Characterization of powdered epidermal vaccine delivery with multiphoton microscopy

Multiphoton laser scanning microscopy (MPLSM) has been adapted to non-invasively characterize hand-held powdered epidermal vaccine delivery technology. A near infrared femtosecond pulsed laser, wavelength at approximately 920 nm, was used to evoke autofluorescence of endogenous fluorophores within ex vivo porcine and human skin. Consequently, sub cellular resolution three-dimensional images of stratum corneum and viable epidermal cells were acquired and utilized to observe the morphological deformation of these cells as a result of micro-particle penetration. Furthermore, the distributional pattern of micro-particles within the specific skin target volume was quantified by measuring the penetration depth as revealed by serial optical sections in the axial plane obtained with MPLSM. Additionally, endogenous fluorescence contrast images acquired at the supra-basal layer reveal cellular structures that may pertain to dendritic Langerhans cells of the epidermis. These results show that MPLSM has advantages over conventional histological approaches, since three-dimensional functional images with sub-cellular spatial resolution to depths beyond the epidermis can be acquired non-invasively. Accordingly, we propose that MPLSM is ideal for investigations of powdered epidermal vaccine delivery.

[1]  P. So,et al.  Confocal microscopy and multi-photon excitation microscopy of human skin in vivo. , 2001, Optics express.

[2]  L. Babiuk,et al.  Gene Gun-Mediated DNA Immunization Primes Development of Mucosal Immunity against Bovine Herpesvirus 1 in Cattle , 2000, Journal of Virology.

[3]  Dexiang Chen,et al.  Epidermal Powder Immunization Induces both Cytotoxic T-Lymphocyte and Antibody Responses to Protein Antigens of Influenza and Hepatitis B Viruses , 2001, Journal of Virology.

[4]  R. E. Kalman,et al.  A New Approach to Linear Filtering and Prediction Problems , 2002 .

[5]  R. Anderson,et al.  Fluorescence excitation spectroscopy provides information about human skin in vivo. , 2000, The Journal of investigative dermatology.

[6]  John White,et al.  Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability , 1999, Nature Biotechnology.

[7]  F. Foster,et al.  A 100-200 MHz ultrasound biomicroscope , 2000, IEEE Transactions on Ultrasonics, Ferroelectrics and Frequency Control.

[8]  R. Yu,et al.  Morphological and quantitative analyses of normal epidermal Langerhans cells using confocal scanning laser microscopy , 1994, The British journal of dermatology.

[9]  J. Fujimoto,et al.  In vivo ultrahigh-resolution optical coherence tomography. , 1999, Optics letters.

[10]  M. Kendall,et al.  Numerical simulation of heat transfer from a transonic jet impinging on skin for needle-free powdered drug and vaccine delivery , 2004 .

[11]  P. Selby,et al.  The response of human dendritic cells to recombinant adenovirus, recombinant Mycobacterium bovis Bacillus Calmette Guerin and biolistic methods of antigen delivery: different induction of contact-dependant and soluble signals. , 2001, Immunology letters.

[12]  L. Keay,et al.  Short term wear of high Dk soft contact lenses does not alter corneal epithelial cell size or viability , 2001, The British journal of ophthalmology.

[13]  R. Wildnauer,et al.  Stratum corneum biomechanical properties. I. Influence of relative humidity on normal and extracted human stratum corneum. , 1971, The Journal of investigative dermatology.

[14]  P. So,et al.  Two-Photon deep tissue ex vivo imaging of mouse dermal and subcutaneous structures. , 1998, Optics express.

[15]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[16]  P. Delaney,et al.  View of normal human skin in vivo as observed using fluorescent fiber-optic confocal microscopic imaging. , 2003, The Journal of investigative dermatology.

[17]  B. R. Masters,et al.  Optical Biopsy of In Vivo Human Skin: Multi-photon Excitation Microscopy , 1998, Lasers in Medical Science.

[18]  H Ermert,et al.  Sonography of the skin at 100 MHz enables in vivo visualization of stratum corneum and viable epidermis in palmar skin and psoriatic plaques. , 1999, The Journal of investigative dermatology.

[19]  J. Fujimoto,et al.  Determination of the refractive index of highly scattering human tissue by optical coherence tomography. , 1995, Optics letters.

