Chlamydia trachomatis is an obligate intracellular, bacterial pathogen. C. trachomatis infections of the human reproductive tract affect approximately 127 million people globally each year. The major concern of C. trachomatis infections is their ability to cause infertility in both men and women, by damaging the upper reproductive tracts. However, we are still lacking information about how Chlamydia travels around the reproductive tract, and reaches the upper tract (ovaries and testes in particular) to cause this damage.
Recent research has shown that macrophages are a key cell type responsible for harbouring and transmitting Chlamydia within the reproductive tract. Macrophages are generally thought of as an immune cell responsible for fighting infections. However, in the case of Chlamydia, this does not always occur. Chlamydia possesses molecular mechanisms that allow it to avoid macrophage-mediated killing and under some circumstances survive and replicate within these cells. Research has shown that during chlamydial infection, cells undergo genetic and transcriptional changes. However, limited information exists about changes that occur within the Chlamydia during growth within immune cells and macrophages.
This project aims to elucidate the infectivity and infection kinetics of chlamydial growth within macrophages.
Hypothesis: Chlamydia that has been grown long-term in macrophages undergoes infectivity and infection kinetic changes that promote its survival.
Aim 1: To determine whether passage one or passage 30 Chlamydia harvested from RAW 264.7 macrophage cells has altered infectivity in macrophages and testicular cells.
Aim 2: To determine whether passage one or passage 30 Chlamydia harvested from RAW 264.7 macrophages has altered inclusion morphology, size, and progeny production in macrophages and testicular cells.
Aim 3: To determine whether passage one or passage 30 Chlamydia harvested from RAW 264.7 macrophages has altered inclusion longevity over a time-course in macrophages and testicular cells.
Approaches/Skills and techniques
Mammalian cell culture, chlamydial cell culture, immunocytochemistry, epifluorescent microscopy, RNA isolation, RNAseq, differential gene analysis, KEGG pathway mapping, biological system analysis, written and oral communication skills.
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