis based on Stop in Flow Cell operation mode has been applied to batch analyzing comparative study of neutrophil activation. The high-sensitivity luminol-dependent chemiluminescence assay has been employed with just 330 μL of cell suspension consumption per sample, and the process has been conducted rapidly and efficiently within 15 min to keep the neutrophils viability. ROS from either unstimulated or substimulated or stimulated salivary polymorphonuclear neutrophils, in comparison with those from autologous circulatory polymorphonuclear neutrophils, have been determined, together with further monitoring the impact of extracellular and intracellular calcium ion (Ca²⁺) on neutrophils ROS generation. Conclusively, the priming state of salivary polymorphonuclear neutrophils during activation has been testified from multiple aspects.