Related Experiment Video
Updated: Jun 14, 2026
In Vitro Establishment of a Genetically Engineered Murine Head and Neck Cancer Cell Line using an Adeno-Associated Virus-Cas9 System
Published on: January 9, 2020
In vitro and in vivo effects of reovirus on HPV16-transformed mice cells
K Figova1, E Sobotkova, M Duskova
1Department of Pediatric Hematology and Oncology, Charles University 2nd Medical School, Prague, Czech Republic.
Unlabelled:
Oncolytic viruses are examined to serve as anticancer therapeutics. It is expected that in addition to direct oncolytic effect their action will also help eliciting a solid antitumor immunity. In presented series of experiments we have employed two HPV16-transformed mouse (strain C57/B6) cell lines, TC-1 and MK16/III/ABC (MK16), and reovirus type 3, strain Dearing (RV). Both cell lines are highly susceptible to RV and produce large amounts of infectious virus in vitro while normal human are not susceptible to RV. Still, some differences were encountered. TC-1 cells produced moderately lesser amounts of infectious virus, but, paradoxically, were more efficient producers of delta1 antigen of RV and as a consequence of virus infection died more rapidly than simultaneously infected MK16 cells. Minor differences between the cell lines were observed in the percentage of cells arrested in theG2/M phase of the cell cycle and in some markers of apoptosis. When inoculating high doses (5x106) of infected cells (MOI 10 PFU/cell) into syngeneic animals their oncogenic activity was strongly suppressed, nearly completely in the case of MK16 cells and somewhat less efficiently in the case of more oncogenic TC-1 cells. Immunizing experiments in which non-oncogenic doses (106) of RV infected TC-1 cells were tested in parallel with the same doses of irradiated cells brought surprising results. When immunized animals were challenged with TC-1 cells, the irradiated cells proved to be a much better immunogen that the infected cells. However, when challenged with MK16 cells the opposite was true. It is believed that this difference was associated with the different biological properties of the cell lines tested.
Keywords:
reovirus type 3, HPV16-transformed mouse cell lines, apoptosis, cell cycle, immunization/challenge experiments.
Insights
Oncolytic reovirus type 3 effectively suppresses tumor growth in mouse models. However, its efficacy as an immunogen depends on the specific cancer cell line used in immunization strategies.
Area of Science:
- Oncolytic virotherapy
- Immunology
- Cancer research
Background:
- Oncolytic viruses are promising anticancer therapeutics.
- Reovirus type 3 (RV) shows potential for direct oncolysis and stimulating antitumor immunity.
- HPV16-transformed mouse cell lines (TC-1 and MK16) are susceptible to RV infection.
Purpose of the Study:
- To evaluate the oncolytic activity and immunogenic potential of reovirus type 3 (RV) in HPV16-transformed mouse cell lines.
- To compare the effects of RV infection on TC-1 and MK16 cell lines regarding viral replication, cell death, and cell cycle arrest.
- To assess the efficacy of RV-infected cells versus irradiated cells as immunogens in preclinical models.
Main Methods:
- In vitro infection of TC-1 and MK16 cells with reovirus type 3 (RV).
- Assessment of viral replication, delta1 antigen production, cell cycle arrest (G2/M phase), and apoptosis markers.
- In vivo studies involving inoculation of high doses of RV-infected cells to assess oncogenic suppression.
- Immunization experiments using RV-infected or irradiated TC-1 cells followed by challenge with TC-1 or MK16 cells.
Main Results:
- Both TC-1 and MK16 cells supported RV replication, with TC-1 cells showing faster death despite lower viral yields.
- RV infection led to cell cycle arrest and apoptosis in both cell lines, with minor differences observed.
- High doses of RV-infected cells significantly suppressed tumor formation in vivo.
- Immunization with irradiated TC-1 cells was more effective against TC-1 challenge, while RV-infected TC-1 cells were better against MK16 challenge.
Conclusions:
- Reovirus type 3 demonstrates oncolytic potential and can suppress tumor growth in vivo.
- The immunogenic properties of RV-infected cells are cell-line dependent.
- Differences in biological properties between TC-1 and MK16 cells likely influence the outcome of immunization/challenge experiments.
