Allergen-specific immunoglobulin E (IgE) mediates immediate-type hypersensitivity reactions and plays a central role in allergic diseases. Although antigen-driven B-cell maturation and isotype switching occur within germinal centres (GCs), the role of GCs in IgE production is poorly understood. In view of this, we investigated the development of IgE-expressing cells within GCs in response to an extensively characterized antigen, 2-phenyloxazolone (phOx). The phOx-specific IgE-expressing cells localized within GCs 7 days after immunization, and peaked in number on day 11. Surprisingly, very few IgE-positive cells were found in the T-cell areas of the lymph node. Flow cytometric studies confirmed that IgE was expressed by B cells and was not the result of trapping by follicular dendritic cells. The specificity of the antibody response was confirmed by microdissection and reverse transcription-polymerase chain reaction using phOx-specific IgE primers. IgE-positive cells were primarily found within GCs while, in contrast, many IgG1-positive cells could also be detected outside GCs in the T-cell areas. Taken together, these data highlight the importance of GCs in the production of antigen-specific IgE antibody.