Granulocytes in the inflammatory process of BALB/c mice infected by Leishmania amazonensis. A quantitative approach.

We used previously immunized (partially resistant) and naive (highly susceptible) BALB/c mice infected with Leishmania amazonensis for evaluating the role of granulocytes in the course of murine leishmaniasis. The animals were examined at different times post-infection and granulocytes counted in lesion tissues examined ultra-structurally. Polymorphonuclear granulocytes predominated during the early phase of infection and their number decreased with progression of infection; their number was similar in both groups during the early and intermediate phases of infection, though slightly higher in immunized animals during the late phase. Eosinophils represented approximately 10% of cells in the inflammatory infiltrate, being higher during the intermediate phase, and not differing between the groups. Another approach was the evaluation of granulocyte migration to the peritoneal cavity of susceptible BALB/c mice or resistant C57BL/6 mice under several stimuli. There was no statistically significant difference between resistant and susceptible animals in any of the treatments. Despite the influx of granulocytes to the lesion and its possible role in the initial destruction of injected Leishmania, this aspect does not seem to have an important effect on the outcome of the leishmanial infection.

In vivo protective effect of the lectin from Canavalia brasiliensis on BALB/c mice infected by Leishmania amazonensis.

In vivo administration of Canavalia brasiliensis lectin (at the time of infection, or maintained throughout the infection) reduced the lesions of highly susceptible BALB/c mice infected by Leishmania amazonensis. At the doses used C. brasiliensis lectin (ConBr) does not interfere with penetration or fate of Leishmania in the macrophages in vitro. Since Interferon-gamma (IFN-gamma) is the major macrophage activating factor, and considered a critical element in the successful immune response against leishmaniasis, we explored its participation in this phenomenon. ConBr either in vivo or in vitro induced IFN-gamma production in normal or in leishmania-infected BALB/c mice. However we were unable to change the course of disease by in vivo IFN-gamma administration (although IFN-gamma preparations were effective in inducing a leishmanicidal effect in vitro on L. amazonensis-infected peritoneal macrophages). Additionally, IFN-gamma neutralization with anti-IFN-gamma monoclonal antibody did not alter the protection conferred by ConBr administration. These data show that lectin administration in vivo is protective in the otherwise unchecked L. amazonensis infection of BALB/c mice, and suggest that such effect is not mediated by IFN-gamma.

Leishmania amazonensis infection: a comparison of in vivo leishmanicidal mechanisms between immunized and naive infected BALB/c mice.

In vitro studies have shown that both macrophage activation and destruction of parasitized macrophages lead to leishmania destruction. The relative role played by such mechanisms in vivo have not been properly evaluated. We took advantage of the model of intravenous immunization with solubilized leishmanial antigen which renders partially resistant the otherwise highly susceptible BALB/c mice to address this issue avoiding the interference of different genetic backgrounds. Leishmania destruction occurred in three situations: destruction of the parasitized macrophage, which were in close contact with lymphocytes or eosinophils; extracellular damage, always surrounded by small foci of granulocytes; and parasite damage inside activated macrophages. Destruction of the parasitized macrophages was frequently seen in immunized and protected animals. Our observations suggest that destruction of parasite-loaded macrophages is an important mechanism of host protection in experimental cutaneous leishmaniasis.

Distinct ultrastructural aspects in different biopsies of a single patient with diffuse cutaneous leishmaniasis.

The authors investigated the relation between parasites and host-cells in active and regressed lesions of a patient with diffuse cutaneous leishmaniasis, evaluating the frequency of different cell types, and the location and integrity of amastigotes. No correlation was found between parasite integrity and size of parasitophorous vacuoles. They observed ultrastructural findings characterizing a cell mediated immune response: macrophages lysis, parasitic destruction inside macrophages, close contact between parasitized macrophages and lymphocytes and between parasites and lymphocytes, lymphocytic infiltration and fibrosis. They suggest that in DCL there is a limited cellular immune response, although insufficient to control infection.