title: Transcutaneous immunization in auricle skin induces antigen-specific mucosal and systemic immune responses in BALB/c mice.
abstract: OBJECTIVE
  Transcutaneous immunization (TCI) is a novel route of vaccination through application of a topical vaccine antigen on skin. Phosphorylcholine (PC) is a structural component of a variety of pathogens, and anti-PC immune responses protect mice against invasive bacterial diseases. The purpose of the study was to examine the effect of TCI using PC in back skin or auricle skin in BALB/c mice.
  
  
  METHODS
  TCI was performed in BALB/c mice in back skin or auricle skin using PC-keyhole limpet hemocyanin (KLH) plus cholera toxin (CT). Inoculations were given once each week for six consecutive weeks. Immunogenicity was evaluated by measuring PC-specific IgG and specific IgG1, IgG2a, IgM, IgA, and secretory IgA antibodies by ELISA. IL-4, IL-5, IL-10, IL-12, IL-13 and IFN-γ levels were also measured by ELISA.
  
  
  RESULTS
  Serum IgG after TCI in auricle skin was significantly higher than after TCI in back skin and in controls. Secretory IgA antibodies after TCI in auricle skin were also significantly higher than after TCI in back skin and in controls in nasal, BALF, vaginal and fecal samples. PC-specific IgG1 and IgG2a were significantly higher after TCI in auricle skin compared to controls and compared to TCI in back skin. IgG1 was significantly higher than IgG2a after TCI in auricle skin. Production of IFN-γ, IL-4 and IL-10 from CD4+cells was significantly higher after TCI in auricle skin than after TCI in back skin and in controls, whereas IL-5, IL-12 and IL-13 were not detected in any mice.
  
  
  CONCLUSIONS
  These results suggest that TCI in auricle skin using PC plus CT in BALB/c mice is a simple approach for induction of systemic and mucosal immune responses that are shifted in the Th2 direction.
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan. Electronic address: nagano@m.kufm.kagoshima-u.ac.jp.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Nagano
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Jimura
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Nagano
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Makise
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Miyashita
authors.affiliation: Department of Otolaryngology and Head and Neck Surgery, Kagoshima University Graduate School of Medical and Dental Sciences, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Kurono
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology*; D000279:Administration, Cutaneous; D000818:Animals; D001415:Back; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002772:Cholera Toxin / Q000008:administration & dosage / Q000494:pharmacology*; D022121:Cholera Vaccines / Q000008:administration & dosage / Q000494:pharmacology*; D054644:Ear Auricle; D005260:Female; D006433:Hemocyanins / Q000008:administration & dosage / Q000494:pharmacology*; D000071497:Immunogenicity, Vaccine / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology; D018664:Interleukin-12 / Q000276:immunology; D018793:Interleukin-13 / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D015848:Interleukin-5 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010767:Phosphorylcholine / Q000008:administration & dosage / Q000494:pharmacology*; D014611:Vaccination / Q000379:methods*
chemical_list: D000276:Adjuvants, Immunologic; D022121:Cholera Vaccines; C556062:IL10 protein, mouse; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D018793:Interleukin-13; D015848:Interleukin-5; D010767:Phosphorylcholine; D016753:Interleukin-10; D018664:Interleukin-12; D015847:Interleukin-4; D007371:Interferon-gamma; D002772:Cholera Toxin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin
keywords: Auricle; Phosphorylcholine; Secretory IgA; Transcutaneous immunization