ABSTRACT
Larrea divaricata Cav. (jarilla) is a plant with well-documented applications in Argentinean folk medicine. In order to determine if the treatment with a purified fraction named F1 was capable to maintain a state of priming of macrophages after 15 days of mice infection with Candida albicans. Infected and uninfected mice were used. The effect of F1 on: cytosolic protein levels, apoptosis, phagocytosis, reactive oxygen species production, nitric oxide (NO), cell activity, lysosomal activity and the tissue fungal burden were studied. The results showed that F1 increased macrophages yeast phagocytosis and reactive oxygen species and NO production. All these effects were related to a decrease of cell activity and possible apoptosis. In conclusion, it was observed that F1 could induce a state of long-term activation of macrophages, since we observed increased activity of macrophages 15 days after infection, and it could be related to the elimination of C. albicans. These data may suggest that F1 fraction could be useful against disseminated candidiasis in patients and further studies on this field are desirable.
Subject(s)
Candida albicans , Candidiasis/drug therapy , Larrea/chemistry , Macrophages, Peritoneal/metabolism , Plant Extracts/pharmacology , Animals , Apoptosis/drug effects , Candidiasis/metabolism , Candidiasis/pathology , Cells, Cultured , Female , Macrophages, Peritoneal/microbiology , Macrophages, Peritoneal/pathology , Mice , Nitric Oxide/biosynthesis , Phagocytosis/drug effects , Plant Extracts/chemistry , Time FactorsABSTRACT
Larrea divaricata Cav. (jarilla) is a plant with well-documented applications in folk medicine in Argentina. In this study, we aimed to evaluate functional parameters of peritoneal macrophages isolated from mice injected with three fractions (F1, F2 and F3) of L. divaricata. The response of macrophages against Candida albicans was evaluated. Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test, apoptosis was evaluated using Giemsa, acridine orange/ethidium bromide and ladder assay, oxidative burst was assayed using nitroblue tetrazolium test and nitrite production using Griess assay. Cell stimulation and their ability to kill C. albicans in vitro were measured. The number and cell viability were similar to controls. However, we found that F1 induces pre-activation of macrophages, and this pre-activation is enhanced by C. albicans. The effects exerted by F1 make it more important than F2 and F3 for the treatment of disseminated candidiasis in patients with immunodeficiency diseases such as AIDS and chronic granulomatous disease, among others.
Subject(s)
Candida albicans/immunology , Immunologic Factors/pharmacology , Larrea/chemistry , Macrophages, Peritoneal/drug effects , Macrophages, Peritoneal/immunology , Plant Extracts/pharmacology , Animals , Apoptosis , Cell Survival , Cells, Cultured , DNA Fragmentation , Ethidium/metabolism , Female , Immunologic Factors/isolation & purification , Male , Mice , Nitrites/metabolism , Nitroblue Tetrazolium/metabolism , Plant Extracts/isolation & purification , Respiratory Burst , Staining and Labeling/methods , Tetrazolium Salts/metabolism , Thiazoles/metabolismABSTRACT
BACKGROUND AND AIM: Larrea divaricata Cav. (Zygophyllaceae) is a plant widely used in Argentina. MATERIAL AND METHODS: We isolated different fractions of L. divaricata aqueous extract containing minor amounts of NDGA, and we analyzed these fractions on mouse macrophages. RESULTS: We showed that a fraction without NDGA was capableof activating macrophages, principally through the production of mitochondrial anion superoxide and H(2)O(2). This could be important in the defense of infections. Moreover, this fraction decreased NO level suggesting an anti-inflammatory action. CONCLUSION: These results indicate that NDGA was not the compound responsible for the immunomodulatory action exerted by the aqueous extract from L. divaricata.
Subject(s)
Immunologic Factors/pharmacology , Larrea , Macrophages, Peritoneal/drug effects , Plant Extracts/pharmacology , Animals , Apoptosis/drug effects , Chromatography, Thin Layer , Female , Larrea/chemistry , Lipopolysaccharide Receptors/analysis , Macrophages, Peritoneal/metabolism , Male , Masoprocol/pharmacology , Mice , Nitric Oxide/biosynthesis , Reactive Oxygen Species/metabolismABSTRACT
The anaerobic bacillus Clostridium chauvoei is the causative agent of blackleg, a lethal disease that has an important impact on the sheep and cattle industry worldwide. Immunity to C. chauvoei is considered to be mainly anticellular, and for this reason there is scarce information about the immunogenicity of extracellular proteins. In this work variations in protein profiles, immune response by ELISA and protective capacity of culture supernatants of three C. chauvoei strains, collected at different growth phases, are reported. Sera raised against extracellular antigens also recognised cellular antigens of the same molecular masses. Partially purified cell-free supernatants and those concentrated 10 times by ultrafiltration (C-CFS), obtained at the early stationary phase of growth, induced a strong immunoprotective response, even at low doses, that was more marked for C. chauvoei strain ATCC 10092 (p < or = 0.05). With C-CFS formulations, a clear relationship was observed between IgG titres, protective capacity and concentration of the antigen doses, indicating a specific immune response.
