Influenza virus-like particles coated onto microneedles can elicit stimulatory effects on Langerhans cells in human skin.

Virus-like particles (VLPs) have a number of features that make them attractive influenza vaccine candidates. Microneedle (MN) devices are being developed for the convenient and pain-free delivery of vaccines across the skin barrier layer. Whilst MN-based vaccines have demonstrated proof-of-concept in mice, it is vital to understand how MN targeting of VLPs to the skin epidermis affects activation and migration of Langerhans cells (LCs) in the real human skin environment. MNs coated with vaccine reproducibly penetrated freshly excised human skin, depositing 80% of the coating within 60 s of insertion. Human skin experiments showed that H1 (A/PR/8/34) and H5 (A/Viet Nam/1203/04) VLPs, delivered via MN, stimulated LCs resulting in changes in cell morphology and a reduction in cell number in epidermal sheets. LC response was significantly more pronounced in skin treated with H1 VLPs, compared with H5 VLPs. Our data provides strong evidence that MN-facilitated delivery of influenza VLP vaccines initiates a stimulatory response in LCs in human skin. The results support and validate animal data, suggesting that dendritic cells (DCs) targeted through deposition of the vaccine in skin generate immune response. The study also demonstrates the value of using human skin alongside animal studies for preclinical testing of intra-dermal (ID) vaccines.

[1]  T. Uyeki,et al.  Prevention and control of influenza: recommendations of the Advisory Committee on Immunization Practices (ACIP), 2008. , 2008, MMWR. Recommendations and reports : Morbidity and mortality weekly report. Recommendations and reports.

[2]  Bjoern Peters,et al.  Pre-existing immunity against swine-origin H1N1 influenza viruses in the general human population , 2009, Proceedings of the National Academy of Sciences.

[3]  Anthony Morrissey,et al.  Cutaneous DNA delivery and gene expression in ex vivo human skin explants via wet-etch microfabricated microneedles , 2005, Journal of drug targeting.

[4]  C. Hughes,et al.  Of Mice and Not Men: Differences between Mouse and Human Immunology , 2004, The Journal of Immunology.

[5]  Beatrix Grubeck-Loebenstein,et al.  Vaccination in the elderly: an immunological perspective. , 2009, Trends in immunology.

[6]  J. Birchall,et al.  Gene Delivery to the Epidermal Cells of Human Skin Explants Using Microfabricated Microneedles and Hydrogel Formulations , 2008, Pharmaceutical Research.

[7]  K. Brown,et al.  Genetic Vaccines and Therapy , 2003 .

[8]  G. Leroux-Roels,et al.  Seasonal influenza vaccine delivered by intradermal microinjection: A randomised controlled safety and immunogenicity trial in adults. , 2008, Vaccine.

[9]  N. Cox,et al.  Prevention and Control of Influenza: recommendations of the Advisory Committee on Immunization Practices (ACIP). , 2006, MMWR. Recommendations and reports : Morbidity and mortality weekly report. Recommendations and reports.

[10]  Mark R Prausnitz,et al.  Precise microinjection into skin using hollow microneedles. , 2006, The Journal of investigative dermatology.

[11]  Mark R. Prausnitz,et al.  Coating Formulations for Microneedles , 2007, Pharmaceutical Research.

[12]  Mark R Prausnitz,et al.  Coated microneedles for transdermal delivery. , 2007, Journal of controlled release : official journal of the Controlled Release Society.

[13]  I. Capua,et al.  Ecology, Epidemiology and Human Health Implications of Avian Influenza Viruses: Why do We Need to Share Genetic Data? , 2008, Zoonoses and public health.

[14]  W. Blackwelder,et al.  Safety and immunogenicity of a recombinant hemagglutinin vaccine for H5 influenza in humans. , 2001, Vaccine.

[15]  U. Reichl,et al.  Continuous cell lines as a production system for influenza vaccines , 2009, Expert review of vaccines.

[16]  Mark Wolff,et al.  Safety and immunogenicity of an inactivated subvirion influenza A (H5N1) vaccine. , 2006, The New England journal of medicine.

