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In vitro and in vivo characteristics of human squamous cell carcinoma of the head and neck cells engineered to secrete interleukin-2.
- Source :
-
Cancer gene therapy [Cancer Gene Ther] 1997 Nov-Dec; Vol. 4 (6), pp. 366-76. - Publication Year :
- 1997
-
Abstract
- Two human squamous cell carcinoma of the head and neck (SCCHN) cell lines, PCI-13 and PCI-52, were transduced with the retroviral construct containing human interleukin-2 (IL-2) cDNA and selected for neomycin resistance in G418 medium. Stably transduced SCCHN cells produced and secreted IL-2, which was shown to have biologic activity in a bioassay, using an IL-2-dependent CTLL-2 cell line. By immunohistochemistry, IL-2 gene-transduced PCI-13 cells were strongly positive for IL-2, and by flow cytometry showed both cell surface and intracytoplasmic expression of IL-2 protein. Expression of IL-2 mRNA was measured by quantitative RT-PCR and found to be considerably increased in transduced SCCHN relative to that in parental cells. There was no difference in expression of IL-2R between the parental and IL-2 gene-transduced cells. In vitro proliferation of IL-2 gene-transduced tumor cells was consistently more rapid than that of parental cells. Sensitivity of the parental and IL-2 gene-transduced targets to lysis or apoptosis mediated by purified human natural killer (NK) cells or IL-2-activated NK (A-NK) cells was comparable as measured in 4-hour 51Cr-release and 1-hour [3H]thymidine-release assays, respectively. However, transduced cells were significantly more sensitive than parental cells to these effectors in 24-hour MTT assays, most likely due to IL-2 production by the transduced targets. PCI-52 cells selected for in vivo experiments formed large subcutaneous tumors in immunosuppressed nude mice. Tumors established by subcutaneous injections of 1 x 10(7) IL-2 gene-transduced cells regressed completely by day 25, while those formed by parental or LacZ gene-transduced tumor cells grew progressively. Tumor regression was mediated by numerous mononuclear cells, identified as murine NK cells and macrophages by immunohistochemistry, which accumulated around the IL-2-secreting, but not parental, tumors within 5-6 days after tumor cell injections. Thus, IL-2 gene-transduced SCCHN cells produce functional IL-2 in vivo in amounts sufficient to support the recruitment to the tumor site and antitumor activity of cytotoxic effector cells. IL-2-secreting SCCHN cells may be a useful component of vaccines designed to induce and sustain effector cell activation at the tumor site.
- Subjects :
- Animals
Carcinoma, Squamous Cell therapy
Cell Division
Cell Line
Granulocyte-Macrophage Colony-Stimulating Factor biosynthesis
Head and Neck Neoplasms therapy
Humans
Interleukins biosynthesis
Kinetics
Mice
Mice, Nude
RNA, Messenger biosynthesis
Recombinant Proteins biosynthesis
Transcription, Genetic
Transfection methods
Transforming Growth Factor beta biosynthesis
Transplantation, Heterologous
Tumor Cells, Cultured
Tumor Necrosis Factor-alpha biosynthesis
Carcinoma, Squamous Cell immunology
Carcinoma, Squamous Cell pathology
Cytotoxicity, Immunologic
Head and Neck Neoplasms immunology
Head and Neck Neoplasms pathology
Interleukin-2 biosynthesis
Killer Cells, Natural immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0929-1903
- Volume :
- 4
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Cancer gene therapy
- Publication Type :
- Academic Journal
- Accession number :
- 9408607