Effect of colon carcinoma (Caco-2) derived extracellular vesicles on secretion of pro-inflammatory chemokine, MCP-1, by THP-1 cells
Cardiff Metropolitan University
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Background aims Extracellular vesicles (EVs) derived from human epithelial colorectal adenocarcinoma cells (Caco-2) contribute to immunomodulation of their microenvironment. Evaluation of chemokine production, monocyte chemotactic protein 1 (MCP-1), secreted by THP-1 cells, as monocytic model, upon stimulation by Escherichia Coli lipopolysaccharide has been investigated. Concentration of MCP-1 protein has been compared amongst THP-1 cells and when EVs were present in the sample. The main aim of this study was further understanding of intracellular communication initiated by EVs and their effects on the microenvironment with an interest in recruitment of innate immune cells. Methods Enzyme-linked immunosorbent assay (ELISA) analysis performed to establish MCP-1 concentration amongst samples containing EVs and THP-1 cells at different conditions. Results Caco-2 derived EVs have shown a significant increase in MCP-1 concentration when compared to THP-1 cells alone upon stimulation with LPS. The concentration of MCP-1 protein was not significantly affected after 1 hour incubation at 37°C however the concentration has significantly increased upon 24-hour incubation when LPS added with further increase at 48-hour incubation. Final investigation revealed a change of MCP-1 concentration of 1795.33 pg/ml compared to samples containing THP-1 only, of 24.64 pg/ml (SD ± 0.031). Conclusions Results of this study had provided important insight into intracellular communication between EVs and our immune cells, providing stimulation by external factor, LPS. Among two trials a statistically significant increase in MCP-1 concentration has been detected when compared to THP-1 cells alone. Although this study has shown a strong positive correlation between MCP-1 protein concentration and presence of EVs in the sample, the study had shown significant limitations by sample shortage, lack of EV characterization and values found to be over the range of plate reader in final trial. Further investigation was needed however was not able to performed due to sample unavailability. Nevertheless, EVs have a strong potential in diagnostics and therapeutics, a better understanding of how EVs communicate with other cells to induce changes in their microenvironments would significantly increase our understanding of many processes behind it and their applications in clinical setting.
BSc (Hons) Biomedical Science
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