Mikrobiologie Normal client4 2 2004-01-30T12:27:00Z 2004-01-30T12:27:00Z 1 333 2104 Universität Regensburg 9.2812

RNA and Codon-optimised Synthetic Genes in HIV specific DNA Vaccination: From epidemiology to clinical trials.

Ralf Wagner1,2, Jens Wild1, Kurt Bieler1, Ludwig Deml1, Frank Notka2, Hans Wolf1, 1Institute of Medical Microbiology, University of Regensburg, Franz Josef Strauss Allee 11; 2GENEART GmbH, Josef-Engert-Straße 9, 93053 Regensburg, Germany

e-mail: ralf.wagner@klinik.uni-regensburg.de

As recently demonstrated, extensive codon-usage modifications increases the overall GC content of late HIV transcripts by almost 30% compared to wildtype genome, enhanced the nuclear stability of viral RNAs and resulted in a constitutive export of these RNAs into the cytoplasm. Besides that, codon-usage modifications tremendously supported viral gene expression that turned out to be now independent from cis acting sequences like RNA packaging elements, Rev and otherwise engaged cellular RNA binding factors limiting nuclear translocation of HI-viral RNA.

As we and others showed in immunisation studies in mice with clade B based vector constructs, vaccination with codon optimised naked DNA induces Th1-type humoral and cellular immune responses substantially higher than those obtained from immunisation with the corresponding wt-constructs. Furthermore, those constructs alone as well as in combination with protein- and MVA-booster immunisations induced antiviral cellular immune responses in rhesus macaques leading to stable CD4 cell counts, a containment of virus replication and protection from disease progression after virus challenge (SHIV89.6P).

These encouraging results justified extensions to human trials with analogous vaccine constructs encoding GagPolNef and Env polygenes derived from a clade C molecular clone (CN54) representing the prevalent virus circulating throughout China as shown earlier by intensive epidemiological studies. Currently available delivery systems (GMP grade materials) include plasmid DNA vaccines lacking any antibiotic resistance gene as well as a replication deficient vaccinia virus (NY strain) expressing the analogous set of antigens. Vaccination studies with preGMP plasmid DNA alone as well as prime-boost regiments using NY-vaccinia virus in Balb/c mice revealed the induction of cellular and humoral immune responses.

Following successful toxicology a first phase 1 clinical trial coordinated by the EUROVAC network has started few weeks ago to test an initial set of immunogens in human volunteers.