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Updated: Mar 31, 2026
Growth, Purification, and Titration of Oncolytic Herpes Simplex Virus
Published on: May 13, 2021
Entry of Oncolytic Herpes Simplex Virus into Human Squamous Cell Carcinoma Cells by Ultrasound
Shusuke Okunaga1, Ayako Takasu2, Noritoshi Meshii3
1Department of Oral and Maxillofacial Surgery, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan. migakedentan@mail.goo.ne.jp.
Abstract:
Low-intensity ultrasound is a useful method to introduce materials into cells due to the transient formation of micropores, called sonoporations, on the cell membrane. Whether oncolytic herpes simplex virus type 1 (HSV-1) can be introduced into oral squamous cell carcinoma (SCC) cells through membrane pores remains undetermined. Human SCC cell line SAS and oncolytic HSV-1 RH2, which was deficient in the 134.5 gene and fusogenic, were used. Cells were exposed to ultrasound in the presence or absence of microbubbles. The increase of virus entry was estimated by plaque numbers. Viral infection was hardly established without the adsorption step, but plaque number was increased by the exposure of HSV-1-inoculated cells to ultrasound. Plaque number was also increased even if SAS cells were exposed to ultrasound and inoculated with RH2 without the adsorption step. This effect was abolished when the interval from ultrasound exposure to virus inoculation was prolonged. Scanning electron microscopy revealed depressed spots on the cell surface after exposure to ultrasound. These results suggest that oncolytic HSV-1 RH2 can be introduced into SAS cells through ultrasound-mediated pores of the cell membrane that are resealed after an interval.
Insights
Low-intensity ultrasound facilitates oncolytic herpes simplex virus type 1 (HSV-1) entry into oral cancer cells by creating temporary membrane pores. This sonoporation technique enhances viral delivery for potential cancer therapy.
Area of Science:
- Biomedical Engineering
- Oncology
- Virology
Background:
- Low-intensity ultrasound induces transient cell membrane pores (sonoporation), enabling material entry.
- The efficacy of introducing oncolytic herpes simplex virus type 1 (HSV-1) into oral squamous cell carcinoma (SCC) via sonoporation is not well-established.
Purpose of the Study:
- To investigate the potential of ultrasound-mediated sonoporation for delivering oncolytic HSV-1 into human oral SCC cells.
- To determine the influence of ultrasound exposure timing relative to viral inoculation on virus entry.
Main Methods:
- Human SCC cell line (SAS) and an oncolytic HSV-1 (RH2) were utilized.
- Cells were exposed to low-intensity ultrasound, with and without microbubbles, followed by inoculation with HSV-1.
- Virus entry was quantified by plaque assays; cell surface changes were observed using scanning electron microscopy.
Main Results:
- Ultrasound exposure significantly increased HSV-1 entry into SCC cells, even without a prior adsorption step.
- The enhancement of virus entry was dependent on the interval between ultrasound exposure and viral inoculation, diminishing with longer delays.
- Scanning electron microscopy showed surface depressions on cells post-ultrasound treatment, indicative of pore formation.
Conclusions:
- Ultrasound-mediated sonoporation effectively introduces oncolytic HSV-1 into oral SCC cells.
- The transient pores created by ultrasound allow for virus entry, which is resealed over time.
- This method holds promise for enhancing the delivery of oncolytic viruses in cancer therapy.
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