Like us, most of our cells need “friends” to live happy lives and realize their full potential. In order to stay close to their friends, some cell lines express adhesion molecules that allow them to bind cells expressing suitable ligands. This adhesion can occur between cells of the same type (homotypic adhesion) and cells of different types (heterotypic adhesion).
During a microarray experiment examining gene expressions in lymphoblastoid cell lines (LCLs), which are B-cells transformed through infection with the Epstein-Barr virus (EBV), the Luftig lab discovered that CD226, an adhesion protein traditionally associated with T and NK cell adhesion, was up-regulated. Former Howard Hughes Research Fellow, Lisa Grossman confirmed CD226 expression across a variety of LCLs and demonstrated that CD226 expression correlated with NF-kB activity (a powerful transcription factor manipulated by EBV).
My project this summer focuses on expanding our lab’s knowledge of CD226 in LCLs. In particular, I am interested in the relationship of CD226 with LFA-1, another adhesion molecule known to be essential to LCL adhesion. In T and NK cells, CD226 is known to assist LFA-1 in binding to its ligand, ICAM. In the process of assisting LFA-1, CD226 forms a physical association with LFA-1, which can be verified through immunoprecipitation of either LFA-1 or CD226 from T cell lysate. However, this association has never been verified in an LCL. Furthermore, CD226 is known to co-localize with LFA-1 in the lipid rafts of T-cells (special regions in the cell membrane which are often involved in cell signaling). While was observed in T-cells through immunofluorescence, it has also has never been verified in LCLs. Additionally, I am attempting to characterize the functional role of CD226 in LCLs through the creation CRISPR-Cas knockout cell lines. By examining how LCLs function (or fail to function) without the presence of CD226, I hope to gain insight into its role in EBV infection.
My project offers a unique opportunity for me to learn and experience new techniques while exploring this very exciting topic. I have a lot I need to accomplish but hopefully this summer will lead to significant progress.
