QUANTITATIVE ESTIMATION OF PORCINE ENDOGENOUS RETROVIRUS RELEASE FROM PK15 CELLS
Magdalena Kimsa, Barbara Strzalka-Mrozik, Malgorzata Kimsa, Jolanta Adamska, Joanna Gola, Krzysztof Lopata, Urszula Mazurek Pol J Microbiol 2012; 61 (3): ICID: 1014509 Article type: Original article IC™ Value: 10.00
Abstract provided by Publisher
The present study focuses on the assessment of PERV release from PK15 cells in a time dependent manner. A PK15 cell line (ATCC CCL-33) was obtained from the American Type Culture Collection and routinely maintained in the DMEM medium. The copy number of PERV A, PERV B and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene were evaluated by real-time QPCR and QRT-PCR. Additionally, virus production and release were estimated by measuring reverse transcriptase (RT) activity. The highest number of viral RNA in PK15 cells was observed after 16 and 20 hours for PERV A and PERV B, respectively. In the culture medium, the highest increase of viral RNA for both subtypes was noticed after 32 hours. The peaks of PERV RT activity at the 28th hour of culture in PK15 cells and at the 32nd hour in the culture medium were found. The copy number of PERV A and PERV B RNA, both in PK15 cells and in the culture medium, slightly changed in a time dependent manner. The monitoring of PERV release from a PK15 cell line may be useful for the evaluation of PERV replication properties.