[20]  N. Romani,et al.  Langerhans cells – dendritic cells of the epidermis , 2003, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[21]  J. Fujimoto.  Optical coherence tomography for ultrahigh resolution in vivo imaging , 2003, Nature Biotechnology.

[22]  Nathan J. Quinlan,et al.  Investigations of gas and particle dynamics in first generation needle-free drug delivery devices , 2001 .

[23]  R. Webb,et al.  In vivo confocal scanning laser microscopy of human skin II: advances in instrumentation and comparison with histology. , 1999, The Journal of investigative dermatology.

[24]  G. Schuler,et al.  A strikingly constant ratio exists between Langerhans cells and other epidermal cells in human skin. A stereologic study using the optical disector method and the confocal laser scanning microscope. , 2001, The Journal of investigative dermatology.

[25]  D. Soll,et al.  Changes in the motility, morphology, and F-actin architecture of human dendritic cells in an in vitro model of dendritic cell development. , 2000, Cell motility and the cytoskeleton.

[26]  Y. Maa,et al.  Needle-free epidermal powder immunization , 2002, Expert review of vaccines.

[27]  T. Liesegang.  Short term wear of high Dk soft contact lenses does not alter corneal epithelial cell size or viability. Stapleton F, ∗ Kasses S, Bolis S, Keay L. Br J Ophthalmol 2001;85:143–146. , 2001 .

[28]  M. Kendall,et al.  Effects of relative humidity and ambient temperature on the ballistic delivery of micro-particles to excised porcine skin. , 2004, The Journal of investigative dermatology.

[29]  C. Sheppard,et al.  Practical limits of resolution in confocal and non‐linear microscopy , 2004, Microscopy research and technique.

[30]  P. So,et al.  [29] Multiphoton excitation microscopy, confocal microscopy, and spectroscopy of living cells and tissues; functional metabolic imaging of human skin in vivo , 1999 .

[31]  M. Kendall,et al.  Intradermal ballistic delivery of micro-particles into excised human skin for pharmaceutical applications. , 2004, Journal of biomechanics.

[32]  Mark A. F. Kendall,et al.  The delivery of particulate vaccines and drugs to human skin with a practical, hand-held shock tube-based system , 2002 .

[33]  Y. Kawaoka,et al.  Epidermal immunization by a needle-free powder delivery technology: Immunogenicity of influenza vaccine and protection in mice , 2000, Nature Medicine.

[34]  Mark A. F. Kendall,et al.  A ballistic study of micro-particle penetration to the oral mucosa , 2003 .

[35]  J. Streilein,et al.  On the extraordinary capacity of allogeneic epidermal Langerhans cells to prime cytotoxic T cells in vivo. , 1989, Journal of immunology.

[36]  Roger W. Ainsworth,et al.  Measurements of the gas and particle flow within a converging-diverging nozzle for high speed powdered vaccine and drug delivery , 2004 .

[37]  R. Tyrrell,et al.  New trends in photobiology. The interaction of UVA radiation with cultured cells. , 1990, Journal of photochemistry and photobiology. B, Biology.

[38]  H. Junginger,et al.  Interactions of elastic and rigid vesicles with human skin in vitro: electron microscopy and two-photon excitation microscopy. , 1999, Biochimica et biophysica acta.

[39]  D. Fuller,et al.  Gene gun-based nucleic acid immunization: elicitation of humoral and cytotoxic T lymphocyte responses following epidermal delivery of nanogram quantities of DNA. , 1995, Vaccine.

[40]  C. Hanlon,et al.  DNA immunization protects nonhuman primates against rabies virus , 1998, Nature Medicine.

[41]  B. Bellhouse,et al.  Identification of second harmonic optical effects from vaccine coated gold microparticles. , 2004, Physics in medicine and biology.

[42]  W. Webb,et al.  Nonlinear magic: multiphoton microscopy in the biosciences , 2003, Nature Biotechnology.

[43]  R. Steinman,et al.  Dendritic cells and the control of immunity , 1998, Nature.

[44]  Mary S. Wu,et al.  Induction of antigen-specific CD8+ T cells, T helper cells, and protective levels of antibody in humans by particle-mediated administration of a hepatitis B virus DNA vaccine. , 2000, Vaccine.