title: Specific IgE and IgG responses and cytokine profile in subjects with allergic reactions to biting midge Forcipomyia taiwana.
abstract: BACKGROUND
  Forcipomyia taiwana is a tiny blood-sucking midge whose habitat covers large parts of Taiwan and southern China. Female midges bite during the day, causing intense pruritus and swelling in allergic individuals. In this study, we investigated the immune responses of different allergic reactions to midge bites.
  
  
  METHODS
  F. taiwana (midge)-specific IgE, -IgG and -IgG subclasses were examined by ELISA in 62 human subjects. Peripheral blood mononuclear cells (PBMC) from 6 subjects with solely delayed reactions (SDR) to midge bites and 6 nonallergic controls (NAC) were cultured with midge extract at various time points and assayed. Proliferation of PBMC was measured by MTT assay. Expression of cytokine mRNA was measured by real-time PCR and protein levels by cytometric bead immunoassay or ELISA. Protease activity in midge extract was determined by the Azocoll method.
  
  
  RESULTS
  Midge-specific IgE among subjects with an immediate reaction were significantly elevated compared to SDR and NAC subjects. There were no differences in the level of midge-specific-IgG, -IgG(1), -IgG(2), -IgG(3) and -IgG(4) among subjects with different biting reactions. Midge extract elicited significantly more PBMC proliferation, higher expression of IFN-gamma, IL-10, IL-6 and TNF-alpha in SDR subjects than in NAC. Protease activity was detected in midge extract. Protease inhibitors E64 and pepstatin suppressed midge-extract-induced IL-8 production.
  
  
  CONCLUSIONS
  Our results suggest that an immediate reaction to midge bites is IgE-mediated. IFN-gamma, IL-6 and TNF-alpha are involved in delayed reactions to midge bites. A protease-activated pathway may also be involved in the intense, itchy reactions to midge bites.
authors.affiliation: Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, Taichung, Taiwan.
authors.forename: Yi-Hsing
authors.initials: YH
authors.lastname: Chen
authors.forename: Mey-Fann
authors.initials: MF
authors.lastname: Lee
authors.forename: Jaw-Ji
authors.initials: JJ
authors.lastname: Tsai
authors.forename: Hsin-Ju
authors.initials: HJ
authors.lastname: Wu
authors.forename: Guang-Yuh
authors.initials: GY
authors.lastname: Hwang