In the last few years, serological identification of tumour-associated antigens (TAAs) by recombinant cDNA expression cloning (SEREX) has enabled the mapping of humoral immune responses against TAAs in various types of cancer. The present paper describes the application of SEREX to Burkitt's lymphoma (BL), a malignancy not previously characterized by SEREX. By using a cDNA library from a BL cell line that does not express IgG, technical difficulties related to background immunoglobulin clones were overcome. Screening with sera from three BL patients revealed immunoreactivity against seven different gene products, six of which represent known human genes. Five proteins had previously been characterized by SEREX in other malignancies or identified as targets of autoantibodies in autoimmune disease. Seroreactivity against ATF-2, a member of the AP-1 transcription factor family, was validated by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using recombinant ATF-2 protein. Autoantibody responses against ATF-2 were detected by ELISA in 6 of 8 BL patients, compared with 6 of 13 patients with T-cell non-Hodgkin's lymphoma (T-NHL), 5 of 23 patients with follicular lymphoma and 2 of 27 diffuse large B-cell lymphoma patients. In contrast, reactivity was found in only 1 of 50 healthy volunteers. Next, we showed by immunohistochemistry that the activated form of ATF2 (ATF-2pp) was highly expressed in six different BL samples. We conclude that the SEREX approach with a B-cell cDNA source is applicable in NHL. Furthermore, we identified genes with possible involvement in the pathogenesis of BL using this technique.