1. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD 1 CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD Skin targeting for vaccine efficacy Laboratory of « Immunity and Infection » INSERM Paris, France Behazine COMBADIERE, PhD Director of research INSERM

2. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD FONDAMENTAL QUESTIONS AND NEW HOPES FOR VACCINATION B cells CD4 T cells CD8 T cells Dendritic cells Intensity Quality (T cells: cytokine released, cytotoxicity, B cells: IgG, IgA) Persistence of Memory Localization (mucosa, skin, liver) 1- Vaccination for whom? Defining the target population History of infection and multiplicity of vaccination Other diseases in the population 2- What type of immunity ? Defining the correlates of protection differs depending of the pathogens 3- How? Adapting the route of immunization bio-diversity of antigen-presenting cells (AP Skin, Mucosa sites

3. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD SKIN TARGETING OF APC DICTATES IMMUNOLOGICAL OUTCOMES B cells CD4 T cells CD8 T cells Dendritic cells Lymph nodes Stratum corneum Epidermis Dermis Hypodermis Lymphatic and blood capillary vessels Dermal- epidermal junction HAIR FOLLICLE Blood vessels Leukocytes recruitment(neutrophils, monocytes, pDC) Lymphatic vessels Differential targeting of antigen-presenting cells dictates the immunological outcomes (intensity, quality, localization) Langerhans cells Dermal DC Lymph node Micro- environments Lymph node Micro- environments

4. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD HAIR FOLLICLE TARGETING OF COMPOUNDS TO LANGERHANS CELLS Stratum corneum Hair fibre Follicular cast + cyanoacrylate glue + follicular casts + ca. 30% of stratum corneum Vogt et al. J Invest Dermatology 2006 and Mahe J Invest Dermatology 2008 Stratum corneum Epidermis Dermis Hypodermis Lymphatic and blood capillary vessels

5. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD SUITABLE SIZE OF VACCINE COMPOUNDS INTO FOLLICULAR DUCTS 5 37°C Skin purified human CD1a+ cells (+40nm) 1500-700 nm 200 nm 40 nm Suitable size for Langerhans cells up-take of vaccine : biodegradable or solid nanoparticles, virus like particles, small viruses, DNA, protein/peptide-carrier

6. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD Influenza-specific CD8 PROOF OF CONCEPT: PHASE I TRIAL OF AN INFLUENZA VACCINE Influenza-specific CD4 Vogt et al. J Immunol 2008 and Combadiere et al Plos One 2010 TC routeIM route Hair follicle Langerhans cells favors CD8 responses and also mucosal immunity but not IgG responses

7. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD FROM HUMANIZED MICE TO CLINICAL TRIALS tailor-cut immunization strategies could be proposed for infectious diseases: HIV, Malaria, Tuberculosis, emerging diseases and cancer EU-FP7 large-scale coordination : CUTHIVAC project 2010-2014 Human skin Explants Human immune cells Immunodeficient mice CLINICAL TRIALS Pre-clinical models Humanized mice

8. CEACHRUCNRSCPUINRAINRIAINSERMINSTITUT PASTEURIRD PERSPECTIVES Alternative methods of vaccination against infectious diseases to increase vaccine efficacy and allow a tailor-cut immune response Understanding skin immunological role for initiation and maintenance of immunity to infectious diseases Translation of basic research using innovative preclinical models into clinical trials 5 Phase I/II/IIa clinical trials are planned for 2011-2014: o Preventive and therapeutic HIV vaccine (3 trials): DNA-GTU® technology (FIT-Biotech) by TC, ID, IM or combined routes o Influenza vaccination in adults and in elderly (2 trials) : trivalent influenza vaccine by TC, ID and IM routes