title: The development of a murine model for Forcipomyia taiwana (biting midge) allergy.
abstract: BACKGROUND
  Forcipomyia taiwana (biting midge) allergy is the most prevalent biting insect allergy in Taiwan. An animal model corresponding to the human immuno-pathologic features of midge allergy is needed for investigating the mechanisms and therapies. This study successfully developed a murine model of Forcipomyia taiwana allergy.
  
  
  METHODS
  BALB/c mice were sensitized intra-peritoneally with midge extract on days 0, 7, 14, 21 then intra-dermally on days 28, 31 and 35. Serum midge-specific IgE, IgG1, and IgG2a were measured every 14 days by indirect ELISA. The mice were challenged intradermally with midge extract at day 40 and then sacrificed. Proliferation and cytokine production of splenocytes after stimulation with midge extract were determined by MTT assay and ELISA, respectively. The cytokine mRNA expression in response to midge stimulation was analyzed by RT-PCR.
  
  
  RESULTS
  Serum IgE, total IgG, and IgG1 antibody levels against midge extract were significantly higher in the midge-sensitized mice than in the control mice. After the two-step sensitization, all mice in the midge-sensitized group displayed immediate itch and plasma extravasation reactions in response to challenge with midge extract. Skin histology from midge-sensitized mice showed marked eosinophil and lymphocyte infiltrations similar to that observed in humans. Stimulation of murine splenocytes with midge extract elicited significant proliferation, IL-4, IL-10, IL-13 and IFN-γ protein production, and up-regulation of mRNA in a dose-dependent manner in the midge-sensitized group, but not in the control group.
  
  
  CONCLUSIONS
  A murine model of midge bite allergy has been successfully developed using a two-step sensitization protocol. The sensitized mice have very similar clinical and immunologic reactions to challenge with midge proteins as the reactions of human to midge bites. This murine model may be a useful platform for future research and the development of treatment strategies for insect bite allergy.
authors.affiliation: Department of Medical Research Taichung Veterans General Hospital, Taichung, Taiwan; Department of Life Science, Tunghai University, Taichung, Taiwan.
authors.forename: Mey-Fann
authors.initials: MF
authors.lastname: Lee
authors.affiliation: Institute of Biotechnology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Kai-Jei
authors.initials: KJ
authors.lastname: Yang
authors.affiliation: Institute of Biotechnology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Nancy M
authors.initials: NM
authors.lastname: Wang
authors.affiliation: Department of Medical Research Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Yung-Tsung
authors.initials: YT
authors.lastname: Chiu
authors.affiliation: Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, Taiwan.
authors.forename: Pei-Chih
authors.initials: PC
authors.lastname: Chen
authors.affiliation: Department of Life Science, Tunghai University, Taichung, Taiwan; Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, Taiwan; Faculty of Medicine, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Yi-Hsing
authors.initials: YH
authors.lastname: Chen
mesh_terms: D000180:Acrylic Resins / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use; D000287:Administration, Topical; D000818:Animals; D000918:Antibody Specificity / Q000276:immunology; D001706:Biopsy; D049109:Cell Proliferation / Q000187:drug effects; D002519:Ceratopogonidae / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis; D003907:Dexamethasone / Q000008:administration & dosage / Q000494:pharmacology / Q000627:therapeutic use; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D006967:Hypersensitivity / Q000150:complications / Q000188:drug therapy / Q000276:immunology* / Q000469:parasitology*; D006968:Hypersensitivity, Delayed / Q000150:complications / Q000188:drug therapy / Q000276:immunology / Q000469:parasitology / Q000473:pathology; D007150:Immunohistochemistry; D007249:Inflammation / Q000150:complications / Q000188:drug therapy / Q000473:pathology; D008807:Mice, Inbred BALB C; D011537:Pruritus / Q000188:drug therapy / Q000276:immunology / Q000469:parasitology / Q000473:pathology; D012867:Skin / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D012882:Skin Tests; D013154:Spleen / Q000473:pathology; D014020:Tissue Extracts
chemical_list: D000180:Acrylic Resins; D016207:Cytokines; D014020:Tissue Extracts; C479038:carbomer; D003907:Dexamethasone