Gene-modified Tumor Cells Research Articles

Immuno-gene therapy of rat glioma with TGF-β2 antisense and IL-2 gene modified tumor cells

Immuno-gene therapy of rat glioma with TGF-β2 antisense and IL-2 gene modified tumor cells

The role of cytokines in mediating the bystander effect using HSV-TK xenogeneic cells.

We have previously demonstrated that genetically modified syngeneic murine tumor cells (KBALB) expressing the herpes simplex virus thymidine kinase gene (HSV-STK) can kill nearby unmodified tumor cells in the presence of ganciclovir (GCV). The killing was mediated by a 'bystander effect' as evidenced by the prolonged animal survival when syngeneic HSV-TK gene-modified tumor cells were inoculated into mice with an intraperitoneal tumor. In this study we investigated whether irradiated xenogeneic HSV-TK gene-modified tumor cells, a human colon carcinoma cell line (HCT) transfected with the HSV-TK gene, can mediate the 'bystander effect' when used in vitro and in vivo. In vitro experiments indicate that irradiated HSV-TK gene-modified xenogeneic cells (HCT) can mediate a bystander effect on the adjacent cells when the tumor population consisted of as few as 10% of the HSV-TK expressing HCT tumor cells. In vivo, animal survival experiments demonstrate that the xenogeneic gene-modified tumor cells could generate the 'bystander effect' in mice with intraperitoneal tumors as evidenced by prolonged animal survival. In addition, histologic examination of the tumors from experimental animals showed extensive tumor necrosis 3 days post HSV-TK/GCV treatment in comparison to control animals. To evaluate the cause of necrosis in vivo, we assayed for cytokines, which may be involved in mediating this process, by performing RT-PCR and immunohistochemistry on tumor RNA and tumor cells, respectively. Production of IL-1 alpha and IL-6 mRNA within the experimental tumors was observed by RT-PCR. However, mRNA expression for other cytokines including IFN-gamma, IL-2 and IL-4 was not present. Immunohistochemical analysis for IL-1 alpha protein showed reactivity within the infiltrating mononuclear cells indicating the release of this soluble factor. These results indicate that the bystander effect can be generated using irradiated xenogeneic cells both, in vitro and in vivo. Furthermore, this process is mediated by the release of cytokines such as IL-1 alpha, IL-6 which enhances the bystander effect in vivo by immunostimulation.

Comparison of Gene Therapy with Interleukin-2 Gene Modified Fibroblasts and Tumor Cells in the Murine CT-26 Model of Colorectal Carcinoma

We compared the efficacy of gene therapy mediated by interleukin-2 (IL-2) gene-modified tumor cells to gene therapy mediated by IL-2 transduced fibroblasts in the CT-26 model of murine colorectal carcinoma. We transduced CT-26 tumor cells and BALB/c 3T3 fibroblasts with three different retroviral vectors using three different promoters for the human IL-2 gene: DC/TKIL-2 (thymidine kinase promoter), LXSN-iIL2 (long terminal repeat promoter), and LNCX-iIL2 (cytomegalovirus promoter). These transductions resulted in CT-26 and 3T3 subclones that secreted different amounts of IL-2. Immunization of animals with either CT-26/IL-2 cells or with fibroblast/IL-2 cells mixed with CT-26 induced similar levels of immunity that protected 62-82% of animals against a subsequent tumor challenge with parental CT-26. However, mice developed tumors at the site of inoculation in 46% of the animals immunized with CT-26/IL-2 cells. In a separate experiment, CT-26/IL-2 cells were exposed to 6,000 cGy of gamma irradiation to prevent tumor growth at the site of inoculation. Although the CT-26/IL-2 cells continued to secrete IL-2 after irradiation, they were no longer effective at inducing antitumor immunity. In contrast, both irradiated and nonirradiated fibroblast/IL-2 cells, mixed with irradiated CT-26, were equally effective at inducing antitumor immunity. These data suggest that in the CT-26 model, fibroblast-mediated IL-2 gene therapy has advantages for the induction of antitumor immunity and abrogation of tumorigenic potential at the site of inoculation compared with tumor cell-mediated IL-2 gene therapy.

Molecular characterization of defective antigen processing in human prostate cancer.

