Sand fly species able to support the survival of the protozoan parasite Leishmania have been classified as permissive or specific, based upon their ability to support a wide or limited range of strains and/or species. Studies of a limited number of fly/parasite species combinations have implicated parasite surface molecules in this process and here we provide further evidence in support of this proposal. We investigated the role of lipophosphoglycan (LPG) and other phosphoglycans (PGs) in sand fly survival, using Leishmania major mutants deficient in LPG (lpg1−), and the phosphoglycan (PG)-deficient mutant lpg2−. The sand fly species used were the permissive species Phlebotomus perniciosus and P. argentipes, and the specific vector P. duboscqi, a species resistant to L. infantum development.
The lpg2− mutants did not survive well in any of the three sand fly species, suggesting that phosphoglycans and/or other LPG2-dependent molecules are required for parasite development. In vitro, all three L. major lines were equally resistant to proteolytic activity of bovine trypsin, suggesting that sand fly-specific hydrolytic proteases or other factors are the reason for the early lpg2− parasite killing. The lpg1− mutants developed late-stage infections in two permissive species, P. perniciosus and P. argentipes, where their infection rates and intensities of infections were comparable to the wild type (WT) parasites. In contrast, in P. duboscqi the lpg1− mutants developed significantly worse than the WT parasites.
In combination with previous studies, the data establish clearly that LPG is not required for Leishmania survival in permissive species P. perniciosus and P. argentipes but plays an important role in the specific vector P. duboscqi. With regard to PGs other than LPG, the data prove the importance of LPG2-related molecules for survival of L. major in the three sand fly species tested.
Phlebotomine sand flies are small blood-feeding insects, medically important as vectors of protozoan parasites of the genus Leishmania. Sand flies species can be divided roughly into two groups, termed specific or permissive, depending on their ability to support development of one or a few strains vs. a broad spectrum of these parasites. In this study, we explored the ability of two Leishmania major glycocalyx mutants to survive within these different types of vectors. The lpg1− mutant, which specifically lacks lipophosphoglycan (LPG), was able to survive normally in two permissive species, Phlebotomus argentipes and P. perniciosus, but was only able to survive within the specific species P. duboscqi for a limited time prior to dissolution of the peritrophic matrix. Consistent with its classification as a specific sand fly vector, P. duboscqi was not able to support development of L. infantum. The lpg2− L. major mutant, which is a broader mutant and lacks all phosphoglycans including LPG and proteophosphoglycans, was unable to survive in all the three vector species tested. This study extends the knowledge on the role of Leishmania major surface glycoconjugates to development in three important vector species and gives supporting evidence for the existence of an LPG-independent mechanism for survival in sand flies, as well as the importance of LPG2-dependent glycoconjugates in parasite survival.