Persistent viral infections are a major health concern, with persistently infected (PI) cells being a source of continued shedding of virus and generation of viral mutants. Here, we hypothesized that cells persistently infected with the enveloped virus parainfluenza virus 5 (PIV5) would show altered expression of endoplasmic reticulum (ER) stress proteins and increased resistance to death caused by drug-induced ER stress. To test this, lysates of mock-infected, PIV5 acute-infected, and PIV5 PI human lung A549 cells were collected and levels of ER stress proteins were compared. Western blotting revealed that immunoglobulin heavy chain binding protein (BiP/GRP78) was present in higher levels in acute-infected and PI cells compared to naïve cells, indicating increased ER stress in both acutely infected and PI cells. Interestingly, basal levels of the ER stress-sensing protein IRE1-alpha were upregulated in PI compared to naïve and acutely infected cells, but PI cells showed decreased activation of IRE1-alpha compared to acutely infected cells. Naïve, acute-infected, and PI A549-NLR cells were treated with ER stress-inducing drugs tunicamycin, thapsigargin, and epigallocatechin gallate and monitored in real-time viability assays for drug-induced cell death. PI cells showed lower levels of stress-induced cell death compared to naive cells, whereas acute-infected cells experienced the greatest extent of cell death when challenged with ER stress-inducing drugs. Together, these results support the hypothesis that PIV5 persistently infected cells display altered ER stress response pathways and that PI cells are more resistant to death caused by ER stress-inducing drugs. Additionally, these results suggest that IRE1-alpha plays a key role in the shift from acute to persistent infection. These results have implications for the treatment of persistent viral infections, as well as the potential for these viruses to be used for oncolytic virotherapy in the future.
Bachelor of Science (B.S.)
College of Medicine
Burnett School of Biomedical Sciences
Biomedical Sciences; Biotechnology
Abbitt, Lauren L., "Role of Endoplasmic Reticulum Stress Response in Parainfluenza Virus Acute to Persistent Infections" (2023). Honors Undergraduate Theses. 1321.