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Updated: Jan 27, 2026
Growth, Purification, and Titration of Oncolytic Herpes Simplex Virus
Published on: May 13, 2021
Cytotoxic effect of dual fluorescent-labeled oncolytic herpes simplex virus type 1 on mouse tumorigenic cell lines
Shahriyar Abdoli1, Farzin Roohvand1, Ladan Teimoori-Toolabi2
1Department of Virology, Pasteur Institute of Iran, Tehran, I.R. Iran.
Abstract:
The increasing incidences of cancer at the global scale have recently resulted in the invention of various biotechnology approaches among which the oncolytic virotherapy is a new strategy for the treatment of multiple tumors. Herpes simplex virus (HSV) based vectors are one of the most studied oncolytic agents, worldwide. Moreover, syngeneic animal models are the principal parts of the oncolytic virotherapies investigation. The effects of a dual fluorescent γ34.5 deleted vector-HSV-GR- on three mouse tumor cell lines were studied in this work. We previously generated a dual fluorescent labeled oncolytic HSV-HSV-GR- (both copies of γ34.5 were inactivated by insertion of two distinct fluorescent dyes, GFP and mCherry) in our laboratory; subsequently, they were used as oncolytic viruses. The three 4T1, TC-1, and CT26 cell lines were infected with HSV-GR. The infection efficacy and the elimination potency of HSV-GR were analyzed by photomicrography and flow cytometry methods. HSV-GR showed a significant efficiency to infect the cell lines examined. Flow cytometry analyses demonstrated that HSV-GR infected 89.3%, 86.1%, and 92.4% of 4T1, TC-1, and CT26 cells, respectively. Moreover, propidium iodide (PI) staining of infected cells indicated that HSV-GR could kill 27.9%, 21.2%, and 21.3% of 4T1, TC-1, and CT26 cells, respectively. Interestingly, HSV-GR infected cells were capable of expressing both GFP and mCherry at the same time. The promising effects of the oncolytic virus HSV-GR in the mouse syngeneic tumor cell system have shed more light on the therapeutic potential of this anti-cancer approach.
Insights
Oncolytic virus therapy using a dual fluorescent herpes simplex virus (HSV-GR) effectively infected and killed mouse tumor cells. This shows promise for developing new cancer treatments.
Area of Science:
- Oncology
- Virology
- Biotechnology
Background:
- Cancer incidence is rising globally, driving innovation in biotechnology for cancer treatment.
- Oncolytic virotherapy, using viruses to target and destroy cancer cells, is a promising new strategy.
- Herpes simplex virus (HSV) based vectors are extensively studied for their oncolytic potential.
Purpose of the Study:
- To evaluate the efficacy of a dual fluorescent, γ34.5-deleted HSV vector (HSV-GR) in infecting and eliminating three mouse tumor cell lines.
- To assess the dual-color fluorescence expression (GFP and mCherry) in infected cells.
- To investigate the therapeutic potential of HSV-GR in a syngeneic tumor model.
Main Methods:
- Generation of a dual fluorescent oncolytic HSV (HSV-GR) with inactivated γ34.5 genes expressing GFP and mCherry.
- Infection of 4T1, TC-1, and CT26 mouse tumor cell lines with HSV-GR.
- Analysis of infection efficacy and cell killing using photomicrography and flow cytometry, including propidium iodide (PI) staining.
Main Results:
- HSV-GR demonstrated significant infection efficiency across all tested cell lines: 89.3% (4T1), 86.1% (TC-1), and 92.4% (CT26).
- HSV-GR induced cancer cell death, with killing rates of 27.9% (4T1), 21.2% (TC-1), and 21.3% (CT26).
- Infected cells successfully co-expressed both GFP and mCherry fluorescent markers.
Conclusions:
- The dual fluorescent oncolytic HSV-GR exhibits potent infectivity and tumor cell elimination capabilities in vitro.
- The simultaneous expression of two fluorescent markers in HSV-GR facilitates tracking and evaluation of viral activity.
- These findings support the therapeutic potential of HSV-GR as an oncolytic virus for cancer treatment.
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