Nucleotide and nucleoside involvement in immunomodulation in experimental Chagas disease.

The aim of this study was to evaluate whether Trypanosma cruzi infections cause alterations in the levels of seric purines, which could contribute to host immunomodulation. Twelve mice were divided into two groups identified as control (uninfected) and infected (T. cruzi) groups. The influence of the disease on seric purine levels was verified on day 20 post-infection (PI) by HPLC. Infected mice had circulating trypomastigotes during the experiment, as well as amastigote forms in the heart associated with inflammatory infiltrates. Increases on adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine (ADO), inosine (INO), and uric acid (URIC) levels were observed in the infected animals, while the adenosine monophosphate (AMP) and xanthine (XAN) levels were reduced compared with mice of the control group, indicating a possible impairment on the purinergic system, and consequently, on the immune system during the clinical course of the disease. In summary, the T. cruzi infection alters the seric purine levels, and consequently, modulates the immune system.

Serum and brain purine levels in an experimental systemic infection of mice by Cryptococcus neoformans: Purinergic immunomodulatory effects.

The aim of this study was to evaluate the purine levels in serum and brains of mice experimentally infected by Cryptococcus neoformans. Twenty-four mice were divided into the following groups: a control group (n = 12; Group A) and an infection group with animals that were infected (n = 12; Group B) with a 0.3-mL intraperitoneal injection containing 1.7 × 107C. neoformans cells. Blood and brains were collected on days 20 (n = 6 per group) and 50 (n = 6 per group) post-infection (PI). Histopathology and lung and brain cultures were performed to confirm fungal infection and tissue injuries. The levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine (ADO), inosine (INO), hypoxanthine (HYPO), xanthine (XAN) and uric acid (UA) in brains and serum were measured by high-performance liquid chromatography (HPLC) analysis. At both time points, histopathology analysis revealed inflammatory infiltrates in the brains and lungs of infected mice; clinical signs, such as piloerection and clinical respiratory distress, were also observed. ATP levels were significantly increased on days 20 and 50 PI (P < 0.01) in brains and serum, while brain ADO levels were increased on day 20 PI; brain and serum ADO levels were decreased on day 50 PI. Levels of ADP and AMP did not differ between groups (P > 0.05). Serum levels of INO of infected mice increased only on day 50 PI (P < 0.05). HYPO levels were reduced in the brains of infected animals at both experimental time points and were decreased in serum at day 50 PI (P < 0.05). XAN levels increased in infected mice only in serum on day 50 PI (P < 0.05). The endogenous anti-oxidant uric acid was significantly increased in brain (days 20 and 50 PI) and decreased in serum. It is possible that C. neoformans infection in mice leads to a high ATP/ADO ratio that may improve the brain pro-inflammatory response during both periods, while high ATP levels in serum act as a systemic signal to improve the immune response. Moreover, the anti-oxidant uric acid may increase in the brain to protect inflamed tissue from oxidative stress.

Participation of purines in the modulation of inflammatory response in rats experimentally infected by Cryptococcus neoformans.

The present study was carried out to assess the participation of purines in the activation and modulation of inflammatory response of rats experimentally infected by Cryptococcus neoformans. Twenty four Wistar rats were divided into two groups of 12 animals each: Group A - uninfected control group and Group B - infected by C. neoformans. Blood was collected 20 and 50 days post-infection (PI) from six animals of each group in order to verify purine levels (adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine (ADO), inosine (INO), hypoxanthine (HYPO), xanthine (XAN) and uric acid (URIC)). ATP levels were significantly increased (P < 0.05) in serum from infected animals on days 20 and 50 PI, as well as adenosine levels after 20 days PI on rats. On day 50 PI, levels of adenosine and uric acid were also reduced, but the levels of inosine and xanthine increased in animals infected by the fungus (P < 0.05). Therefore, it was possible to conclude that the purine levels in serum were altered and that these changes may be able to influence the pathogenesis of the disease caused by C. neoformans due the participation of purines (ATP and adenosine main) in the activation and modulation of inflammatory response.

Hepatic and seric levels of purines in rats experimentally infected by Fasciola hepatica.

