Six hundred sixteen women with a history of hysterectomy were examined for the presence of human papillomavirus deoxyribonucleic acid (DNA) in vaginal smears using filter in situ hybridization. One hundred twenty patients had had hysterectomy for cervical intraepithelial neoplasia and invasive neoplasia, 54 for noncervical anogenital cancer, and 442 for benign uterine disorders. Human papillomavirus DNA was detected in 18% of all vaginal smears. A history of cervical neoplasia was associated with a significantly higher human papillomavirus infection rate, compared with patients with benign disease (33 versus 14%). Human papillomaviruses 16 and 18 were the most common types detected in patients with a history of cervical carcinoma, whereas the majority of patients with benign uterine disease were infected with human papillomavirus 6/11. Colposcopy identified lesions in more than half of the patients; these appeared in more than 90% as "condylomatous vaginitis." In 5% of the human papillomavirus-positive patients, human papillomavirus 16-positive vaginal intraepithelial neoplasia could be diagnosed. Increased risk of vaginal intraepithelial neoplasia after hysterectomy is associated with vaginal human papillomavirus infection. Virologic results explain the higher risk for vaginal neoplasia in patients with a history of cervical neoplasia.