title: TH1-promoting DNA immunization against allergens modulates the ratio of IgG1/IgG2a but does not affect the anaphylactic potential of IgG1 antibodies: no evidence for the synthesis of nonanaphylactic IgG1.
abstract: BACKGROUND
  In mouse, IgG1 has been reported to make up 2 functionally distinct phenotypes that also differ in their induction requirements. One of these phenotypes lacks anaphylactic activity.
  
  
  OBJECTIVE
  We hypothesized that nonanaphylactic IgG1 could modulate allergic reactions and investigated whether such antibodies are induced by DNA immunization.
  
  
  METHODS
  Mice were immunized with allergen-encoding plasmid DNA or with recombinant allergens and alum. Sera were analyzed for IgG subclasses by ELISA for anaphylactic IgE by rat basophil degranulation, and after heat inactivation of IgE for anaphylactic IgG by passive cutaneous anaphylaxis assay. IFN-gamma and IL-5 from in-vitro restimulated spleen cells were quantitated by ELISA.
  
  
  RESULTS
  After protein immunization, mice produced IgG1 and IgE, whereas DNA immunization elicited IgG1 and IgG2a but no IgE. However, all sera were positive for non-IgE-mediated passive cutaneous anaphylaxis. In the presence of anaphylactic IgG1, the additional occurrence of nonanaphylactic IgG1 cannot be strictly ruled out. To circumvent this problem, we immunized IL-4 receptor-deficient mice against Bet v 1a, because anaphylactic but not nonanaphylactic IgG1 has been reported to depend on IL-4. These animals produced only low amounts of IgG1, but sera were again positive for non-IgE-mediated anaphylactic activity.
  
  
  CONCLUSIONS
  Our results revealed no evidence for the production of nonanaphylactic IgG1. Furthermore, our data indicate that the development of non-IgE-mediated anaphylaxis does not require IL-4 receptor signaling.
mesh_terms: D000485:Allergens / Q000008:administration & dosage / Q000235:genetics; D000707:Anaphylaxis / Q000209:etiology* / Q000276:immunology* / Q000628:therapy; D000818:Animals; D052179:Antigens, Plant; D015550:Cell Degranulation; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D007114:Immunization; D007074:Immunoglobulin G / Q000097:blood*; D007167:Immunotherapy; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D010940:Plant Proteins / Q000008:administration & dosage / Q000235:genetics; D019948:Receptors, Interleukin-4 / Q000378:metabolism; D015398:Signal Transduction; D018417:Th1 Cells / Q000276:immunology*; D014407:Tumor Cells, Cultured; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology / Q000494:pharmacology*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C096695:Api g 1 protein, Apium graveolens; D016207:Cytokines; D007074:Immunoglobulin G; D010940:Plant Proteins; D019948:Receptors, Interleukin-4; D019444:Vaccines, DNA; C061312:Bet v 1 allergen, Betula