THE INVESTIGATION OF CYTOKINE SECRETION IN THP-1 DERIVED MICROGLIA
Cardiff Metropolitan University
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The aim of the research was to determine whether cytokines would be produced when stimulating THP-1 cells with different concentration of LPS. The cytokines, TNF-α, IL-6 and IL-1β are expressed as a response to bacterial lipopolysaccharide (LPS) and through the production of mediators activate several host defence functions. IL-1β, IL-6 and TNF-α are inflammatory cytokines with many roles within the immune system. TNF- α is a cytokine that causes a pro-inflammatory response which is essential for the removal of pathogens when they cross the blood brain barrier. Similarly, IL-1β as well as IL-6 is a pro –inflammatory mediator that’s involved in cell proliferation, cell apoptosis etc. The hypothesis of the research was that by stimulating the cells with LPS, it would induce the expression of these inflammatory cytokines. In order to investigate this, THP-1 cells were differentiated into microglia like cells by manipulating the cells in a specific neural media, then they were treated with different concentration of LPS (1μg, 100ng and 10ng). Supernatants from the media was collected and analysed through a duoset ELISA method where three ELISA plates were incubated for 6 hours (each plate for a different cytokine). The results obtained was highly counterintuitive towards the hypothesis of the study. TNF-α was hypothesized to be expressed within 6 hour of incubation as well as IL-1β and IL-6 at high concentration of LPS. Conversely, TNF-α and IL-1β was significantly expressed more from the microglia like cells than the monocytes at low concentration of LPS, whereas no IL-6 was expressed. In conclusion, the hypothesis was disproved as the cytokines that were produce reacted to LPS at a lower concentration rather than a high concentration of LPS. Evidence have suggested that TNF- α and IL-1β optimum incubation time is around 6 hours, therefore the expression of cytokines should have had peaked at this time.
BSc Biomedical Science
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