Leishmania tropica: protective response in C3H mice vaccinated with excreted factor crosslinked with the synthetic adjuvant, muramyl dipeptide.
Excreted factor, an immunosuppressive, acidic polysaccharide released by promastigotes of Leishmania tropica major in culture, was chemically crosslinked to the synthetic adjuvant muramyl dipeptide via the bifunctional imidoester dimethyladipimidate and poly-L-lysine. This conjugate, an uncrosslinked mixture of the components, or each of the components alone were injected one to three times into different groups of 8- to 12-week-old C3H mice. The mice were challenged 2 weeks after the last injection with 2 X 10(6) promastigotes of L. t. major in the base of the tail. For the next 5 weeks, the animals were monitored for number of parasites and size of the lesion which developed at the site of the challenge. Mice receiving one intraperitoneal injection of the conjugate were partially protected against challenge. Treated animals had higher initial parasite numbers but showed a more rapid clearing of the parasites. Furthermore, the treated animals developed smaller lesions that healed quicker than did those of the control groups. Multiple injections, or injection into a footpad, rather than intraperitoneally, reduced the ability to elicit a protective response. On the other hand, muramyl dipeptide injected into a footpad was partially protective. Antibody production to excreted factor, which was measured by indirect hemagglutination of sensitized erythrocytes, was detected after challenge in mice which had received conjugate or conjugate components. A delayed hypersensitivity reaction (measured by skin testing) was not detected in any of the groups prior to challenge.