RESUMO
Visceral Leishmaniasis is a chronic zoonosis and, if left untreated, can be fatal. Infected dogs have decreased cellular immunity (Th1) and develop a potent humoral response (Th2), which is not effective for elimination of the protozoan. Immune response can be modulated by microRNAs (miRNAs), however, characterization of miRNAs and their possible regulatory role in the spleen of infected dogs have not been done. We evaluated miRNA expression in splenic leukocytes (SL) from dogs naturally infected with Leishmania infantum and developing leishmaniasis (CanL; n = 8) compared to healthy dogs (n = 4). Microarray analysis showed increased expression of miR 21, miR 148a, miR 7 and miR 615, and downregulation of miR 150, miR 125a and miR 125b. Real-time PCR validated the differential expression of miR 21, miR 148a and miR 615. Further, decrease of miR 21 in SL, by means of transfection with a miR 21 inhibitor, increased the IL-12 cytokine and the T-bet/GATA-3 ratio, and decreased parasite load on SL of dogs with CanL. Taken together, these findings suggest that L. infantum infection alters splenic expression of miRNAs and that miR 21 interferes in the cellular immune response of L. infantum-infected dogs, placing this miRNA as a possible therapeutic target in CanL.
Assuntos
Doenças do Cão/diagnóstico , Interleucina-12/metabolismo , Leishmaniose Visceral/diagnóstico , Leucócitos/metabolismo , MicroRNAs/metabolismo , Baço/metabolismo , Animais , Antagomirs/metabolismo , Anticorpos Monoclonais/imunologia , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Regulação para Baixo , Fator de Transcrição GATA3/metabolismo , Imunidade Celular , Interleucina-12/antagonistas & inibidores , Leishmania infantum/imunologia , Leishmania infantum/fisiologia , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/parasitologia , Leucócitos/citologia , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , Baço/imunologia , Proteínas com Domínio T/metabolismo , Células Th1/citologia , Células Th1/imunologia , Células Th1/metabolismo , Células Th2/citologia , Células Th2/imunologia , Células Th2/metabolismo , Regulação para CimaRESUMO
Leptospira genus contains species that affect human health with varying degrees of pathogenicity. In this context, we aimed to evaluate the differences in the modulation of host gene expression by strains of Leptospira varying in virulence. Our data showed a high number of differentially expressed transcripts in murine macrophages following 6h of infection. Leptospira infection modulated a set of genes independently of their degree of virulence. However, pathway analysis indicated that Apoptosis, ATM Signaling, and Cell Cycle: G2/M DNA Damage Checkpoint Regulation were exclusively regulated following infection with the virulent strain. Taken together, results demonstrated that species and virulence play a role during host response to Leptospira spp in murine macrophages, which could contribute to understanding the pathogenesis of leptospirosis.
Assuntos
Interações Hospedeiro-Patógeno , Leptospira/patogenicidade , Macrófagos/microbiologia , Transcriptoma , Animais , Apoptose/genética , Ciclo Celular/genética , Linhagem Celular , Leptospira/genética , Macrófagos/metabolismo , Camundongos , Virulência/genéticaRESUMO
Visceral leishmaniasis (VL) in humans is a chronic and often fatal disease if left untreated. Dogs appear to be the main reservoir host for L. infantum infection, however, in many regions other canids such as jackals, foxes, wolves and other mammals, such as hares or black rats, have been implicated as wild reservoirs. Most dogs cannot form an effective immune response against this infection, and this could be modulated by small non-coding RNAs, called microRNAs, responsible for post-transcriptional control of gene expression. Here, we evaluated the expression of miRNAs in peripheral blood mononuclear cells (PBMC) of symptomatic dogs naturally infected with Leishmania (L.) infantum (n = 10) and compared to those of healthy dogs (n = 5). Microarray analysis revealed that miR-21, miR-424, miR-194 and miR-451 had a 3-fold increase in expression, miR-192, miR-503, and miR-371 had a 2-fold increase in expression, whereas a 2-fold reduction in expression was observed for miR-150 and miR-574. Real-time PCR validated the differential expression of miR-21, miR-150, miR-451, miR-192, miR-194, and miR-371. Parasite load of PBMC was measured by real-time PCR and correlated to the differentially expressed miRNAs, showing a strong positive correlation with expression of miR-194, a regular positive correlation with miR-371 expression, and a moderate negative correlation with miR-150 expression in PBMC. These findings suggest that Leishmania infection interferes with miRNAs expression in PBMC, and their correlation with parasite load may help in the identification of therapeutic targets in Canine Visceral Leishmaniasis (CVL).