Gene-modified tumor cell vaccines have shown efficacy in animal models of malignancy, including prostate cancer. Class I major histocompatibility complex (MHC) assembly and function in the cellular targets of such therapies is pivotal in determining the efficacy of specific cytokine-secreting tumor vaccines. To help guide development of genetically engineered vaccine therapy for human prostate cancer, potential immune resistance pathways were evaluated by analysis of class I MHC assembly in prostate cancer cells. Class I MHC assembly in metastasis-derived human prostate cancer cell lines (LNCaP, PPC-1, DU-145, PC-3, and TSU) and a normal prostate-derived cell line (TP-2) were characterized by phenotypic, molecular, and functional assays. Assembled class I MHC and antigen was measured by flow cytometry; mRNA levels of assembly components (class I MHC heavy chain, beta 2-microglobulin, and the antigen transporter gene product TAP-2) were determined; and antigen processing was measured with a chimeric reconstituted system using vaccinia vectors. Restoration of antigen processing was attempted by interferon gamma stimulation and by transfection with mouse class I MHC heavy-chain cDNA. Assembled class I MHC was underexpressed in two (LNCaP and PPC-1) of five prostate cancer cell lines compared with normal prostate-derived controls. PPC-1 cells underexpressed TAP-2 mRNA despite abundant class I MHC and beta 2-microglobulin message. Induction of TAP-2 by interferon gamma indicated that coding sequences for TAP-2 message were present in PPC-1. Resistance to cytotoxic T lymphocytes (CTL) lysis showed a functional defect in antigen transport by PPC-1 cells; reversal of the molecular defect with interferon gamma led to restoration of functional antigen processing. In contrast, LNCaP cells had competent antigen transport but deficient class I MHC heavy-chain function despite abundant class I MHC RNA; though refractory to stimulation by interferon gamma, this defect responded to transfection of class I MHC heavy-chain cDNA. Metastatic prostate cancer cells can escape T-cell recognition via divergent mechanisms of defective class I MHC assembly. The specific underexpression of TAP-2 gene product in PPC-1 cells contrasts with prior studies of TAP gene underexpression in lung cancer (which concurrently underexpressed class I MHC heavy chain) and provides evidence for a regulatory pathway controlling TAP-2 gene expression in human cancers that may not affect class I MHC heavy-chain expression. In clinical application of gene therapy for prostate cancer, these findings provide a rationale for focusing on strategies that can circumvent sole reliance on class I MHC-mediated tumor cell recognition by CTL.

CYTOKINE GENE-MODIFIED VACCINES FOR IMMUNOTHERAPY OF CANCER (REVIEW)

Cytokine gene therapy has recently become a new prospective modality in the treatment of cancer. Active immunization with gene-modified tumour cells in experimental systems employs either live Vaccines at subthreshold, non-tumorigenic doses, or irradiated cytokine-producing vaccines. The effects exerted on tumour cells by the vaccine-produced cytokines are usually pleiomorphic, either direct (cytolysis, upregulation of MHC, TAA, or adhesion molecule expression, terminal differentiation) or indirect (recruitment, amplification and activation of APC, helper and defence effector cells, damage of the blood supply) or both. The aim of this article is to review and summarize recent results of the cytokine gene therapy in experimental tumour systems, as well as to discuss the prospects and limitations of this novel approach in the management of cancer patients. The review is primarily focused on the therapeutic vaccination; however, when relevant, the results obtained with preventive vaccination are also briefly discussed.

The role of IL-2 secreted from genetically modified tumor cells in the establishment of antitumor immunity.

Although the importance of CD4+ T cells for the induction of an effective CD8+ cytolytic T cell response is well documented, the mechanism by which MHC class II-negative tumor cells recruit CD4+ T help is not well understood. We have previously shown that IL-2 or IFN-gamma gene-modified CMS-5 tumor cells do not grow in syngeneic mice; however, mice which rejected the cytokine-secreting tumor cells develop a protective immune response against a challenge with parental tumor cells. Here we show that rejection of IL-2-secreting CMS-5 cells is not mediated by T cells. However, establishment of a protective immune response against CMS-5 tumor cells requires the presence of both CD4+ and CD8+ T cell subsets during the period of immunization with IL-2-secreting CMS-5 cells as well as during the effector phase. Extensive histologic analysis has failed to detect the presence of T cells at the site of immunization with either IL-2- or IFN-gamma-secreting CMS-5 cells. The main infiltrate at the site of inoculation with IL-2-secreting CMS-5 cells consisted of NK cells that appeared to play a role in their rejection. The predominant infiltrate at the site of inoculation with IFN-gamma-secreting CMS-5 cells consisted of macrophages. These observations argue against a direct role for the intact tumor cell in presenting either T helper or CTL epitopes to the immune system, and support the view that specialized APC are responsible for the in vivo priming of a T cell response against MHC class I-restricted Ag.

