Human immunodeficiency virus (HIV) affects more than 40 million people worldwide and more than 5 million in South Africa alone. There is no cure for the disease yet, and novel effective drugs need to be discovered to make any progress in combating the disease. Twelve extracts from indigenous South African plants were analysed, of which Elaeodendron croceum showed potent inhibition of transcription factors and a recombinant HIV strain in an MT-2 VSV-pseudotyped recombinant virus assay at 100 ng mL(-1). Bioassay guided isolation of an ethanolic extract of E. croceum yielded a well-known digitoxigenin-glucoside as the only active compound. It showed significant inhibition (90%) at 0.2 µM. The in vitro toxicity of digitoxigenin-glucoside proved to be quite low, and its therapeutic index was 250. This observation indicates that digitoxigenin-glucoside could represent a potential pharmacophore for the treatment of HIV from natural sources.