Identification of a 30 kDa antigen from leishmania (L.) chagasi amastigotes implicated in protective cellular...

An antigen of apparent molecular mass of 30 kDa, termed p30, was purified from Leishmania (L.) chagasi amastigotes after separation of parasite extracts by sodium dodecyl sulfate-polyacrylamide gel eletroctrophoresis followed by electroelution. The use of the purified antigen in lymphocyte cultures from BALB/c mice previously immunised with L. (L.) chagasi amastigotes led to high levels of proliferation. Animal immunisation with p30 plus complete Freund's adjuvant either by subcutaneous or intraperitoneal route led to comparable antigenic stimulation. Similar stimulation indices induced by p30 were also obtained when animals were immunised with Corynebacterium parvum as adjuvant by the intraperitoneal route. Detection of IL-2 and IFN-gamma in the supernatants from lymphocytes stimulated by p30 and inhibition of the production of these lymphokines in the presence of anti-CD4 strongly indicated the involvement of the Th1 subset in the responses elicited by p30 antigen. Immunisation of BALB/c mice with p30 provided partial protection against challenge with L. (L.) chagasi amastigotes, indicating a protective role for p30 and that Th1 can be related to accquired resistance to visceral leishmaniasis in a murine model. Further characterisation studies were performed by the use of a monoclonal antibody directed to a cysteine proteinase of 30 kDa from L. (L.) amazonensis amastigotes. Despite the cross-reactivity presented by p30 from both Leishmania species, the p30 from L. (L.) chagasi amastigotes lacks proteolytic activity.