Differential protein expression of PK-15 cells infected with PCV2 encoding full-length or truncated ORF3 protein in response to swine interferon gamma treatment

Name: Pattama Mutthi

LCSH: Kasetsart University -- Dissertations. Ph.D. (Genetic Engineering) 2018

Classification :.LCCS: SF977.V57

LCSH: Kasetsart University. -- Interdisciplinary Graduate Program -- Dissertations

LCSH: Swine -- Virus diseases

LCSH: Circoviruses

LCSH: Porcine reproductive and respiratory syndrome

LCSH: Interferon

LCSH: Gene expression

Abstract: Porcine circovirus type 2 (PCV2) is a causative agent of post-weaning multisystemic wasting syndrome (PMWS) or porcine circovirus associated disease (PCVAD). One of the key points is the imbalance of pig immune status or cytokines which involved in immunostimulation or immunosuppression cause of the PCV2 outbreak. Here in, we have determined the growth kinetics of PCV2 encoding truncated-ORF3 and wild type PCV2 with full-length ORF3 protein in the response to swine interferon gamma (swIFNγ). Furthermore, we have explored the differential expression of cellular proteins that are related to the enhanced PCV2 replication in PK-15 cells using mass spectrometry. The results showed that the wild type PCV2 had higher replication rate than PCV2 with the truncated-ORF3. The differential expressed proteins included those involved in cell growth, cell division and cell survival. Level of cdk3 and map3K7 gene expression were increased significantly (P<0.05) whereas the expression of other genes (traf5, bcl-2, hsp90, c-myc, casp3, casp9 and 14-3-3 epizilon, ras-GTPase and ubb2B) were not significantly different. However, swIFNγ effect was determined only in the wild type PCV2 expressing full-length ORF3 protein. The results showed that replication rate of PCV2 in the presence or absence of swIFNγ at 48 hours post swIFNγ treatment were not significantly different in both conditions. Both PCV2 presented a similar growth pattern. The differential protein expressions between the two groups were predominantly involved in cell apoptosis, cell growth and cell survival. The proteomics data was confirmed by comparison of the relative gene expression level using quantitative RT-PCR. The results of RT-qPCR showed that three of nine genes including traf5, c-myc and ras-GTPase were increased significantly (P<0.05) but only map3K7 gene was significantly decreased. However, other genes (bcl-2, hsp90, casp3, casp9 and 14-3-3 episilon) were not statistically significant. The overall study showed that ORF3 protein had a strong direct effect of PCV2 replication while swIFNγ might have an indirect effect to increase virus replication in vitro. The ORF3 protein may be important for protein-protein interaction between PCV2 and host cellular proteins resulting in the production of cellular biological substances that enrich for PCV2 replication. Also, swIFNγ had an indirect effect on the increased PCV2 replication that helps extending cell survival and induces cell proliferation to support viral replication in infected cells resulting in the production of new PCV2 progeny.

Kasetsart University. Office of the University Library

Address: Bangkok

Email: tdckulib@ku.ac.th

Name: Porntippa Lekcharoensuk

Role: Thesis Advisor

Name: Sittiruk Roytrakul

Role: Thesis CO-Advisor

Created: 2018

Modified: 2025-06-29

Issued: 2025-06-29

วิทยานิพนธ์/Thesis

application/pdf

URL: https://www.lib.ku.ac.th/KUthesis/2561/pattama-mut-all.pdf

CallNumber: SF977.V57 .P37

eng

DegreeName: Doctor of Philosophy

Level: Doctoral Degree

Descipline: Genetic Engineering

Grantor: Kasetsart University

©copyrights Kasetsart University

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