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Production and characterization of anti-DEC205 antibodies fused to the E7 protein of human papillomavirus

Grant number: 11/20660-0
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): January 01, 2012
Effective date (End): December 31, 2012
Field of knowledge:Biological Sciences - Microbiology
Principal Investigator:Silvia Beatriz Boscardin
Grantee:Tamara Silva Frias Garcia
Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

In the last decade, a strategy for targeting antigens to dendritic cells (DCs) in vivo has been developed successfully in animal models. DCs are central cells in the process of inducing immunity against various pathogens and tolerance. Our strategy is to use a monoclonal antibody against a receptor on the surface of DC fused with the antigen of interest. The administration of low doses of chimeric antibodies, in the presence of DC maturation stimuli, is able to activate antigen-specific T cells and induce the production of high titers of antibodies. The CD8+DEC205+ DC subpopulation expresses the endocytic receptor DEC205 and also the CD8 molecule alpha chain. A large number of recent studies demonstrated that immunization of animals with anti-DEC205 fused with different proteins in the presence of a maturation stimulus for DCs is capable of inducing a strong immune response (both cellular and humoral), and in some cases, protection against challenges with the pathogen. The human papillomavirus (HPV) is the main etiological agent of cervical cancer and other cancers. The protein E7 encoded by HPV is involved in cell cycle control. Recently, an experimental strategy based on a DNA vaccine encoding the E7 protein of HPV-16 (the most relevant viral type epidemiologically) was successfully developed. The vaccine contains the E7 gene of HPV-16 fused to glycoprotein D (gD) of the herpes simplex virus type 1 (HSV-1).In this project, we intend to clone the E7 protein and a protein formed by the E7 protein and gD of herpes simplex virus type 1 fused with the sequence of the anti-DEC205 antibody, produce the hybrid antibody and test its binding capacity to cells that express the DEC205 receptor. Also, we intend to produce an isotype control that is unable to bind to the DEC205 receptor. (AU)

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