Toxoplasma gondii: Comparing Vesicles from Healthy and Infected CellsÂ
Standing
Undergraduate
Type of Proposal
Oral Research Presentation
Challenges Theme
Open Challenge
Faculty Sponsor
Dr. Sirinart Ananvoranich
Proposal
Toxoplasma gondii is a protozoan parasite with a successfully high infection rate in humans. T. gondii infection can cause a clinical illness called toxoplasmosis when a person ingests contaminated food and water or from mother to fetus during pregnancy as it can cross the placenta. Approximately 1/3rd of the current general population is infected with toxoplasmosis. Typically, toxoplasmosis presents as asymptomatic and dormant in immunocompetent individuals. However, for immunocompromised individuals, toxoplasmosis can present as a serious disease that can lead to various negative effects such as infertility, blindness, encephalitis, acute respiratory failure, and occasionally, death. There is no vaccine for toxoplasmosis, although recent studies show that small extracellular vesicles (EVs) relate to disease protection and the induction of an immune response. T. gondii changes the content of vesicles within cells that it infects, which in turn changes the way cell-to-cell communication occurs between non-infected and infected cells. To test this hypothesis, human foreskin fibroblast (HFF) cells will be cultured in D10 media until fully confluent. Afterwards, using osmotic vesiculation, the EVs of both non-infected and infected cells are excreted and isolated through centrifugation. Then, the EVs go through a Bligh and Dyer procedure to separate proteins embedded on the plasma membrane surface to allow better lipid characterization. Once the proteins are separated, the EVs undergo staining with fluorescent dyes which bind onto different lipid phase regions thus allowing us to characterize the lipid content. Comparison of the vesicle content that is derived from both non-infected cells and infected cells will help identify significant differences which will help current researchers gain a more thorough understanding of how toxoplasmosis affects cell-to-cell communication.
Grand Challenges
Viable, Healthy and Safe Communities
Toxoplasma gondii: Comparing Vesicles from Healthy and Infected CellsÂ
Toxoplasma gondii is a protozoan parasite with a successfully high infection rate in humans. T. gondii infection can cause a clinical illness called toxoplasmosis when a person ingests contaminated food and water or from mother to fetus during pregnancy as it can cross the placenta. Approximately 1/3rd of the current general population is infected with toxoplasmosis. Typically, toxoplasmosis presents as asymptomatic and dormant in immunocompetent individuals. However, for immunocompromised individuals, toxoplasmosis can present as a serious disease that can lead to various negative effects such as infertility, blindness, encephalitis, acute respiratory failure, and occasionally, death. There is no vaccine for toxoplasmosis, although recent studies show that small extracellular vesicles (EVs) relate to disease protection and the induction of an immune response. T. gondii changes the content of vesicles within cells that it infects, which in turn changes the way cell-to-cell communication occurs between non-infected and infected cells. To test this hypothesis, human foreskin fibroblast (HFF) cells will be cultured in D10 media until fully confluent. Afterwards, using osmotic vesiculation, the EVs of both non-infected and infected cells are excreted and isolated through centrifugation. Then, the EVs go through a Bligh and Dyer procedure to separate proteins embedded on the plasma membrane surface to allow better lipid characterization. Once the proteins are separated, the EVs undergo staining with fluorescent dyes which bind onto different lipid phase regions thus allowing us to characterize the lipid content. Comparison of the vesicle content that is derived from both non-infected cells and infected cells will help identify significant differences which will help current researchers gain a more thorough understanding of how toxoplasmosis affects cell-to-cell communication.