Certain types of human papillomavirus (HPV) are associated with anogenital carcinomas, including carcinomas of the anal canal. Whereas several serological studies have found an association between papillomavirus antibody responses and cervical carcinoma, the antibody response against papillomavirus antigens among patients with anal carcinoma has not been investigated. The present study has examined the antibody responses to a panel of papillomavirus-derived antigens and compared the serological profile with the histology and HPV carrier state of the tumor, as well as with the stage and prognosis of the disease. Sera from 64 patients with anal cancer and from 79 healthy blood donors were studied in ELISA for the presence of IgA and IgG antibodies to 5 previously described HPV16-derived synthetic peptide antigens. Serum IgA antibodies to a peptide antigen derived from the E2 region of HPV16 were found in 89% of patients with anal cancer as compared to 24% of controls (p = 0.0001). The IgA reactivity to the 4 other antigens showed only low and non-significant increases in mean titer. Serum IgG responses were similar among patients and controls. Among patients who had progressive disease, 21/21 were seropositive for IgA anti-E2 at diagnosis, as compared to 36/43 patients who were in remission after a mean follow-up of 41 months (p = 0.05). Forty-seven cases of anal carcinoma were also studied for the presence of HPV by in situ hybridization using a probe mix of 7 anogenital HPV types. Sixteen patients (35%) carried HPV in their anal cancer and one patient had an HPV-positive benign lesion adjacent to the tumor. Patients with HPV-carrying anal cancer were significantly younger than those with HPV-negative anal cancers (mean age: 57 and 68 years, respectively, p = 0.03). No differences in seroreactivity or HPV carrier state were seen depending on the stage or histological type of the tumor.