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Interferon-alfa genetherapy for colorectal cancer control

Grant number: 14/05609-6
Support Opportunities:Regular Research Grants
Duration: September 01, 2014 - August 31, 2016
Field of knowledge:Biological Sciences - Immunology - Cellular Immunology
Principal Investigator:Ramon Kaneno
Grantee:Ramon Kaneno
Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Associated researchers:Bryan Eric Strauss ; Graziela Gorete Romagnoli


Interferon alpha (IFN-alpha) is a type I IFN with great therapeutic potential, since it is able to directly fight tumor cells and enhance the maturation of dendritic cells (DCs), the main antigen-presenting cells, required for an effective antitumor response. However, the systemic administration of cytokines can induce severe collateral effects. Therefore, the induction of cytokine secretion in situ should represent a more adequate approach for cytokine-based immunotherapy. Thus, the goal of this study is to induce IFN-alpha secretion by colon cancer cells by transfection with an lentivirus vector carrying human IFN-alpha genes, followed by analysis of its lytic and immunomodulatory potential. For this purpose, non-replicant lentivirus carrying human IFN-alpha genes will be used to transfect tumor cells. Such cells will be co-cultured with monocyte-derived DCs from healthy donors, previously loaded with wild type tumor lysate. Twenty-four hours later, DCs will be evaluated for their phenotype (activation/maturation markers) by flow cytometry, their ability to induce allogeneic response (MLR) and effectiveness to induce cytotoxic T cells (evaluated by cytotoxicity assay and IFN-a production). Since this IFN is required for inducing IL-12 producing DCs (alpha-DC1 type), secretion of this cytokine by DCs will be analyzed. IFN-alpha is also a potent activator of NK cells and so the generation of these cells in vitro will also be analyzed. (AU)

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