Assuntos
Doenças do Cão , Regulação da Expressão Gênica , Leishmania infantum , Leishmaniose Visceral , Leucócitos Mononucleares/metabolismo , MicroRNAs/biossíntese , Animais , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Leishmaniose Visceral/sangue , Leishmaniose Visceral/veterinária , Leucócitos Mononucleares/parasitologiaRESUMO
Leptospirosis is a bacterial zoonosis, caused by Leptospira spp., that leads to significant morbidity and mortality worldwide. Despite considerable advances, much is yet to be discovered about disease pathogenicity. The influence of epigenetic mechanisms, particularly RNA-mediated post-transcriptional regulation of host immune response has been described following a variety of bacterial infections. The current study examined the microtranscriptome of macrophages J774A.1 following an 8h infection with virulent, attenuated and saprophyte strains of Leptospira. Microarray analysis revealed that 29 miRNAs were misregulated following leptospiral infection compared to control macrophages in a strain and virulence-specific manner. Pathway analysis for targets of these differentially expressed miRNAs suggests that several processes involved in immune response could be regulated by miRNAs. Our data provides the first evidence that host miRNAs are regulated by Leptospira infection in macrophages. A number of the identified miRNA targets participate in key immune response processes. We suggest that post-transcriptional regulation by miRNAs may play a role in host response to infection in leptospirosis.
Assuntos
Leptospira/fisiologia , Leptospirose/genética , Macrófagos/microbiologia , Transcriptoma , Animais , Cricetinae , Humanos , Leptospira/classificação , Leptospira/genética , Leptospira/patogenicidade , Leptospirose/metabolismo , Leptospirose/microbiologia , Macrófagos/metabolismo , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Filogenia , VirulênciaRESUMO
This paper contains data on differentially expressed miRNAs in peripheral blood mononuclear cells (PBMC) of dogs naturally infected by Leishmania (L.) infantum compared to healthy dogs. In recent years, studies with miRNAs have shown that these molecules play a critical role in the regulation and function of immune response.Differentially expressed miRNAs were identified by microarray, validated by real time PCR and compared with parasite load in the dogs. Targets and pathways were analyzed using the Ingenuity Pathway Analysis program.
RESUMO
The datasets reported herein provide information about microarray experiment of macrophage cell line J774A.1 infected with three different strains of Leptospira spp. Transcriptomic profiles were generated using Affymetrix® Mouse Gene 2.1 ST Array Strip. Data was normalized and statically process, p-value < 0.01, FDR < 0.05 and log2 fold change (± 2). The microarray raw data are available in Gene Expression Omnibus (GEO) under accession number GSE105141.
