RESUMEN
Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.
Asunto(s)
Adenosina Trifosfato , Antineoplásicos , Cisplatino , Vaciamiento Gástrico , Condicionamiento Físico Animal , Ratas Wistar , Receptores Purinérgicos P2X7 , Animales , Cisplatino/farmacología , Masculino , Adenosina Trifosfato/metabolismo , Vaciamiento Gástrico/efectos de los fármacos , Vaciamiento Gástrico/fisiología , Receptores Purinérgicos P2X7/metabolismo , Condicionamiento Físico Animal/fisiología , Antineoplásicos/farmacología , Ratas , Antagonistas del Receptor Purinérgico P2X/farmacologíaRESUMEN
Patients undergoing chemotherapy with cisplatin commonly present gastrointestinal effects such as constipation and gastric emptying (GE) delay. Both the purinergic system and physical exercise modulate the gastrointestinal (GI) tract. In the current study, we investigated the role of ATP, physical exercise, and P2X7 receptor blocking on GE delay induced by cisplatin in rats. Male rats were divided into the following groups: control (C), cisplatin (Cis), exercise (Ex), Brilliant Blue G (BBG), ATP, Cis+Ex, Cis+ATP, Cis+BBG, Cis+Ex+BBG, Cis+Ex+BBG+ATP, and Cis+ATP+BBG. GE delay was induced by treatment with 1 mg/kg cisplatin (1 time/week for 5 weeks, ip). The moderate physical exercise was swimming (1 h/day, 5 days/week for 5 weeks). At the end of the treatment or exercise and 30 min before the GE assessment, some groups received BBG (50 mg/kg, sc) or ATP (2 mg/kg, sc). Then, GE was assessed after a 10-min postprandial period. Chronic use of Cis decreased GE delay (P<0.05) compared to the control group. Both exercise and ATP prevented (P<0.05) GE delay compared to Cis. The pretreatment with BBG significantly inhibited (P<0.05) the effect of exercise and ATP. On the other hand, the association between exercise and ATP reversed (P<0.05) the effect of the BBG and prevented GE delay. Therefore, we suggest that both exercise and treatment with ATP activate P2X7 receptors and prevent GE delay induced by cisplatin in rats.
RESUMEN
Schistosoma mansoni is a blood fluke parasite responsible for schistosomiasis. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. In this study, we cloned, expressed and purified SmTSP-2 fused to the N- and C-terminal halves of Sm29 and tested these chimeras as vaccine candidates using an adjuvant approved to be used in humans. The results demonstrated that vaccination with SmTSP-2 fused to N- or C-terminus of Sm29-induced reduction in worm burden and liver pathology when compared to control animals. Additionally, we detected high levels of mouse-specific IgG, IgG1 and IgG2a against both chimeras and significant amounts of IFN-γ and TNF-α and no IL-4. Finally, studies with sera from patients resistant to infection and living in schistosomiasis endemic areas revealed high levels of specific IgG to both chimeras when compared to healthy individuals. In conclusion, SmTSP-2/Sm29 chimeras tested here induced partial protection against infection and might be a potential vaccine candidate.
Asunto(s)
Antígenos Bacterianos/inmunología , Antígenos Helmínticos/inmunología , Proteínas Bacterianas/inmunología , Proteínas del Helminto/inmunología , Glicoproteínas de Membrana/inmunología , Schistosoma mansoni , Esquistosomiasis mansoni/inmunología , Esquistosomiasis mansoni/prevención & control , Tetraspaninas/inmunología , Vacunas/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Antihelmínticos/sangre , Antígenos Bacterianos/administración & dosificación , Antígenos Helmínticos/administración & dosificación , Proteínas Bacterianas/administración & dosificación , Islas de CpG , Citocinas/sangre , Femenino , Proteínas del Helminto/administración & dosificación , Humanos , Inmunoglobulina G/sangre , Hígado/patología , Glicoproteínas de Membrana/administración & dosificación , Ratones , Ratones Endogámicos C57BL , Oligodesoxirribonucleótidos/administración & dosificación , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunología , Tetraspaninas/administración & dosificación , Vacunas/administración & dosificaciónRESUMEN
Schistosomiasis is a parasitic disease with more than 200 million people infected worldwide. The formation of granulomas around eggs trapped in the liver is the main cause of disease morbidity. Therefore, the aim of this investigation was to characterize the immunopathological response induced by the recombinant (r) IPSE/alpha-1 egg protein in mice. Herein, we have shown that splenocytes from mice immunized with rIPSE/alpha-1 produced IFN-gamma, TNF-alpha, IL-4, IL-5 and IL-13 characterizing a mixed Th1/Th2 type of immune response. Pathological analysis of the liver revealed that there was no alteration in the number of eggs and granulomas; however, we observed an increase in granuloma area in immunized mice. Furthermore, eosinophil peroxidase assay showed that there was no alteration in the eosinophil infiltration in the liver; however, n-acetyl-beta-glucosaminidase measurement revealed an increase in macrophage activity. Despite the alteration in the profile of liver inflammatory cells in rIPSE immunized mice, the production of chemokines such as CCL2, CCL3, CCL5 and CCL11 was unaltered compared with the control group. In conclusion, IPSE/alpha-1 immunization induces a mixed Th1/Th2 type of immune response and enlargement of hepatic granuloma caused by an increased macrophage activity, but does not alter Th2 cytokines following infection.
