Prophylactic HPV vaccines: next generations
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1 New Trends in Tumor Virology: Annecy Jan 19, 2010 Prophylactic HPV vaccines: next generations Lutz Gissmann
2 Conflict of interest LG is a consultant to GSK and Sanofi Pasteur MSD and, due to existing IP, receives royalties from sales of Gardasil and Cervarix TM 2 Gissmann
3 1 st generation prophylactic vaccines: HPV L1 VLPs N wildtyp L1 C recombinant baculovirus, yeast 5x 72x monomer capsomer 100nm neutralizing antibodies capsid (VLP) 3 Gissmann
4 Prophylactic HPV vaccines Gardasil (MSD) Cervarix (GSK) composition HPV 6/11/16/18 HPV 16/18 adjuvant AAHS 1 AS04 2 scheme i.m. 0,2,6 months i.m. 0,1,6 months availability June 2006 October AAHS: Amorphous Aluminum Hydroxyphosphate 2 ASO4: Aluminum hydroxide + 3-deacetylated monophosphoryl Lipid A (MPL): detoxified lipopolysaccharide prepared from Salmonella minnesota 4 Gissmann
5 Strengths and weaknesses of the existing HPV vaccines are safe and highly efficient in clinical trials have the potential to significantly reduce the burden of cervical and other HPV-related cancers don t prevent all cases of cervical cancer and precursors continuation of screening don t treat existing infections or lesions are not particularly suited for use in areas of highest need cost, stability, multiple and invasive immunization 5 Gissmann
6 Future HPV Vaccines: Basic and Applied Research conducting research accompanying the introduction of the HPV vaccine exploiting existing vaccine in off-label trials modifying existing vaccines supplementing prophylactic vaccines developing HPV vaccines for other targets 6 Gissmann
7 Future HPV Vaccines: Basic and Applied Research conducting research accompanying the introduction of the HPV vaccine exploiting existing vaccine in off-label trials modifying existing vaccines supplementing prophylactic vaccines developing HPV vaccines for other targets 7 Gissmann
8 Future HPV Vaccines: Basic and Applied Research conducting research accompanying the introduction of the HPV vaccine exploiting existing vaccine in off-label trials modifying existing vaccines supplementing prophylactic vaccines developing HPV vaccines for other targets 8 Gissmann
9 Research accompanying the introduction of the HPV vaccine unresolved issues need to be address in population-based trials duration of protection immune correlate for protection replacement by uncovered HPV types efficacy in real life reduction in cancer incidence comparison of the different products analysis of bonus effects (protection against other tumors) 9 Gissmann
10 Future HPV Vaccines: Basic and Applied Research conducting research accompanying the introduction of the HPV vaccine exploiting existing vaccine in off-label trials modifying existing vaccines supplementing prophylactic vaccines developing HPV vaccines for different targets 10 Gissmann
11 Modifying the existing vaccines Special need for developing countries multivalent long-protecting without boosting immunogenic by non-invasive application 11 Gissmann
12 conserved N-terminus of PV L2 induces cross-neutralizing yet low titer abs 12 Gissmann Buck et et al 2008 J Virol 82:5190
13 Multimerization of L2 epitopes displayed by thioredoxin 13 Gissmann
14 Multimerized HPV 16 L2 epitope (20-38) induces high-titer neutralizing abs 14 Gissmann
15 Needle-less single-dose immunization: recombinant Adeno-Associated Virus (AAV) AAVs are apathogenic in humans AAVs induce persistent infections 15 Gissmann
16 HPV 16 L1 raav induces L1-specific serum antibodies after i.n. immunization 16 Gissmann Kuck et al. 2006
17 Single-dose immunization (intranasal) induces long-lasting immune response 17 Gissmann Kuck et al. 2006
18 Modifying the existing vaccines Special need for developing countries AFFORDABLE AND ROBUST multivalent long-protecting without boosting immunogenic by non-invasive application 18 Gissmann
19 HPV 16 L1 capsomeres versus VLPs: costly, >stable, = immunogenic N D 1-10 C E. coli 5x monomer capsomer 19 Gissmann
20 GST renders L1 soluble: purification of immunogenic L1 capsomeres Thrombin cleavage site GST HPV 16 L1 L1 GST lysate + ATP, MgCl 2 + 3,5 M urea centrifugation supernatant dialysis overnight gel filtration Binding to Glutathione- sepharose beads Thrombin cleavage Elution L1 100nm 20 Gissmann
21 assembly of capsomeres into VLPs ph = 5.4 1M NaCl D , 428 Cys Ser D (Dh4) T=7 T=7, T=4? capsomeres 21 Gissmann
22 The ability of L1 capsomeres to assemble into VLPs correlates with immunogenicity Dilution in PBS 37 C Stable particles immunogenicity Instable particles Capsomeres Capsomeres 22 Gissmann
23 HPV 16 L1-specific IgG titer Immunogenicity of HPV 16 L1 capsomeres correlates with VLP formation HPV16 L1-specific antibody titers L1DN10DC29 L1D N10 dcys DC29-E7 L1DN10Dh4 DC29-E7 E7-L1D N10 Dh4DC29 CVLPs E7 pep 23 Gissmann
24 Immunogenicity of HPV 16 L1 capsomeres can be enhanced by adjuvants 6 rhesus macaques/group, 10 g HPV 16 capsomeres +/- week 0 and 8 24 Gissmann
25 Future HPV Vaccines: Basic and Applied Research conducting research accompanying the introduction of the HPV vaccine exploiting existing vaccine in off-label trials modifying existing vaccines supplementing prophylactic vaccines developing HPV vaccines for different targets 25 Gissmann
26 Supplementing prophylactic vaccines therapeutic vaccines treatment of precursors combi vaccines with prophylactic and therapeutic features post-exposure vaccination 26 Gissmann
27 HPV Vaccines: different options for attack HPV tumor cell E6/E7 epithelial cell kill APC E6/E7 E6/7-specific CTL preventing infection (disease) neutralizing antibodies curing disease E6/E7-specific CTLs 27 Gissmann
28 Supplementing prophylactic vaccines therapeutic vaccines treatment of precursors combi vaccines with prophylactic and therapeutic features post-exposure vaccination 28 Gissmann
29 HPV-specific vaccines: windows of attack Prophylaxis Therapy pre- post-exposure INFECTION persistence HSIL cancer age [y] ? antibodies +? T cells (Th1) 29 Gissmann
30 HPV chimeric capsomeres L1 E7/E6 5x monomer capsomere prophylactic and therapeutic features in mice? treating persistent infections and preventing reinfection in a post-exposure scenario? suitable for developing countries: phase I clinical trial 30 Gissmann
31 Regression of HPV 16 E7 pos tumors 31 Gissmann
32 Summary The current HPV vaccines, although of very good performance, are far from being perfect. Several lines of research are needed to fully exploit the potential of the existing vaccines and to generate vaccines for different applications either following the existing concept (VLP) or developing novel strategies. 32 Gissmann
33 Acknowledgements Thanx Birgit Aengeneyndt Corinna Klein Yvonne Rubio Lysann Schädlich Tilo Senger Martin Müller Jürgen Kleinschmidt Robert L. Garcea Massimo Tommasino 33 Gissmann
34 34 Gissmann
Conflict of interest
Helsinki 2012 HPV vaccines for developing countries Lutz Gissmann l.gissmann@dkfz.de Conflict of interest LG is a consultant to GSK and Sanofi Pasteur MSD and, due to existing IP, receives royalties from
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