Regression of bladder tumors in mice treated with interleukin 2 gene-modified tumor cells.

This study explored the use of interleukin 2 (IL-2) and interferon gamma (IFN-gamma) gene-modified tumor cells as cellular vaccines for the treatment of bladder cancer. The mouse MBT-2 tumor used is an excellent model for human bladder cancer. This carcinogen-induced tumor of bladder origin resembles human bladder cancer in its etiology and histology, and responds to treatment in a manner similar to its human counterpart. Using retroviral vectors, the human IL-2 and mouse IFN-gamma genes were introduced and expressed in MBT-2 cells. The tumor-forming capacity of the cytokine gene-modified MBT-2 cells was significantly impaired, since no tumors formed in mice injected intradermally with either IL-2- or IFN-gamma-secreting cells, using cell doses far exceeding the minimal tumorigenic dose of parental MBT-2 cells. Furthermore, mice that rejected the IL-2- or IFN-gamma-secreting tumor cells became highly resistant to a subsequent challenge with parental MBT-2 cells, but not to 38C13 cells, a B cell lymphoma of the same genetic background. To approximate the conditions as closely as possible to the conditions prevailing in the cancer patient, inactivated cytokine-secreting cells were used to treat animals bearing tumors established by orthotopic implantation of MBT-2 cells into the bladder wall of the animal. Treatment of mice carrying a significant tumor burden with IL-2-secreting MBT-2 cells had a significant inhibitory effect on tumor progression with extended survival. Moreover, in 60% of the mice the tumor regressed completely and the animals remained alive and free of detectable tumor for the duration of the observation period. Treatment of tumor-bearing animals with IL-2-secreting MBT-2 cells was superior to the use of cisplatin, a chemotherapeutic agent used in the treatment of bladder cancer. The therapeutic effect of IFN-gamma-secreting cells was minimal and treatment with unmodified MBT-2 cells had no effect on tumor growth or survival, showing that the parental MBT-2 cells were nonimmunogenic in this experimental setting. Most importantly, mice that exhibited complete tumor regression after treatment with IL-2-secreting MBT-2 cells became resistant to a subsequent challenge with a highly tumorigenic dose of parental MBT-2 cells, indicating that long-term immunological memory was established in the "cured" mice.

A Pilot Study of Immunization with HLA-A2 Matched Allogeneic Melanoma Cells That Secrete Interleukin-2 in Patients with Metastatic Melanoma

