Related Experiment Video
Updated: May 20, 2026
Ex Vivo Infection of Murine Epidermis with Herpes Simplex Virus Type 1
Published on: August 24, 2015
[Generation of a herpes simplex virus-permissive mouse melanoma cell line B16RHSV]
Xiu-fen Zhuang1, Ai-ping Zhou, Gui-lan Shi
1Cancer Institute & Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.
Objective:
To generate an oncolytic herpes simplex virus (oHSV) permissive mouse melanoma cell line B16RHSV, preserving the tumorigenic ability in syngeneic mice.
Methods:
The herpes simplex virus entry mediator (HVEM) gene was amplified by PCR from human melanoma cell line A375, and cloned into pGEM-T Easy vector for sequencing. The HVEM gene was then cloned into pcDNA3 vector to generate pcDNA3-HVEM for transfection of mouse melanoma cell line B16-F10 cells. After that, the putative transfected cells were selected in full growth medium containing G418. The HVEM-expressing cells were isolated by immunomagnetic bead separation. The mouse melanoma cell line expressing oHSV receptor-HVEM, designated as B16RHSV, was generated. The permissibility of B16RHSV cells to oHSV infection was examined with green fluorescence protein (GFP)-expressing oHSV (oHSVGFP). To investigate the tumorigenic ability of both cells in vivo, 2×10(5) cells in 100 µl were subcutaneously inoculated into the right flanks of C57/BL mice.
Results:
In vitro, the B16RHSV mouse melanoma cells were shown by fluorescence microscopy capable of being infected by oHSVGFP. In vivo, the B16RHSV cells, like their wild type counterpart, grew to form melanoma in syngeneic mice.
Conclusion:
A herpes simplex virus-permissive mouse melanoma cell line was established. Its tumorigenicity remained unchanged.
Insights
Researchers created a new mouse melanoma cell line, B16RHSV, that is permissive to oncolytic herpes simplex virus (oHSV) infection. This cell line retains its ability to form tumors in mice, making it a valuable tool for melanoma research.
Area of Science:
- Oncology
- Virology
- Cell Biology
Context:
- Melanoma is a significant form of skin cancer.
- Oncolytic herpes simplex virus (oHSV) shows promise as a cancer therapy.
- Developing virus-permissive cell lines is crucial for studying oHSV efficacy.
Purpose:
- To establish a mouse melanoma cell line (B16RHSV) that is permissive to oHSV.
- To ensure the generated cell line retains its tumorigenic potential in vivo.
- To facilitate research into oHSV-based melanoma treatments.
Summary:
- The herpes simplex virus entry mediator (HVEM) gene was introduced into mouse melanoma B16-F10 cells.
- Transfected cells expressing HVEM (B16RHSV) were confirmed to be permissive to oHSV infection in vitro.
- In vivo studies demonstrated that B16RHSV cells maintain tumorigenicity in syngeneic mice.
Impact:
- Provides a novel, virus-permissive melanoma cell line for preclinical oHSV research.
- Enables detailed investigation of oHSV interactions with melanoma cells.
- Supports the development of targeted oHSV therapies for melanoma.
