Alpha Omega Alpha Honor Medical Society

2010 Research Abstract

The Effect of Rab5 GTPase Protein Knockdown in Papillomavirus Cell Trafficking and Infection in Human Keratinocyte Cells

Investigator: Nina L. Gazanfari, Chicago Medical School - Rosalind Franklin University of Medicine and Science

Mentor: Patricio I. Meneses PhD, Rosalind Franklin University of Medicine and Science, Department of Microbiology and Immunology

Human papillomavirus is a double-stranded DNA virus from the papillomaviridae family designated by the CDC as the most common sexually transmitted infection. Although the association between human papillomavirus type 16 and 18 infection and the development of cervical cancer has been clearly established, the process of intracellular trafficking of the virus remains widely undetermined. Studies have shown that the host cell Rab5 GTPase protein may be involved in the docking and fusion of vesicles during early trafficking of the virus. The objective of this study was to investigate the involvement of the Rab5 GTPase in cellular trafficking of the human papillomavirus, and to confirm or deny the role of the Rab5 protein in papillomavirus infection of human keratinocyte cells. We used various transfection agents to insert siRNA complementary to Rab5 mRNA, decreasing expression of the Rab5 protein with intent to observe if human papillomavirus is capable of infecting Rab5 null cells. To transfect the cells with siRNA, we used Genejet, the Amaxa nucleofector kit, and Interferin along with the Odyssey Imaging system to visualize the western blots. We infected control and experimental Rab5 null cells with HPV pseudovirion and assessed infection rates by microscopy and FACS (fluorescence activated cell sorter). The data are preliminary but suggest a decrease in infection rate when the HaCaT cells are transfected with Rab5 siRNA, i.e. when Rab5 protein knockdown occurs. These experiments in transfection methods lay the groundwork for future experiments regarding the cellular trafficking of the human papillomavirus. Further experiments to support or refute our hypothesis will be helpful in the discovery of new drug targets to prevent the onset of HPV infection, and thus HPV related disease including invasive cervical carcinoma.

Supported in part by an Alpha Omega Alpha Carolyn L Kuckein Student Research Fellowship.

Updated on June 16, 2011.

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