The aim of this study was to evaluate hepatic and seric levels of purines, as well as their breakdown products in rats infected by Fasciola hepatica on days 15 and 87 post-infection (PI). Rats were divided into two groups: uninfected (n = 10) and infected (n = 20). On day 15 (n = 5 for uninfected group and n = 10 for infected group) and 87 PI (n = 5 for uninfected group and n = 10 for infected group), animals were euthanized for sampling to evaluate levels of purines by high-performance liquid chromatography. In serum, ATP increased (P < 0.05) and ADP decreased (P < 0.05) on days 15 and 87 PI, while AMP increased (P < 0.05) only on day 15 PI. Hypoxanthine levels increased (P < 0.05) on days 15 and 87 PI, while adenosine and xanthine levels decreased and increased (P < 0.05), respectively, on day 87 PI. No difference was observed regarding seric inosine and uric acid (P > 0.05). Hepatic ATP, adenosine, and uric acid levels decreased (P < 0.05) on days 15 and 87 PI. AMP levels decreased (P < 0.05) on day 87 PI, while xanthine levels increased (P < 0.05) on day 15 PI in the liver. Also in the liver, hypoxanthine levels increased (P < 0.05) on day 15 PI and decreased (P < 0.05) on day 87 PI. On the other hand, there was no difference on hepatic ADP and inosine levels (P > 0.05). Therefore, it is possible to conclude that F. hepatica infection can change purine levels, which may be associated with an inflammatory process, and these alterations may influence fasciolosis pathogenesis.

Changes in purine levels associated with cellular brain injury in gerbils experimentally infected with Neospora caninum.

The present study was carried out in order to assess the possible alterations in purine levels of brain, associated neuronal lesions in gerbils experimentally infected with Neospora caninum. For that, gerbils (Meriones unguiculatus) were inoculated with Nc-1 strain of N. caninum, composing two different experiments: Experiment I (EI) and experiment II (EII), where purine levels were measured along with the histopathologic study, on days 7 (EI), 15 and 30 (EII), post-infection (PI). As a result, it was possible to observe that the purine levels (ATP, ADP, AMP, adenosine, inosine and xanthine) in brain in EI are significantly reduced (p < 0.05), while in EII we faced a different pattern, since in the majority the purine levels were significantly increased (p < 0.05) on days 15 (ATP, AMP, adenosine, hypoxanthine and xanthine) and 30 PI (ATP, ADP, AMP, adenosine, and uric acid). Results of brain histopathology did not show histological lesion in animals of EI; however, in gerbils of EII it was possible to verify that the alterations (lesions) were more pronounced in gerbils evaluated on day 30 PI when compared to day 15 PI. Therefore, it was possible to conclude that the purine levels in brain were altered in both experiments, concomitant with the histopathological injuries observed in EII.

Differential expression of nucleotide pyrophosphatase/phosphodiesterases by Walker 256 mammary cancer cells in solid tumors and malignant ascites.

AIMS: Expression of ectoenzymes responsible for nucleotide phosphohydrolysis to form adenosine may represent a mechanism that facilitates the proliferation and spread of malignancy. In this study, we have identified and characterized the ectonucleotide pyrophosphatase/phosphodiesterase (E-NPP) family members expressed during the subcutaneous tumor growth and in the ascitic form of Walker 256 mammary tumor cells. MAIN METHODS: The biochemical characteristics in ascitic forms and expression of NPP 1, 2, and 3 in both solid and ascitic forms of Walker 256 tumor were investigated using RT-PCR and real-time PCR. KEY FINDINGS: Walker 256 tumor cells demonstrate E-NPP activities that are associated with extracellular hydrolysis of p-Nph-5'-TMP, and define the biochemical characteristics. The K(m) and maximal velocity for the hydrolysis of p-Nph-5'-TMP in the ascitic tumor cells were in accordance with the NPP reaction. The mRNA expression in the cells of the ascitic form of Walker 256 tumor revealed transcripts for NPP2 and NPP3, whereas elevated expression of NPP3 was observed in solid tumor, after 6, 10, and 15days of inoculation. The dominant gene expressed in both forms of the tumor was the NPP3 enzyme. However, this enzyme was expressed more during tumor development in vivo, when compared with the ascitic cells. SIGNIFICANCE: We have previously demonstrated that Walker 256 tumor cells express mRNA for ecto-5'-nucleotidase and E-NTPDases. Thus, coexistence with NPP3 suggests an ectonucleotidase "enzyme chain" that is responsible for the sequential hydrolysis of ATP to adenosine, which may be an important therapeutic target in anticancer therapy.