Evaluation of peripheral blood mononuclear cells (PBMC) for detection of interleukin-12 production using luminex / Areena Balqis Mohd Fuad

PBMCs embody the cellular part of the blood organ containing all blood cells with a round nucleus which mainly constitute of monocytes, dendritic cells, natural killer (NK) cells, B cells, T cells, and which plays a crucial role in the immune system and also respond in an inflammatory manner. The populations of PBMC are of which the immune cells that are often recognized by as buffy coat of which where the cells are collected during the method of Ficoll fractionation. The role of the interleukin-12 (IL-12) is greatly essential towards the ability of adaptive and innate immune systems in order to work together and communicate. Bacterial lipopolysaccharide (LPS) is vastly implemented in models for the studying of inflammation due to it impersonate numerous inflammatory response of the cytokines. The findings from this research is to evaluate on the benefits and also the ability of PBMC in the detection of IL-12 production in ex-vivo culture using Luminex. The Luminex cytokine assay implement high-throughput screening of compounds in primary cells using cytokine profiles in biological samples derived from cell culture, animals or patients as physiologically relevant readouts.Whole blood sample is isolated into PBMC where cell viability was performed before proceeding with cell culture then later analyzed with Luminex. The stimulated PBMC is tested against the non-stimulated PBMC in identifying the difference of IL-12 expression between stimulated and non-stimulated PBMC in ex-vivo culture. The outcome of this study supports previous studies’ claims where the LPS induces the pro-inflammatory cytokine release in this case of which is IL-12.