Related Experiment Video
Updated: Aug 5, 2026
Handling of the Cotton Rat in Studies for the Pre-clinical Evaluation of Oncolytic Viruses
Published on: November 24, 2014
Assessment of the Novel rVSV-PD-1-4-1BBL Oncolytic Activity on Mouse and Human Cancer Cell Lines
Margarita Zinovieva1, Anastasia Ryapolova1, Ilnaz Imatdinov2
1Department of Gene Therapy, Sirius University of Science and Technology, 1 Olympic Avenue, 354340 Sochi, Russia.
Abstract:
Background: Oncolytic viruses (OVs), a promising anti-cancer therapeutic, replicate more efficiently in cancer cells rather than in healthy cells due to the alterations in antiviral response mechanisms and dysregulation of signaling pathways. Vesicular stomatitis virus (VSV) is known for low pathogenicity, tropism to various cancer cells, and the ability to lyse cells in the hypoxic tumor microenvironment (TME). Targeted delivery of immune checkpoint and co-stimulatory molecules can enhance the anti-tumor immune response and remodel the immunosuppressive TME. The aim of this study was to compare the activity of rVSV-GFP with rVSV, encoding the programmed cell death protein 1 (PD-1) and tumor necrosis factor ligand superfamily member 9 (4-1BBL). Methods: The oncolytic efficacy of these rVSV variants used at 105, 106, and 107 TCID50 was evaluated at 24 and 48 h post-infection by flow cytometry in a panel of mouse and human cancer cell lines. Quantitative real-time polymerase chain reaction (qPCR) was used to evaluate mRNA expression levels of certain genes at 12 and 48 h post-infection. Results: Murine hepatocellular carcinoma (H22) and human melanoma (A375) or human lung carcinoma (A549) were the most sensitive to rVSV therapy cell lines. The higher relative expression of the antiviral response genes RIG-I and IFIT1 within each biological species (mouse or human) correlated with lower sensitivity to rVSV. No such effect was observed for the type I interferons (IFNs), despite their proposed key role in resistance to OV therapy. Conclusions: H22, A375, and A549 are more susceptible to the oncolytic activity of the novel rVSV-PD-1-4-1BBL.
Insights
This study shows that modified vesicular stomatitis virus (rVSV) encoding PD-1 and 4-1BBL effectively targets cancer cells. Certain cancer cell lines, like H22, A375, and A549, are more sensitive to this oncolytic virus therapy.
Area of Science:
- Oncolytic virotherapy
- Cancer immunotherapy
- Molecular virology
Background:
- Oncolytic viruses (OVs) show preferential replication in cancer cells due to altered antiviral responses.
- Vesicular stomatitis virus (VSV) exhibits low pathogenicity and tropism for cancer cells, effective in hypoxic tumor microenvironments.
- Engineering OVs to express immune checkpoint and co-stimulatory molecules can enhance anti-tumor immunity and modify the tumor microenvironment.
Purpose of the Study:
- To compare the oncolytic efficacy of rVSV-GFP with a novel rVSV variant encoding programmed cell death protein 1 (PD-1) and tumor necrosis factor ligand superfamily member 9 (4-1BBL).
- To evaluate the sensitivity of various mouse and human cancer cell lines to different doses of these rVSV variants.
Main Methods:
- Oncolytic efficacy was assessed using flow cytometry in cancer cell lines treated with rVSV variants (10^5 to 10^7 TCID50) at 24 and 48 hours post-infection.
- Quantitative real-time polymerase chain reaction (qPCR) was employed to measure mRNA expression of antiviral response genes at 12 and 48 hours post-infection.
Main Results:
- Murine hepatocellular carcinoma (H22), human melanoma (A375), and human lung carcinoma (A549) cell lines demonstrated the highest sensitivity to rVSV therapy.
- Higher expression of antiviral genes RIG-I and IFIT1 correlated with decreased sensitivity to rVSV within species, while type I IFNs showed no such correlation.
- The rVSV-PD-1-4-1BBL variant showed promising oncolytic activity in susceptible cell lines.
Conclusions:
- H22, A375, and A549 cell lines are particularly susceptible to the oncolytic effects of the engineered rVSV-PD-1-4-1BBL.
- Antiviral gene expression, specifically RIG-I and IFIT1, influences cancer cell sensitivity to VSV-based oncolytic therapy.
- The novel rVSV-PD-1-4-1BBL represents a potential therapeutic candidate for enhancing anti-tumor immune responses.