RESUMO
Scarabaeinae (Coleoptera: Scarabaeidae), comprises the beetles popularly called dung beetle, which are important group of animal detritivorous and biomarkers of environmental impacts on ecosystems. Pampa Biome areas are natural ecosystems with high diversity of plant and animal species, but are however, little known compared to other biomes of Brazil. Data on escarabeinofauna the Pampa Biome is scarce. Therefore, the aim of this study was to inventorize the fauna of these beetles in areas located in the far south of the Rio Grande do Sul, as well as learning their habits and eating behaviors. The sampling areas were located in the cities of Herval and Arroio Grande. 216 traps baited pitfall trap type with human dung and pig and beef decaying were placed distributed in six areas. They recorded 2,243 individuals, organized into six tribes, 13 genera and 35 species. The species were classified as coprophagous, general and scavenger. Greater number of subjects were caught in traps baited with human and pig manure than in rotting meat. For the first time Trichillum morelli Verdú & Galante, 1997 was found in Brazil and Canthidium taurinum Harold, 1867 is a new record for the state.(AU)
Scarabaeinae (Coleoptera: Scarabaeidae), compreende compreende os besouros popularmente chamados de rola-bosta, que constituem importante grupo de animais detritívoros e bioindicadores de impactos ambientais nos ecossistemas. Áreas do Bioma Pampa são ecossistemas naturais com alta diversidade de espécies vegetais e animais, no entanto, pouco conhecidas em comparação com outros biomas do Brasil. Dados sobre a escarabeinofauna do Bioma Pampa é escassa. Portanto, os objetivos deste trabalho foram inventariar a fauna destes besouros em áreas localizadas no extremo-sul do Rio Grande do Sul, bem como, conhecer seus hábitos e comportamentos alimentares. As áreas de amostragem se localizaram nos municípios de Herval e Arroio Grande. Foram dispostas 216 armadilhas do tipo pitfall trap iscadas com esterco humano/suíno e carne bovina em decomposição, distribuídas em seis áreas. Registraram-se 2.243 indivíduos, organizados em 6 tribos, 13 gêneros e 35 espécies. As espécies foram classificadas em coprófagas, generalistas e necrófagas. Foi capturado um maior número de indivíduos em armadilhas iscadas com esterco suíno e humano do que em carne apodrecida. Pela primeira vez Trichillum morelli Verdú & Galante, 1997 foi encontrada no Brasil e Canthidium taurinum Harold, 1867 é um novo registro para o estado.(AU)
Assuntos
Animais , Besouros , Grupos de População Animal , EcossistemaRESUMO
Scarabaeinae (Coleoptera: Scarabaeidae), comprises the beetles popularly called dung beetle, which are important group of animal detritivorous and biomarkers of environmental impacts on ecosystems. Pampa Biome areas are natural ecosystems with high diversity of plant and animal species, but are however, little known compared to other biomes of Brazil. Data on escarabeinofauna the Pampa Biome is scarce. Therefore, the aim of this study was to inventorize the fauna of these beetles in areas located in the far south of the Rio Grande do Sul, as well as learning their habits and eating behaviors. The sampling areas were located in the cities of Herval and Arroio Grande. 216 traps baited pitfall trap type with human dung and pig and beef decaying were placed distributed in six areas. They recorded 2,243 individuals, organized into six tribes, 13 genera and 35 species. The species were classified as coprophagous, general and scavenger. Greater number of subjects were caught in traps baited with human and pig manure than in rotting meat. For the first time Trichillum morelli Verdú & Galante, 1997 was found in Brazil and Canthidium taurinum Harold, 1867 is a new record for the state.
Scarabaeinae (Coleoptera: Scarabaeidae), compreende compreende os besouros popularmente chamados de rola-bosta, que constituem importante grupo de animais detritívoros e bioindicadores de impactos ambientais nos ecossistemas. Áreas do Bioma Pampa são ecossistemas naturais com alta diversidade de espécies vegetais e animais, no entanto, pouco conhecidas em comparação com outros biomas do Brasil. Dados sobre a escarabeinofauna do Bioma Pampa é escassa. Portanto, os objetivos deste trabalho foram inventariar a fauna destes besouros em áreas localizadas no extremo-sul do Rio Grande do Sul, bem como, conhecer seus hábitos e comportamentos alimentares. As áreas de amostragem se localizaram nos municípios de Herval e Arroio Grande. Foram dispostas 216 armadilhas do tipo pitfall trap iscadas com esterco humano/suíno e carne bovina em decomposição, distribuídas em seis áreas. Registraram-se 2.243 indivíduos, organizados em 6 tribos, 13 gêneros e 35 espécies. As espécies foram classificadas em coprófagas, generalistas e necrófagas. Foi capturado um maior número de indivíduos em armadilhas iscadas com esterco suíno e humano do que em carne apodrecida. Pela primeira vez Trichillum morelli Verdú & Galante, 1997 foi encontrada no Brasil e Canthidium taurinum Harold, 1867 é um novo registro para o estado.