Assessment of Inflammatory Response on Pyrogenic Induction by In vitro and In vivo Methods

Abstract

The incidence of pyrogenic contaminations creates a paramount safety concern in the field of biotherapeutics, as it compromises the normal well-being of an individual. After exposure to pyrogenic agents, the febrile response can create a severe reaction from shock and further complications leading to death. So it is imperative that the medical aids, as well as the parenteral drugs, must be pyrogen-free. The possible reliability of continuous supply of healthy human blood can be overcome by implementing a novel approach by pooling the blood samples for checking the pyrogenicity. In this study, an indigenously developed sandwich ELISA method to quantify pro-inflammatory cytokine (IL-1β) activated after pyrogenic exposure was used. The study unraveled the possibility of using the human pooled blood system to detect the endogenous pyrogen (IL-β) after exposure to endotoxins from Gram-positive and negative bacteria and chemical toxicants such as phytohemagglutinin (PHA) and 2,4,6-trinitrophenol (TNP). The results indicate that upon exposure to the high concentration of endotoxin such as LPS (5EU/ml) and LTA (1µg/ml), the maximum release of IL-1β was observed within 2h of post-stimulation. The stimulation with chemical toxicants PHA and TNP triggered a sudden release of IL-1β within 2h in both cases after 30µg/ml treatment. The study conveys a better platform for providing a continuous supply of healthy human blood, thus minimizing personal variations for the detection of IL-1β for evaluating in vitro pyrogenic response. It also depicts understanding the possible underlying mechanism of the pyrogenic response level of inflammatory cytokines and the possible rise in biochemical parameters leading to complications that affect the normal homeostasis by lipopolysaccharide-induced in rabbits.