RESUMO
Protozoans of the Leishmania genus are the etiological agents of a wide spectrum of diseases commonly known as leishmaniases. Lipid organization of the plasma membrane of the parasite may mimic the lipid organization of mammalian apoptotic cells and play a role in phagocytosis and parasite survival in the mammal host. Here, we analyzed the phospholipid dynamics in the plasma membrane of both the L. (Leishmania) and the L. (Viannia) subgenera. We found that the activity and substrate specificity of the inward translocation machinery varied between Leishmania species. The differences in activity of inward phospholipid transport correlated with the different sensitivities of the various species towards the alkyl-phospholipid analogue miltefosine. Furthermore, all species exhibited a phospholipid scramblase activity in their plasma membranes upon stimulation with calcium ionophores. However, binding of annexin V to the parasite surface was only detected for a subpopulation of parasites during the stationary growth phase and only marginally enhanced by scramblase activation.
Assuntos
Membrana Celular/metabolismo , Leishmania/metabolismo , Fosfolipídeos/metabolismo , Anexina A5/metabolismo , Transporte Biológico , Ionomicina/farmacologia , Leishmania/efeitos dos fármacos , Bicamadas Lipídicas/metabolismo , Proteínas de Transferência de Fosfolipídeos/metabolismo , Ligação ProteicaRESUMO
In addition to its role as a protein component in Leishmania, serine is also a precursor for the synthesis of both phosphatidylserine, which is a membrane molecule involved in parasite invasion and inactivation of macrophages, and sphingolipids, which are necessary for Leishmania to differentiate into its infective forms. We have characterized serine uptake in both promastigote and amastigote forms of Leishmania (Leishmania) amazonensis. In promastigotes, kinetic data show a single, saturable transport system, with a Km of 0.253+/-0.01 mM and a maximum velocity of 0.246+/-0.04 nmol/min per 10(7) cells. Serine transport increased linearly with temperature in the range from 20 degrees C to 45 degrees C, allowing the calculation of an activation energy of 7.09 kJ/mol. Alanine, cysteine, glycine, threonine, valine and ethanolamine competed with the substrate at a ten-fold excess concentration. Serine uptake was dependent on pH, with an optimum activity at pH 7.5. The characterization of the serine transport process in amastigotes revealed a transport system with a similar Km, energy of activation and pH response to that found in promastigotes, suggesting that the same transport system is active in both insect vector and mammalian host Leishmania stages. This could constitute an evolutionary mechanism that guarantees the provision of such an essential molecule during host change events, such as differentiation into amastigotes and macrophage invasion, as well as to ensure that the parasite maintains the infection in the mammalian host.
Assuntos
Leishmania/metabolismo , Serina/metabolismo , Animais , Transporte Biológico , Concentração de Íons de Hidrogênio , CinéticaRESUMO
A case of feline cutaneous leishmaniasis is reported in a domestic cat (Felis catus) as an apparently natural infection in a non-endemic area. Amastigotes were seem in smears of a nodular lesion on the cat's nose. No parasite could be seen in cytological preparations of liver or spleen but DNA obtained from a sample of the spleen produced the expected fragment in a Leishmania specific rDNA based PCR assay. The PCR product, a 520 bp fragment, was sequenced and the nucleotide sequence was identical to that of Leishmania (Leishmania) infantum chagasi. These results are surprising since no autochthonous human or canine cases of visceral leishmaniasis have ever been reported from this region. This case suggests that natural transmission of this disease is occurring in this area, and that cats could act as a reservoir of L. (L.) infantum chagasi.