Exploration of laboratory techniques relating to Cryptosporidium parvum propagation, life cycle observation, and host immune responses to infection.

摘要

Cryptosporidium causes cryptosporidiosis, a self-limiting diarrheal disease in healthy people, but causes serious health issues for immunocompromised individuals. Cryptosporidiosis has been observed in humans since the early 1970s and continues to cause public health concerns. Cryptosporidium has a complicated life cycle making laboratory study challenging. This project explores several ways of studying Cryptosporidium parvum, with a goal of applying existing techniques to further understand this life cycle. Utilization of a neonatal mouse model demonstrated laser microdissection as a tool for studying host immune response to infeciton. A cell culture technique developed on FrameSlides(TM) enables laser microdissection of individual infected cells for further analysis. Finally, the hypothesis that the availability of cells to infect drives the switch from asexual to sexual parasite reproduction was tested by time-series infection. The results suggest this isn't accurate. These experiments open the door to several avenues of Cryptosporidium study and the host response to cryptosporidiosis.