Topical application of ozonated sunflower oil accelerates the healing of lesions of cutaneous leishmaniasis in mice under meglumine antimoniate treatment.

Besides being scarce, the drugs available for treating cutaneous leishmaniasis have many adverse effects. Ozone is an option to enhance the standard treatment due to the wound-healing activity reported in the literature. In this study, we evaluated the efficiency of ozonated sunflower oil as an adjuvant in treating cutaneous lesions caused by Leishmania amazonensis. BALB/c mice were infected with L. amazonensis, and after the lesions appeared, they were treated in four different schedules using the drug treatment with meglumine antimoniate (Glucantime®), with or without ozonated oil. After thirty days of treatment, the lesions' thickness and their parasitic burden, blood leukocytes, production of NO and cytokines from peritoneal macrophages and lymph node cells were analyzed. The group treated with ozonated oil plus meglumine antimoniate showed the best performance, improving the lesion significantly. The parasitic burden showed that ozonated oil enhanced the leishmanicidal activity of the treatment, eliminating the parasites in the lesion. Besides, a decrease in the TNF levels from peritoneal macrophages and blood leukocytes demonstrated an immunomodulatory action of ozone in the ozonated oil-treated animals compared to the untreated group. Thus, ozonated sunflower oil therapy has been shown as an adjuvant in treating Leishmania lesions since this treatment enhanced the leishmanicidal and wound healing effects of meglumine antimoniate.

Human MHC class I molecule, HLA-A2.1, mediates activation of CD8+ T cell IFN-γ production and the T cell-dependent protection against reactivation of cerebral Toxoplasma infection.

To examine whether the HLA-A2.1, one of the most common MHC class I molecules in humans, activates the protective immunity against reactivation of cerebral infection with Toxoplasma gondii, HLA-A2.1-transgenic and wild-type (WT) mice were infected and treated with sulfadiazine to establish chronic infection in their brains. One month after discontinuation of sulfadiazine, which initiates reactivation of the infection, mRNA levels for tachyzoite (the acute stage form)-specific SAG1 and numbers of the foci associated tachyzoites were significantly less in the brains of the HLA-A2.1-transgenic than WT mice. Greater numbers of IFN-γ-producing CD8+ T cells were detected in the spleens of infected transgenic than WT mice, and CD8+ T cells from the former produced markedly greater amounts of IFN-γ than the T cells from the latter in response to tachyzoite antigens in vitro. When their CD8+ T cells were systemically transferred to infected immunodeficient NSG mice expressing the HLA-A2.1, the CD8+ T cells from HLA-A2.1-transgenic mice inhibited reactivation of the cerebral infection in the recipients more efficiently than did the WT T cells. Furthermore, the inhibition of reactivation of the infection by CD8+ T cells from the transgenic mice was associated with increased cerebral expression of IFN-γ and effector molecules against tachyzoites in the recipients when compared to the WT CD8+ T cell recipients. Thus, the human HLA-A2.1 is able to effectively activate IFN-γ production of CD8+ T cells against T. gondii tachyzoites and confer a potent protection against reactivation of cerebral infection with this parasite through the CD8+ T cells activation.

Leishmanicidal, Trypanocidal and Antioxidant Activity of Amyrin-Rich Extracts from Eugenia pyriformis Cambess.

AIMS: This study aimed to characterize and evaluate leishmanicidal and trypanocidal action as well as cytotoxicity on macrophages and antioxidant ability of extracts, obtained by supercritical CO2 and ultrasound-assisted extractions of Uvaia (Eugenia pyriformis) leaves. METHODS: Leaves from E. pyriformis were submitted to supercritical CO2 (E1) and ultrasound-assisted (E2) extractions. The characterization of extracts was done using GC-MS and HPLC. L. amazonensis (promastigotes) and T. cruzi (epimastigotes and trypomastigotes) were treated with crescent concentrations of E1 and E2. After this, parasites were counted and the percentage of inhibition and IC50/LC50 was calculated. Murine macrophages were treated with both extracts for 48 h and after that, the cellular viability was determined and CC50 was calculated. DPPH method was used to determine the antioxidant capacity of both extracts. RESULTS: The results of identification showed a great amount of α and ß-amyrin in E1 and E2. Both extracts showed growth inhibition of L. amazonensis with an IC50 of 5.98 and 9.38 µg/mL to E1 and E2, showing a selectivity index > 30. In trypanocidal tests, an LC50 of 16.69 and 7.80 µg/mL (trypomastigotes) and IC50 of 5.56 and 34.34 µg/mL (epimastigotes) was reached by E1 and E2. Both extracts showed no toxicity to macrophages and an antioxidant capacity similar to the positive control (tocopherol). CONCLUSIONS: This is the first study demonstrating the activity of an amyrin rich-extract against microorganisms that cause Chagas disease and leishmaniasis, as well as its antioxidant capacity, justifying further studies for future in - vivo tests.