Skin Vaccination against Cervical Cancer Associated Human Papillomavirus with a Novel Micro-Projection Array in a Mouse Model

Kendall MAF (2010) Skin Vaccination against Cervical Cancer Associated Human Papillomavirus with a Novel Micro-Projection Array in a Mouse Model. PLoS ONE 5(10): e13460. doi:10.1371/journal.pone.0013460 Skin Vaccination against Cervical Cancer Associated Human Papillomavirus with a Novel Micro-Projection Array in a Mouse Model Holly J. Corbett 1 Germain J. P. Fernando 1 Xianfeng Chen 1 Ian H. Frazer 0 1 Mark A. F. Kendall 0 1 Sotirios Koutsopoulos, Massachusetts Institute of Technology, United States of America 0 ), Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane, Queensland , Australia , 2 Diamantina Institute, Princess Alexandra Hospital, The University of Queensland , Woolloongabba, Queensland , Australia 1 1 Delivery of Drugs and Genes Group (D Background: Better delivery systems are needed for routinely used vaccines, to improve vaccine uptake. Many vaccines contain alum or alum based adjuvants. Here we investigate a novel dry-coated densely-packed micro-projection array skin patch (NanopatchTM) as an alternate delivery system to intramuscular injection for delivering an alum adjuvanted human papillomavirus (HPV) vaccine (GardasilH) commonly used as a prophylactic vaccine against cervical cancer. Methodology/Principal Findings: Micro-projection arrays dry-coated with vaccine material (GardasilH) delivered to C57BL/6 mouse ear skin released vaccine within 5 minutes. To assess vaccine immunogenicity, doses of corresponding to HPV-16 component of the vaccine between 0.4360.084 ng and 3006120 ng (mean 6 SD) were administered to mice at day 0 and day 14. A dose of 5566.0 ng delivered intracutaneously by micro-projection array was sufficient to produce a maximal virus neutralizing serum antibody response at day 28 post vaccination. Neutralizing antibody titres were sustained out to 16 weeks post vaccination, and, for comparable doses of vaccine, somewhat higher titres were observed with intracutaneous patch delivery than with intramuscular delivery with the needle and syringe at this time point. Conclusions/Significance: Use of dry micro-projection arrays (NanopatchTM) has the potential to overcome the need for a vaccine cold chain for common vaccines currently delivered by needle and syringe, and to reduce risk of needle-stick injury and vaccine avoidance due to the fear of the needle especially among children. - Funding: Cancer Council of Queensland, the University of Queensland, National Health and Medical Research Council (Australia) Grants ID# 569726, ID# 456150, Australian Research Council Grant ID# DP077464 and the Queensland Smart State Scheme funded this research. Professor Mark A. F. Kendall was the recipient of an Australian Research Council Future Fellowship. Professor Ian H. Frazer was the recipient of a Queensland Government Premiers Fellowship. The University of Queensland and Professor Ian H. Frazer derive royalty income from the sale of HPV VLP vaccines. The funders had no role in the study design, data collection and analysis, decision to publish, or the preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. Most vaccines are currently delivered by needle and syringe. However as a vaccine delivery device, the needle and syringe has many important shortcomings. These include potential transmission of blood borne diseases through needle-stick injuries [1] and needle reuse approximately 30% of injections for the purpose of vaccination in developing nations are unsafe [2], and that needlestick injuries cause more than 500,000 deaths per year [3]. Needlephobia and the pain associated with an intramuscular injection are also downsides it is estimated that needle phobia is present in at least 10% [4] of the population, or higher [5]. The muscle is also a highly inefficient site for vaccination, as it does not have a high density of antigen presenting cells. In contrast, the skin is an attractive alternative site for vaccination due to its dense network of potent antigen presenting cells (APCs) including Langerhans Cells (LCs) [6], and many sub-sets of dermal dendritic cells (dDCs) [7]. The close proximity of these cells to the skin surface means it could be possible to target them in ways which may reduce pain and potential of transmission of blood borne pathogens. While cutaneous delivery has great potential, the closest method used currently in the clinic intradermal injection is technically difficult, necessitating development of advanced targeting methods as reviewed in [8,9]. In this study a novel skin patch called the NanopatchTM is used to target these skin immune cells. The NanopatchTM is a microprojection array with uniquely dense projection packing (.20,000/ cm2) and short projections (110 mm in length). This needle density was designed such that delivered vaccine has been co-localized with 50% skin immune cells in both epidermis and dermis upon cutaneous application without relying on diffusion (see Figure 1) [10]. Previous studies with NanopatchTM immunization have utilized ovalbumin and split influenza vaccine as antigens without addition of an adjuvant. Crichton et. al [11] demonstrated high antibody titers after one immunization with under 2 mg via NanopatchTM using the model antigen ovalbumin in C57BL/6 mice without a boost using 65 mm long Nanopatch projections. Fernando et. al. [10] demonstrated induction of protective levels of functional antibodies against influenza in mice with 110 mm long NanopatchTM projections (same as used in this study) using a split virus, unadjuvanted trivalent influenza vaccine (Fluvax 2008H); with a factor of 100 in delivered dose-sparing, compared to the needle and syringe. In these previous studies, vaccines were delivered without adjuvant. In the current study we extend to explore the utility of the NanopatchTM in delivering an alum adjuvant. This is important, because many vaccines are adjuvanted with Alum in the most widely used [12]. Indeed, until the recent licensure of AS04, alum was the only adjuvant to be licensed by the FDA [13]. AS04 is alum based, with the addition of a lipid based toll-like receptor 4 agonist 3-Odesacyl-49-monophosphoryl lipid A (MPL) [14]. Alum is chemically either aluminum oxyhydroxide, or aluminum hydroxyphosphate. For new technologies to take advantage of currently licensed vaccines, ideally one should work with alum-adjuvanted vaccines. So far, solid formulation work has been performed with alum adjuvant for epidermal powder immunization (EPI) with hepatitis B [15,16,17], and diphtheria and tetanus toxoids [17]. Extensive alum gel coagulation during drying is suspected to inhibit the release of antigen such that it is not recoverable upon rehydration [16], and losses of efficacy have been reported after lyophilization or freezing [18,19,20]. To minimize these losses without significantly reducing the amount of alum in the total solid (,1% [18]), ra (...truncated)