Asunto(s)
Proteínas del Huevo/inmunología , Proteínas del Helminto/inmunología , Schistosoma mansoni/inmunología , Schistosoma mansoni/patogenicidad , Esquistosomiasis mansoni/patología , Esquistosomiasis mansoni/parasitología , Células Th2/inmunología , Factores de Virulencia/inmunología , Acetilglucosaminidasa/metabolismo , Animales , Citocinas/metabolismo , Proteínas del Huevo/genética , Eosinófilos/inmunología , Femenino , Granuloma/inmunología , Granuloma/parasitología , Granuloma/patología , Proteínas del Helminto/genética , Leucocitos Mononucleares/inmunología , Hígado/parasitología , Hígado/patología , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Peroxidasa/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Esquistosomiasis mansoni/inmunología , Bazo/inmunología , Factores de Virulencia/genéticaRESUMEN
Proteins associated with the schistosome tegument are of great importance for the development of new intervention strategies since they may be exposed on the surface of the parasite. Herein, we have isolated a cDNA clone encoding for the Schistosoma mansoni SmIg and its recombinant protein was tested as a potential vaccine candidate. Initially, its amino acid sequence was analysed by bioinformatics and shown to possess an N-terminal signal peptide, a C-terminal transmembrane helix, 4 glycosylation sites, an immunoglobulin conserved domain and 73% similarity with a hypothetical S. japonicum protein of unknown function. SmIg was produced by E. coli as a recombinant protein (rSmIg) and its protective effectiveness was evaluated against S. mansoni infection with 100 cercariae in a murine model. Mice immunized with rSmIg induced an immune response characterized by dominant IgG1 isotype and significant levels of IFN-gamma, TNF-alpha, IL-10 and IL-4. Although immunogenic, the recombinant vaccine failed to induce worm burden reduction when compared to the infected control group. However, rSmIg-immunized mice had significant reductions of liver granuloma volume and fibrosis content by 31.8% and 49%, respectively. In conclusion, SmIg is a new tegument protein from S. mansoni that plays an important role in reducing pathology induced by parasite infection.
Asunto(s)
Antígenos Helmínticos/administración & dosificación , Proteínas del Helminto/administración & dosificación , Hígado/inmunología , Hígado/patología , Schistosoma mansoni/inmunología , Esquistosomiasis mansoni/prevención & control , Vacunas de ADN/administración & dosificación , Secuencia de Aminoácidos , Animales , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/genética , Antígenos Helmínticos/inmunología , Femenino , Proteínas del Helminto/química , Proteínas del Helminto/genética , Proteínas del Helminto/inmunología , Inmunización , Inmunoglobulina G/sangre , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Esquistosomiasis mansoni/inmunología , Vacunas de ADN/genética , Vacunas de ADN/inmunologíaRESUMEN
Members of the leucine-rich repeat protein family are involved in diverse functions including protein phosphatase 2-inhibition, cell cycle regulation, gene regulation and signalling pathways. A novel Schistosoma mansoni gene, called SmLANP, presenting homology to various genes coding for proteins that belong to the super family of leucine-rich repeat proteins, was characterized here. SmLANP was 1184bp in length as determined from cDNA and genomic sequences and encoded a 296 amino acid open reading frame that spanning from 6 to 894bp. The predicted amino acid sequence had a calculated molecular weight of 32kDa. Analysis of the predicted sequence indicated the presence of 3 leucine-rich domains (LRR) located in the N-terminal region and an aspartic acid rich region in the C-terminal end. SmLANP transcript is expressed in all stages of the S. mansoni life cycle analyzed, exhibiting the highest expression level in males. The SmLANP protein was expressed in a GST expression system and antibodies raised in mice against the recombinant protein. By immunolocalization assay, using adult worms, it was shown that the protein is mainly present in the cell nucleus through the whole body and strongly expressed along the tegument cell body nuclei of adult worms. As members of this family are usually involved in protein-protein interaction, a yeast two hybrid assay was conducted to identify putative binding partners for SmLANP. Thirty-six possible partners were identified, and a protein ATP synthase subunit alpha was confirmed by pull down assays, as a binding partner of the SmLANP protein.