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Laboratory of Molecolar Virology  

Group leader
Antonia Radaelli, Associate Professor

Team
Carlo De Giuli Morghen, Contract Professor carlo.degiulimorghen@unimi.it
Carlo Zanotto, Researcher
Elena Illiano, Research associate elena.illiano@unimi.it
Massimiliano Bissa, PhD student massimiliano.bissa@unimi.it
Sole Maria Pacchioni, Student sole.pacchioni@studenti.unimi.it

 
Research interests
Construction of recombinant vaccines for the prevention and therapy of infectious diseases, also caused by bioterroristic viral agents

The goal of our research program is the construction of DNA and viral immunogens, based on the genetic background of avipoxviruses, for vaccine purposes. At present, we are preparing recombinants expressing either HIV-1 gag/pol and env genes for the prevention/therapy of AIDS, or HPV-16 E6/E7 oncogenes against HPV-related tumours. These candidate vaccines are evaluated for the expression, innocuity and immunogenicity, and efficacy in adequate animal models. In the context of the deliberate release of an infectious agent for bioterrorism, we are also developing different genetic and viral recombinants, coding for heterologous genes for the protection against smallpox and other orthopoxviruses, which can also be used as bioterristic agents. The most important aspect of our research is the use of safe immunogens and viral vectors unable to replicate in mammalian cells, but expressing the transgenes at high levels and able to elicit a complete immune response. The use of prime-boost protocols, where genetic vaccines are combined with fowlpox and protein recombinants, can increase their immunogenicity.

Techniques

The main techniques used in our laboratory regard:
immunology: ELISA, Western Blot, immunoprecitation, immunofluorescence
cell biology: culture of permanent cell lines, preparation and culture of primary cells, primary cell immortalization by SV40
molecular biology: PCR, RT-PCR, real-time PCR, cloning and amplification of viral and cell genes
microbiology: competent cell preparation, bacterial cell transformation, plasmid amplification, antibiogram and sensitivity tests of anti-bacterial compounds
virology: culture and titration of RNA and DNA viruses, in-vitro recombination, gradient and plaque purification, in-vitro virus neutralization assays
electron microscopy: preparation and analysis of biological samples for the ultrastructural analysis of cells and virus identification

 

Selected publications

1. Radaelli A (2007) Prime boost immunization with DNA, recombinant fowlpox virus and VLPSHIV elicit both neutralizing antibodies and IFNg producing T cells against the HIV envelope protein in mice that control env bearing tumour cells. Vaccine 25:2128-2138.

2. Pozzi E (2009) MHC-restricted CTL assay: an improved method based on naïve and SV40-immortalized rabbit epidermal target cells. J. Virol. Methods 155:77-81.

3. Pozzi E (2009) Construction and characterization of recombinant fowlpox viruses expressing human papilloma virus E6 and E7 oncoproteins. J. Virol. Methods 158: 184-189.

4. Pacchioni S Canarypox and fowlpox viruses as recombinant vaccine vectors: an ultrastructural comparative analysis. Arch. Virol. 155: 915-924.

5. Radaelli A (2010) Fowlpox virus recombinants expressing HPV-16 E6 and E7 oncogenes for the therapy of cervical carcinoma elicit humoral and cell-mediated responses in rabbits. J. Transl. Medicine 8: 40-51.

6. Zanotto C (2010) Canarypox and fowlpox viruses as recombinant vaccine vectors: a biological and immunological comparison. Antiviral Res. 88:53-63.

7. Zanotto C (2011) Construction and characterisation of a recombinant fowlpox virus that expresses the human papilloma virus L1 protein. J. Transl. Medicine 9: 190-200.

8. Radaelli A. (2012) A prime/boost strategy by DNA/fowlpox recombinants expressing a mutant E7 protein for the immunotherapy of HPV-associated cancers. Virus Res. 170: 44-52.

9. Bissa M (2013) GFP co-expression reduces the A33R gene expression driven by a fowlpox vector in replication permissive and non-permissive cell lines. J. Virol. Methods 187: 172-176.

10. Pacchioni S (2013) L1R, A27L, A33R, and B5R vaccinia virus genes expressed by fowlpox recombinants as putative novel orthopoxvirus vaccines. J. Transl. Medicine 11: 95-104

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