The build-up of long-term immunological memory in the gut mucosal immune system may explain long-lasting protection against new attacks of cholera in convalescents from natural disease. We have looked for gut mucosal antitoxin immunological memory and memory cells in mice after oral immunization with cholera toxin. Our results show that mice that were orally primed with cholera toxin and then boosted 2 years later with a single oral antigen dose mounted a rapid and vigorous IgA antitoxin response in the intestinal lamina propria. A specific secondary antitoxin response could also be elicited without any in-vivo boosting by in-vitro stimulation of isolated lymphocytes from the lamina propria, Peyer's patches, mesenteric lymph nodes, and spleen. The results provide evidence for the almost life-long persistence of anti-cholera toxin memory B cells (and perhaps also T cells) in the intestine and probably also recirculating cells, after oral immunization with cholera toxin. A functional antitoxic immune response may be boosted rapidly on renewed enteral exposure to cholera toxin by the stimulation of memory cells both in the lamina propria and in the Peyer's patches.