Combined suicide gene and immunostimulatory gene therapy using AAV-mediated gene transfer to HPV-16 transformed mouse cell: decrease of oncogenicity and induction of protection
Language English Country Greece Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
12579310
Knihovny.cz E-resources
- MeSH
- B7-1 Antigen genetics MeSH
- Antiviral Agents therapeutic use MeSH
- Chemokine CCL2 genetics MeSH
- Defective Viruses genetics MeSH
- Dependovirus genetics MeSH
- Granulocyte-Macrophage Colony-Stimulating Factor genetics MeSH
- Ganciclovir therapeutic use MeSH
- Genetic Therapy * MeSH
- Genetic Vectors genetics therapeutic use MeSH
- HeLa Cells MeSH
- Immunotherapy methods MeSH
- Lac Operon MeSH
- Kidney MeSH
- Humans MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Cell Line, Tumor transplantation MeSH
- Oncogene Proteins, Viral physiology MeSH
- Papillomaviridae physiology MeSH
- Papillomavirus E7 Proteins MeSH
- Repressor Proteins * MeSH
- Simplexvirus enzymology genetics MeSH
- Thymidine Kinase antagonists & inhibitors genetics MeSH
- Transduction, Genetic MeSH
- Cell Line, Transformed MeSH
- Neoplasm Transplantation MeSH
- Cell Transformation, Viral * MeSH
- Xenograft Model Antitumor Assays MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- B7-1 Antigen MeSH
- Antiviral Agents MeSH
- Chemokine CCL2 MeSH
- E6 protein, Human papillomavirus type 16 MeSH Browser
- Granulocyte-Macrophage Colony-Stimulating Factor MeSH
- Ganciclovir MeSH
- oncogene protein E7, Human papillomavirus type 16 MeSH Browser
- Oncogene Proteins, Viral MeSH
- Papillomavirus E7 Proteins MeSH
- Repressor Proteins * MeSH
- Thymidine Kinase MeSH
To test the effects of combined transduction of a suicide gene and genes coding for various immunostimulatory factors on the oncogenicity and immunogenicity of TC-1 cells (HPV-16 transformed C57BL/6 mouse cells), several bicistronic recombinant adeno-associated viruses (rAAV) were constructed. Each of these constructs carried, and in infected cells expressed, the herpes simplex type 1 thymidine-kinase gene (HSV-TK) and the gene of one of the following immunostimulatory factors: human monocyte chemoattractant protein 1 (MCP-1), mouse B7.1 costimulatory molecule (B7.1), or mouse granulocyte-macrophage colony-stimulating factor (GM-CSF). For control purposes, an rAAV carrying the HSV-TK gene and neomycin resistance gene (neo) and an rAAV containing the lacZ gene were used. All of these constructs proved functional both in mouse TC-1 and human 293T cells. For experiments in mice, TC-1 cells were infected in vitro with the AAV recombinants at an input multiplicity of 50 particles/cell; these cells were then administered to 5-week-old mice. As from day 5, half of the animals were given ganciclovir (GCV) (2.5 mg/day) for 10 days. With a single exception, none of the mice inoculated with cells treated with rAAV expressing HSV-TK + B7.1 or HSV-TK + MCP-1 developed tumour irrespective of GCV treatment. The tumour suppressive effect was less marked in animals inoculated with TC-1 cells infected with rAAV expressing HSV-TK + GM-CSF, and among these it was somewhat more pronounced in GCV-untreated animals. A clear antitumour effect of GCV treatment was only observed in mice inoculated with TC-1 cells transduced with rAAV expressing HSV-TK but no immunostimulatory factor. Mice that remained tumour-free on day 54 were challenged with untreated TC-1 cells. The tumour resistance rates found were related not only to the immunostimulatory gene used for the transduction, but also to GCV treatment. The best protection was recorded in mice pre-inoculated with TC-1 cells transduced with either B7.1 or MCP-1-expressing rAAV and not given GCV.