Human Gene TherapyVol. 3, No. 6 News and CommentsA Pilot Study of Immunization with HLA-A2 Matched Allogeneic Melanoma Cells That Secrete Interleukin-2 in Patients with Metastatic MelanomaPRINCIPAL INVESTIGATORS Bernd Gansbacher, CO-PRINCIPAL INVESTIGATORS Alan Houghton, Philip Livingston, CO-INVESTIGATORS Lori Minasian, Felicia Rosenthal, Eli Gilboa, Herbert Oettgen, Tom Steffens, Soo Young Yang, and George WongPRINCIPAL INVESTIGATORS Bernd GansbacherSearch for more papers by this author, CO-PRINCIPAL INVESTIGATORS Alan HoughtonSearch for more papers by this author, Philip LivingstonSearch for more papers by this author, CO-INVESTIGATORS Lori MinasianSearch for more papers by this author, Felicia RosenthalSearch for more papers by this author, Eli GilboaSearch for more papers by this author, Herbert OettgenSearch for more papers by this author, Tom SteffensSearch for more papers by this author, Soo Young YangSearch for more papers by this author, and George WongSearch for more papers by this authorPublished Online:13 Jun 2008https://doi.org/10.1089/hum.1992.3.6-677AboutSectionsPDF/EPUB ToolsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail FiguresReferencesRelatedDetailsCited ByNucleic Acid-Based Approaches for Tumor Therapy9 September 2020 | Cells, Vol. 9, No. 9Are Vaccinations for Prostate Cancer Realistic?Methoden der GenübertragungTherapeutic Vaccination against Metastatic Carcinoma by Expression-Modulated and Immunomodified Autologous Tumor Cells: A First Clinical Phase I/II Trial6 July 2004 | Human Gene Therapy, Vol. 12, No. 3Molecular Strategies Interfering with Tumor Progression of Melanoma and Improving Anti-Tumor ImmunityCOMPARISON OF NON-VIRAL TRANSFECTION METHODS IN MELANOMA CELL PRIMARY CULTURESCytokine, Vol. 12, No. 6Establishment and Characterization of Colon Carcinoma and Renal Cell Carcinoma Primary Cultures29 January 2009 | Cancer Biotherapy & Radiopharmaceuticals, Vol. 15, No. 2Disabled Infectious Single-Cycle Herpes Simplex Virus as an Oncolytic Vector for Immunotherapy of Colorectal Cancer6 July 2004 | Human Gene Therapy, Vol. 10, No. 17Cancer VaccinesJournal of Clinical Oncology, Vol. 17, No. 3Gentherapie — Aktueller Forschungsstand und PerspektivenInterleukin-2 Gene-Modified Allogeneic Tumor Cells for Treatment of Relapsed Neuroblastoma20 March 2008 | Human Gene Therapy, Vol. 9, No. 9GENE THERAPY FOR MELANOMA IN HUMANSHematology/Oncology Clinics of North America, Vol. 12, No. 3Retroviral Vectors From Laboratory Tools to Molecular MedicinesCytokine Gene Transduction in the Immunotherapy of CancerMethoden der GenübertragungGene therapy in surgeryEuropean Surgery, Vol. 29, No. 1Gene TherapySomatic Gene Therapy in CancerActive Immunotherapy for Solid TumoursA Novel Herpes Vector for the High-Efficiency Transduction of Normal and Malignant Human Hematopoietic CellsBlood, Vol. 89, No. 1A Novel Herpes Vector for the High-Efficiency Transduction of Normal and Malignant Human Hematopoietic CellsBlood, Vol. 89, No. 1Gene therapy for cancer, the course aheadCancer and Metastasis Review, Vol. 15, No. 3Immunotherapy I: Cyclosine gene transfer strategiesCancer and Metastasis Reviews, Vol. 15, No. 3High-Efficiency Transduction of Freshly Isolated Human Tumor Cells Using Adenoviral Interleukin-2 Vectors20 March 2008 | Human Gene Therapy, Vol. 7, No. 10Retroviral vectorsMolecular Biotechnology, Vol. 5, No. 2Transcriptionally Targeted Gene TherapyA B7-1-transfected human melanoma line stimulates proliferation and cytotoxicity of autologous and allogeneic lymphocytesEuropean Journal of Immunology, Vol. 25, No. 10European school of Oncology position paper. 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Immunization of Cancer Patients Using Autologous Cancer Cells Modified by Insertion of the Gene for Interleukin-2 (National Institutes of Health)

When tumor is resected from patients as part of the natural course of their treatment, an attempt will be made to establish a tissue culture line of the tumor. The gene coding for interleukin-2 will be introduced into these tumor cells and the integration and expression of this gene will be tested. Patients will become eligible for this study only if they develop metastatic cancer that has failed all standard effective treatment and have no other effective treatment options available to them. Tumor cells will be injected intradermally and subcutaneously into the thigh of these patients. The amount of tumor injected will be less than 1/50th the total tumor burden of that patient. In previous studies we have shown that these gene modified tumor cells are more immunogenic than the native unaltered tumor. Attempts will then be made to grow immune lymphocytes either from the tumor site or from the draining lymph nodes of these patients in order to use these lymphocytes for adoptive immunotherapy as detailed in previous protocols. The direct effect of the immunization with these immunogenic tumor cells will also be measured by assessing the impact on established tumor at other sites. Fifty patients receiving tumor inoculation will be included in this study.

Immunization of Cancer Patients Using Autologous Cancer Cells Modified by Insertion of the Gene for Tumor Necrosis Factor (National Institutes of Health)

When tumor is resected from patients as part of the natural course of their treatment, an attempt will be made to establish a tissue culture line of the tumor. The gene coding for tumor necrosis factor will be introduced into these tumor cells and the integration and expression of this gene will be tested. Patients will become eligible for this study only if they develop metastatic cancer that has failed all standard effective treatment and have no other effective treatment options available to them. Tumor cells will be injected intradermally and subcutaneously into the thigh of these patients. The amount of tumor injected will be less than 1/50th the total tumor burden of the patient. In previous studies we have shown that these gene-modified tumor cells are more immunogenic than the native unaltered tumor. Attempts will then be made to grow immune lymphocytes either from the tumor site or from the draining lymph nodes of these patients in order to use these lymphocytes for adoptive immunotherapy as detailed in previous protocols. The direct effect of the immunization with these immunogenic tumor cells will also be measured by assessing the impact on established tumor at other sites. Fifty patients receiving tumor inoculation will be included in this study.