[17]  C. Griffiths,et al.  Cytokines and Langerhans cell mobilisation in mouse and man. , 2005, Cytokine.

[18]  M. Prausnitz,et al.  Transdermal Influenza Immunization with Vaccine-Coated Microneedle Arrays , 2009, PLoS ONE.

[19]  M. Prausnitz,et al.  Stabilization of Influenza Vaccine Enhances Protection by Microneedle Delivery in the Mouse Skin , 2009, PloS one.

[20]  M. Boes,et al.  Roles for IL-1 and TNFalpha in dynamic behavioral responses of Langerhans cells to topical hapten application. , 2007, Journal of dermatological science.

[21]  C. Davis,et al.  Induction of Long-Term Protective Immune Responses by Influenza H5N1 Virus-Like Particles , 2009, PloS one.

[22]  Malbea A Lapete,et al.  Prevention and Control of Influenza Recommendations of the Advisory Committee on Immunization Practices ( ACIP ) , 2004 .

[23]  A. Kamen,et al.  Development of a simple and high-yielding fed-batch process for the production of influenza vaccines. , 2009, Vaccine.

[24]  M. Prausnitz,et al.  Improved influenza vaccination in the skin using vaccine coated microneedles. , 2009, Vaccine.

[25]  W. Keitel,et al.  Safety and immunogenicity of inactivated, Vero cell culture-derived whole virus influenza A/H5N1 vaccine given alone or with aluminum hydroxide adjuvant in healthy adults. , 2009, Vaccine.

[26]  R. Compans,et al.  Kinetics of Immune Responses to Influenza Virus-Like Particles and Dose-Dependence of Protection with a Single Vaccination , 2009, Journal of Virology.

[27]  H. Koprowski,et al.  Smallpox subunit vaccine produced in planta confers protection in mice , 2007, Proceedings of the National Academy of Sciences.

[28]  Xiu-Feng Wan,et al.  Evolution of Highly Pathogenic H5N1 Avian Influenza Viruses in Vietnam between 2001 and 2007 , 2008, PloS one.

[29]  Christopher Allender,et al.  Development of an ex vivo human skin model for intradermal vaccination: tissue viability and Langerhans cell behaviour. , 2009, Vaccine.

[30]  松本 慶蔵,et al.  季節性インフルエンザ(Seasonal Influenza)の総括 , 2009 .

[31]  Estanislao Nistal-Villán,et al.  Attacking the flu: New prospects for the rational design of antivirals , 2009, Nature Medicine.

[32]  T. Tumpey,et al.  Mucosal Delivery of Inactivated Influenza Vaccine Induces B-Cell-Dependent Heterosubtypic Cross-Protection against Lethal Influenza A H5N1 Virus Infection , 2001, Journal of Virology.

[33]  Sion A. Coulman,et al.  Microneedles and other physical methods for overcoming the stratum corneum barrier for cutaneous gene therapy. , 2006, Critical reviews in therapeutic drug carrier systems.

[34]  Mark R Prausnitz,et al.  Formulation and coating of microneedles with inactivated influenza virus to improve vaccine stability and immunogenicity. , 2010, Journal of controlled release : official journal of the Controlled Release Society.

[35]  Michel Cormier,et al.  Microneedle-based vaccines. , 2009, Current topics in microbiology and immunology.

[36]  M. Prausnitz,et al.  Immunization by vaccine-coated microneedle arrays protects against lethal influenza virus challenge , 2009, Proceedings of the National Academy of Sciences.

[37]  R. Compans,et al.  Virus-Like Particle Vaccine Induces Protective Immunity against Homologous and Heterologous Strains of Influenza Virus , 2007, Journal of Virology.

[38]  Yu Wang,et al.  Safety and immunogenicity of an inactivated adjuvanted whole-virion influenza A (H5N1) vaccine: a phase I randomised controlled trial , 2006, The Lancet.

[39]  J. Jones,et al.  Early Assessment of Anxiety and Behavioral Response to Novel Swine-Origin Influenza A(H1N1) , 2009, PloS one.