Porcine Circovirus Cap-Induced Apoptosis of Non-Infected PK15 Cells

Rowell, Jared S.

Porcine Circovirus Cap-Induced Apoptosis of Non-Infected PK15 Cells

dc.contributor.advisor	Czub, Markus
dc.contributor.author	Rowell, Jared S.
dc.contributor.committeemember	Zheng, Xilong
dc.contributor.committeemember	Jirik, Frank Robert
dc.contributor.committeemember	Knight, Cameron G.
dc.contributor.committeemember	Corcoran, Jennifer A.
dc.date	2019-11
dc.date.accessioned	2019-07-18T20:09:11Z
dc.date.available	2019-07-18T20:09:11Z
dc.date.issued	2019-07-16
dc.description.abstract	Porcine Circovirus 2 (PCV2) is a pathogen of major importance for swine production around the world. Despite development of several vaccines and robust vaccination programs, PCV2 continues to persistently transmit within and between swine herds causing infections marked by immunosuppression via lymphocyte depletion. This study aims to improve understanding of porcine circovirus diseases (PCVD) pathogenesis caused by PCV capsid cytotoxicity which activates apoptosis in non-infected bystander cells. Putatively non-pathogenic Porcine Circovirus 1 (PCV1) was also included to help understand why there is a difference in pathogenicity between PCV1 and PCV2. Porcine Circovirus 3 (PCV3) has recently emerged worldwide as a possible etiological agent of clinical syndromes observed in swine that are similar to PCVD caused by PCV2. The capsid protein of PCV3 (PCV 3 Cap) was also included in this study to provide initial information regarding the cytotoxicity of this protein. Results were obtained through flow cytometric analysis of apoptosis using Annexin V-FITC and 7-AAD to measure apoptosis markers and cell death respectively. Flow cytometry results were reinforced by also utilizing a TUNEL assay in conjunction with confocal imaging to detect cells undergoing DNA fragmentation leading to apoptosis. Results show that monomeric PCV2 Cap, 1% formaldehyde inactivated and dialyzed PCV2, PCV1 virus-like particles (VLPs), PCV2 VLPs, and PCV3 VLPs are all able to activate apoptosis in porcine kidney (PK15) cells at a rate of ~%30 after a 24-hour exposure; this apoptotic similar to apoptosis caused by 50 µM etoposide, which is a strong chemical inducer of apoptosis. These results coupled with preliminary data obtained by pre-treating PK15 cells with caspase-8 or caspase-9 inhibitors suggest the involvement of caspase-8 and caspase-9 in PCV2 Cap induced apoptosis and lend weight to the idea of PCV2 Cap as a primary virulence factor during infections. In contrast, PCV1 Cap induced apoptosis does not appear to involve caspase-8 or caspase-9 indicating an alternate pathway to PK15 cell apoptosis, while PCV3 Cap appears to induce PK15 cell apoptosis with some involvement of caspase-9.	en_US
dc.identifier.citation	Rowell, J. S. (2019). Porcine Circovirus Cap-Induced Apoptosis of Non-Infected PK15 Cells (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.	en_US
dc.identifier.doi	http://dx.doi.org/10.11575/PRISM/36752
dc.identifier.uri	http://hdl.handle.net/1880/110647
dc.language.iso	eng	en_US
dc.publisher.faculty	Cumming School of Medicine	en_US
dc.publisher.institution	University of Calgary	en
dc.rights	University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.	en_US
dc.subject	Porcine circovirus, baculovirus, apoptosis, immunnosuppression, virus-like-particles	en_US
dc.subject.classification	Virology	en_US
dc.title	Porcine Circovirus Cap-Induced Apoptosis of Non-Infected PK15 Cells	en_US
dc.type	master thesis	en_US
thesis.degree.discipline	Medicine – Microbiology & Infectious Diseases	en_US
thesis.degree.grantor	University of Calgary	en_US
thesis.degree.name	Master of Science (MSc)	en_US
ucalgary.item.requestcopy	true	en_US