Open Access Senior Honors Thesis
The murine alveolar macrophage cell line, MH-S, constitutively expresses the immunoregulatory antigen CD40. Previous experiments have demonstrated that CD40 expression is upregulated following infection with the DNA virus, mouse adenovirus 1 1 Obua (MA V-1 ), suggesting a role for CD40 in MA V-1 infection. The purpose of this experiment is to investigate the effect ofCD40 silencing on MA V-1 replication in MH-S cells. To elucidate the importance ofCD40 signaling in MA V-1 replication, CD40 expression was reduced using a short hairpin RNA (shRNA) RNAi system. Five CD40-silenced lines were generated using shRNA and showed between 26-79 percent reduction in cell surface CD40 expression from wild type. To assess the effect of MA V-1 infection on costimulatory markers, CD40 expression was compared between the silenced cell line that showed the greatest level of CD40 suppression (designated NPBS) and wild type cells at I-day post infection. Flow cytometry revealed CD40 expression was increased in both cell lines following infection with MA V-1. Infected CD40-silenced cultures expressed approximately 50% the level of CD40 as infected wild type cultures. Further characterization of CD40-deficient MH-S cells may reveal novel mechanisms through which MA V-1 infection alters host cell metabolism.
Obua, Andre, "Effect of CD40 Silencing in MAV-1 Infected MH-S Cells" (2017). Senior Honors Theses & Projects. 550.