Approximately 2 million South Africans are HIV/TB coinfected, and many develop skeletal disease. The resurgence of spinal tuberculosis, including atypical forms, is due largely to HIV-associated immune suppression. We investigated the impact of HIV coinfection on the histological features of the disease and the occurrence of atypical opportunistic organisms in infectious spondylitis in an HIV/TB endemic region. We analyzed blood and tissue biopsies from 60 patients with tuberculous spondylitis. Investigations included full blood counts, CD4/CD8 counts, HIV-1 serology and RNA quantification (tissue and plasma), acid-fast bacilli localization and routine TB culture, histopathologic evaluation of biopsies, and bacterial genotyping using the 16S rDNA gene. Twenty-two patients (37%) were HIV positive with a mean age of 29 years (range, 2-65 years). Forty-one (68%) tissue specimens were culture negative for Mycobacterium tuberculosis (Mtb), although nontuberculous mycobacteria (NTM) were identified in three HIV-negative patients. Histopathologic features were characteristic of TB infection in 91.4% of all specimens tested and 100% of the HIV-infected group. Genotyping of 10 culture-positive isolates identified Mtb (3/10), NTMs (2/10), and environmental bacilli (3/10). Our observations suggest HIV-induced immune suppression impacts the histological and clinical features of infectious spondylitis but has no impact on the incidence of NTMs in this setting.