NON-COMMUNICABLE DISEASES / Immunology
DNA vaccines, antibody engineering, anti-idiotypic Abs, dendritic cells, membrane-IgE, Wiskott-Aldrich syndrome, molecular biology of rotavirus.
Description of Research
DNA vaccines and Dendritic cells
The Group is interested in implementing different strategies for immunisation with appropriate DNA constructs, against both tumours and viruses. An effort has been made in the design of immunogens, an essential aspect to achieve availability of Ag and efficient priming of cellular and humoral responses. A B-cell lymphoma DNA vaccine is already being tested, in close collaboration with a team at the University of Pisa. The Group has also developed vaccines against a murine, breast tumour model based on the tumour-associated antigen HER2. The design comprises specific targeting of internalising receptors in Dendritic cells to improve antigen delivery and uptake by antigen presenting cells to achieve robust cytotoxic T-cell responses against tumours. Our experience in DNA vaccines has been extended to the design of DNA vaccines against the envelope protein of Dengue virus, a high impact human pathogen. Results in mice have shown that our novel genetic constructs are able to induce strong antibody responses.
Igs and antibody engineering
Engineered antibodies of different isotypes are being used to achieve sustained passive immunisation in mice against a tumour model. Recombinant AAV based constructs have been obtained and shown to efficiently express full antibody molecules for several months, as well as molecules on SIP format. This strategy has the potential to be widely used in many different cases.
ER to cytosol retro-translocation
We have recently developed a method to specifically target molecules within the secretory pathway, to induce their degradation. We made use of proteins involved in the ERAD (ER associated degradation) pathway that removes misfolded proteins from the ER, by inducing their ER to cytosol retro-translocation and subsequent proteasome degradation. A patent on these newly developed molecules, termed degradins, has been filed. In addition, our interest in the retro-translocation mechanism itself has led us to describe a novel method to specifically detect retro-translocated molecules using an in vivo biotinylation strategy (see Figure).
Molecular Biology of Rotavirus
Many different aspects of the biology of rotavirus replication remain obscure. This dsRNA virus has a complete cytoplasmic replication within well defined structures called viroplasms. We have found that in order for the viroplasms to assemble, it is required that cells have a totally functional proteasome. In addition, the virus is capable of inducing phosphorylation of Erk1/2 in a non-conventional activation pathway. While inserting exogenous viral genome segments into Rotavirus particles remains a difficult challenge, we have recently been able to produce a virus containing a recombinant biotinylated viral capsid protein (VP6).
Campagna M., Marcos-Villar L., Arnoldi F., de la Cruz-Herrera C.F., Gallego P., González-Santamaría J., González D., Lopitz-Otsoa F., Rodríguez M.S., Burrone, O.R., Rivas, C. 2012. Rotavirus Viroplasm Proteins Interact with the Cellular SUMOylation System: Implications for Viroplasm-Like Structures Formation. J Virol In press
Contin, R., Arnoldi, F Mano, M. and Burrone, O.R. 2011. Rotavirus replication requires a functional proteasome for effective assembly of viroplasms. J Virol 85, 2781-2792 PubMed link
Petris, G., Vecchi, L., Bestagno, M., Burrone, O.R. 2011. Efficient Detection of Proteins Retro-Translocated from the ER to the Cytosol by In Vivo Biotinylation. PlosOne 6, e23712 PubMed link
Contin R., Arnoldi, F., Campagna, M., Burrone O.R. 2010. Rotavirus NSP5 orchestrates recruitment of viroplasmic proteins. J Gen Virol 91, 1782-1793 PubMed link
Pulecio J., Petrovic, J., Prete, F., Chiaruttini, G., Lennon-Dumenil A.M., Desdouets C., Gasman, S., Burrone, O.R., Benvenuti, F. 2010. Cdc42-mediated MTOC polarization in dendritic cells controls targeted delivery of cytokines at the immune synapse. J Exp Med 207, 2719-2732 PubMed link
Cesco-Gaspere, M., Zentilin, L., Giacca, M., Burrone, O.R. 2008. Boosting anti-idiotype immune response with recombinant AAV enhances tumour protection induced by Gene Gun vaccination. Sc. J. Imm. 68, 58-66 Pubmed link