Surface expression of CD14, TLR4 and TLR2 and cytokine release by Monomac-6 cells in response to Lipopolysaccharide and Pam3 stimulation
Cardiff Metropolitan University
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Sepsis still remains the most common cause of death of the patients in the intensive care unit. Therefore, investigation of the mechanisms behind the recognition of bacterial structural components can be used to determine potential therapeutic targets for septic shock. In this study, the surface expressions of CD14, TLR4 and TLR2, as well as TNF- and IL-8 releases by MM6 cells in response to LPS and Pam3 were studied. The surface expressions of CD14, TLR4 and TLR2 were determined by flow cytometry while the concentrations of TNF- and IL-8 released were determined by ELISA. The CD14 surface expression of MM6 cells peaked at 24 hours of LPS stimulation while the TLR4 surface expression peaked between 1 and 2 hours of stimulation. The study also compared the effects of three different LPS strains. The rough LPS was found to show the least responses in comparison to the smooth forms. The cellular responses to LPS were therefore, found to be dependent on the concentration and type of LPS (rough or smooth) and the length of exposure. Pam3 stimulation had also induced an up-regulation of the surface CD14 expression. However, Pam3 did not cause any significant change to the TLR2 expression at the time-points tested. The Pam3-induced TNF- and IL-8 releases were found to be concentration-dependent and were comparatively lesser than the LPS-induced responses.
MSc. Biomedical Science (Immunohaematology)
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