The focus of the experiment that was conducted during the summer of 2021 was to determine the effects of chlamydia on human host cells, specifically in regards to apoptosis. Apoptosis, commonly referred to as “programmed cell death”, is triggered in high stress environments in order to stop pathogens from spreading without causing inflammation in the host. In order to initiate apoptosis, host cells must release ceramide, a messenger molecule. The hypothesis was that chlamydia was sequestering ceramide from the host cell to inhibit apoptosis. An MTT assay was used to measure decreased mitochondrial function, which typically is a good indicator of apoptosis. For experimentation, Chlamydia infected and uninfected cells were exposed to various concentrations of ceramide. It was anticipated that infected cells exposed to the same concentration of ceramide as uninfected cells would exhibit lower levels of apoptosis since the ceramide used to trigger this action was being uptook by the bacterium. After several trials, it could be concluded that the MTT assay was not the most accurate test in confirming apoptotic activity because chlamydia interferes with mitochondrial function and gives false readings. Moving forward, experimentation will be done with a caspase assay that measures protein activation in regards to apoptosis to give a more accurate portrayal of the impacts of chlamydia on host cell apoptosis. This experience was relevant for my intended career as a researcher as it gave me the opportunity to grow my technical skills in the lab, learn how to develop a procedure, and plan an experiment.
Author: Maddie Canter
Advisor: Joseph Carlin, Microbiology
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